Friday, March 15, 2024

Sero Prevalence of Bovine Foot and Mouth Diseases in Bale Zone Selected Districts Sinana and Agarfa Oromia, Ethiopia - Juniper Publishers

 Journal of Dairy & Veterinary Sciences - Juniper Publishers


Abstract

Ethiopia is country with the most abundant livestock population in Africa with an estimated domestic animal number of 56.71 million cattle. The Foot and mouth disease (FMD) virus is a highly contagious and economically devastating transboundary disease of cloven hooved domestic and wild animals. The Foot and mouth disease virus, the causative agent of foot and mouth disease belongs to the genus Pithovirus and the family Picornaviridae. It has seven sero types, namely: O, A, C, South African Territories (SAT) SAT1, SAT 2, SAT 3, and Asia.

The study was conducted from January 2022 to February 2022 in two selected districts of Bale zone namely Sinana and Agarfa which are in the administrative zone of Oromia Region, Ethiopia. The study populations were local and cross breed of cattle above the age of six months having no clinical symptom of any disease were included. A cross sectional study design was taking place to determine the sero-prevalence of FMD. The present study considered 50% expected prevalence, 95% confidence level and 5% absolute precision or marginal error. Whole blood samples each amounting to 8-10ml were collected from jugular vein of cattle using disposable needles and 10 ml non-heparinized vacutainer tube and 21 Gauge needle. The collected sera were tested by commercially available FMD NSP competitive ELISA kit (Non-structural proteins used as DIVA test) (ID Screen(r) FMD NSP Competition, ID-VET, Grabels, France) for the detection of antibody to 3ABC polyprotein. A total of 461 serum samples were collected and tested for FMD. Over all 103 samples gave positive results yielding a prevalence of 22.34%. Of the samples collected from Agarfa 25.97 % (60/231) were positive whereas of the 18.70% (43/230) samples collected from Sinana were found positive. The presence of this disease in the country is a major obstacle to the development of livestock resources because of its adverse effects on production and their product exports. An extensive regular surveillance and serotyping of FMD isolates throughout the country should be conducted to check the introduction and circulation of new serotypes in the country.

Keywords:Foot and mouth disease (FMD); sero-prevalence; competitive ELISA kit.

Abbreviations:FMD: Foot and Mouth Disease; SAT: South African Territories; FMDV: Foot and mouth disease virus; NSP: Non-Structural Proteins

Introduction

Ethiopia is country with the most abundant livestock population in Africa with an estimated domestic animal number of 56.71 million cattle [1]. The Foot and mouth disease (FMD) virus is a highly contagious and economically devastating transboundary disease of cloven hooved domestic and wild animals [2]. FMD is also among the most important livestock diseases that affects production and trade of animal and animal products in Ethiopia [3]. Sero-surveys done in different parts of Ethiopia reported FMD with different degrees of prevalence reaching up to 26% (6-9). Outbreak incidence studies have also indicated that FMD occurs throughout the country with significant variation in geography and production systems [3]. Among the seven serotypes of FMDV, four of them (O, A, SAT 2, and SAT 1) have been reported in Ethiopia in recent times [1].

The Foot and mouth disease virus (FMDV), the causative agent of foot and mouth disease (FMD), belongs to the genus Aphthovirus and the family Picornaviridae. It has seven serotypes, namely: O, A, C, South African Territories (SAT) SAT1, SAT 2, SAT 3, and Asia [4]. The highly infectious nature of the virus, the generation of high titers in respiratory secretions, the prolonged survival of the virus in secretions, the fast replication cycle, and the brief incubation time contribute to the virus’s rapid dissemination to fully susceptible populations. In addition to the animal-to-animal transmissions, FMDV is easily transmitted mechanically via fomites such as clothes, shoes, vehicles, and veterinary instruments [5]. Moreover, its spread is aggravated by uncontrolled movements of infected animals across geographic boundaries [6]. In Ethiopia, the disease has been affecting mainly cattle, while also causing problems in small ruminants at infrequent intervals [7].

Historically, the disease was first identified in 1957, but it had certainly been present in the country before that, since most livestock keepers were familiar with it and some were using traditional methods of immunization against it, such as “mouthing [7]. According to the report of [8] FMD serotypes O, A, and C were responsible for FMD outbreaks from 1957 to 1979. A separate report on the genetic characterization of FMDV from 1981 to 2007 disclosed additional serotypes such as SAT1 and SAT2. The occurrence of the disease leads to loss of production, restriction of exports, and other socio-economic problems in the area [9].

Direct impact of FMD includes meat and milk production losses, loss of drought power, lower weight gains, fertility problems, changes in herd structure, delay sale of cattle and products, and death of cattle, while the indirect impacts include additional cost of treatment, vaccination, vaccine delivery, movement control, diagnostic tests, culled cattle, and denied access to both local and international markets [10]. FMD currently is widely prevalent and distributed in all areas of Ethiopia across the different farming systems and agro ecological zones of the country. Previously, the disease used to frequently occur in the pastoral herds of the marginal low-land areas of the country. However, this trend has changed, and the disease is frequently noted in the central/ highland parts of the country [11]. Sero-prevalence investigations undertaken so far in the different parts of the country reported the prevalence that ranges from 5.6% to 53.6% in cattle [12]. In the current study area, knowing the status of the disease is very important because of a high cattle population and cattle marketing activities, the practice of communal grazing and watering and cattle movement.

FMD is endemic to most countries in sub-Saharan Africa and will not be eradicated from southern East Africa while infected buffalo are present except for few countries Southern Africa, where the disease is controlled by the separation infected wildlife from susceptible livestock as well as by vaccination. Largely due to the endemicity of the disease and the fact that FMD does not cause high mortality rates in adults’ animals which is (2%) and 20% in young animals. Several countries now realize that FMD is one of the transboundary diseases that should be controlled to ensure economic stability and access to lucrative international export markets for animals and animal products. Furthermore, they recognize that a regional approach would be needed to succeed [13]. Lack of movement control within countries and across international borders for both wildlife and domestic animals aggravates the problem and gives credence to the fact that FMD will remain a problem on the sub-continent for the foreseeable future [14]. Countries free of FMD impose strict import regulation on animals and animal products and potential viral contaminated fomites from FMD free countries. Greater loss can result from refusal from FMD free countries to import livestock and livestock products from endemic regions [15].

Several studies have been conducted on the sero-prevalence and associated risk factors of FMD in cattle in different parts of the country and still there is a scarcity of information in the study area. Implementing both sero-prevalence and associated risk factors investigation is crucial to generate baseline information about the disease in the selected Zone.
 Therefore, the objective of the present survey was to estimate the sero-prevalence and to assess associated risk factors of FMD sero-prevalence in Sinana and Agarfa districts.

Materials and Methods

Description of the Study Areas

The study was conducted from January 2022 to February 2022 in two selected districts of Bale zone namely Sinana and Agarfa which are in the administrative zone of Oromia Region, Ethiopia. Sinana is in Oromiya Regional State, Bale Administrative Zone. It is about 460 Km southeast of Addis Ababa and it has an altitude of 2400 m above sea level. It’s geographical from 07° 06’29’’ northern latitude and from 40° 12’52’’ eastern longitude. It has a bimodal rainfall pattern with the first peak from April to May and the second from August to October. The mean annual maximum temperature is 21.06°C and monthly values range between 19.39°C in October and 22.94°C in February. The mean annual minimum temperature is 9.32°C and monthly values range between 7.12°C in December and 10.81°C in April. The coldest month is December whereas February is the hottest month.

Agarfa district is found in the Bale Administrative Zone of Oromia Regional State, in Southeastern part of Ethiopia. It lays between 7°8’N to 7°28’N latitude and 39°31’E to 40°5’E longitude. The elevation of Agarfa district ranges from 1400 m to 3800 m above mean sea level (a.m.s.l). About 61% of the district is plain with slopes ranging from 0 to 8 degrees and most of this area lies in the southeastern and western parts of the study area. Wabe Shabelle river gorges and related rugged terrains make up about 31% of the district. Agarfa district falls within three traditional agro-climatic zones, vernacularly termed as Gamoji (hot), Badadare (temperate), and Bada (cold). Mean maximum and mean minimum temperatures are 25°C and 10°C respectively. The amount of maximum and minimum rainfall received in the area ranges between 1200 mm and 400 mm, respectively.

Study Populations

The study populations were local and cross breed of cattle above the age of six months having no clinical symptom of any disease were included. In addition, herd sizes considered were small, medium, and large as some of research handle the same way.

Study Design

A cross sectional study design was taking place to determine the sero-prevalence of FMD and associated risk factors in two selected districts of Oromia region and different herd sizes were included in the study based on the inclusion criteria. Semistructured questionnaires were administered to herd owners for the assessments of animal and herd level risk factors.

Sampling technique and Sample Size determination

Study districts were purposively selected based on higher study population, access to transportation and history of outbreaks for sero-prevalence determination and assessment of potential risk factors of FMDV. Individual animals from each herd were selected randomly as sampling unit to draw the required sample size. Since there was no previous study conducted on FMD in cattle found in the selected areas, the present study was considered 50% expected prevalence, 95% confidence level and 5% absolute precision or marginal error. Based on these assumptions, the total number of animals to be included in the study was determined using Thru field (2007) formula form the two Districts selected. The sample size was determined using the formula given as follows:
Where, N = required sample size, Pexp = expected prevalence, d2 = desired absolute precision.

Sample Collection and Transportation

A total of 768 whole blood samples each approximately amounting to 8-10ml were collected from jugular vein of cattle using disposable needles and 10 ml non-heparinized vacutainer tube and 21 Gauge needle. Following whole blood sample collection, vacutainer tubes were labeled and transported to around veterinary clinic and kept overnight at room temperature to allow the blood clot at slant position. Correspondingly, each sample was identified along with sex, age, and district. Then, serum samples are transferred from vacutainer tubes to cryogenic vials and stored in -20°C refrigerator at Asela veterinary regional laboratory. The samples were tested using the FMD non-structural protein ELISA to determine if animals in the herd had been recently infected with FMD virus thereby estimating the sero-prevalence in the two selected districts.

Open and closed ended questionnaires were administered to herd owners to assess potential risk factors of the disease alongside sample collection. Respondents from each district were randomly selected and interviewed to assess potential risk factors of the disease. Study populations’ sex, age, herd size and district are considered as hypothesized risk factors for the occurrence of FMDV. Herd owners having cattle are the sampling units for questionnaire survey. All necessary epidemiological information was collected, tabulated, coded, and analyzed using suitable statistical analysis on individual animal bases.

Serological Diagnostic Test

The collected sera were tested by commercially available FMD NSP competitive ELISA kit (Non-structural proteins used as DIVA test) (ID Screen(r) FMD NSP Competition, ID-VET, Grabels, France) for the detection of antibody to 3ABC polyprotein which is a useful indicator of past FMDV infection regardless of the serotype involved. The 3-ABC-ELISA was used according to the manufacturer’s instructions. The test principle is the blocking of plate bound NSP antigen by antibodies present in the serum samples. Any antibody specific for 3ABC binds to the antigen in the wells and forms an antigen/antibody complex on the plate well surface. The antibody to the assay was performed according to manufacturer’s instruction and results were analyzed and interpreted.

Data Management and Analysis

Data generated from laboratory analysis and questionnaire survey were recorded and coded using Microsoft Excel spreadsheet (Microsoft Corporation) and analyzed using STATA version 13.0 and R studio. Descriptive statistics (frequency and percentage) were employed to calculate the proportion of risk factors for FMD. Individual level animal prevalence was calculated by dividing the number of animals with positive ELISA tests by the total number of tested animals. In all the analyses, confidence levels at 95% were calculated, and a P < 0.05 was used for statistics.

Results

Prevalence of FMD in the samples

total of 461 serum samples were collected and tested for FMD. Over all 103 samples gave positive results yielding a prevalence of 22.34 %. Of the samples collected from Agarfa 25.97 % (60/231) were positive whereas of the 18.70% (43/230) samples collected from Sinana were found positive as described in table 1 below. At the kebele/Peasant Association level 18.25%, 19.23%, 26.78% and 25.21%of the samples collected from Alemgena, Kasoshek maro, Makora chafe, kasomaro, respectively gave positive results for FMD as shown in table 2 below. From tested samples for FMD cross breeds were 26.40% and local breeds were 30.04% as shown in table 3 below. Risk factors such as breed, sex, age, and body conditions were considered. Higher sero-prevalence was recorded in local breed cattle with prevalence of 30.04 %. Cattle with medium body condition have higher sero-prevalence 23.3%. Sex and age show almost nearly the same result.

Discussion

The overall sero-prevalence rate of 22.34% reported in this study agreed with the previous finding from Ethiopia [16] in which sero-positivity of 26.5% was reported. Compared to the present findings lower prevalences of 5.6% [17], 8.01% [18] and 9% [19] were reported from Afar Regional State, Dire Dawa and western Ethiopia, respectively. On the other hand, relatively higher seroprevalence was previously reported in samples from the Eastern zone of Tigray with 41.5%; followed by the Guji zone of Oromia and Yeka district of Addis Ababa city, with 32.7% and 30% respectively [20]. [21] in Sudan, [22], in Saudi Arabia and Uganda also reported sero-prevalences of 16%, 53% and 77% respectively, from FMD virus infected cattle. The observed prevalence variation may be resulted from differences in individual animals breed, age, and sex and production system.

Conclusion and Recommendations

FMD is prevalent in the study Districts as conformed serologically and reported to be endemic in Ethiopia. The presence of this disease in the country is a major obstacle to the development of livestock resources because of its adverse effects on production and their product exports. In Ethiopia, factors such as the presence of high numbers of susceptible domestic animals, herd composition or the involvement of multiple hosts (cattle, sheep and goats), herd size and individual animal age variability, lack of prophylactic vaccination, absence of regulation for prohibition of animal movement, high contact of animals at marketing and common grazing place as well as at watering points could contribute to the occurrence of FMD and create the difficulty in controlling the FMD Disease..

Therefore, the followings are offered as recommendations:

An extensive regular surveillance and serotyping of the FMD isolates throughout the country should be conducted to check the introduction and circulation of new serotype in the country and to ensure that circulating viruses are protected by existing vaccines and/or look for alternative vaccines.
 Government strategy in FMD control through regular vaccination and movement control should be implemented.
 Further detailed studies on the molecular characterization of the viruses and their genetic closeness with the existing vaccines need to be conducted to establish the nature of their diversity on the tree.


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Wednesday, March 13, 2024

A Proteomic Approach for The Study of Macrotyloma Uniflorum (Horsegram) Seed Proteins for Kidney Stone Degradation: Conceptual Study in Cultivars of Himachal Pradesh, India - Juniper Publishers

 Advances in Biotechnology & Microbiology- Juniper Publishers




Abstract

Proteomics is one of the complex and significant methodologies to know the genetic functions of proteins. The study of plant proteins having antilithiatic activity is an unexplored area and can be hit through proteomics. The present study was framed for in-depth identification and characterization of antilithiatic proteins from horse gram (Macrotyloma uniflorum). Total protein of Macrotyloma uniflorum seeds from six different location of Himachal Pradesh, India was isolated in protease inhibitor buffer. The identification of cultivar seed proteins was done by Matrix-assisted laser desorption/ionization-time of flight Mass spectrometry (MALDI-TOF MS). The results were framed by database search with the MASCOT server. The protein sample of Rampur location contained maximum number of peptides followed by Sundernagar, Palampur and Kullu. Seeds of Chamba location showed the presence of very few peptides. The quantification of Macrotyloma uniflorum seed proteins was done with Bradford method and further protein with high oxalate inhibition activity (15µg) was loaded on 12% Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gel. In the proteomic study of loaded protein, a total of three major low molecular weight protein bands were observed (~27KDa, ~23.1 and ~17KDa respectively). Seed protein SDS-PAGE may be used to investigate genetic diversity and relatedness in cultivars at a low cost. Furthermore, the proteomic analysis of the seed protein of M. uniflorum aids in the creation of a protein-based marker that can be used to identify cultivars in the future.

Keywords: Proteomics; Macrotyloma uniflorum; MALDI-TOF MS; MASCOT server; Antilithiatic proteins

Abbreviations: MALDI-TOF MS: Matrix-Assisted Laser Desorption/Ionization-Time of Flight-MASS Spectrometry; SDS-PAGE: Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis; TFA: Trifluoroacetic Acid; HCCA: Hydroxycinnamic Acid; CDPK: Calcium-dependent protein kinase 29-like

Introduction

Our urinary system is susceptible to a variety of diseases, which affects its functioning. Urolithiasis is one the third most frequent urological problems among people worldwide [1]. Stone illness influences 2-20% of populace around the world with a predominance rate of 15% in India [2,3]. The majority of Ca-Ox crystallizes in the urinary tract under physiological conditions and is then excreted easily in urine. If the crystals are allowed to remain in the kidney, they will expand and develop into stones, causing damage to the renal epithelial cells and providing a site for the development of a stone [4]. Renal cells become acidic when exposed to elevated concentrations of oxalate ions and/or Ca-Ox crystals under pathological conditions. Harm to renal cells causes changes in cell surface properties, revealing the attachment site for crystal adhesion and/or internalization by renal epithelial cells [5,6]. Various medicinal plants have been used as herbal medicines since they have been documented to have antilithiatic activity [7], and they have been a part of traditional medicine since ancient times. The development of plant-protein dependent medicinal drugs is currently the subject of numerous pharmaceutical industries. These plant proteins could be mass-produced and modified with recombinant DNA technology. Because of the presence of acidic amino acids and/or calcium binding domains (EF Hand motifs), the majority of plant-based antilithiatic proteins known to date are anionic [8].

These acidic amino acids bind to calcium, stopping it from interacting with oxalate and causing renal cell adherence [9]. The present study aimed to assess the bioactivity of anionic anti-calcifying proteins from dried seeds of Macrotyloma uniflorum (Horsegram) on Ca-Oxalate crystals to provide objective support for M. uniflorum's anti-urolithiatic ability. This Fabaceae-family plant is widely used in herbal medicine to treat several ailments. This crop, which belongs to the Fabaceae family, is commonly used in traditional medicine for a variety of cardiovascular disorders. It also has antioxidant, anti-inflammatory, and moderate diuretic properties [10]. M. uniflorum has also been shown to suppress CaOx crystallization and crystal adhesion to renal epithelial cells in vitro [11-13]. The aim of this analysis was to isolate and classify antilithiatic proteins from M. uniflorum seeds, as well as determine their impact on different stages of CaOx inhibition.

Materials and Methods

Plant material

Macrotyloma uniflorum seeds were collected from different locations of Himachal Pradesh (north-western states of India) such as Kullu, Chamba, Rampur, Sundernagar, Dharampur and Palampur. The seeds (HPK-4, HPKM-317, and HPKM-249) from the Palampur region were procured from the Department of Agricultural Biotechnology, CSK, Himachal Pradesh Krishi Vishwavidyalaya H.P., India. All the seeds were authenticated Macrotyloma uniflorum (HP-MU) by Dr. RK Chahota from the Department of Agriculture Biotechnology, CSK Himachal Pradesh Agricultural University, Palampur. The seeds were stored at room temperature.

Protein extraction

Surface-sterilized seeds of M. uniflorum from all locations were soaked in water for 72 h. all the seeds were then crushed and extracted with Tris-buffer [1 gm seed powder was mixed in 3 mL of buffer (pH 8.0) consisting Tris base (0.05 M), polyethylene glycol (1.0%-H 4000), cysteine hydrochloride (0.1%), ascorbic acid (0.1%), citric acid (0.007 M) and β-mercaptoethanol (1 mM)] under cold conditions. The extracted samples were filtered and centrifuged at 12,000 rpm for 20 min at 4°C [Ranjan et al. 2012], and supernatants were assayed for calcium oxalate inhibition potential along with their respective controls. The proteins in all the samples were also estimated by the Bradford method (Bradford 1976).

Electrophoretic techniques

The protein sample showing highest calcium oxalate inhibition activity was then separated by electrophoresis in 12% polyacrylamide gels in the presence of Sodium Dodecyl Sulphate (SDS-PAGE). Extracted and lipolyzed protein samples were added 3:1 to 4X SDS-PAGE sample buffer. The gels were then stained with Coomassie brilliant blue R-250 staining solution. The gels were imaged in a gel documentation system.

Trypsin digestion

A total of 50 μg of proteins from each sample was diluted in 100 mM ammonium bicarbonate to final volume of 100 μl, reduced with 10 mM DTT and alkylated with 55m Miodoacetamide followed by trypsin digestion (Promega, India) for 12-16 h at 37°C. The dried peptide mixture was suspended in 50% trifluoroacetic acid (TFA) containing 0.1% acetonitrile (ACN) buffer. The peptides obtained were mixed with α-cyano-4-hydroxycinnamic acid (HCCA) matrix in 1:1 ratio and resulting 2μL was spotted on the MALDI TOF/TOF ULTRAFLEX III instrument and further analysis was done with FLEX ANALYSIS SOFTWARE for obtaining the Peptide.

Peptide mass fingerprinting

Digested peptide samples were lyophilized (Labconco, USA) and dis- solved in 0.1% triflouro acetic acid /acetonitrile and added to 0.5μl of matrix [α-Cyano-4-hydroxycinnamic acid (Bruker; 20 mg/ ml in 0.1% trifluoroacetic/30% (v/v) ACN (1:2)], dried at room temperature and subjected to MALDI-TOF/TOF proteomics analyzer (UltrafleXtreme TM MS; Bruker Daltonics, Germany). A mass standard starter kit (Bruker Daltonics, Germany) and a standard tryptic BSA di- gest (Bruker Daltonics, Germany) were used for MS and MS/MS cali- b rations. A combined MS and MS/MS were performed using Bio Tools 3.0 software (Bruker Daltonics, Germany). The TOF spectra were recorded in positive ion reflector mode with a mass range from 700 to 3500Da. The two most abundant peptide ions were subjected to fragmentation analysis to determine the peptide sequence. Database search was performed using MASCOT search engine (v.2.1; Matrix Science, London, U.K.). The search parameters in NCBInr were as follows: taxonomy, Viridiaplantae (green plants 25779625 sequences); enzyme, trypsin; fixed modification, carbamidomethyl (C); variable modification, Glu-> pyro-Glu (N-term Q) and oxidation(M); missed cleavage, one; parent ion mass tolerance, 100 ppm; MS/MS mass tolerance, 0.7 Dalton; and MS/MS peak filtering, monoisotropic and M+H+. The identified proteins which met three criteria viz., 1) be among the 5 top hits on the search report, 2) having significant ions scores/sequence coverage or extensive homology (p < 0.05) and 3) proteins matched by a minimum of two peptide sequences were considered as positive identification.

Gene ontology using Blast2Go software

The identified proteins were grouped into functional categories using Blast2GO (v. 2.6.0) [14].

Results and Discussion

MALDI-TOF Mass spectrometric analysis and identification of purified proteins

The crude proteins obtained after extraction of seed samples of Macrotyloma uniflorum from different locations were subjected to MALDI-TOF MS and MASCOT search engine analysis. The mass over charge ratio data obtained from the MALDI-TOF of the peaks A1, B1, B2, C1 matched significantly with nuclear pore anchor, DEAD Box ATP dependent RNA helicase 45, Lon protease homolog 1 and Heat shock protein 90-3, respectively. The amino acid sequence of these respective proteins obtained from MASCOT search was used as an input to search for the presence of active domain using Scan Prosite. Merchant et al., [15] used MS-MALDI and ESI-MS to compare to a Swiss Prot human protein database and a translated human genome database, respectively. They discovered 158 proteins in the stones of five patients. They also discovered large levels of myeloperoxidase and osteopontin, indicating that proteolytic action is involved in stone formation [15] (Table1).

Functional classification

Figure1A depicted the biological and molecular function annotated using Blast2GO. MALDI-TOF-MS identification showed a total of 31 proteins using NCBI green plants. A total of 31 proteins were identified which could be functionally characterized into developmental process, anatomical structure development, tube development and multicellular development. 1-Aminocyclopropane-1-carboxylate synthase (ACC) noticed in Chamba sample. ACC is an enzyme mainly involved in the synthesis of ethylene (Oz, Gulen, & Eris, 2010) and is likely to be part of the sequence of ripening events, with ethylene being a well-recognized trigger for fruit softening and cell wall breakdown, produced during the climacteric [16]. RNA polymerase II is a general transcription factor, present in nucleus and mainly in protein biosynthesis, transcript and transcription regulation. Ribosomal protein L16 (chloroplast) was found in Bharmour sample. Chloroplast r-proteins are encoded within two genetic compartments, the chloroplast and the nucleus and are much conserved structures. Ribosomal protein L16 (chloroplast) protein is mainly involved in mitochondrial translational network.

Genes mainly involved in ribosomal proteins are mainly present in chloroplast genome [17]. Putative tRNA pseudo uridine synthase Pus10-like, partial is metal ion binding protein prominently present in Kullu protein sample which involved in primary miRNA processing and pseudo uridine synthases are responsible for installation of pseudo uridine modification in RNA [18]. Molecular processes involved transporter activity, ion transmembrane transporter activity and carboxylic/ dicarboxylic acid transmembrane activity. Calcium ions (Ca2+) play essential roles in plant growth and development (Figure 1B). Calcium-dependent protein kinase 29-like (CDPK)mainly identified commonly in plants, are a kind of vital regulatory protein deciphering calcium signals triggered by various developmental and environmental stimuli [19]. CDPKs have commonly presented in plants, protists, oomycetes and green algae, but not in animals and fungi [20].

Number of peptides generated in the protein samples of different locations using MALDI-TOF

The protein extracts of seeds of Macrotyloma uniflorum from different locations of Himachal Pradesh were also analyzed for number of peptides generated in MALDI-TOF. The protein sample of Rampur location contained maximum number of peptides followed by Sunder Nagar, Palampur and Kullu. Seeds of Chamba location showed the presence of very few peptides (Figure 2). The kidney stones are known to be composed of mainly calcium oxalate and Macrotyloma uniflorum seed extracts provide a good preface for these stone degradations. The seed protein of Macrotyloma uniflorum with highest calcium oxalate inhibition potential was lipolyzed and then separated through SDS-PAGE technique. The quantification of Macrotyloma uniflorum seed proteins was done with Bradford method and further protein with high oxalate inhibition activity (15µg) was loaded on 12% Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gel according to Laemmli discontinuous system (Figure 3). In the proteomic study of loaded protein, a total of three major low molecular weight protein bands were observed (~27KDa, ~23.1 and ~17KDa respectively). These results will help in correlating the protein composition and bring us closer to treatment for kidney stone degradation. SDS-PAGE technique can be used to identify genotype in the future, as well as to broaden genetic diversity and relatedness in cultivars, as well as to distinguish mutants from their parent genotype based on seed protein [21].

Summary

In plants, MALDI-TOF MS profiling has been limited to metabolite profiles and has only been documented in a few cases [22]. As a result, no archive containing spectra from protein profiles in plant tissues or organs in physiological or altered states exists to date. However, this technique's use in plant proteomic studies may have a big effect, helping with sample differentiation and/or protein marker discovery in agriculture and industry. The discovery of biological markers is valuable areas of research that can help breeders pick cultivars that are best suited to a variety of biotic and abiotic stresses, as well as different stages of growth. Overall, MALDI-TOF MS profiling is an important tool with many possible applications in plant proteomics, such as protein marker discovery, and can contribute significantly to genetic breeding programs and biotechnology. While efforts must be taken to adapt the technique for various plant tissues, our research has demonstrated that this technology is feasible [23]. Also, seed protein SDS-PAGE may be able to also be used to investigate genetic diversity and relatedness in cultivars at a low cost. Furthermore, the proteomic analysis of the seed protein of M. uniflorum aids in the creation of a protein-based marker that can be used to identify cultivars in the future.




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Tuesday, March 12, 2024

Weight Loss in Diabetes: Pharmacological Management and Other Strategies - Juniper Publishers

 Current Research in Diabetes & Obesity - Juniper Publishers


Abstract

Diabetes and obesity are prevalent metabolic disorders that present a significant global public health challenge. This review article provides a comprehensive overview of pharmacological and surgical management strategies for weight loss in diabetes. Diabetes, characterized by high blood glucose levels, and obesity, resulting from excessive body fat accumulation, are closely linked, making effective management essential. Pharmacological interventions for weight loss in diabetes include metformin, which can lead to modest weight reduction in some patients. GLP-1 receptor agonists, such as Semaglutide, liraglutide, and exenatide, offer effective glycemic control and weight loss options. Dual GIP/GLP-1 agonists, particularly Tirzepatide, show promise in treating diabetes and obesity with significant weight loss and glycemic control observed in clinical trials. SGLT-2 inhibitors, like dapagliflozin and empagliflozin, not only manage diabetes but also offer cardiovascular benefits and weight reduction. Amylin mimetics, exemplified by pramlintide, regulate post-meal glucose levels, suppress glucagon secretion, and promote satiety, leading to sustained weight reduction in diabetic patients. Gastric bypass surgery, an option for select individuals with obesity and diabetes, shows significant weight loss and improved metabolic profiles. By understanding these treatment options, healthcare professionals can develop personalized management plans combining lifestyle modifications, pharmacotherapy, and surgery where appropriate. Such approaches can lead to better patient outcomes, including weight loss and improved metabolic control. Ongoing research is essential to explore the long-term efficacy and safety of these interventions to optimize diabetes and obesity management in diverse patient populations.

Keywords: Diabetes mellitus; Obesity, Weight loss; Pharmacological management; Surgical management; Metformin; GLP-1 receptor agonists; Semaglutide; Liraglutide; Exenatide; Dual GIP/GLP-1 agonists; Tirzepatide; SGLT-2 inhibitors; Dapagliflozin; Empagliflozin; Amylin mimetics; Pramlintide; Gastric bypass surgery

Abbreviation: DM2: Type 2 Diabetes Mellitus; GLP-1 RA: Glucagon-like Peptide 1 Receptor Agonist; FDA: U.S. Food and Drug Administration; CKD: Chronic Kidney Disease; SGLT2i: Sodium-Glucose Cotransporter 2 Inhibitors; T2DM: Type 2 Diabetes Mellitus; BMI: Body Mass Index; GLP-1: Glucagon-like Peptide 1; DPP-4: Dipeptidyl Peptidase-4; LRYGB: Laparoscopic Roux-en-Y Gastric Bypass; HDL - High-Density Lipoprotein

Introduction

Diabetes is a metabolic disorder characterized by high blood glucose levels due to inadequate insulin production or impaired insulin action. It can lead to severe complications affecting various organs and systems in the body, such as the heart, kidneys, nerves, and eyes. Obesity, in contrast, is a condition in which excessive body fat accumulates, leading to an increased risk of various health problems, including diabetes [1,2]. The risk factors for diabetes include genetics, sedentary lifestyle, unhealthy eating habits, obesity, and age. Type 2 diabetes mellitus (DM2), the most common form, is closely linked to obesity, as excess body fat can lead to insulin resistance and impair the body’s ability to regulate blood sugar levels [1-3]. The epidemiology of obesity and diabetes is concerning, with both conditions showing a significant increase worldwide. In the United States, approximately 34.2 million people have diabetes, representing about 10.5% of the population.

Obesity in the US is even higher, affecting around 42.4% of adults [3]. Diabetes and obesity rates also rise in other countries, contributing to a global public health challenge [4]. The pathogenesis of weight gain in diabetes is multifactorial. In type 2 diabetes, insulin resistance plays a key role, as the body’s cells do not respond adequately to insulin, leading to decreased glucose uptake and increased fat storage. Moreover, certain medications used to treat diabetes, such as insulin and sulfonylureas, can promote weight gain by stimulating fat storage and reducing appetite control [5]. Complications of obesity in diabetes can be severe and affect various organ systems. Cardiovascular complications include an increased risk of heart disease, stroke, and high blood pressure. Diabetes-related obesity can also lead to complications such as diabetic retinopathy, diabetic neuropathy, kidney disease, and non-alcoholic fatty liver disease [4-6]. Treating obesity in diabetes involves a combination of lifestyle modifications, pharmacotherapy, and sometimes bariatric surgery.

Lifestyle changes, including a healthy diet, regular physical activity, and behavior therapy, are the cornerstone of management. Pharmacological interventions may include medications that promote weight loss or reduce appetite, such as GLP-1 receptor agonists and SGLT-2 inhibitors [1-8]. This article aims to provide a comprehensive overview of pharmacological management and other strategies for weight loss in diabetes. By understanding the definition, risk factors, epidemiology, pathogenesis of weight gain, complications, and treatment options for obesity in diabetes, healthcare professionals can develop practical approaches to managing these interconnected conditions and improve patient outcomes.

Metformin

Metformin is an oral anti-diabetic medication commonly prescribed for managing DM2. Its mechanism of action involves reducing hepatic glucose production, enhancing peripheral insulin sensitivity, and increasing glucose uptake by skeletal muscle. Metformin does not stimulate insulin secretion from the pancreas [9-11]. The typical dose ranges from 500mg to 2,000 mg per day, divided into two or three doses [10,11]. It is usually taken with meals to minimize gastrointestinal side effects. The duration of treatment is generally long-term, as metformin is considered a foundational therapy for type 2 diabetes [9-13]. Metformin is indicated for patients with type 2 diabetes, particularly those who have not achieved glycemic control with diet and exercise alone. It can be used as monotherapy or in combination with other oral anti-diabetic medications or insulin. The drug is generally welltolerated, but common adverse effects include gastrointestinal disturbances such as nausea, diarrhea, and abdominal discomfort.

Metformin is contraindicated in patients with renal impairment, significant hepatic disease, or a history of lactic acidosis [11-13]. Regarding its efficacy for weight loss, metformin may lead to modest weight reduction in some individuals with type 2 diabetes, especially those who are overweight or obese. However, the extent of weight loss is generally mild, and it should not be prescribed solely for weight management [10,11]. The FDA approves metformin and has been widely used for many years. It is generally considered safe in pregnancy, but monitoring blood glucose levels closely during pregnancy is crucial, as the dosing may need adjustment. It is not typically used in children younger than 10 years old; safety and efficacy in pediatric populations require careful consideration [11]. In terms of cost, metformin is relatively affordable and available in generic formulations, making it a cost-effective option for diabetes management.

GLP-1 Agonists

Semaglutide

Semaglutide is a 31-amino acid peptide hormone that mimics native GLP-1. It resists degradation by dipeptidyl peptidase-4 (DPP- 4) and has reduced renal clearance, leading to a prolonged plasma half-life. As a GLP-1 receptor agonist (GLP-1 RA), Semaglutide regulates glucose homeostasis by increasing insulin secretion and decreasing glucagon secretion in a glucose-dependent manner. It also delays gastric emptying and lowers both fasting and postprandial blood glucose levels, resulting in weight reduction. Semaglutide activates sites in the hypothalamus to control energy intake and reduce food cravings. The recommended dose for weight management is 2.4mg injected subcutaneously once weekly. However, it carries a black box warning against use in patients with a history of medullary thyroid carcinoma or pancreatitis, and it should not be used during pregnancy [14,15].

Liraglutide

Liraglutide is a modified GLP-1 receptor agonist that binds to serum albumin non-covalently, leading to a longer half-life of 11 to 15 hours. It enables 24-hour glycemic control with once-daily dosing. Liraglutide improves fasting and post-prandial glycemic control through increased insulin secretion, reduced glucagon levels, and minor gastric emptying delays. Additionally, it reduces appetite and energy intake and has favorable effects on postprandial lipid profiles. The FDA approved liraglutide for treating type 2 diabetes with a daily injectable dose of 1.8 mg and for chronic weight management with a 3.0 mg daily injectable dose. Liraglutide was found to reduce fasting blood glucose, hemoglobin A1c, and systolic blood pressure [16].

Exenatide

Exenatide is an incretin mimetic agent, a synthetic analog of GLP-1, derived from salivary secretions of the lizard Heloderma. As an adjunctive therapy for type 2 diabetes, exenatide improves glycemic control by stimulating glucose-dependent insulin secretion, suppressing elevated glucagon levels, delaying gastric emptying, and reducing food intake. It mainly undergoes elimination through glomerular filtration. The recommended dosing is 1.0 mg subcutaneously twice daily. Adverse events, including mild to moderate nausea, have been reported in clinical trials. Exenatide has shown significant improvements in glycemic control and modest weight loss in patients receiving add-on therapy with metformin or sulfonylurea [17,18]. Overall, these GLP-1 receptor agonists, including Semaglutide, liraglutide, and exenatide, offer effective options for the management of type 2 diabetes and weight reduction in appropriate patients, with varying dosing regimens and administration routes.

Dual GIP/GLP-1 Agonists

Tirzepatide

Tirzepatide is a synthetic peptide that acts as a dual gastric inhibitory polypeptide (GIP) and glucagon-like peptide 1 (GLP- 1) receptor agonist. It consists of 39 amino acids and stimulates insulin release from the pancreas, reducing hyperglycemia in individuals with type 2 diabetes mellitus (T2DM). Additionally, it increases adiponectin levels and lowers appetite, making it more effective in controlling hyperglycemia and inducing weight reduction compared to GLP-1 agonists alone. The drug is administered via subcutaneous injection and is available in different dosages.

The standard dosing is once weekly, starting with an initiation dose of 5 mg/0.5 mL, which can be adjusted based on efficacy and adverse effects. The most common adverse drug reactions are gastrointestinal, such as abdominal discomfort and nausea, which may influence the dosing titration based on patient tolerance. Tirzepatide is FDA-approved for T2DM treatment but not for type 1 diabetes or pancreatitis. It can also be used off-label for obesity management, similar to GLP-1 medications like Semaglutide [19- 21].

Regarding safety, Tirzepatide has been shown to be generally well-tolerated, with gastrointestinal adverse effects being the most common. Nausea, diarrhea, and decreased appetite are frequently reported, and constipation and vomiting are reported less frequently. It can also cause delayed gastric emptying, potentially affecting the absorption of other oral medications and leading to reduced efficacy of oral contraceptives. Other reported adverse effects include sinus tachycardia, acute kidney injury, hypersensitivity reactions at the injection site, and worsening of diabetic retinopathy in some patients. There is a risk of hypoglycemia, especially in patients on insulin therapy or sulfonylureas, and the drug is contraindicated in individuals with a personal or family history of medullary thyroid carcinoma or multiple endocrine neoplasia syndrome type 2 [22-29].

Tirzepatide offers significant potential in treating T2DM and obesity, with double-digit weight loss and glycemic control seen in clinical trials. A growing body of evidence from various clinical trial programs, such as SURPASS and SURMOUNT, supports the drug’s efficacy and safety. Further research is needed to assess its long-term cardiovascular safety and effectiveness and its impact on other cardiometabolic complications in people with T2DM and obesity. The FDA has approved Tirzepatide as Mounjaro® for improving glycemic control in adults with T2DM. Still, it is not indicated for type 1 diabetes and is not recommended for use in children under 18 years old [30,31].

Regarding use during pregnancy and lactation, there is insufficient data to determine the drug-related risk for pregnant women, but animal data suggests that Tirzepatide may cause fetal harm. Therefore, its use in pregnant women is not recommended. As for breastfeeding, there is limited information on the clinical use of Tirzepatide during lactation. The drug is a large peptide molecule with low absorption potential in infants, but more data is needed to fully understand its safety during breastfeeding. Until further evidence becomes available, caution should be exercised when using Tirzepatide during breastfeeding, especially in nursing newborns or preterm infants [32].

SGLT-2 Inhibitors

Sodium-glucose cotransporter 2 inhibitors (SGLT2i) represent a novel class of hypoglycemic drugs that includes dapagliflozin, empagliflozin, canagliflozin, ertugliflozin, and Sotagliflozin. These drugs work by lowering the renal glucose threshold and increasing urinary glucose excretion, thereby improving glycemic control [33]. Besides their antidiabetic effects, SGLT2i has demonstrated benefits in cardiovascular conditions such as heart failure, myocardial infarction, hypertension, cardiomyopathy, and arrhythmia [34]. Moreover, evidence supports their use in diabetic individuals with chronic kidney disease (CKD) and nondiabetic populations with CKD [35,36].

The mechanism of action of SGLT2i involves their competitive binding to SGLT2 proteins in the proximal segments (S1) of the renal tubules. The drugs reduce glucose reabsorption by inhibiting these cotransporters, increasing urinary glucose excretion and decreasing plasma glucose levels. Unlike other antidiabetic medications, SGLT2i does not stimulate pancreatic β cells to secrete insulin, making them insulin-independent and associated with a low risk of hypoglycemia. However, it’s worth noting that their efficacy may decrease in individuals with reduced renal function [37-39].

In addition to their antidiabetic effects, SGLT2i have shown beneficial effects on endovascular function by indirectly reducing oxidative stress and inhibiting proinflammatory mediators [40]. These drugs also improve myocardial metabolism by inhibiting sodium-hydrogen exchanger 1 isoform in the myocardium, reducing calmodulin-dependent kinase II activity, and enhancing mitochondrial calcium levels, ultimately improving cardiac efficiency [39-41]. Furthermore, SGLT2i have been found to improve renal function, reduce adipose-mediated inflammation and sympathetic overdrive, modulate the intrarenal reninangiotensin system, and increase erythropoietin levels, which positively impacts vascular progenitor cells [41].

Sodium-glucose cotransporter 2 inhibitors have also demonstrated significant efficacy in promoting weight loss in addition to their antidiabetic and cardiovascular benefits. This weight loss effect is particularly advantageous for individuals with obesity or overweight, making SGLT2i a favorable option for those aiming to reduce body weight [42]. The weight loss mechanism of SGLT2i is attributed to their ability to increase urinary glucose excretion. As glucose is excreted in the urine, it leads to the loss of calories, resulting in a reduction in body weight [10]. Studies have shown that treatment with SGLT2i is associated with clinically meaningful weight loss in diabetic and non-diabetic patients with obesity [42,43].

In clinical trials, SGLT2 inhibitors, such as dapagliflozin and empagliflozin, have significantly reduced body weight compared to other antidiabetic medications or placebo [43,44]. This weight loss effect can be particularly beneficial for individuals with type 2 diabetes who often struggle with overweight or obesity, as it can contribute to improved glycemic control and reduced cardiovascular risk factors [42,43]. The combination of glycemic control, cardiovascular benefits, and weight loss makes SGLT2i a valuable therapeutic option for patients with type 2 diabetes and obesity. However, it’s essential to consider individual patient characteristics and potential side effects when determining the most suitable treatment approach.

Close monitoring and personalized management are crucial to optimize the outcomes and safety of SGLT2 inhibitors in diverse patient populations [43-45]. Overall, SGLT2 inhibitors have proven to be promising medications not only for the management of diabetes but also for their beneficial effects on cardiovascular, renal, and weight loss conditions. Their unique mechanisms of action set them apart from other antidiabetic drugs and offer potential advantages in terms of safety and cardiovascular risk reduction. Further research is ongoing to explore their full potential and role in treating various metabolic and cardiovascular disorders.

Amylin Mimetics

Pramlintide

The drug is a synthetic analog of amylin, a peptide hormone produced by the pancreas. It differs from native amylin in three amino acid substitutions and is a stable and soluble analog administered via subcutaneous injection at mealtime. Pramlintide regulates post-meal blood glucose levels by slowing gastric emptying, suppressing abnormal postprandial glucagon secretion, and promoting satiety, which leads to reduced caloric intake [46-50]. The glucose-dependent mechanism of pramlintide prevents hypoglycemia without therapies that cause it and helps exogenous insulin therapy better match physiologic needs.

The drug is only approved for use in patients with type 1 and type 2 diabetes who are taking prandial insulin. Dosing varies depending on the type of diabetes, and patients may require adjustments in pre-meal insulin doses to achieve euglycemia. The primary adverse events associated with pramlintide are nausea, vomiting, and anorexia, which are more common in patients with type 1 diabetes and tend to diminish over time. There is an increased risk of severe hypoglycemia, especially in patients with type 1 diabetes, when pramlintide is started at full doses without reducing insulin doses. Pramlintide has also been associated with migraine-like symptoms, likely due to the activation of amylinresponsive receptors in various body parts [47-54].

One significant benefit of pramlintide therapy is its association with weight loss. Clinical studies have shown that improved glycemic control with pramlintide is correlated with sustained and significant reductions in body weight. The drug induces satiety and decreases caloric intake, possibly contributing to weight loss. In combined data from pramlintide clinical trials, patients with type 1 diabetes experienced weight reductions of approximately 1.5 kg at 25 weeks, while patients with type 2 diabetes saw reductions of 1.2 kg compared to placebo-treated patients, who had slight weight gains.

Weight reductions were sustained for up to 52 weeks in longterm trials for both types of diabetes [50-56]. Regarding lactation, pramlintide has a high molecular weight and a short half-life, making it unlikely to pass into breast milk in clinically significant amounts. Additionally, it is a peptide likely to be digested in the infant’s gastrointestinal tract, further reducing the potential for reaching clinically important levels in the infant’s serum. However, since there is limited information on using pramlintide during breastfeeding, an alternate drug may be preferred to ensure the safety of the nursing infant [57,58].

Gastric Bypass Surgery

Gastric bypass surgery involves creating a small stomach reservoir and a gastrojejunoanastomosis using a defunctionalized Roux Y loop to alter digestion. It is recommended as an option for the treatment of type 2 diabetes in selected surgical candidates with a BMI ≥40 kg/m² (or ≥37.5 kg/m² in Asian individuals) and in adults with BMIs between 35.0 and 39.9 kg/m² (or 32.5– 37.4 kg/m² in Asian Americans) who have not achieved lasting weight loss or improvement in comorbidities with non-surgical methods. Metabolic surgery may also be considered for adults with a BMI of 30.0–34.9 kg/m² (or 27.5–32.4 kg/m² in Asian American individuals) who have not succeeded with non-surgical methods [59]. While there are no absolute contraindications to bariatric surgery, relative contraindications exist. These include severe heart failure, unstable coronary artery disease, end-stage lung disease, active cancer treatment, portal hypertension, drug/ alcohol dependency, impaired intellectual capacity, and Crohn’s disease, in the case of LRYGB. Additionally, gastric bypass surgery is generally contraindicated in patients with uncontrolled medical conditions such as severe cardiovascular disease, uncontrolled hypertension, or uncontrolled diabetes. Patients with active substance abuse and severe mental health disorders may not be suitable candidates due to hindrances in post-operative recovery and adherence to lifestyle changes. Pregnant or planning pregnant women, individuals with previous gastrectomy or bowel resection, young adolescents, and patients at high surgical risk may also not be ideal candidates for the procedure [60].

Studies have demonstrated the efficacy of gastric bypass surgery in weight loss for individuals with a BMI greater than 35 kg/m² and those with a lower BMI. Randomized clinical trials and observational studies have shown significant weight loss in gastric bypass groups compared to non-surgical treatment groups. The surgery has also been associated with superior glycemic control, cardiovascular risk reduction, decreased incidence of microvascular diseases, improved quality of life, and reduced risk of cancer and other associated risks. Gastric bypass surgery has been found to lower triglyceride levels, increase high-density lipoprotein (HDL) cholesterol, and reduce the need for insulin in type 2 diabetes patients [61-64]. In some cases, metabolic surgery has shown potential benefits for individuals with type 1 diabetes, but larger and longer studies are needed to confirm its role in these cases. Despite the high initial costs, some analyses have suggested that metabolic surgery may be cost-effective for people with type 2 diabetes, depending on assumptions about its long-term efficacy and safety [65]. Overall, gastric bypass surgery appears to be a valuable treatment option for selected individuals with obesity and type 2 diabetes, offering significant weight loss, improved metabolic profiles, and reduced cardiovascular risks [66].

Conclusion

The treatment of weight loss in individuals with diabetes requires a comprehensive approach. Pharmacological interventions, such as metformin, GLP-1 receptor agonists, SGLT- 2 inhibitors, and amylin mimetics, can play a significant role in weight reduction and glycemic control. Additionally, dual GIP/ GLP-1 agonists, such as tirzepatide, demonstrated significant potential in treating type 2 diabetes and obesity, offering considerable weight loss and glycemic control. However, further research is needed to assess their long-term cardiovascular safety and the impact on other cardiometabolic complications.

Gastric bypass surgery is a valuable treatment option for selected individuals with obesity and type 2 diabetes, offering significant weight loss and improved metabolic profiles. It has been associated with superior glycemic control, cardiovascular risk reduction, and improved quality of life. However, it is essential to carefully select suitable candidates for surgery and consider potential contraindications and risks. Personalized treatment plans, lifestyle modifications, and continued research are needed to optimize outcomes and improve management strategies for weight loss in individuals with diabetes.

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Monday, March 11, 2024

Uncovering the Effects of the Southwest Monsoon on Fishing Activity in the Indian Ocean with VIIRS Boat Detection Data - Juniper Publishers

 Oceanography & Fisheries - Juniper Publishers


Abstract

In 2015, the Earth Observation Group (EOG) created the VIIRS Boat Detection (VBD) product, which is now employed by various fishery agencies to oversee fishing activities and ensure compliance with closures. VBD data offer real-time insights into the movements of fishing vessels, aiding in the detection of illegal fishing practices within restricted areas and intrusions into exclusive economic zones (EEZs). The Indian Ocean, a critical hub for global fisheries contributes over 14% of the world’s wild-caught fish production. The fishing grounds in the Indian Ocean region are amorphous clusters, which depict cyclical patterns in the number of boat detections, corresponding to the monsoon season from June to September, and fishing restrictions imposed by the government. An exponential increase in boat detections was also observed since October 2016. Contrarily, the vessels detected at the anchorage are predominantly shipping vessels, and their temporal pattern shows less pronounced cyclical variation compared to fishing grounds. Employing the ‘Pelt’ algorithm to detect change points in monthly boat counts for the anchorage from 2012 to 2022 revealed a significant change in the average number of boat detections per month in October 2018. This study highlights the utility of VIIRS boat detection data in monitoring fishing activity and its response to seasonal and regulatory influences in the Indian Ocean, ultimately contributing to more effective fisheries management and conservation efforts.

Keywords: VBD; Fishing grounds; Anchorages; Monsoon; Indian Ocean

Abbreviations: EOG: Earth Observation Group; VBD: VIIRS Boat Detection; EEZs: Exclusive Economic Zones; DMSP: Defense Meteorological Satellite Program; OLS: Operational Linescan System; VIIRS: Visible Infrared Imaging Radiometer Suite; JPSS: Joint Polar Satellite System; USAID: United States Agency for International Development; VMS: Vessel Monitoring System; AIS: Automatic Identification System; VNF: VIIRS Night fire

Introduction

Since the 1970s, it has been established that fishing boats emitting strong light can be identified using nighttime visible low-light imaging data obtained by polar-orbiting meteorological sensors [1]. Among the sensors capable of low-light imaging, two notable ones are the U.S. Air Force Defense Meteorological Satellite Program (DMSP) Operational Linescan System (OLS) and the NASA/NOAA Visible Infrared Imaging Radiometer Suite (VIIRS). These sensors employ light enhancement techniques to enable the detection of moonlit clouds during the night, meeting the requirements of meteorologists for both daytime and nighttime visible and thermal cloud imagery.

The OLS digital archive spans from 1992 to the present, offering relatively coarse resolution (2.7km ground sample distance) global data. On the other hand, the VIIRS sensor, in operation since 2012, provides significant advancements in low-light imaging. It boasts finer pixel resolution (742m × 742m) and in-flight calibration to radiance units [2]. In 2015, the Earth Observation Group (EOG) developed the VIIRS boat detection (VBD) algorithm with support from NOAA’s Joint Polar Satellite System (JPSS) with aid from the United States Agency for International Development (USAID) [3]. VBD data are generated in near real-time, with nightly records available for Southeast Asia since April 2012, and globally from 2017 onwards. In addition to the nightly data, EOG also produces monthly and annual summary grids.

Numerous fishery agencies, including those in the Philippines, Indonesia, Thailand, Japan, and South Korea, rely on VIIRS boat detection (VBD) data for monitoring fishing activities and identifying vessels operating in restricted areas [4]. VBD provides valuable maritime domain awareness, especially when it comes to identifying locations of “dark” vessels which lack Vessel Monitoring System (VMS) or the Automatic Identification System (AIS) [5]. To delineate fishing grounds and assess usage patterns over time, the monthly and annual summary grids are very useful. Recently, a global long-term VBD cumulative detection grid, spanning 2012-2021 has been developed and the findings will be shared in this report.

Methods

Besides, the monthly and annual summary grids, all the nightly VBD data have been stored in a database designed for easy retrieval based on criteria specified by users. Each pixel in the VBD data is assigned a label based on its clarity and location, often referred to as quality flags or QF. To construct a multiyear cumulative detection grid, pixels labeled QF 1, 2, 3, 8, and 10, corresponding to strong boat detection, weak detection, blurry detection, recurring detection, weak and blurry detections have been selected [4,6].

The accumulations of these detections have been aggregated into a global grid with 15 arc-second grid cells, which are approximately half a kilometer on each side at the equator. These grids provide information on the number of detections, the count of valid VIIRS observations, and the average radiance for each grid cell. To account for differences in data availability over various time periods in Southeast Asia and other parts of the world, the percentage frequency of detection is calculated by dividing the number of detections by the count of valid observations. This normalization ensures a fair comparison between these regions.

Vector polygons of ‘fishing grounds’ and ‘anchorages’ were drawn on the percentage frequency VBD grid. The anchorages have definite shapes, like a square or rectangle, whereas fishing grounds are closer to those anchorages and structureless. Some of the fishing grounds also coincide with ‘upstream’ gas flare locations detected in the VIIRS Night fire (VNF) dataset which have been eliminated before extracting temporal profiles [6]. EOG has developed methods for extracting temporal profiles from the database for specific spatial features identified in the summary grids.

Results

The Indian Ocean hosts some of the world’s most significant fisheries, contributing to more than 14% of the global catch of wild-caught fish [6]. Fishery agencies have the potential to depend on VIIRS boat detection (VBD) data to enforce regulations and implement management measures effectively. VBD data can offer real-time insights into the actions of fishing vessels, including signs of unlawful fishing practices within restricted zones and intrusions into exclusive economic zones (EEZs) [7]. Figure 1 shows the percent frequency of VBD detection in the northern Indian Ocean. The fishing grounds show up as large light gray features. The fishing grounds in the Indian Ocean seems to be in amorphous clusters [8]. Although fishing grounds encompass the largest expanse with VBD detections, the percentage of detection for an individual half-kilometer grid cell within these fishing grounds is quite low, seldom surpassing 1%. However, there are concentrated groups of non-fishing VBD features, primarily located near the shoreline, which exhibit higher percentage frequencies of detection, typically ranging from 1% to 3%. These non-fishing features comprise anchorages, platforms, and aquaculture sites. Figure 2 shows the temporal profile of fishing ground seen in figure 1.

The equation of the exponential curve shown in Figure 2 is:

y = 4E −15e0.0009x

The temporal profile of the fishing ground clearly demonstrates a cyclical seasonal pattern and a steady increase of fishing activity since October 2016. The decline in VBD in the fishing grounds during the monsoons (June -September) is not simply the effects of cloud obscuration but aversion of the fishing boats to the inclement weather and rough seas. Besides, fishing communities cease their fishing operations in concordance with government regulations, which ban commercial fishing during the monsoons to allow breeding and restoration of fishing stock [9-11]. The monsoon is the spawning season for fish and the ban helps in replenishing the fish stock and protect the fishermen from the rough sea. It is interesting to see the differences in the temporal profiles of the fishing ground in Figure 2 and the anchorage in Figure 3. The boats detected from the largest anchorage (Figure 3) are shipping vessels - not fishing boats. The anchorage’s temporal profile indicates that shipping activities are not heavily impacted by the monsoon. The cyclical pattern, although apparent, is not as distinct as it is for the temporal profile of the fishing ground.

To identify potential variations in the average number of anchored boats, a kernel-based offline multiple change point detection algorithm known as ‘Pelt’ was employed, which stands for “Pruned Exact Linear Time” [12]. By applying the ‘Pelt’ algorithm to the time series data of monthly boat counts for the anchorage from 2012 to 2022, only one significant change in the average number of boats was detected. This change occurred in October 2018, where the average number of boats per month shifted from 66 to 111. The average line for 66 is shown in red, and the average line for 111 is shown in green.

Conclusion

At night, the ocean presents itself as an extensive, dark canvas where any form of illumination becomes easily detectable by the VIIRS day/night band. However, the VIIRS boat detection data, despite being generated on a nightly basis, do not provide any indication of these sharply defined structures. The true nature of these structures becomes apparent when we examine them over a longer timeframe, accumulating detections across multiple years. In the Indian Ocean, due to south-west monsoon we see seasonal variations in fishing grounds, but the anchorages do not indicate the same. Moreover, the ban on fishing activity during the monsoons also has a significant effect and thus a distinct cyclical pattern of detections are noticed for the fishing grounds. Also, an exponential growth of fishing activity was noticed since October 2016. As for the profile of the largest anchorage, a rise in the average number of fishing boats was noticed in October 2018. It will be interesting to compare and study fishing grounds around the world which show similar rhythms.

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Thursday, March 7, 2024

Characterization of Headache in Patients with Temporomandibular Joint Disorders - Juniper Publishers

Dentistry & Oral Health - Juniper Publishers


Abstract

Objective: The incidence and characteristics of headache in patients with temporomandibular joint disorder (TMJD) had not been described previously. The aim of the current study was to discuss this issue.

Materials and methods: The study involved 200 patients with TMJD and 200 control subjects. The incidence and characteristics of headache, as well as associated symptoms were compared between the two groups.

Results: The incidence of chronic headache was significantly higher in the TMJD patient group (88%) than in the control group (37%); (P=0.001). The headache patterns included unilateral throbbing migraine headache, as well as bilateral regional headache.

Conclusion: The results lend support to the theory of overactivity of pericranial muscles in TMJD, as well as extracranial –intracranial cross-talk in sensory nociceptive nerve fibers.

Keywords: Headache; Temporomandibular Joint Disorder; Chronic pain

Abbreviations: TMJD: Temporomandibular joint disorder; CNS: Central nervous system; TTH: Tension-type headache; TMD: Temporomandibular disorder; TMJs: Temporomandibular joints; DC: Diagnostic Criteria; MRI: Magnetic resonance imaging; ICHD: International Classifications of Headache Disorders; CT: Computed tomography

Introduction

The pathogenesis of headaches, especially primary headaches, is probably multifactorial. Various structures in the cranium have been implicated in the causation of headache; with central nervous system (CNS) plastic changes contributing to the chronicity of the condition [1,2]. The cranial calvarial bones are covered from outside by skin and pericranial myofascial structures. Myofascial trigger points are prevalent in both tension-type headache (TTH) and migraine [3,4]. Underneath the calvarial bones is the dura mater. The dura mater has a rich supply of nociceptors and mechanoreceptors; both of which are implicated in the pathogenesis of headaches [5,6]. Animal studies, as well as human imaging studies, have stressed the importance of sterile dural inflammation as a universal finding in primary headaches; thereby sensitizing the receptors [7,8]. The dural inflammation is associated with an increased barrage of action potentials, along nociceptive fibers to the trigeminal ganglion [9,10]. Furthermore, non-trigeminal nociceptive afferents from the occipital dura mater and cervical structures contribute to headaches; being mediated through the upper three cervical nerves [11,12]. Nociceptive afferents from head and neck structures converge onto the subnucleuscaudalis in the brain stem and upper cervical spinal cord [13].

Temporomandibular disorder (TMD) is an umbrella term for pain and dysfunction involving the masticatory muscles and the temporomandibular joints (TMJs) [14]. Most studies on TMD patients report that headache is a frequently associated symptom [15-17]. The Diagnostic Criteria for Temporomandibular Disorders (DC / TMD) considers headache as one of the diagnostic categories of TMD; in addition to TMJ and masticatory muscle disorders [18]. Temporomandibular joint disorder (TMJD) represents a major category of TMD; although, occasionally, it is not associated with painful symptoms [19]. Radiological studies, especially magnetic resonance imaging (MRI) have confirmed the various clinical signs elicited in patients with TMJD [20]. Pathophysiologically, a dysfunctional, abnormal TMJ cannot be considered as an isolated disorder. Many factors, including bruxism, malocclusion, lateral pterygoid muscle dysfunction (which is centrally-mediated) contribute to the joint dysfunction [21,22]. These factors cause abnormal stresses on the TMJ, which are associated with ligament laxity, a low-grade joint inflammation, and deterioration in joint lubrication; all of which predispose to a dysfunctional joint [23,24]. Pain in TMJD is an intringuing issue, and there is often a poor correlation between the degree of pain over the joint and the degree of joint pathology [25,26]. A recent systematic review on structural and functional brain MRI studies revealed aberrant CNS changes in TMJD with pain. The CNS aberrations involved sensory, motor, and affective regions in the brain. Interestingly, the imaged brain aberrations were frequently resolved after intra-oral splint therapy [27].

From a clinical perspective, headache disorders tend to be under-recognized and under-treated, especially in developing populations [28]. The reason for this is that the diagnosis of primary headaches, which is clinically based, frequently does not fulfill the strict criteria provided by the International Classifications of Headache Disorders (ICHD). In these situations, the diagnoses of (probable TTH) and (probable migraine) are often made [29,30]. Furthermore, TTH and migraine frequently coexist [31]. In contrast to the rather diffuse head pain in primary headaches, TMJD pain is localized infront of the ear, being frequently associated with joint noises and functional limitations [32,33]. Additionally, TMJD is commonly associated with otoneurological and alleged sinus symptoms; known as Costen’s syndrome [34-36]. Although the headache associated with masticatory myofascial pain had been previously characterized, headache associated with TMJD had not been previously specifically addressed. Notably, headache associated with masticatory myofascial pain most commonly resembles TTH with a bilateral temporal location [37-39]. On the other hand, the TMJ contains; in addition to nociceptors, mechanoreceptors (proprioceptors), which guide mandibular movements through their inputs to the central pattern generator in the brain stem [40]. The distribution of the proprioceptors in the joint is altered in cases with joint pathology [41]. According to the (gate control theory), altered proprioceptive input to the CNS can alter pain sensation experienced in patients with TMJDs [42]. In addition to exaggerated nociceptive input from the TMJ in TMJD, altered proprioceptive input to the CNS may also contribute to CNS plastic changes causing a peculiar type of pain, in addition to abnormal jaw mechanical behavior [43]. From these perspectives, the aim of the current study was to characterize the incidence and pattern of headache in patients with documented TMJD. As far as the author is aware, and after a Pub Med literature search from 1970 till 2023, this issue had not been previously addressed.

Materials and Methods

The current study was a clinical study conducted at the craniofacial pain clinic at El-Sahel Teaching Hospital in Cairo, Egypt. The study period was 6 months, starting on the 1st of January 2023. Ethical approval was obtained from the Ethics Committee of the General Organization for Teaching Hospitals and Institutes in Cairo (Approval number: 83- 2022). Informed consent was obtained from the patients and control subjects to participate in the study. The patient group consisted of consecutive adult subjects with chronic jaw pain, who reported the experience of joint sounds during mandibular movements. The presence of joint sounds was later confirmed on examination. This criterion allowed objective evidence of TMJD for inclusion in the study. Patients who had jaw pain (arthralgia and/or myalgia) without joint sounds were not included in the patient group. The control group consisted of adult relatives of the patients presenting to the clinic, who experienced no sounds elicited on movement of the mandible. The absence of joint sounds in the control group was later confirmed on examination, revealing clinical evidence of normal TMJs. Exclusion criteria in both groups consisted of any acute disorder in the head and neck, as well as previous macrotrauma to the jaw.

A questionnaire and examination findings sheet were filled in by the author. The main criterion investigated in the current study was the reporting of chronic headache for at least 6 months in both cohorts of participants. In the participants suffering from headache, the characteristics of the head pain were noted according to the description of the subject (pressing or throbbing). Furthermore, the laterality of the head pain was noted (unilateral versus bilateral). The most commonly invloved region in the head was addressed (diffuse, frontoorbital, temporal, occipital, or at the vertex). The presence of otalgia, facial pain, and/or neck pain in association with the headache was reported. Questioning also included the presence or absence of autonomic symptoms accompanying the headache, including nasal congestion, rhinorrhea, and/or lacrimation. Additionally, conditions possibly predisposing to TMJD were asked about (bruxism and dental status). Relevant to TMD, a diagnosis of depression was noted if present. For the reporting of depressive symptoms, the patient had to have been diagnosed by a psychiatrist or reported daily weeping. A previous history of open-lock was addressed. Otological symptoms pertaining to Costen’s syndrome (subjective hearing loss, subjective tinnitus, and/or vertigo) were explored.

Examination of the subjects followed a standard protocol. The TMJ was palpated, noting for any exquisite tenderness (by observing a palpebral reflex). Tenderness over the TMJ was assigned as arthralgia. Any TMJ noises elicited during opening and closing the mouth were classified as click (snapping noise), or crepitus (grating noise). An audible noise was confirmed by palpation of the joint. The presence of joint noises was used to categorize the subject as a TMJD patient. Next, the temporalis and masseter muscles were palpated, noting for any exquisite tenderness (eliciting a palpebral reflex), and any referral pattern for the pain (denoted as myalgia). The oral cavity was observed for the dental status; and accordingly, the subjects were classified as having normal dentition, being partly edentulous, or totally edentulous. While the mouth was fully opened, any trismus was noted (inter-incisal distance less than 40 mm). While the mouth was closed, any open-bite deformity was observed. Any gross deviation of the mandible during opening and closing the mouth was noted. Following examination of the stomatognathic system, examination of the ears, nose, pharynx, and eyes was performed.

Statistical analysis

Results were expressed as mean ± standard deviation or number percent (n %). Comparison between categorical data (n %) was performed using Chi square test. Comparison between mean values of age in the two groups was performed using unpaired t test. Statistical analysis was performed using SPSS computer program (version 19 windows). P value ≤ 0.05 was considered significant.

Results

The current study comprised 200 TMJD patients and 200 control subjects. The mean age of the patient group was 40.34 ± 12.36 years, while the mean age of the control group was 39.1 ± 12.38 years (P= 0.319). There was no significant difference in the male-to-female ratio between the two groups (Table 1). The incidence of chronic headache in the patient group (88%) was significantly higher than in the control group (37%); (P= 0.001). Migraine headache was unilateral and throbbing. Its incidence was significantly higher in the patient group (13%) than in the control group (6%); (P= 0.017). The incidence of bilateral regional headache was significantly higher in the patient group than in the control group for temporal, fronto-orbital, and occipital locations (Table 2). Table 3 reveals clinical features pertinent to TMJD in patients versus controls. Notably, clicking was noted in 85 % of TMJD patients, while crepitus was elicited in 15 % of TMJD patients. The control subjects showed clinically normal TMJs. All other symptoms and signs related to the stomatognathic system had higher incidence in TMJD patients versus control subjects (Table 3). Table 4 reveals the incidence of symptoms related to Costen’s syndrome in patients and control subjects. All otological and alleged sinus symptoms had a higher incidence in TMJD patients versus controls.

Data are expressed as mean ± SD or number (%).

TMJD= temporomandibular joint disorder.

p> 0.05= not significant.

Discussion

Temporomandibular joint disorder (TMJD) involves a set of functional and pathological changes affecting the TMJ [44]. As the two TMJs cannot function independently during mandibular movements, TMJD is very frequently a bilateral disorder [45]. The current concept is that the pathogenesis of TMJD involves excessive or abnormal stresses on the TMJ, that exceed the adaptive capacity of the joint tissues [46]. These abnormal stresses acting on the TMJ lead to laxity of the joint ligaments, thereby predisposing to disc displacement [47]. Furthermore, an associated low-grade inflammation in the TMJ has been reported to initiate a cascade of events, including impaired lubrication and increased friction coefficient in the joint [48,49]. The result of increased friction in the joint is a disturbance in disc-condyle complex harmony and predisposition to disc displacement and degenerative joint disease [50,51]. Moreover, increased tension in the lateral pterygoid muscle had been postulated to account for the most common form of disc displacement (anteromedially) [52]. Clinically, reducing disc displacement manifests as clicking, whereas degenerative joint disease manifests as crepitus [53]. In the current study, the TMJD patient group demonstrated clicking in 85% of the patients, whereas crepitus was present in 15% of the patients. In the control group, there was absence of joint noises during mandibular movement. A painful, tender TMJ is designated as TMJ arthralgia. The current view is that the degree of TMJ arthralgia is correlated with the levels of inflammatory mediators and neuropeptides, which sensitize and activate afferent nociceptive nerve fibers in the joint [54,55]. The current study revealed TMJ arthralgia in 75.5% of patients, versus 9% of control subjects (P= 0.001).

A continuous barrage of action potentials from the sensitized and activated TMJ nociceptors contribute to the chronicity of TMJ pain [56]. In pain literature, chronic pain is defined as a multidimensional experience involving negative sensory, affective, and cognitive aspects [57]. These aspects are associated with corresponding abnormal structural and functional brain regions, seen on neuroimaging [58]. Currently, there are two proposed theories for these brain changes in chronic painful TMJD. The first theory involves sensitization and activation of CNS pathways relevant to chronic pain in its three dimensions, consequent on a continuous barrage of action potentials from the peripheral TMJ nociceptors [59,60]. The alternative hypothesis is the activation of a pre-set brain pain neuromatrixby stress and peripheral pain [61-64]. In both theories, the result of chronic painful TMJD is the unwanted experience of pain, alteration in homeostatic mechanisms; and importantly, an altered motor output to the masticatory and cranial muscles [65-67].

The altered motor output to these muscles contributes to trigger points, commonly observed in patients with TMJD associated with headache [68,69]. Temple headaches, frontoorbital headaches, and occipital headaches represent common presentations of headache disorders. Various studies propose that these regional headache disorders primarily arise due to entrapment of respective sensory nerve fibers by the overcontracting pericranial muscles. These nerve fibers include various trigeminal and upper cervical nerve fibers which course through the muscles [70-72]. In the current study, 26% of patients with TMJD had headache at the temples; 21% had headache with a fronto-orbital location; and 12% had headache at the occipital region (Table 2). In the control subjects, 12.5% had headache at the temples, 10.5% had headache at the fronto-orbital region; and 2% had headache at the occipital region. These regional headaches had a pressing, tightening quality and were of bilateral location. The difference in incidence of these respective headaches between the TMJD patient group and the control group was statistically significant (Table 2).

Data are expressed as number (%).

TMJD= temporomandibular joint disorder.

p> 0.05= not significant; p≤ 0.05= significant.

Data are expressed as number (%).

TMJD= temporomandibular joint disorder.

p≤ 0.05= significant

TMJD may act as a migraine trigger in genetically predisposed individuals [73]. A recent large longitudinal population study had suggested that the relation between TMJD and migraine is bidirectional [74]. A biological explanation for the association between TMJD and migraine is that extracranial and intracranial trigeminal innervation forms a functional unit [75]. This concept is supported by numerous animal studies, which reveal that dural afferents project through the calvarial bones to innervate extracranial structures [76]. It is hypothesized that there would be a physiological and pathological cross-talk between trigeminal nociceptive nerve fibers in both the intracranial and extracranial compartments [77,78]. Activation of the meningeal and meningovascular nociceptive afferents, with the release of various neuropeptides, results in a sterile dural inflammation [79]. Activation of dural mechanoreceptors, in the context of dural inflammation, explains the throbbing pain, experienced in some forms of headache, especially migraine [5]. Migraine headache is typically a unilateral throbbing headache, associated with photophobia, phonophobia and nausea/vomiting [80]. In the current study, headache typical of migraine was experienced in 13% of TMJD patients versus 6% of controls (P=0.017).

Totally, 88% of TMJD patients in the current study reported chronic headache, in contrast to only 37% of control subjects (P=0.001). Importantly, 66% of TMJD patients also reported chronic neck pain versus 45% of control subjects (P=0.001). Neck pain is mediated via the upper cervical nerves, and it is commonly associated with headache and TMD [12]. Indeed, the cranio-mandibular-cervical complex has been considered as an anatomical and functional unit, with neuromuscular restoring forces stabilizing this unit against the force of gravity [81,82]. Epidemiological, clinical, and imaging studies have revealed a significant association between TMJD and neck pain [83-86]. It is postulated that this association might be due to the convergent nociceptive input from the TMJ and neck structures onto the subnucleuscaudalis in the brain stem [87]. An alternative theory that was also postulated involves abnormal proprioceptive afferents from the TMJ that activate the trigemino-cervical reflex, resulting in abnormal neck muscle contraction and neck pain [88]. The trigemino-cervical reflex could also be implicated in migraine and TTH [89,90], thereby providing a putative explanation for the association of TMJD, headache, and neck pain.

Further adding to the morbidity in patients with TMJD, is the frequent association of symptoms pertaining to Costen’s syndrome. In 1934, James Costen reported (a syndrome of ear and sinus symptoms dependent upon disturbed function of the temporomandibular joint) [91]. These symptoms have recently been validated by audiometric tests and computed tomography (CT) scans of the paranasal sinuses [34-36]. In the current study, hearing loss, subjective tinnitus, and vertigo were reported in 39%, 40.5%, and 56.5% of TMJD patients, respectively. The incidence of these symptoms in control subjects was 20.5%, 21%, 32%, respectively (Table 4). The difference in incidence of these symptoms between the two groups is statistically significant. The origin of these symptoms in TMJD patients was postulated to be due to an increased tension on the malleus of the middle ear from a displaced TMJ disc, leading to abnormal position of the stapes at the oval window [92-94]. This, in turn, would lead to altered firing patterns from inner-ear hair cells [95]. Additionally, inputs from higher brain centers onto relevant brain stem nuclei may exaggerate these symptoms in conditions of pain, stress, anxiety, and lack of sleep, via top-down mechanisms [96,97]. Alleged sinus symptoms, especially facial pain, are also common in patients with TMJD [36]. In the current study, facial/eye pain was present in 52% of TMJD patients, versus 29% of control subjects (P=0.001). Facial/ eye pain had been attributed to the convergence of nociceptive branches of the trigeminal nerve onto the subnucleuscaudalis in the brain stem [98]. Jones described (midfacial segment pain), as a condition analogous to tension-type headache, with a similar pathophysiology, in patients with normal nasal endoscopy and clear sinus CT scans [99-101]. He postulated that this condition was attributed to abnormal nociceptive afferents from a regional source. Interestingly, Costen, in his original reports, mentioned that the otologic and alleged sinus symptoms were alleviated by correction of dental bite, in his patients with malocclusion [91]. Correction of the dental bite probably also ameliorates TMJD pain and headache [102,103]. The incidence of malocclusion in the current study was 30.5% in TMJD patients, versus 23% in control subjects (P=0.001).

Data are expressed as number (%).

TMJD= temporomandibular joint disorder.

p≤ 0.05= significant

Physical and emotional stress appear to be a common denominator in patients with headache, neck pain, and TMJD [104,105]. The stress response involves a pronounced sympathetic outflow and the release of stress hormones, and it is associated with a documented overactivity of head and neck muscles [106,107]. Moreover, the stress response is associated with peripheral and central sensitization in pain pathways, thereby contributing to the chronicity of headache, neck pain, and jaw pain [108,109]. Biomechanically, overactivity of masticatory muscles, as in bruxism, causes abnormal stresses on the TMJ, leading to its deterioration [110]. Overactivity of the lateral pterygoid muscle leads to the most common form of disc displacement (anteromedially) [111]. Clinically, bruxism, as a model of stress, is associated with TMJD [112,113]. In the current study, a report of bruxism was found in 50% of TMJD patients, versus 39% of control subjects (P=0.027).

The main strength of the current study was that it shedded light on the patterns of headache and associated symptoms in patients with TMJD, a previously undiscussed topic. The presence or absence of TMJ noises allowed an objective clinical distinction between TMJD patients and control subjects, respectively. Furthermore, the cohorts of TMJD patients and control subjects in the current study were of comparable socio-economic conditions, thereby allowing reliable distinctions to be made between both groups. The major limitation of the current study was that the reporting of symptoms depended entirely on the recalling of symptoms by the participants, which was totally subjective. From this view, the diagnosis of headache depends mainly on the verbal description of the patient. Apart from migraine, which has a classic presentation, no attempt was made to classify the headache from a diagnostic point of view. Rather, a description of headache sites was used in the current study, which was generally of a pressing, tightening quality and of bilateral location. The variable locations of headache in the study lends support to the theory of sensory nerve entrapment by respective over-contracting muscles. This notion emphasizes the recently-documented fact that extracranial and intracranial nociceptive nerve fibers act as a functional unit. A further limitation of the current study was that the author was not blinded, as regards the participant’s clinical features. However, the author has several publications regarding TMJD, which may validate the clinical signs elicited in the current study, as well as the recording of the relevant symptoms.

Conclusion

The current study revealed a higher incidence of headache in patients with TMJD, compared to control subjects. In addition to the typical migraine unilateral, throbbing headache, the patterns of headache frequently revealed a bilateral regional distribution with a pressing, tightening quality. This lends support to the theory of overactivity of pericranial muscles in TMJD, causing entrapment of isolated sensory trigeminal or cervical nerve branches, thereby explaining the regional headache patterns. Moreover, the connections between the extracranial and intracranial sensory nerve fibers account for the frequency of migraine in several participants.



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