Prevalence of vitamin D deficiency and the effect of vitamin D3 supplementation on response to anti-tuberculosis therapy in patients with extrapulmonary tuberculosis.

BACKGROUND: We aimed to assess serum 25-hydroxyvitamin D3 (25(OH)D3) concentrations in extrapulmonary tuberculosis (EPTB) patients and to evaluate the effect of vitamin D3 supplementation on their treatment course. METHODS: Serum 25(OH)D3concentrations were measured in 47 newly diagnosed EPTB patients and 42 controls. Vitamin D-deficient EPTB patients were randomly assigned to receive 50,000 IU of vitamin D3 (cholecalciferol) orally once a week for 6 weeks (total 300,000 IU), followed by maintenance doses of 1000 IU a day besides anti-TB drugs or the first line anti-TB treatment only. Follow up serum 25(OH)D3 concentrations were measured after 3 months of starting vitamin D3 supplementation. Both groups were evaluated for clinical, laboratory, and radiological outcomes after treatment. RESULTS: Serum 25(OH)D3 concentrations were significantly lower among TB cases (17.1 ± 5.5 nmol/L) compared to healthy controls (51.8 ± 27.3 nmol/L), and vitamin D deficiency was observed in all EPTB patients (n = 47). Patients in VD3 supplementation group had significantly higher weight gain and serum albumin level at 2 months and end of treatment, higher hemoglobin concentration at the end of treatment, significantly lower CRP and ESR at 2 months and at the end of treatment. In cases with TB pleurisy, a significant higher rate of full resolution of pleural fluid after 6 months of anti-TB treatment and shorter treatment duration were noted compared to the other group. CONCLUSIONS: Vitamin D deficiency is prevalent in EPTB patients, in whom, vitamin D supplementation is a useful adjunctive therapy to anti-TB drugs and improves treatment course.

A Systematic Review and Network Meta-Analysis of the Outcomes of Patients With Total Knee Arthroplasty Using Cemented, Uncemented, or Hybrid Techniques.

In this study, we aim to explore the differences among the three types of fixation methods for the components of the knee joint in patients who underwent total knee arthroplasty (TKA). These methods are cemented, uncemented, and hybrid fixation. Cemented fixation means that a special type of grout is used to attach the components to the bone. Uncemented fixation means that the components are designed to fit tightly into the bone and allow new bone growth to secure them. Hybrid fixation means that a combination of cemented and uncemented fixation is used for different components. We searched four online databases to find studies relevant to our research question. We use the R program (R Foundation for Statistical Computing, Vienna, Austria) for network meta-analysis (NMA) to analyze the data from the studies. We calculate the mean difference (MD) and the 95% confidence interval (CI) for each outcome, which are statistical measures of the difference and the uncertainty between methods. We use these measures for continuous outcomes, meaning they can have any value. For dichotomous outcomes, meaning they can only have two values, we use the risk ratio (RR) and the 95% CI, which are statistical measures of the relative risk and the uncertainty between methods. We assess the quality of randomized controlled trials, which are studies that randomly assign participants to different methods, using the Cochrane Risk of Bias Assessment Tool 1, a tool that evaluates the potential biases in the studies. We include 21 studies, and our analysis shows that cemented TKA reveals a statistically significant decrease in pain with hybrid TKA (MD = -2.82). That said, we find no significant differences between uncemented and cemented or hybrid (MD = -0.80 and -2.02, respectively). The results show that there is no significant difference between uncemented TKA and cemented TKA or hybrid technique (RR = 0.87, 95% CI 0.35-2.14; RR = 0.73, 95% CI 0.22-2.39, respectively). Also, we find no significant difference between cemented TKA and hybrid TKA (RR = 0.84, 95% CI 0.24-2.93). Cemented TKA is associated with a lower risk of deep vein thrombosis (DVT) incidence rate. Moreover, it shows a significant decrease in pain compared to hybrid TKA. Future research is needed to compare among the three interventions.

A Systematic Review and Meta-Analysis of Conventional Versus Robotic-Assisted Total Knee Arthroplasty.

This study aims to compare the outcomes and advantages of total knee arthroplasty (TKA) performed using conventional surgical techniques with those conducted using robotic-assisted methods in terms of operation time, Oxford knee score, range of motion, tourniquet time, and Western Ontario and McMaster Universities Arthritis index. We performed a literature search through five databases, namely, PubMed, Cochrane Central, Scopus, Embase, and Web of Science, from inception until July 3, 2023. Randomized clinical trials (RCTs) and cohorts comparing conventional TKA with robotic-assisted TKA were included. The risk of bias of the included RCTs was determined using the Cochrane risk of bias tool and the National Institutes of Health tool for cohort studies. We conducted a meta-analysis using Review Manager 5.4. To analyze continuous data, we calculated the mean difference (MD) along with its corresponding 95% confidence interval (CI). By synthesizing data from a comprehensive analysis, the study unveiled noteworthy distinctions between robotic-assisted arthroplasty and conventional arthroplasty across critical parameters. First, a substantial alteration in the hip-knee-ankle (HKA) angle was observed, with the robotic-assisted approach demonstrating a significant difference (MD = 0.84, 95% CI = 0.25-1.43, p = 0.005). Second, in terms of operative time, a notable reduction in surgical duration was noted with conventional TKA (MD = 16.85, 95% CI = 8.08-25.63, p = 0.0002). The assessment of tourniquet time exhibited a significantly longer duration for robotic-assisted arthroplasty (MD = 35.70, 95% CI = 27.80-43.61, p < 0.001). Our findings indicate that conventional TKA outperforms robotic-assisted TKA, primarily due to its shorter operative and tourniquet times, along with a more favorable change in the HKA angle. However, it is worth noting that robotic-assisted TKA showed a slight advantage in pain outcomes, although this advantage was not statistically significant. To gain a more comprehensive understanding, we recommend conducting a large-scale randomized controlled trial that directly compares both TKA methods. This trial should evaluate costs and long-term outcomes while ensuring consistent follow-up durations among studies. Such an approach would greatly assist orthopedic decision-making and contribute to improved TKA outcomes.

Robotic Versus Conventional Unicompartmental Knee Surgery: A Comprehensive Systematic Review and Meta-Analysis.

Robotic-assisted surgery is a computer-controlled technique that may improve the accuracy and outcomes of unicompartmental total knee arthroplasty (TKA), a partial knee replacement surgery. The purpose of a meta-analysis about robotic-assisted versus conventional surgery for unicompartmental TKA is to compare the effectiveness of these two methods based on the current evidence. Our meta-analysis can help inform clinical decisions and guidelines for surgeons and patients who are considering unicompartmental TKA as a treatment option. We searched four online databases for studies that compared the two methods until March 2023. We used RevMan software to combine the data from the studies. We calculated the mean difference (MD) and the 95% confidence interval (CI) for each outcome, which are statistical measures of the difference and the uncertainty between the two methods. We included 16 studies in our analysis. We found that robotic-assisted surgery had a better hip-knee-ankle angle, which is a measure of how well the knee is aligned, than conventional surgery (MD = 0.86, 95% CI = 0.16-1.56). We also found that robotic-assisted surgery had a better Oxford Knee score, which is a measure of how well the knee functions, than conventional surgery (MD = 3.03, 95% CI = 0.96-5.110). This study compared the results of conventional and robotic-assisted unicompartmental knee arthroplasty in 12 studies. We concluded that robotic-assisted surgery may have some benefits over conventional surgery in terms of alignment and function of the knee. However, we did not find any significant difference between the two methods in terms of other outcomes, such as pain, range of motion, health status, and joint awareness. Therefore, we suggest that more research is needed to confirm these results and evaluate the long-term effects and cost-effectiveness of robotic-assisted surgery.

Efficacy and Safety of Therapeutic Hypothermia as an Adjuvant Therapy for Percutaneous Coronary Intervention in Acute Myocardial Infarction: A Systematic Review and Meta-Analysis.

The study aims to compare the use of hypothermia in patients with myocardial infarction (MI) undergoing percutaneous coronary intervention (PCI) with control. We systematically searched four electronic databases until March 2022. The inclusion criteria were any study design that compared hypothermia in patients with MI undergoing PCI with control. The risk of bias assessment of the included randomized controlled trials was conducted through Cochrane Tool, while the quality of the included cohort studies was assessed by the NIH tool. The meta-analysis was performed on RevMan. A total of 19 studies were entered. Regarding the mortality, there were nonsignificant differences between hypothermia and control (odds ratio [OR] = 1.06, 95% confidence interval [CI] 0.75 to 1.50, p = 0.73). There was also no significant difference between the control and hypothermia in recurrent MI (OR = 1.21, 95% CI 0.64 to 2.30, p = 0.56). On the other hand, the analysis showed a significant favor for hypothermia over the control infarct size (mean difference = -1.76, 95% CI -3.04 to -0.47, p = 0.007), but a significant favor for the control over hypothermia in the overall bleeding complications (OR = 1.88, 95% CI 1.11 to 3.18, p = 0.02). Compared with the control, hypothermia reduced the infarct size of the heart, but this finding was not consistent across studies. However, the control had lower rates of bleeding problems. The other outcomes, such as death and the incidence of recurrent MI, were similar between the two groups.

Safety and efficacy of edaravone in patients with amyotrophic lateral sclerosis: a systematic review and meta-analysis.

AIM: The study aims to increase understanding of edaravone's efficacy and safety as an amyotrophic lateral sclerosis (ALS) treatment and provide significant insights regarding this field's future research. METHODS: We conducted a comprehensive search of the Embase, PubMed, Cochrane Library, Web of Science, and Scopus databases for randomized controlled trials and observational studies up until September 2022. We evaluated the studies' quality using the Cochrane risk of bias tool and the National Institutes of Health tool. RESULTS: We included 11 studies with 2845 ALS patients. We found that edaravone improved the survival rate at 18, 24, and 30 months (risk ratio (RR) = 1.03, 95% confidence interval (CI) [1.02 to 1.24], P = 0.02), (RR = 1.22, 95% CI [1.06 to 1.41], P = 0.007), and (RR = 1.17, 95% CI [1.01 to 1.34], P = 0.03), respectively. However, the administration of edaravone did not result in any significant difference in adverse effects or efficacy outcomes between the two groups, as indicated by a P value greater than 0.05. CONCLUSION: Edaravone improves survival rates of ALS patients at 18, 24, and 30 months with no adverse effects. However, edaravone does not affect functional outcomes. In order to ensure the validity of our findings and assess the results in accordance with the disease stage, it is essential to carry out additional prospective, rigorous, and high-quality clinical trials. The current study offers preliminary indications regarding the effectiveness and safety of edaravone. However, further comprehensive research is required to establish the generalizability and sustainability of the findings.

Formulation of sustained release bioadhesive minitablets containing solid dispersion of levofloxacin for once daily ocular use.

This study aimed to increase ocular residence time of levofloxacin by formulation into zero-order sustained release mucoadhesive minitablets for once daily administration using a hydrophobic-hydrophilic polymeric matrix. Levofloxacin was first formulated into solid dispersion with different ratios of Eudragit® RS then the resulting solid dispersion was mixed with different concentrations of Carbopol® and other excipients to be finally compressed into minitablets. A 24 full factorial design was employed to estimate the effects and interactions of two formulation factors, and to establish their relationships with selected responses in the developed minitablets. The studied factors were: drug to Eudragit® RS ratio, and percent of Carbopol® in the minitablets. Sixteen ocular minitablets formulations were prepared and evaluated for the cumulative percentages drug release at 6, 12, and 24 h, as well as mucoadhesion time, mucoadhesive strength, and swelling index as response variables. After optimizing the responses, the optimized formulation was found to be stable on sterilization using gamma-irradiation and storage at 40 °C/75% RH for six months. In vivo testing of the optimized formulation showed that the minitablets extended levofloxacin release up to 24 h without causing any ocular irritation. The optimized formulation exhibited superior microbiological activity compared to the commercial product.

Low Dermatitis Potential of a Powder-Free, “Accelerator-Free” Non Natural Rubber Latex Gloves Using Modified Draize Study

Introduction: The escalated demand for protective rubber glove in the healthcare industries has resulted in increased prevalence of glove related skin problem, irritant and allergic contact dermatitis and latex sensitivity. The industry has recently introduced a new nitrile glove product using a novel patented non-sulphur system to effect co-valent bond crosslinking to provide the desired elasticity of the gloves. This glove also has ionic crosslinking provided by the zinc oxide used in the formulation and the carboxylic group of the nitrile latex. The main objective of this study is to prove that residual chemical additives at a level that may induce Type IV allergy in the unsensitized general user population are not present in this rubber glove and to compare it with a powder free latex examination glove. Methods: In collaboration with the Islamic University of Gaza, we conduct modified test on a specially formulated and powder free, accelerator free LOW DERMA™ enhanced nitrile rubber glove that has physical properties and barrier integrity similar to that of NRL gloves. This glove does not contain sulphur or sulphur related compound. Two sets of Powder free, accelerator free LOW DERMA™ Nitrile Patient Examination Gloves*, white and blue colour were tested using the modified draize-95’ test. Filter paper soaked in normal saline and powder free latex examination glove were used as control. Results: A total of 209 subjects, 149 subjects, Caucasian (71.29%), 30 subjects, Afro Caribbean (14.35%) and 30 subjects, Asiatic (14.35%) were recruited. All 209 subjects had a final patch testing scoring of not more than 1.5 during both the induction phase and the challenge phase for both types of Powder Free Nitrile Patient Examination Gloves (white and blue) and to the negative control, normal filter paper and the powder free NRL control glove. Conclusion: The skin sensitization test (‘Modified Draize-95’ Test) of Powder Free Nitrile Patient Examination Gloves (white and blue) and the powder free NRL examination glove were negative. There was no clinical evidence on the presence of residual chemical additives at the level that may induce Type IV allergy in unsensitized general user population for both Powder Free Nitrile Patient Examination Gloves, blue and white colored, non-sterile. Both gloves qualify for “Low dermatitis Claim”.

High expression of matrix metalloproteinases: MMP-2 and MMP-9 predicts poor survival outcome in colorectal carcinoma.

AIM: To evaluate the expression pattern of matrix metalloproteinases (MMPs); MMP-2, MMP-7 and MMP-9 in colorectal cancer (CRC) and determine its prognostic potential. PATIENTS & METHODS: CRC samples of 127 patients were studied. Protein expressions of MMP-2, -7 and -9 were analyzed by immunohistochemistry and association with clinicopathological variables was statistically analyzed. RESULTS: Overexpressions of MMP-2 and MMP-9 correlated with poor outcome as evaluated by univariate Kaplan-Meier for disease-free survival (p = 0.04, p = 0.0001) and disease-specific survival (p = 0.01, p = 0.01), respectively. Cox analysis of MMP-2 and -9 were significant independent predictors of disease-free survival (p = 0.006, p = 0.018) and disease-specific survival (p = 0.004, p = 0.049), respectively. CONCLUSION: MMPs expression patterns provide useful prognostic information in CRC, while predicting the patients at high risk for recurrent disease.

ß-carotene and retinol contents in the meat of herbivorous ungulates with a special reference to their public health importance.

The aim of this study was to estimate total carotenoids, ß-carotene and retinol concentrations in the livers and muscles of some ungulates (cattle, buffalo, sheep, goats and horses) in comparison with the Wistar rats as a control. Cattle and horses had the highest contents of total carotenoids and ß-carotene. Unexpectedly, sheep was the highest accumulator of retinol with a mean concentration of 203 ± 23.34 µg/g, while the least accumulator was buffalo, having a mean value of 58.28 ± 13.77 µg/g. Livers had higher contents of the examined phytochemicals than muscles. Consumption of these tissues may provide human with some needs from these important phytochemicals, though ingestion of livers, especially that of the sheep, is not advisable for the pregnant women.

Genetic diversity of the pandemic influenza A (H1N1) virus in Saudi Arabia.

INTRODUCTION: Pandemic influenza A (H1N1) virus emerged and spread globally in the spring of 2009. Saudi Arabia also witnessed a severe H1N1 pandemic virus epidemic with considerable morbidity and mortality in different parts of the kingdom beginning in June 2009. The influenza A(H1N1)pdm09 virus was detected in samples collected between May 2009 and November 2010 from Makkah region. This study provides data on the viral diagnosis and genetic diversity of hemagglutinin (HA) and neuraminidase (NA) genes of influenza A (H1N1)pdm09 virus from Saudi Arabia. METHODOLOGY: Nasopharyngeal swabs from 100 clinically infected patients in the peak of the outbreak were collected from Makkah region and processed for viral diagnosis by viral culture and real-time polymerase chain reaction (PCR). HA and NA genes of 10 selected samples were sequenced and analyzed. RESULTS: A total of 100 samples were collected; only 10 samples were found to be positive for influenza A virus infection by real-time PCR. Nucleotide sequence analysis of the HA and NA genes of influenza A (H1N1) from Saudi Arabia showed significant similarities with selected isolates. The phylogenetic tree constructed for both HA and NA genes formed close clusters with selected reference isolates. CONCLUSIONS: Nucleotide sequence analysis and phylogenetic relationships of the HA and NA genes of influenza A (H1N1) virus from Saudi Arabia with selected reference isolates indicates that they were genetically close and most probably originated from influenza A(H1N1)pdm09.

Two-stage PCR assay for detection of human brucellosis in endemic areas.

BACKGROUND: Brucellosis is a common zoonosis that can cause a severe febrile illness in humans. It constitutes a persistent health problem in many developing countries around the world. It is one of the most frequently reported diseases in Saudi Arabia and incidence is particularly high in the Central region, and around the city of Riyadh. The aim of this study was to evaluate a two-stage PCR assay for detection of human brucellosis particularly in endemic areas. METHODS: A total of 101 serum samples were collected from patients with acute febrile illness (AFI) of unknown cause from two different locations in the Western region of Saudi Arabia. The first location (Northern) is characterized by a nomadic rural population while the second (Central) is a modern urban city. All samples were subjected to DNA extraction and Brucella genus-specific PCR amplification using B4/B5 primers of the bcsp31 gene. Positive B4/B5 samples were subjected to multiplex species-specific Brucella PCR amplification. RESULTS: In the Northern location, 81.9% of the AFI samples were confirmed Brucella positive, while all the samples collected from the Central region proved to be Brucella negative. Samples positive for Brucella were subjected to multiplex species-specific Brucella amplification. B. abortus was detected in 10% and B. melitensis in 8% of the samples, while the majority (82%) of samples showed both B. abortus and B. melitensis. As expected, B. suis was not detected in any of the samples. CONCLUSIONS: This study concluded that a two-stage PCR assay could be useful as a rapid diagnostic tool to allow the consideration of brucellosis as a possible cause of AFI, particularly in non-urban locations. It also recommends the collection of epidemiological data for such patients to obtain further information that may help in rapid diagnosis.

A molecular survey on cystic echinococcosis in Sinnar area, Blue Nile state (Sudan) / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cystic echinococcosis (CE) is a zoonosis caused by the cestodes of the Echinococcus species. Its life cycle involves dogs and other canids as definitive hosts for the intestinal tapeworm, as well as domestic and wild ungulates as intermediate hosts for the tissue-invading metacestode (larval) stage. The disease has a special impact on disadvantaged pastoralist communities and is listed now among the three top priority neglected tropical disease (NTD). Therefore, CE is a neglected disease even in high endemicity regions. This study aimed at investigation of the prevalence of CE in different animals slaughtered for food consumption in Sinnar area, Blue Nile states in Sudan.</p><p><b>METHODS</b>A survey of CE in livestock was conducted from April 2009 to March 2011 in Sinnar area, Blue Nile state in Sudan. Location, parasitological status and fertility conditions were determined. In addition, 120 hydatid cysts (30 from camels, 62 from cattle and 28 from sheep) were examined by polymerase chain reaction (PCR) and mitochondrial gene sequencing for the genetic allocation of Echinococcus strains or species</p><p><b>RESULTS</b>The prevalence of CE was 29.7% (30/101) in camels, 2.7% (62/2310) in cattle and 0.6% (26/4378) in sheep. It was shown that infection rates increased with age in camels, cattle and sheep. In camels, 67% (20/30) of the infected animals were aged between 2 - 5 years whereas 58% (36/62) of the infected cattle were > 5 years. In sheep, the prevalence rate was distributed equally between animals ranging 2 - 5 years and > 5 years. Even though multiple cysts were found in some animals, the average number of cysts per animal was close to 1 in all examined species. Lungs were found to be the predilection sites for the parasite in both camels and cattle, while most of the cysts found in sheep were located in the liver. About 63.4% of cysts encountered in camels were considered as large (5 - 7 cm), whereas those in cattle and sheep were medium (2 - 4 cm) and small (< 2 cm) respectively. The highest fertility rate was found in camel cysts with 85.4% (35/41) followed by cattle (50.0%, 32/64) and sheep (39.0%, 11/28). All examined cysts belonged to Echinococcus canadensis G6, which was confirmed to be the overwhelmingly predominant species in that area.</p><p><b>CONCLUSION</b>The epidemiological situation in Sinnar area, Blue Nile state is characterized by intense transmission of Echinococcus canadensis G6, thereby closely resembling the situation in most other regions of Sudan.</p>

Early predictors of microvascular complications in type 1 diabetic patients.

OBJECTIVES: To investigate the possibility of depending on adiponectin and leptin as early predictors of microvascular complications in type 1 diabetic subjects. DESIGN AND METHODS: We studied 63 type 1 diabetic subjects from the National Institute of Diabetes (30 normoalbuminuric and 33 microalbuminuric). Clinical, demographic characteristics and kidney function tests were monitored. Plasma levels of adiponectin, leptin, interlukein-6 (IL-6), and the high sensitive C-reactive protein (CRP) were measured in these subjects. RESULTS: Microalbuminuric subjects showed a significant elevation in adiponectin levels and a significant decrease in leptin levels as compared to normoalbuminuric subjects. Adiponectin showed a significant positive correlation with microalbuminuria concentrations while leptin showed a significant negative correlation with both fasting blood glucose and glycated hemoglobin A(1c). CONCLUSION: The results of this study introduced the possibility of depending on adiponectin and leptin as early, reliable, and sensitive predictors for the microvascular complications monitored by microalbuminuria concentration and glycemic control indices.

Adiponectin, leptin, and lipid profile in type 1 diabetic children and adolescents.

BACKGROUND: Adipose tissue is known to produce and secrete a variety of bioactive substances known as adipocytokines. Adiponectin and leptin are considered to be among the most important adipocytokines: OBJECTIVES: We sought to explore the relationships between adipocytokines (adiponectin and leptin), plasma lipoprotein lipid, and diabetic control indices in type 1 diabetic subjects. SUBJECTS AND METHODS: In this study 63 clinically diagnosed type 1 diabetic subjects and 30 age- and sex-matched healthy control subjects were analyzed. Age, sex, diabetic duration, family history of diabetes, daily insulin dose, weight, height, body mass index, and systolic and diastolic blood pressure were recorded. Fasting blood glucose, glycated hemoglobin A(1c), total hemoglobin, plasma lipoprotein, lipid and plasma concentrations of adiponectin and leptin were measured in type 1 diabetic subjects and control subjects. RESULTS: In this study a significant increase in triglycerides and high-density lipoprotein cholesterol in plasma of type 1 diabetics was found as compared with normal control subjects. In type 1diabetic subjects, plasma adiponectin was significantly elevated, whereas leptin showed a significant decrease as compared to a normal control group. Leptin concentrations showed a positive correlation with body mass index and systolic blood pressure but a negative correlation with both fasting blood glucose and glycated hemoglobinA(1c). CONCLUSION: The results of this study suggest that blood leptin but not adiponectin concentrations have a significant correlation with indices of glycemic control.

The Wolbachia genome of Brugia malayi: endosymbiont evolution within a human pathogenic nematode.

Complete genome DNA sequence and analysis is presented for Wolbachia, the obligate alpha-proteobacterial endosymbiont required for fertility and survival of the human filarial parasitic nematode Brugia malayi. Although, quantitatively, the genome is even more degraded than those of closely related Rickettsia species, Wolbachia has retained more intact metabolic pathways. The ability to provide riboflavin, flavin adenine dinucleotide, heme, and nucleotides is likely to be Wolbachia's principal contribution to the mutualistic relationship, whereas the host nematode likely supplies amino acids required for Wolbachia growth. Genome comparison of the Wolbachia endosymbiont of B. malayi (wBm) with the Wolbachia endosymbiont of Drosophila melanogaster (wMel) shows that they share similar metabolic trends, although their genomes show a high degree of genome shuffling. In contrast to wMel, wBm contains no prophage and has a reduced level of repeated DNA. Both Wolbachia have lost a considerable number of membrane biogenesis genes that apparently make them unable to synthesize lipid A, the usual component of proteobacterial membranes. However, differences in their peptidoglycan structures may reflect the mutualistic lifestyle of wBm in contrast to the parasitic lifestyle of wMel. The smaller genome size of wBm, relative to wMel, may reflect the loss of genes required for infecting host cells and avoiding host defense systems. Analysis of this first sequenced endosymbiont genome from a filarial nematode provides insight into endosymbiont evolution and additionally provides new potential targets for elimination of cutaneous and lymphatic human filarial disease.

Construction of bacterial artificial chromosome libraries from the parasitic nematode Brugia malayi and physical mapping of the genome of its Wolbachia endosymbiont.

The parasitic nematode, Brugia malayi, causes lymphatic filariasis in humans, which in severe cases leads to the condition known as elephantiasis. The parasite contains an endosymbiotic alpha-proteobacterium of the genus Wolbachia that is required for normal worm development and fecundity and is also implicated in the pathology associated with infections by these filarial nematodes. Bacterial artificial chromosome libraries were constructed from B. malayi DNA and provide over 11-fold coverage of the nematode genome. Wolbachia genomic fragments were simultaneously cloned into the libraries giving over 5-fold coverage of the 1.1 Mb bacterial genome. A physical framework for the Wolbachia genome was developed by construction of a plasmid library enriched for Wolbachia DNA as a source of sequences to hybridise to high-density bacterial artificial chromosome colony filters. Bacterial artificial chromosome end sequencing provided additional Wolbachia probe sequences to facilitate assembly of a contig that spanned the entire genome. The Wolbachia sequences provided a marker approximately every 10 kb. Four rare-cutting restriction endonucleases were used to restriction map the genome to a resolution of approximately 60 kb and demonstrate concordance between the bacterial artificial chromosome clones and native Wolbachia genomic DNA. Comparison of Wolbachia sequences to public databases using BLAST algorithms under stringent conditions allowed confident prediction of 69 Wolbachia peptide functions and two rRNA genes. Comparison to closely related complete genomes revealed that while most sequences had orthologs in the genome of the Wolbachia endosymbiont from Drosophila melanogaster, there was no evidence for long-range synteny. Rather, there were a few cases of short-range conservation of gene order extending over regions of less than 10 kb. The molecular scaffold produced for the genome of the Wolbachia from B. malayi forms the basis of a genomic sequencing effort for this bacterium, circumventing the difficult challenge of purifying sufficient endosymbiont DNA from a tropical parasite for a whole genome shotgun sequencing strategy.

Test-retest validity of acoustic pharyngometry measurements.

BACKGROUND: Acoustic reflectometry is a relatively new technique that quantifies upper airway obstruction. The oropharyngeal airway is geometrically complex and variable; therefore establishing a standard operating protocol and understanding the possible sources of artifacts are of great importance in obtaining reliable results. This work aims at assessing the repeatability of pharyngeal cross-sectional area measurements obtained from normal and snoring individuals. METHODS: Twenty adult normal volunteers (16 men and 4 women; mean age, 35.9 years) and 10 adult snorers (9 men and 1 woman; mean age, 36.4 years) were examined by acoustic reflectometry following the developed standard operating protocol. RESULTS: Measurements of pharyngeal cross-sectional area are analyzed in 2 groups. In normal subjects where mean pharyngeal cross-sectional area in the first session was 3.187 cm(2), in the second session (same-day test-retest), the mean pharyngeal cross-sectional area was 3.239 cm(2), and in the third session 7 to 10 days later (day-to-day test-retest), it was 3.245 cm(2) (P > 0.4). In a second group of snoring patients where mean pharyngeal cross-sectional area in the first session was 2.244 cm(2), in the second session, mean pharyngeal cross-sectional area was 2.237 cm(2), and mean pharyngeal cross-sectional area in the third session (7 to 10 days later) was 2.238 cm(2) (P > 0.9). CONCLUSIONS: These results show that repeatability of acoustic reflection results can be achieved following the standard operating protocol. SIGNIFICANCE: The study results add to the reliability of this technique in assessing the pharyngeal airway in patients with snoring and obstructive sleep apnea.

Acoustic pharyngometry patterns of snoring and obstructive sleep apnea patients.

OBJECTIVE: Anatomic narrowing of the pharyngeal airway increases the relative negative pressure generated during inspiration, thus affecting the dynamic behavior of the upper airway. The aim of this work was to measure pharyngeal area in snoring patients with and without obstructive sleep apnea (OSA), as categorized by polysomnography and by acoustic pharyngometry, and to analyze the different curve patterns obtained from patients of both groups. METHODS: We examined 50 snorers who were divided into 2 groups matched for age, gender, and body mass index. RESULTS: Mean Apnea Index (AI) in nonapneic snorers (group 1) was 4, and mean pharyngeal area was 2.41 cm(2). In snoring patients with OSA (group 2), mean AI was 25.9 with a mean pharyngeal area of 1.589 cm(2) (P < 0.001). In both groups, the dependent variable (AI) can be predicted from a linear relation with the independent variable (pharyngeal area) with normality and constant variance tests passed. In group 1, the resulting curve can be 1 of 2 types depending on the relative area of the pharyngeal segment to the velopharyngeal area. In group 2, the curve pattern can be categorized into 3 patterns depending on the possible pathology of pharyngeal obstruction. CONCLUSION: The acoustic reflection technique is reproducible, noninvasive, and free from potential side effects. The good correlation between AI and pharyngeal area adds to the potential of acoustic pharyngometry. Careful study of the pharyngeal cross-sectional area and curve topography may give a good idea about the site of upper airway obstruction.

An aspartate aminotransferase of Wolbachia endobacteria from Onchocerca volvulus is recognized by IgG1 antibodies from residents of endemic areas.

Wolbachia are intracellular alpha-proteobacteria, closely related to Rickettsia, that infect various arthropods and filarial parasites. In the present study, the cDNA encoding the aspartate aminotransferase (AspAT) of Wolbachia from the human pathogenic filarial parasite Onchocerca volvulus (Ov-WolAspAT) was identified. At the amino acid level, the identity of the Ov-WolAspAT was 56% to Rickettsia prowazekii AspAT and 54% to the AspAT of the nitrogen-fixing bacterium Sinorhizobium meliloti, but the highest degree of identity was found to the putative AspAT of Wolbachia from Brugia malayi and Drosophila melanogaster (85%). All of these bacterial AspATs are members of the AspAT subclass Ib. A 35 kDa fragment of the Ov-WolAspAT was expressed in Escherichia coli, and immunolocalization using polyclonal antibodies against this antigen revealed that Ov-WolAspAT is present in a considerable proportion of the Wolbachia from O. volvulus, as well as in the endobacteria of several other filarial parasites. Western blot analysis using recombinant Ov-WolAspAT as antigen showed that IgG1 antibodies were present in 70 (51%) individuals living in areas endemic for O. volvulus, B. malayi or Wuchereria bancrofti and no IgG4 or IgE antibodies were found. Among 40 sera of persons from Uganda and Liberia who were putatively not infected with human filarial parasites, 11 (28%) individuals presented IgG1 antibodies, while none of the 33 sera from healthy Europeans and none of the 14 sera from patients with proven Rickettsia or Brucella infections reacted with the antigen. These results also show that an intracellular protein of Wolbachia endobacteria (WolAspAT) acts as antigen in human filariasis.