Identification of <Emphasis Type="Italic">moa</Emphasis>A3 gene in patient isolates of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> in Kerala,which is absent in <Emphasis Type="Italic">M. tuberculosis</Emphasis>H37Rv and H37Ra

Background  

Tuberculosis is endemic to developing countries like India. Though the whole genome sequences of the type strain M. tuberculosis H37Rv and the clinical strain M. tuberculosis CDC1551 are available, the clinical isolates from India have not been studied extensively at the genome level. This study was carried out in order to have a better understanding of isolates from Kerala, a state in southern India.     

Infection of total hip prosthesis by <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> and <Emphasis Type="Italic">Mycobacterium chelonae</Emphasis> in a patient with rheumatoid arthritis

Atypical mycobacterial infections of the musculoskeletal system are very rare and are generally associated with predisposing factors, such as trauma, use of corticosteroids, or an immunocompromised state. There have only been three reports of Mycobacterium chelonae prosthetic infection of which two cases were associated with total hip arthroplasty and one with total knee arthroplasty and no reports of both Mycobacterium tuberculosis and M. chelonae occurring in the same joint. We report a case of a patient with rheumatoid arthritis treated with low-dose methotrexate (15 mg/week) who developed infection with both M. tuberculosis and M. chelonae after the revision of a prosthetic hip. Joint infections by mycobacteria are clinically indistinguishable from those caused by more common bacterial pathogens and, therefore, diagnosis is often delayed. Recurrent prosthetic hip infections, particularly in immunosuppressive patients, should alert the physician to consider the possibility of both tuberculous and atypical mycobacterial infections. Obtaining appropriate cultures can be critical in making the diagnosis and directing treatment. With the increasing use of immunosuppressive agents, including TNF alpha inhibitors, it is likely that there will be an increase in the number of mycobacterial infections complicating arthroplasties.     

Predominant <Emphasis Type="Italic">porB1A</Emphasis> and <Emphasis Type="Italic">porB1B</Emphasis> genotypes and correlation of gene mutations with drug resistance in <Emphasis Type="Italic">Neisseria gonorrhoeae</Emphasis> isolates in Eastern China

Background  

Variations of porB1A and porB1B genes and their serotypes exist in Neisseria gonorrhoeae isolates from different geographical areas, and some site mutations in the porB1B gene correlate with drug resistance.     

Genetic diversity of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> Complex in Jos,Nigeria

Background  

Nigeria has a high tuberculosis incidence, and genotyping studies of Mycobacterium tuberculosis Complex (MTC) in the country are necessary in order to improve our understanding of the epidemic.     

Identification of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> clinical isolates in Bangladesh by a species distinguishable multiplex PCR

Background  

Species identification of isolates belonging to the Mycobacterium tuberculosis complex (MTC) seems to be important for the appropriate treatment of patients, since M. bovis is naturally resistant to a first line anti-tuberculosis (TB) drug, pyrazinamide, while most of the other MTC members are susceptible to this antimicrobial agent. A simple and low-cost differentiation method was needed in higher TB burden countries, such as Bangladesh, where the prevalence of M. bovis among people or cattle has not been investigated.     

<Emphasis Type="Italic">hSNF5</Emphasis><Emphasis Type="Italic">/INI1</Emphasis> mutation analysis in acute myeloid leukemia

Previous studies indicated that region 11.2 of the long arm of chromosome 22 (22q11.2) might be a locus encoding a tumor suppressor gene, since its deletion is a recurrent genetic characteristic of aggressive pediatric cancer. This region is found in the human immunodeficiency virus integrase interactor 1 (hSNF5/INI1) gene. To investigate whether the hSNF5/INI1 gene is involved in leukemogenesis, mutation analysis of the hSNF5/INI1 gene was performed in the present study using 5 hematopoietic cell lines, acute myeloid leukemia (AML) specimen and normal control. We found two single nucleotide polymorphisms at the hSNF5/INI1 gene in exon 4 and exon 9. The results of this study suggest that the hSNF5/INI1 gene does not play an important role in the leukemogenesis of AML.     

Clinical characteristics of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> infection among rheumatoid arthritis patients

To evaluate the clinical characteristics of Mycobacterium tuberculosis infection in rheumatoid arthritis (RA) patients, we examined the clinical manifestations and radiography/computed tomography (CT) findings in RA patients with tuberculosis (RA+/TB+). A total of 1121 tuberculosis patients were admitted to our hospital from 1995 to 2003, with the RA patients among them comprising 1.8% (20 cases; 9 men and 11 women). This is approximately three times as high as the prevalence of RA in the entire population in Japan. In addition, the RA+/TB+ patients were older and had a longer history of RA than the 140 outpatients in our RA clinic who did not have tuberculosis (RA+/TB–). Half of the RA+/TB+ patients had no symptoms (e.g., cough, sputum, pyrexia), and their tuber-culosis was detected accidentally by radiography/CT. The positive rates of the bacilli in the smear and culture of the sputum from the RA+/TB+ patients were lower than those from 143 patients randomly selected from among 1091 tuberculosis patients without any collagen disease including RA (RA–/TB+). The RA+/TB+ patients had a higher incidence of extrapulmonary tuberculosis (30%), including four cases (20%) of miliary tuberculosis, an incidence seven times higher than among the general population of tuberculosis patients. Among 14 cases of pulmonary tuberculosis patients with RA, bilateral lesions and noncavitary lesions were found in 71.4% and 64.3%, respectively, which tended to be a higher incidence than in the RA–/TB+ patients. The mortality rate and sputum conversion time of the RA+/TB+ patients were no different from those of the RA–/TB+ patients. The prevalence of tuberculosis in RA patients is expected to increase after introduction of anticytokine therapy in Japan, and careful observation should be done to avoid this complication in RA patients.     

Association of <Emphasis Type="Italic">Lewis</Emphasis> and <Emphasis Type="Italic">Secretor</Emphasis> gene polymorphisms and <Emphasis Type="Italic">Helicobacter pylori</Emphasis> seropositivity among Japanese-Brazilians

Background Secretor (Se) and Lewis (Le) genes are involved in the synthesis of Lewis b (Le^b) and type I antigens throughout the body, especially in the epithelial cells of gastric mucosa. Helicobacter pylori can attach to the gastric epithelial cells with the blood group antigen-binding adhesin, which binds to Le^b or H type I carbohydrate structures. In a previous study, a marked association between H. pylori seropositivity and polymorphism of the Se and Le genes was observed among Japanese outpatients of a gastroenterology clinic. The present work aims to investigate the associations between Se and Le gene polymorphisms and H. pylori infection among Japanese-Brazilians.Methods The subjects consisted of 942 healthy volunteer Japanese-Brazilians, who were tested for the presence of anti-H. pylori IgG antibodies and genotyped for Se and Le polymorphisms.Results The sex-age-adjusted odds ratios (aORs) for H. pylori seropositivity were 0.99 for the Sese genotype relative to the SeSe genotype (95% confidence interval [CI], 0.73–1.33), and 1.03 for sese relative to SeSe (95% CI, 0.71–1.48). On the other hand, the aOR for the subjects with the le allele (Lele or lele) relative to the LeLe genotype was 1.48 (95% CI, 1.07–1.79). When the Se and Le genotypes were analyzed in combination according to risk group, no statistically significant association was observed.Conclusions These results are inconsistent with previous work and may have been modulated by an external factor or some other unidentified factor. Japanese-Brazilians are genotypically the same as Japanese, but their lifestyle is adapted to that of Brazil. Further investigations are necessary to clarify this influence on susceptibility to H. pylori infection.     

Mutational analysis of serotonin receptor genes: <Emphasis Type="Italic">HTR3A</Emphasis> and <Emphasis Type="Italic">HTR3B</Emphasis> in fibromyalgia patients

The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) has been implicated in numerous human disorders. Dysfunction of serotonergic neurotransmission is thought to play a major role in the pathophysiology of the fibromyalgia syndrome (FMS) which is characterised by non-restorative sleep and severe pain. In our study, both serotonin receptor subunit genes, HTR3A and HTR3B, have been investigated for sequence variations in FMS patients in order to reveal a possible involvement in the aetiology of FMS. We examined DNA samples from 48 patients with FMS representing sporadic cases by single-strand conformation polymorphism (SSCP) and denaturing high-performance liquid chromatography (dHPLC) analysis, sequenced samples with conspicuous patterns and performed statistical calculations. HTR3A mutational analysis revealed one novel as well as five known sequence variations. Investigating HTR3B, we detected seven formerly described mutations and one novel sequence variant. Statistical computation rated all variants as probably non-disease-related polymorphisms. Nevertheless, one might speculate about an effect of the respective sequence variants on the severity of the disease. Sequence variants of the serotonin receptor subunit genes HTR3A and HTR3B indicate no obvious significance in the aetiology of fibromyalgia, yet they represent the basis for future studies on their pharmacogenetic relevance.     

Identification of a predominant isolate of <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> using molecular and clinical epidemiology tools and <Emphasis Type="Italic">in vitro</Emphasis>cytokine responses

Background  

Tuberculosis (TB) surveillance programs in Canada have established that TB in Canada is becoming a disease of geographically and demographically distinct groups. In 1995, treaty status aboriginals from the province of Manitoba accounted for 46% of the disease burden of this sub-group in Canada. The TB incidence rates are dramatically high in certain reserves of Manitoba and are equivalent to rates in African countries. The objective of our study was to identify prevalent isolates of Mycobacterium tuberculosis in the patient population of Manitoba using molecular epidemiology tools, studying the patient demographics associated with the prevalent strain and studying the in vitro cytokine profiles post-infection with the predominant strain.     

<Emphasis Type="Italic">In vivo</Emphasis> expression of innate immunity markers in patients with <Emphasis Type="Italic">mycobacterium tuberculosis</Emphasis> infection

Background  

Toll-like receptors (TLRs), Coronin-1 and Sp110 are essential factors for the containment of Mycobacterium tuberculosis infection. The purpose of this study was to investigate the in vivo expression of these molecules at different stages of the infection and uncover possible relationships between these markers and the state of the disease.     

False-Positive <Emphasis Type="Italic">Mycobacterium tuberculosis</Emphasis> Culture Revealed by Restriction Fragment Length Polymorphism Analysis

Background: The microbiological analysis of respiratory specimens is the most reliable approach to diagnose active pulmonary tuberculosis. Patient and Methods: We report a 60-year-old female patient (index patient) who underwent diagnostic bronchoscopy for chronic cough. No acid-fast bacilli were detected in bronchial washings. Although cough subsided with symptomatic treatment, Mycobacterium tuberculosis grew on egg-based media after 12 weeks. A false-positive culture result was suspected. Chart review and DNA fingerprinting were carried out. Results: The bronchoscope used to examine the index patient was previously used for a 30-year-old patient (source patient) with smear- and culture-positive pulmonary tuberculosis. Restriction fragment length polymorphism (RFLP) analysis based on the IS6110 element confirmed that the two strains were identical. Conclusion: Cross-contamination is a reason for falsepositive cultures with M. tuberculosis and should be suspected in patients with a low clinical probability for active tuberculosis. Received: March 28, 2002 · Revision accepted: July 5, 2002 O.D. Schoch (corresponding author)     

<Emphasis Type="Italic">T-</Emphasis>Scores and <Emphasis Type="Italic">Z</Emphasis>-Scores

Bone mineral density (BMD) can be measured by a variety of techniques at several skeletal sites. Once measured, the manufacturers’ software uses the BMD to calculate a T-score and/or Z-score. Both T-scores and Z-scores are derived by comparison to a reference population on a standard deviation scale. The recommended reference group for the T-score is a young gender-matched population at peak bone mass, while the Z-score should be derived from an age-matched reference population. T-scores and Z-scores are widely quoted in scientific publications on osteoporosis and BMD studies, and are the values used for DXA diagnostic criteria and current clinical guidelines for the management of osteoporosis. Errors in BMD measurement, differences in reference populations, and variations in calculation methods used, can all affect the actual T-score and Z-score value. Attempts to standardize these values have made considerable progress, but inconsistencies remain within and across BMD technologies. This can be a source of confusion for clinicians interpreting BMD results. A clear understanding of T-scores and Z-scores is essential for correct interpretation of BMD studies in clinical practice.     

Novel mutations in <Emphasis Type="Italic">WWOX</Emphasis>, <Emphasis Type="Italic">RARS2</Emphasis>, and <Emphasis Type="Italic">C10orf2</Emphasis> genes in consanguineous Arab families with intellectual disability

Intellectual disability is a heterogeneous disease with many genes and mutations influencing the phenotype. Consanguineous families constitute a rich resource for the identification of rare variants causing autosomal recessive disease, due to the effects of inbreeding. Here, we examine three consanguineous Arab families, recruited in a quest to identify novel genes/mutations. All the families had multiple offspring with non-specific intellectual disability. We identified homozygosity (autozygosity) intervals in those families through SNP genotyping and whole exome sequencing, with variants filtered using Ingenuity Variant Analysis (IVA) software. The families showed heterogeneity and novel mutations in three different genes known to be associated with intellectual disability. These mutations were not found in 514 ethnically matched control chromosomes. p.G410C in WWOX, p.H530Y in RARS2, and p.I69F in C10orf2 are novel changes that affect protein function and could give new insights into the development and function of the central nervous system.     

<Emphasis Type="Italic">DI*A</Emphasis> and <Emphasis Type="Italic">DI*B</Emphasis> Allele Frequencies Among Southern Thai Blood Donors

Diego (DI) blood group genotyping is clinically important in Asian populations. Data of Diego blood type among southern Thais is still unknown. This study aimed to report DI*A and DI*B allele frequencies in southern Thai blood donors and to estimate potential risk of Di^a incompatibility and alloimmunization in Thai populations. DNA samples obtained from 427 southern Thai blood donors were genotyped for DI*A and DI*B alleles by polymerase chain reaction with sequence-specific primer. DI*A and DI*B allele frequencies among southern Thais were 0.0047 and 0.9953. Their frequencies were similar to those among American Native, Italian, Filipino, Alaska Native/Aleut and Hawaiian/Pacific Islander populations; while, the frequencies significantly differed from central and northern Thai, Southeast Asian, Brazilian, Southern Brazilian, Brazilian Japanese descendants, Japanese, Han Chinese, Chinese, and Korean populations (P < 0.05). The Di^a incompatibility among southern Thais (0.93%) was lower than among central Thais (3.49%), corresponding to a significantly lower probability of Di^a alloimmunization (P < 0.05). This is the first report of DI*A and DI*B allele frequencies among southern Thais, which is beneficial for not only creating information for estimating risk of alloimmunization, but also providing antigen-negative red cell donors to prevent both alloimmunization and adverse transfusion reactions.     

<Emphasis Type="Italic">Anisakis</Emphasis> spp. burden in <Emphasis Type="Italic">Trachurus trachurus</Emphasis>

Anisakis is a parasite of marine mammals that uses a great number of fish species as intermediate or paratenic hosts. It is common in commercially important marine fishes and its presence is of great concern for both human health and economic reasons. Horse mackerels (Trachurus trachurus) originated from the Northern Aegean Sea were examined for the presence of Anisakis spp. larvae. The prevalence of Anisakis spp. was found 98.8 %. The number of parasites was significantly related to the host’s length but was not related to the fish gender. The month of sampling affected the size of the fishes and consequently the number of parasites. The length of larvae was not related to the host’s length. The present study resulted in the design of a prediction model for the number of existing parasites in the fish by measuring only its Fixed Length.     

Molecular differentiation of Turkey cattle isolates of <Emphasis Type="Italic">Fasciola hepatica</Emphasis> and <Emphasis Type="Italic">Fasciola gigantica</Emphasis>

The most common and widespread liver flukes of the genus Fasciola are Fasciola hepatica and F. gigantica. Adults of both species occur in many domestic ruminants and in humans and can cause serious disease. The differential diagnosis of these flukes infection is very important because of their different transmission and epidemiological characteristics. A simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the common restriction enzymes AluI and RsaI, is described to distinguish between both fasciolid species. After the digestion of the mitochondrial cytochrome c oxidase 1 (CO1) PCR product with the restriction enzyme AluI, the RFLP profile obtained from F. hepatica revealed two fragments, whereas F. gigantica was not cut. The RsaI digestion generated two fragments from F. gigantica, whereas it did not cut the PCR product from F. hepatica. Results were confirmed with CO1 sequence analysis of both F. hepatica and F. gigantica. The present study suggests that the PCRRFLP method described here can be used for the proper identification of Fasciola species.