Flow cytometric analyses of the lymphocyte subsets in peripheral blood of children with untreated active juvenile dermatomyositis.

Juvenile dermatomyositis (JDMS) is a vasculopathy affecting primarily skin and muscle. Although the etiology is unknown, immunopathogenetic mechanisms appear to play a role in both the susceptibility to the disease and its progression. We measured the percentage and absolute numbers of B cells and T-cell subsets in the peripheral blood of untreated JDMS patients with active early disease and compared the results with those obtained from a study of peripheral blood obtained from a heatlhy age-related control group. The absolute number of total lymphocytes in the peripheral blood of the JDMS patients was significantly lower (P < 0.002) than that observed in the healthy control population, with an associated decrease in the absolute number of all T-cell subsets. No concomitant decrease in the absolute number of B lymphocytes was observed in the JDMS patients. In contrast, the percentage of B lymphocytes and the T-helper/T-suppressor cell ratio were significantly higher in the JDMS group than in the control group (P < 0.001 and P < 0.002, respectively). Retrospective analysis of JDMS patients' serum samples obtained within 1 month of the flow cytometric evaluation indicated that 79% of the sera contained an antinuclear antibody and 46% had immunoglobulin G values above age-adjusted reference ranges. The increased percentage of B cells, the increased T-helper/T-suppressor cell ratio, the positive antinuclear antibody results, and the increased concentration of serum immunoglobulin suggest that humoral immune dysregulation may contribute to the pathogenesis of JDMS.     

Molecular genetic studies of major histocompatibility complex genes in children with Juvenile dermatomyositis: Increased risk associated with HLA-DQA10501

Juvenile dermatomyositis (JDMS) is an inflammatory disease associated with HLA-DR3. We therefore undertook molecular genetic studies of HLA region genes to determine whether HLA-DR3 itself confers susceptibility to JDMS or whether susceptibility is conferred by alleles in linkage disequilibrium with HLA-DR3. Our results indicate that JDMS is associated with the HLA-DQA1 allele DQA1^*0501 on non-DR3 haplotypes in Caucasian JDMS. Furthermore, the reported of association between the C4A gene deletion and JDMS is likely due to linkage disequilibrium with HLA-DR3. Human Immunology 32, 235–240 (1991)     

Ten-year experience of juvenile dermatomyositis: a retrospective study.

BACKGROUND AND PURPOSE: Juvenile dermatomyositis (JDMS) is a rare multisystemic disease of unknown etiology that primarily affects muscle and skin. This study aimed to evaluate the initial laboratory data, clinical manifestations, complications, and clinical outcomes of patients with JDMS in Taiwan. METHODS: We reviewed medical charts of patients younger than 18 years with a diagnosis of JDMS at the pediatric department of National Taiwan University Hospital between 1994 and 2004. RESULTS: A total of 21 patients were included. The female-to-male ratio was 4.25:1. The mean age at onset of symptoms was 6.4 +/- 3.7 years (range, 2 to 14.2 years). The mean age at diagnosis was 6.9 +/- 3.9 years (range, 2.2 to 15 years). Among the initial cutaneous features, Gottron's rash (62%) and facial rash (including malar rash, 62%) were the most common findings. Gottron's rash was also the most common sign (81%) at any time during the disease course. Among other systemic features at disease onset, myalgia (33%) was the most common symptom, followed by arthralgia (19%) and dysphagia (19%). Interstitial pneumonitis was a complication in 2 patients, one of whom was a mortality case. Gastrointestinal tract vasculitis was highly suspected in one patient complicated with duodenal perforation and retroperitoneal abscess. Calcinosis developed in 6 patients (28.5%), and one of these patients presented with disseminated calcinosis. Five of 21 patients achieved drug-free remission for 28 to 94 months from the time they discontinued therapy to the end of the study. Two patients with diagnosis of amyopathic dermatomyositis were also included in this study. CONCLUSIONS: The long-term outcomes in this study are thought to be good. Because the understanding of the pathogenesis of JDMS is limited, certain complications are still troublesome clinically.     

Oligoclonal expansion of circulating and tissue-infiltrating CD8+ T cells with killer/effector phenotypes in juvenile dermatomyositis syndrome

Although triggering by infectious agents and abnormal immune responses may play some role in the pathogenesis of juvenile dermatomyositis syndrome (JDMS), the precise mechanism of muscle destruction and vascular damage is largely unknown. In this study, we tried to elucidate the role of cytotoxic T cells in two patients with JDMS, who were diagnosed based on the characteristic symptoms, laboratory data, MRI findings and electromyographic patterns. Peripheral blood T cell phenotypes were determined by flow cytometry, using mAbs against specific T cell receptor (TCR) Vbetas. Complementarity-determining region3 (CDR3) size analysis was performed by gene scanning of CDR3 polymerase chain reaction (PCR) amplification products specific for each Vbeta. Subsequently, CDR3 nucleotide sequences were obtained after cloning of the predominant products. The distribution of lymphocytes infiltrating the muscle tissue was analysed by immunohistochemistry. In both patients examined, a unique combination of TCR Vbeta repertoires was increased within the CD8+ T cells. These subpopulations expressed a characteristic phenotype, indicating that they are memory/effector T cells with killer functions. At the same time, immunohistological and molecular biological examinations of the biopsied muscle samples revealed that identical CD8+ T cell clones with identical phenotypes/TCR Vbeta infiltrated within the inflammatory tissue, in particular around vessels. These findings indicate that oligoclonal expansion of CD8+ T cells plays a central role in the pathogenesis of muscle injury in the juvenile form of dermatomyositis syndrome and may provide a useful clinical parameter of disease activity and responsiveness to anti-inflammatory therapy.     

High heterogeneity of apolipoprotein E gene frequencies in South American Indians

The apolipoprotein E (APOE) polymorphism was investigated in 186 individuals from six South American Indian tribes, and the results integrated with those previously presented for this ethnic group. The three APOE alleles commonly reported in other populations were also observed in South Amerindians with a highly heterogeneous distribution. As in other populations, APOE*3 was the most common allele (51-98%) followed by APOE*4 (2-47%). These two isoforms were identified in all tribes, but APOE*2 was observed among the Wai Wai (2%) and Mataco (4%) only. No previous indications of inter-ethnic admixture were observed among the Wai Wai, but the introduction of this allele among the Mataco through non-Indian sources cannot be excluded.     

Acute nonlymphocytic leukemia after therapy with alkylating agents for ovarian cancer: a study of five randomized clinical trials

We evaluated the occurrence of acute nonlymphocytic leukemia (ANL) among 1399 women with ovarian cancer who were treated in five randomized clinical trials. Of the 1399 women, 998 had been treated with alkylating agents, and among these, 12 cases of ANL were observed; the expected number was 0.11. Ten patients with ANL had received melphalan, and two chlorambucil. ANL was not observed in 401 women who had been treated with surgery or radiation or both, without alkylating agents. The excess risk of ANL that was associated with alkylating-agent therapy was 5.8 cases per 1000 women per year, and the cumulative seven-year risk of ANL among patients who were treated with chemotherapy alone was indistinguishable from that observed in patients receiving both radiation and chemotherapy. A positive correlation between initial drug dose and the risk of ANL was suggested. These data underscore the need to assess other cytotoxic agents and regimens of drug administration to identify those that do not have harmful late effects.     

Isoelectric focusing studies in Brazilian Indians--uncovering variation of ORM, AHSG and IF

Allele frequencies for the orosomucoid 1 (ORM1), orosomucoid 2 (ORM2), alpha-2-HS-glycoprotein (AHSG) and complement component I (IF) loci were studied in 393 individuals of three Brazilian Indian tribes. In the ORM1 locus only two alleles were observed among the Urubu-Kaapor, while five were found among the Pacaás Novos. The frequency of ORM1*1 was similar in these two tribes (0.734 and 0.715, respectively) but departed more markedly among the Parakan? (0.870). Variation for ORM2 locus was found among the Pacaás Novos only, with ORM2*3 being observed in just three individuals. A new variant (AHSG*PN) was found in the AHSG system. Frequency for AHSG*1 was unexpectedly low in the three tribes, especially, among the Pacaás Novos, where the prevalence (0.145) is the lowest considering other data reported thus far. For IF locus, variability was also restricted to only one trible (Urubu-Kaapor) and attributed to a new polymorphic allele, IF* A3.     

High in vitro spontaneous histamine release in long-term hemodialyzed patients.

Adverse reactions, such as skin flush and pruritus, have been observed among hemodialyzed (HD) patients. A potential relationship between hemodialysis and allergic phenomena prompted us to investigate some allergologic parameters. Total IgE, specific IgE antiethylene oxide and anti-isocyanate toluene diisocyanate, and in vitro spontaneous histamine release (SHR) were measured in 105 long-term stable HD patients. Thirty-two of the patients had occasionally developed skin flush reactions and/or pruritus during dialysis sessions. Blood samples were taken at the start of dialysis, sera were kept frozen until assay, and in vitro SHR was performed on washed leukocytes at 37 degrees C for 40 minutes. A high (range, 20% to 80%) and reproducible in vitro SHR was observed among 24% of patients. No correlation was found between SHR and the type of dialysis membrane used (copolymer of acrylonitrile-methallyl-sodium sulfonate, cuprophan, and polysulfone). High SHR (i.e., greater than 20%) was observed mainly among patients with detectable antiethylene oxide IgE antibodies (p less than 0.001). High levels of IgE (greater than 300 ng/ml), specific IgE antibodies, and a high in vitro SHR were all observed concomitantly in 26% of the patients with skin flush reactions and/or pruritus, whereas all three of these parameters together were present in only 3% of the symptom-free patients. To our knowledge, this is the first time that a high in vitro SHR has been described among HD patients, and putative correlations with in vivo phenomena are discussed.     

Observations and comments on the psychosocial determinants of depressive illness among Nigerian adults

The psychosocial characteristics of 142 depressive patients seen for the first time at the psychiatric hospital at Enugu were studied. Women (54.9 percent) outnumbered men (45.1 percent). Whereas neurotic depression occurred more commonly in those under 30 years of age, psychotic depression was more often seen in those aged over 30 years. Married status was more common among female psychotic depressives, but was not significant among neurotic depressives. Psychotic depression occurred more often among rural dwellers, whereas neurotic depression was diagnosed more often among urban residents. The majority of the patients had a low level of education. A significant incidence of neurotic depression was observed among students and housewives. The probable sociocultural factors responsible for the observed differences are discussed. A suggestion is made for a central control of all available alternative forms of treatment (orthodox, novel, and traditional) for improved delivery of mental health care.     

Sleep duration and overweight among elementary schoolchildren: a population-based study in Japan

Although a number of studies have investigated the relationship of sleep duration to overweight and obesity, studies conducted among population-based elementary schoolchildren have been limited in Japan. The aim of the present study was to investigate the relationship between sleep duration and overweight among elementary schoolchildren in Japan. The study subjects were all fourth-grade schoolchildren (9 or 10 years of age) in Ina-town, Saitama Prefecture, Japan from 1999 to 2008. Information concerning each subject's sex, age, and lifestyle was obtained using a self-administered questionnaire, while measurements of his or her height and weight were carried out. Childhood overweight was determined according to the definition established by the International Obesity Task Force. Data from 3,433 children were analyzed. In logistic regression analysis, a statistically significant dose-response relationship was observed between sleep duration and overweight among boys (p for trend = 0.014) but not among girls (p for trend = 0.149). Short sleep duration was associated with childhood overweight, and the sex difference in the association was observed. These findings suggested that it is important to consider sleep duration as part of any program to prevent overweight among elementary schoolchildren, especially among boys.     

Polymorphisms in the sialic acid-binding immunoglobulin-like lectin-8 (Siglec-8) gene are associated with susceptibility to asthma

Sialic acid-binding immunoglobulin-like lectin-8 (Siglec-8) promotes the apoptosis of eosinophils and inhibits FcɛRI-dependent mediator release from mast cells. We investigated the genetic association between sequence variants in Siglec-8 and diagnosis of asthma, total levels of serum IgE (tIgE), and diagnosis of eosinophilic esophagitis (EE) in diverse populations. The effect of sequence variants on Siglec-8 glycan ligand-binding activity was also examined. Significant association with asthma was observed for SNP rs36498 (odds ratios (OR), 0.69, P=8.8 × 10⁻⁵) among African Americans and for SNP rs10409962 (Ser/Pro) in the Japanese population (OR, 0.69, P=0.019). Supporting this finding, we observed association between SNP rs36498 and current asthma among Brazilian families (P=0.013). Significant association with tIgE was observed for SNP rs6509541 among African Americans (P=0.016), and replicated among the Brazilian families (P=0.02). In contrast, no association was observed with EE in Caucasians. By using a synthetic polymer decorated with 6′-sulfo-sLe^x, a known Siglec-8 glycan ligand, we did not find any differences between the ligand-binding activity of HEK293 cells stably transfected with the rs10409962 risk allele or the WT allele. However, our association results suggest that the Siglec8 gene may be a susceptibility locus for asthma.     

Genetic polymorphisms among Iranian Jews in Israel

Iranian Jews represent a very ancient Jewish community with a high frequency of inbreeding. A sample of Iranian Jews, mainly unrelated students, was tested for genetic markers of red blood cells and serum. The frequency of glucose-6-phosphate dehydrogenase deficiency was not uniform among Jews who had lived in different areas of Iran; it was lower among those from central Iran (6.7%) than in those from southern and western Iran (16.7% and 20.6%, respectively). The frequencies of B, CDe, cDE, S, and K alleles were among the highest recorded in Jewish ethnic groups. Iranian Jews were similar to Iraqi Jews with respect to the frequencies of the blood markers B, CDe, cde, cDe, ACP, PGM1, ADA, and Hp; however, the B and CDe markers occur with similar frequencies among indigenous Iranians. The presence of the cDe allele and the Gm1,5,13,14,17 haplotype in low frequencies indicates black admixture. Mongoloid admixture is indicated by the polymorphism of the Gm1,13,15,16,17 haplotype. The very rare phenotype Gm(3,5,13,14,17) was observed in 4.8% of 167 individuals tested. This phenotype has not been previously observed among Jews.     

Lung cancer--differences of incidence between the sexes

During the last decade increasing incidence of lung cancer among women have been observed in Poland. The aim of the study was to demonstrate differences among men and women with lung cancer. Lung cancer was diagnosed in 785 female and 4619 male in 1995 in Pulmonary Outpatients Departments. Women were younger than man when all histologic types of lung cancer were analysed (59.7 vs 61.9 years p. < 0.001). Particularly younger subjects were those with adenocarcinoma and small cell lung cancer (56.9 and 57.4 years for women and for men respectively 60.2 and 59.6 years, p < 0.001). Although squamous lung cancer was the most prevalent histological type among men (43.7%) and women (24.7%), about two times higher percentage of men had this neoplasm (p. < 0.001). Adenocarcinoma (18% vs 6.6%, p. < 0.001) and small cell lung cancer (18.5% vs 15.5% p. < 0.001) were prevalent in significantly higher percentage among female than male. Nonsmokers were more frequently noticed among women then men (20.4% vs. 1.9%, p. < 0.001), particularly those with adenocarcinoma. Also women smoked less intensively (33.6 pack/years vs. 42.3 pack/years, p < 0.001) except those with squamous cancer. The higher incidence of cancer was observed among mothers (7% vs 3.8% p. < 0.001) and fathers (7.1% vs 5.6%, p. < 0.001) of women than men with lung cancer.     

HLA A*02 allele frequencies and B haplotype associations in Western Indians

The population of Western India is described as Australoid or Proto-Australoid elements with admixture from Caucasian, Mongoloid, and Aryan races. We investigated the frequencies of human leukocyte antigen (HLA) A*02 alleles and their B* allele haplotype associations among 664 healthy unrelated Western Indians. Fifty-one of 204 serologically typed A2 individuals were analyzed for 56 molecular A*02 subtypes using high resolution polymerase chain reaction-reverse line strip-sequence-specific oligonucleotide hybridization (PCR-RLS-SSOP). A total of seven different A*02 alleles were identified, A*02011 (16%), A*0203 (16%), A*0205 (2%), A*0206 (2%), A*0211 (52.9%), A*0222 (4%), and A*0236 (8%). The most common HLA B allele associated with A*02 was B*40. Among the 42 subtypes HLA B*4006 (37.22%) was the most frequent subtype. HLA A*0211 was highly frequent in this population, A*0206 and A*0203 are common alleles observed among Asian populations, whereas A*0205 occurs in Caucasians and Africans and A*0222 has been observed among Hispanics. A*0236 has been observed among the western Indians exclusively. Most of the HLA A*02 subtypes observed were associated with B*4006 haplotype, although A*0236 was with B*0702 or B*1302 among the western Indians. The prevalence of A*0211 at high frequencies, A*0222, A*0236 novel alleles along with A*02 related haplotypes, demonstrate a substantial heterogeneity, which may be a consequence of founder effect, racial admixture, or selection pressure due to environmental factors.     

Prevalence of congenital anomaly syndromes in a Spanish gypsy population.

We analysed the sample of gypsies included in the Spanish Collaborative Study of Congenital Malformations (ECEMC), a hospital based, case-control study and surveillance system. Special emphasis was placed on the birth prevalence of recessive multiple congenital anomaly syndromes, comparing their frequency in the gypsy population with that observed among non-gypsies. We observed an increased prevalence of birth defects, mostly because of groups of children with patterns of multiple anomalies and with autosomal recessive syndromes. The latter were approximately seven times more frequent in gypsies than in non-gypsies. We also estimated the carrier frequency in both groups (gypsy and non-gypsy). We consider that the frequent occurrence of the conditions observed reflects the high rate of consanguineous couples among the Spanish gypsy population.     

Time trends in waiting time to pregnancy among Danish twins

BACKGROUND: Little is known about time trends in fecundity because few population-based data are available. In a survey among female twins born from 1953 to 1976, their time to pregnancy did not differ from singletons and can be considered to represent the fecundity of the general population. METHODS: Information was collected by interview about waiting time to first pregnancy (TTP) and any periods of subfecundity among both male and female twins born between 1931 and 1952. Trends were analysed by considering the year of birth of the index person (birth cohort effect) or year at which the first attempt started (period effect). RESULTS: Eighty-five percent of male and 81.3% of female eligible twins participated. A total of 1598 male twins and 1653 female twins reported a TTP value and 1671 men and 1715 women had a value for their first attempt to conceive including unsuccessful attempts. No overall trends in either male or female TTP were observed with increasing year of birth or of starting time, after adjustment for confounders, but for attempts fecundity increased among female twins by year of birth or of starting the attempt. A decreasing risk of severe infertility with increasing year of birth or year for start of the attempt was observed among male twins, but this observation was based on only 81 men and was not seen among female twins. CONCLUSION: No decreasing trend in fecundity was observed among Danish twins born between 1931 and 1952 who had completed their reproduction. Female twins had a slight increase in fecundity, and men a decrease in severe infertility. In addition, TTP was apparently well reported and recalled for up to 50 years among both male and female twins.     

HLA A*02 allele and B-associated haplotype diversity in Indians

Human leucocyte antigen (HLA) A2 is the most heterogeneous allele at the HLA A locus, with approximately 56 different subtypes. Substantial heterogeneity in A*02 distribution has been observed in populations worldwide. HLA A2 allele distribution varies (4-25%) in different Indian populations and castes. HLA B40 is the most common allele associated with A2 haplotypes. In this study, molecular A*02 subtyping in Maratha (western Indian--Aryan) and Nadar (south Indian--Dravidian) caste groups using high-resolution polymerase chain reaction/reverse line strip/sequence-specific oligonucleotide hybridisation (PCR-RLS-SSOP) indicates the presence of 10 subtypes (A*02011, A*0203, A*0205, A*0206, A*0207, A*0208, A*0209, A*0211, A*0222 and A*0236) in variable frequencies. Moreover, A*0211-B*4006 was commonly observed among the north Indian Maratha caste and A*02011-B*4006 among the south Indian Nadar caste groups. HLA A*0211 is found frequently in this population and in Asian Indians, and has been reported with low frequencies in many other populations worldwide. A*0222 has been observed among Hispanics, while A*0236 has been observed exclusively among members of the Maratha caste. The findings demonstrate a substantial heterogeneity among the groups studied that may be a consequence of founder effect, racial admixture or selection pressure due to environmental factors.     

Associations between the use of antimicrobial agents for growth promotion and the occurrence of resistance among Enterococcus faecium from broilers and pigs in Denmark, Finland, and Norway

This study compares the susceptibility of Enterococcus faecium isolated from pigs and poultry in Denmark, Finland, and Norway to antimicrobial agents used for growth promotion. E. faecium was isolated from 211 broilers and 55 pigs in Denmark in 1997, from Norwegian 55 poultry farms (turkey and broiler farms) and 4 swine farms between 1995 and 1997, and Finnish poultry (52) and swine (43) in 1996 and examined for susceptibility to avilamycin, avoparcin, bacitracin, flavomycin, monensin, salinomycin, spiramycin, tylosin, and virginiamycin. Only a limited number of isolates were categorized as resistant to monensin or salinomycin. In general, an association between the usage of antimicrobial agents in the respective countries and the occurrence of associated resistance was observed. Resistance to avilamycin was frequently observed among isolates from broilers in Denmark, where avilamycin has been used, whereas all isolates from Finland and Norway, where these drugs have not been used, were susceptible. The same phenomenon could be observed for avoparcin, bacitracin, tylosin, and virginiamycin; resistance was frequently observed among isolates from where these antimicrobials have been widely used, but rarely among isolates from where the use has been limited. Also for avoparcin and bacitracin, an association between usage and occurrence of resistance was observed. All isolates categorized as avoparcin resistant contained the vanX gene; isolates from broilers had the T variant in position 8,234 and isolates from pigs the G variant. Three (1%) of the 222 isolates resistant to tylosin contained the ermA gene and 196 (88%) ermB. Sixteen (11%) of the 146 virginiamycin-resistant isolates from broilers and two (7%) of the 27 virginiamycin-resistant isolates from pigs in Denmark contained the satA gene, whereas satA was not observed among any of the virginiamycin-resistant isolates from Finland. A total of 72% of the virginiamycin-resistant E. faecium from broilers in Denmark and all nine virginiamycin-resistant E. faecium from Finland contained satG. This gene was also observed among two (7%) of the virginiamycin-resistant isolates from pigs in Denmark. This study indicates that the use of antimicrobial agents for growth promotion in Denmark, Finland, and Norway have selected for resistance to most of these drugs among E. faecium in food animals.     

Spatial and habitat distribution of Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) in Banambani village, Mali

We studied the larval distribution and composition of Anopheles arabiensis Patton, An. gambiae s.s. Giles, and its forms, among local habitats; and their association with the adults between these habitats in Banambani village, Mali during the mid-rainy seasons of 1997-1999. For species and form identification we used polymerase chain reaction (PCR) and PCR-restriction fragment-length polymorphism (RFLP). Differences among species in the distribution of larvae were observed in 1998, but not in 1997 or 1999, although they were on the borderline of statistical significance. Differences among the M and S molecular forms were statistically significant in 1999 when rainfall was high, but not in the two prior, drier sampling periods. Combining all information into the Fisher multiple comparisons test, there were statistically significant differences between species and molecular forms during the 3-yr study period. Hybrid larvae between the M and S forms were observed (0.57%), the first such observation to our knowledge. In spite of differences among larval distribution, no differences of adult species composition were observed among habitats. Factors that influence the distributions of An. gambiae larval populations are discussed.