A Search for Candidate Viruses in Type 1 Diabetic Pancreas Using the Polymerase Chain Reaction

In order to investigate a possible viral aetiology for Type 1 diabetes the polymerase chain reaction (PCR) technique was used. Pancreatic tissue from Type 1 diabetic subjects was examined for the presence of a panel of common viruses. Primers specific for mumps, measles, cytomegalovirus, and Epstein Barr virus, as well as primers located in a highly conserved region of the enterovirus genome which are capable of detecting all of the following family members: Coxsackie B, echovirus, polio, mengovirus, and encephalomyocarditis virus were used to screen 18 Type 1 diabetic subjects of whom 3 had proven insulitis, 12 Type 2 diabetic subjects and 18 non-diabetic controls. Epstein Barr virus was detected in two Type 1 (13%), two Type 2 (22%), and three of the normal non-diabetic pancreases (20%), and the DNA sequences confirmed by direct sequencing. Cytomegalovirus was detected in one of the normal pancreases only and no evidence of any of the other viruses was found. It is concluded that the Type 1 diabetic pancreatic samples studied did not show persistence of infection with any of the above viruses. Non-persistent acute infection of the pancreas by the above viruses cannot be excluded in the aetiology of Type 1 diabetes from this study.     

Detection of Common Viruses Using the Polymerase Chain Reaction to Assess Levels of Viral Presence in Type 1 (Insulin-dependent) Diabetic Patients

The polymerase chain reaction was used to detect a range of common viruses in the peripheral blood of Type 1 diabetic and non-diabetic control patients in order to identify any abnormal viral presence, with possible roles in the pathogenesis of Type 1 diabetes. Peripheral blood from 17 newly diagnosed Type 1 diabetic patients, 38 Type 1 diabetic patients with disease of longer duration, and 43 age and sex matched non-diabetic controls was obtained. Samples were screened for cytomegalovirus, Epstein-Barr virus, enterovirus (including coxsackie), and mumps virus. Cytomegalovirus was detected in control patients only (5 %), Epstein-Barr virus was detected equally in newly diagnosed and control patients (12%), and enterovirus was detected slightly more frequently in diabetic than non-diabetic patients (41% and 31%, respectively). Mumps virus was not detected in any of the samples. It is concluded that Type 1 diabetic individuals are neither more prone to persistence of common viruses nor to more frequent acute infections with the viruses tested for than non-diabetic individuals. If common viruses are involved in the pathogenesis of Type 1 diabetes then they act either as non-specific agents to which the host has abnormal immune responses, or, the diabetogenic viruses are eliminated from the body by the time of disease diagnosis.     

用PCR方法检测婴儿肝炎综合征患婴的HCMV感染情况

两年来收集到婴儿肝炎综合征患者（15－180天）共62例。用Polymerase Chain reactin(PCR)方法检测尿标本中的HCMV DNA。结果显示：阳性率为45．2％（28／62），并且1月内患婴的阳性率（60％）明显高于1－6月患婴（31．3％）。提示Cytomagalovirus(HCMV)是引起婴儿肝炎综合征的主要病原，并且在1月内患婴中是首要病原。     

TaqMan探针实时荧光PCR方法检测粪便中微小隐孢子虫卵囊

以微小隐孢子虫(Cryptosporidium parvum)特异性DnaJ-like蛋白基因的保守序列为模板,设计和合成特异性引物和荧光标记探针,通过检测微小隐孢子虫卵囊DNA和加标模拟样品进行敏感性分析,建立标准曲线,并对其特异性和干扰性进行评价。结果显示,该方法只对微小隐孢子虫卵囊进行特异性扩增,其他常见的肠道原虫和肠道病原菌均不能扩增;微小隐孢子虫纯卵囊基因组DNA检测的灵敏度为26个/ml;对加标粪样可检测至2 600个/ml卵囊。提示本研究建立的实时荧光PCR检测微小隐孢子虫卵囊方法具有快速、特异性和敏感性高等优点。     

Analysis of Clonal Relationship Using Single-Cell Polymerase Chain Reaction in a Patient with Concomitant Mantle Cell Lymphoma and Multiple Myeloma

We report a case of concomitant mantle cell lymphoma (MCL) and multiple myeloma (MM) in which we investigated the possibility of a clonal relationship. A 76-year-old man was diagnosed with MCL [immunoglobulin (Ig)M,D-kappa; stage IVB] and MM (IgG-kappa; stage I). Ig heavy chain (IgH) gene complementarity-determining region 3 in DNA from both the MCL tumor and from single MM cells from bone marrow smears was amplified to investigate whether there was a clonal relationship between MCL and MM. Sequence analysis revealed no clonal relationship between MCL and MM in our patient.     

用PCR方法检测粪便难辨梭状芽胞杆菌

目的：采用PCR方法检测粪便难辨梭状芽胞杆菌。方法：北京协和医院的36例应用抗生素后腹泻的病人粪便标本,采用PCR方法对难辨梭状芽胞杆菌（以下简称C．D）的毒素B基因和毒素A基因进行扩增,同时进行厌氧细菌培养。结果：PCR法毒素B基因的阳性率为46．7％（16／36）,毒素A基因为36．1％（13／36）;用厌氧菌培养的阳性率为19．2％（7／36）。PCR检测C．D毒素基因方法与厌氧培养法的一致性为100％。结论：用PCR方法检测C．D比厌氧培养法检出率明显增高（P＜0．05）,与厌氧菌培养法的一致性高。     

Patient Education Provided to Asthmatic Children: A Historical Cohort Study of the Implementation of NIH Recommendations

NIH guidelines for treatment of childhood asthma emphasize educating both patients and family about avoiding triggers and providing information to support self-management of asthma. To determine the extent to which primary care providers had implemented these recommendations, we examined the patient education provided to a cohort of asthmatic children (n = 331) between January and December 1994.

During 1994 education of any type was documented for less than half the children. Provision of education was associated with asthma severity: An action plan for exacerbations was discussed with the majority with moderate or severe asthma (61%). Avoiding triggers (aOR: 2.38, 95% CI: 1.37-4.12) and treatment goals (aOR: 3.14, 95% CI: 1.46-6.75), were more likely to be discussed with children who were prescribed inhaled anti-inflammatory medication, after adjustment for asthma severity and age.

Limited implementation of the NIH recommendations by primary care providers in our study may have reduced their impact on the management of childhood asthma.     

地里纤恙螨红、白体色体内恙虫病立克次体的聚合酶链反应技术检测及其传病作用

本文应用聚合酶链反应技术（ＰＣＲ）检测了红、白体色地里纤恙螨亲代成虫及子代幼虫体内恙虫病立克次体ＤＮＡ的动态变化,结果表明：恙虫病立克次体能在红、白体色恙螨体内生长繁殖,且至少持续２７０天,并可以经卵传递给子代。叮咬试验证明两体色恙螨幼虫均具有叮咬和传病能力。     

Failure to Detect Hepatitis A Virus RNA in Factor VIII Concentrates Using the Polymerase Chain Reaction: Results of a Preliminary Study

The transmission of hepatitis A virus (HAV) infection to recipients of some batches of Factor VIII has recently been reported. The polymerase chain reaction (PCR) was used for the detection of HAV RNA in factor VIII concentrates. Primer sequences used were derived from a consensus of published sequences in the 5' non coding region; a nested PCR was used to increase sensitivity and specificity and the resulting fragment was 151 base pairs in length. The PCR was initially validated in clinical samples and only IgM anti-HAV positive patient samples and a sample of liver tissue from a patient who required liver transplantation for fulminant hepatitis A were HAV PCR positive. Other samples tested included those that were IgG anti-HAV positive; these were found to be PCR negative. In an investigation of coagulation factor VIII concentrates by HAV PCR, 40 batches of solvent/detergent-treated high-purity concentrate from four different manufacturers, including one batch of factor VIII possibly implicated in HAV transmission, and a further 3 batches of monoclonal antibody purified factor VIII were all HAV PCR negative. Gel chromatography material, before and after use in factor VIII purification, and eluates from this material were also negative for HAV RNA. Our preliminary results therefore suggest that either the contamination of factor VIII concentrates by HAV RNA is an extremely rare event or that the PCR is insufficiently sensitive to detect an infective HAV dose since each batch of factor VIII concentrate would have been derived from a plasma pool consisting of 10,000 donations, or more and the resulting concentration of virus may be 10² or less.     

Low Prevalence of Asthma in Westernizing Countries—Myth or Reality? Prevalence of Asthma in Estonia—A Report from the “FinEsS” Study

Aim: The aim of this study was to examine the prevalence of asthma among adults in Estonia by using different diagnostic methods and criteria for the disease. Methods: In 1995-1996, a postal questionnaire was randomly distributed in three areas of Estonia to a representative sample of 22,579 subjects aged 15-64 years. The participation rate was 78%. A clinical follow-up study including structured interview, skin prick test, lung function, and methacholine test, was performed from 1997 to 2000 among randomly selected responders to the postal questionnaire. Of 2676 subjects, 53% participated. Results: The prevalence of physician-diagnosed asthma for the postal questionnaire (PQ) was 2.7%, and in the same subjects for the structured interview (SI) 3.8%. Respiratory symptoms, except recurrent wheeze, were more common in the SI than PQ. Combinations of symptoms, except wheezing with breathlessness apart from cold, were more prevalent in the PQ responders. The prevalence of asthma defined by different symptom combinations varied from 5.4% to 8.2%. Among responders, 71-87% demonstrated bronchial hyperreactivity defined as methacholine reactivity ≤ 8 mg/mL. The symptom combinations used as surrogate variables for asthma were strongly associated with a positive skin test. Conclusion: The low prevalence of physician-diagnosed asthma probably reflects a considerable underdiagnosis of asthma in Estonia. Disease criteria for asthma based on symptom combinations together with hyperreactivity yielded a prevalence of 5%-8%, which is similar to the prevalence of asthma among adults in neighboring Western countries.     

Quantitative Assessment of Minimal Residual Disease in Childhood Lymphoid Malignancies Using an Allele-Specific Oligonucleotide Real-Time Quantitative Polymerase Chain Reaction

We developed an assay using a real-time quantitative polymerase chain reaction (RQ-PCR) for the quantitative assessment of minimal residual disease (MRD) in childhood lymphoid malignancies by using a consensus V-region probe combining a allele-specific oligonucleotide (ASO) reverse primer. Our strategy employs a set consisting of a consensus V-region probe, an ASO reverse primer, and a patient-specific forward primer for clonal antigen-receptor (IgH, immunoglobulin heavy chain; TCR, T-cell receptor) gene rearrangements (IgH-ASO and TCR-ASO RQ-PCR assays). The limit of detection in both assays was 5 copies of the target/10(5) cell equivalents. We tested the assays in 17 childhood malignancies (14 cases of acute lymphoblastic leukemia and 3 of non-Hodgkin's lymphoma). High correlation coefficients of the standard curves (>0.980) and PCR efficiency (>0.95) were achieved with all primer/probe sets. In 2 (12%) of the 17 patients, ASO primers could not be designed because there was no junctional N-sequence. The quantitative data suggest that the copy number of clonal antigen receptors markedly decreased after induction therapy in 15 of 17 patients and that 1 patient relapsed and died of the disease. Consensus probes make it possible to examine a large number of patients with only a limited number of probes. The strategy used for IgH-ASO and TCR-ASO RQ-PCR assays is accurate and reliable in the clinical prospective study of MRD in childhood lymphoid malignancies.     

A Strategy for the Identification of Candidate Genes for Alcohol-Related Phenotypes and Other Human Disorders Using Rapid Polymerase Chain Reaction Mapping of Gene-Based Sequence-Tagged Sites

We describe a method for the rapid identification and mapping of human genes, including those possibly contributing to disease and alcohol-related phenotypes. New human genes are identified from cDNA libraries through single-pass sequencing into the 3' untranslated (3'UT) regions of human brain cDNAs. Primers derived from the 3'UT region sequences [representing gene-based, sequence-tagged sites (STSs)] are used for polymerase chain reaction (PCR) analyses of the CEPH megabase insert yeast artificial chromosome (YAC) DNA pools. With this approach, ∼18,000 megabase YACs can be screened and a single YAC identified using only 52 PCR reactions. The YAC localization in conjunction with other mapping techniques, such as PCR mapping to human chromosomes using somatic cell hybrids, allows identification of chromosomal band locations. In this manner, each gene can be associated with its own STS, which in turn specifies both a corresponding genomic clone and specific location in the genome. These locations can be compared with the purported locations of disease genes. The locations of the STSs can also be compared with those of Quantitative Trait Loci implicated for quantitative traits (e.g., alcohol-related phenotypes) on the basis of synteny between the mouse and human genes. Using this strategy, we found candidates for 78 human disease/syndrome genes among the first 220 genes mapped.     

Prevalence of asthma,allergic rhinitis and pollinosis in a city of Brazil: A monitoring study

BackgroundAllergic respiratory diseases such as asthma and allergic rhinitis have increased considerably in the last decades.ObjectiveThe present study estimates prevalence trends of asthma, allergic rhinitis and pollinosis in the population of a city of Southern Brazil, without restriction of age, from 2011 to 2018, using the ISAAC standardized questionnaire.MethodsData was collected from March to June of 2011 and during the same months in 2018, in order to verify trends in the prevalence of these allergic conditions. The total sample consisted of 3132 individuals of both sexes living in the municipality of Santo Ângelo, in the state of Rio Grande do Sul, Brazil.ResultsNo differences were observed in the prevalence of asthma diagnosis (15.1% in 2011 and 13.8% in 2018), however the prevalence of current wheeze was significantly reduced from 24.7% in 2011 to 21.2% in 2018 (p < 0.05). Regarding allergic conditions in 2011 and in 2018, a significant reduction was observed (p < 0.001) in reported current rhinitis (63.3% vs. 50.5%), rhinoconjunctivitis (48.9% vs. 38.8%), hay fever (52.0% vs. 43.3%), and pollinosis (29.0% vs 17.0%). Moreover, we observed an inverse relation between age and rhinoconjunctivitis and hay fever, and all symptoms were more frequent in females. Rhinoconjunctivitis and hay fever, as well as current rhinitis and pollinosis were highly prevalent among 30–39 years-old individuals, whereas current wheeze affected mainly the age group 10–19 years-old.ConclusionWhile the prevalence of asthma remained similar after seven years, allergic rhinitis and pollinosis declined between 2011 and 2018.     

Study of the Prevalence of Asthma in Adults in North India Using a Standardized Field Questionnaire

Prevalence of asthma was calculated using a Hindi adaptation of the International Union Against Tuberculosis and Lung Disease (IUATLD)-1984 questionnaire. The questionnaire was administered to patients with and without asthma to select specific items to define asthma. Questionnaire validity was tested against physician-diagnosed asthma in 753 individuals in the field, with sensitivity and specificity of 63.1% and 98.8%, respectively. The field prevalence study was conducted among 958 men and 1158 women. True population prevalence, calculated from the observed prevalence using specificity and sensitivity of the questionnaire, was 3.94%. in urban men, 3.99% in rural men, and 1.27% in both urban and rural women.     

Study of the Prevalence of Asthma in Adults in North India Using a Standardized Field Questionnaire

Prevalence of asthma was calculated using a Hindi adaptation of the International Union Against Tuberculosis and Lung Disease (IUATLD)-1984 questionnaire. The questionnaire was administered to patients with and without asthma to select specific items to define asthma. Questionnaire validity was tested against physician-diagnosed asthma in 753 individuals in the field, with sensitivity and specificity of 63.1% and 98.8%, respectively. The field prevalence study was conducted among 958 men and 1158 women. True population prevalence, calculated from the observed prevalence using specificity and sensitivity of the questionnaire, was 3.94%. in urban men, 3.99% in rural men, and 1.27% in both urban and rural women.     

Knowledge, Attitudes, and Behavior of the Parents of Asthmatic Children in Maputo

Asthma is increasing in frequency worldwide. The education of affected children and their parents is fundamental for the management of the disease. The aim of our study was to describe the knowledge, attitudes, and behavior of the parents of asthmatic children. We studied 152 adults accompanying asthmatic children to consultations in the pediatrics department of Maputo Central Hospital (Mozambique). In general, knowledge about asthma was poor: 11% of the adults thought that asthma was contagious and transmitted from person to person, and 4% thought that it was transmitted by contaminated food. More than half the parents thought that the child could not lead a normal life even during the periods between attacks. A large proportion of the parents thought that asthma could be cured by medical treatment in 7% of cases and by alternative treatment in 43% of cases. The precipitating factors were well identified by the adults, but they had an inaccurate perception of the symptoms of an asthma attack. The actions of the various classes of drugs used were poorly understood. Education programs for both the children and their parents should be developed to improve the management of asthmatic children. Surveys of this type facilitate the targeting of such programs.     

Knowledge,Attitudes, and Behavior of the Parents of Asthmatic Children in Maputo

Asthma is increasing in frequency worldwide. The education of affected children and their parents is fundamental for the management of the disease. The aim of our study was to describe the knowledge, attitudes, and behavior of the parents of asthmatic children. We studied 152 adults accompanying asthmatic children to consultations in the pediatrics department of Maputo Central Hospital (Mozambique). In general, knowledge about asthma was poor: 11% of the adults thought that asthma was contagious and transmitted from person to person, and 4% thought that it was transmitted by contaminated food. More than half the parents thought that the child could not lead a normal life even during the periods between attacks. A large proportion of the parents thought that asthma could be cured by medical treatment in 7% of cases and by alternative treatment in 43% of cases. The precipitating factors were well identified by the adults, but they had an inaccurate perception of the symptoms of an asthma attack. The actions of the various classes of drugs used were poorly understood. Education programs for both the children and their parents should be developed to improve the management of asthmatic children. Surveys of this type facilitate the targeting of such programs.