Screening of Serum Biomarkers for Distinguishing between Latent and Active Tuberculosis Using Proteome Microarray

Objective To identify potential serum biomarkers for distinguishing between latent tuberculosis infection（LTBI） and active tuberculosis（TB）. Methods A proteome microarray containing 4,262 antigens was used for screening serum biomarkers of 40 serum samples from patients with LTBI and active TB at the systems level. The interaction network and functional classification of differentially expressed antigens were analyzed using STRING 10.0 and the TB database, respectively. Enzyme-linked immunosorbent assays（ELISA） were used to validate candidate antigens further using 279 samples. The diagnostic performances of candidate antigens were evaluated by receiver operating characteristic curve（ROC） analysis. Both antigen combination and logistic regression analysis were used to improve diagnostic ability. Results Microarray results showed that levels of 152 Mycobacterium tuberculosis（Mtb）-antigenspecific IgG were significantly higher in active TB patients than in LTBI patients（P 〈 0.05）, and these differentially expressed antigens showed stronger associations with each other and were involved in various biological processes. Eleven candidate antigens were further validated using ELISA and showed consistent results in microarray analysis. ROC analysis showed that antigens Rv2031 c, Rv1408, and Rv2421 c had higher areas under the curve（AUCs） of 0.8520, 0.8152, and 0.7970, respectively. In addition, both antigen combination and logistic regression analysis improved the diagnostic ability. Conclusion Several antigens have the potential to serve as serum biomarkers for discrimination between LTBI and active TB.

Association of Helicobacter pylori IgA antibodies with the risk of peptic ulcer disease and gastric cancer

AIM: To compare the prevalence of Helicobacter pylori (Hpylori) IgG and IgA antibodies between adult subjects,with defined gastric diseases, nondefined gastric disorders and those representing the population.METHODS: Data on H pylori IgG and IgA antibodies,determined by enzyme immunoassay, were analyzed in 3 252 subjects with DGD including 482 patients with gastric ulcer, 882 patients with duodenal ulcer, 1 525patients with chronic gastritis only and 363 subjects with subsequent gastric cancer, 19 145 patients with NoDg and4 854 POPUL subjects. The age-adjusted prevalences were calculated for 1- and 20-year age cohorts.RESULTS: The prevalences of IgG antibodies were equally high (89-96%) in all 20-year age cohorts of the DGD groups, whereas the prevalences of IgG antibodies were lower and increased by age in the POPUL and NoDg groups. The prevalences of IgA antibodies were also higher in the DGD groups; among them CA (84-89%) and GU groups (78-91%) showed significantly higher prevalences than DU (68-77%) and CG patients (59-74%) (OR 2.49, 95%CI 1.86-3.34 between the GU and DU groups). In the CA, GU, and DU groups, the IgA prevalences showed only minor variation according to age, while they increased by age in the CG, POPUL, and NoDg groups (P≤0.0001). The IgA response, but not the IgG response, was associated with an increased risk of CA (OR 2.41, 95%CI 1.79-3.53) and GU (OR 2.57,95%CI 1.95-3.39) in comparison with CG patients.CONCLUSION: An IgA antibody response during H pylori infection is significantly more common in CA and GU patients as compared with CG patients.

Inflammation and immunosuppression in severe acute pancreatitis

Acute pancreatitis(AP) is a common disease,which usually exists in its mild form.However,in a fifth of cases,the disease is severe,with local pancreatic complications or systemic organ dysfunction or both.Because the development of organ failure is the major cause of death in AP,early identification of patients likely to develop organ failure is important.AP is initiated by intracellular activation of pancreatic proenzymes and autodigestion of the pancreas.Destruction of the pancreatic parenchyma first induces an inflammatory reaction locally,but may lead to overwhelming systemic production of inflammatory mediators and early organ failure.Concomitantly,anti-inflammatory cytokines and specific cytokine inhibitors are produced.This anti-inflammatory reaction may overcompensate and inhibit the immune response,rendering the host at risk of systemic infection.At present,there is no specific treatment for AP.Increased understanding of the pathogenesis of systemic inflammation and development of organ dysfunction may provide us with drugs to ameliorate physiological disturbances.

Immunogenetic characteristics of patients with autoimmune gastritis

AIM:To explore whether predisposition to autoimmune gastritis (AIG) is found in human leukocyte antigen (HLA), cytokine or killer cell immunoglobulin-like receptor (KIR) gene variations.METHODS: Twelve Finnish patients with autoimmunetype severe atrophy of the gastric corpus were included. The patients' serum was analyzed for pepsinogen-interleukin (IL)-1 gene cluster, IL-2, IL-4, IL-6, IL-10, IL-12, interferon γ, transforming growth factor β, and tumor necrosis factor α. Variation in KIR genes was also explored. The results were compared with prevalence of the polymorphisms in Finnish or European populations.RESULTS: All patients had pepsinogen-CONCLUSION: As explored with modern DNA-based methods, HLA-DRB1*04 and DQB1*03 alleles, but not HLA-B8-DRB1*03, may predispose to AIG.

Association of autoimmune type atrophic corpus gastritis with Helicobacter pylori infection

AIM:To study the association between Helicobacter pylori(H.pylori)infection and autoimmune type atrophic gastritis. METHODS:Twenty-three patients with different grades of atrophic gastritis were analysed using enzyme immunoassay-based serology,immunoblot-based serology,and histology to reveal a past or a present H.pylori infection.In addition,serum markers for gastric atrophy(pepsinogenⅠ,pepsinogenⅠ/Ⅱand gastrin)and autoimmunity[parietal cell antibodies(PCA), and intrinsic factor(IF),antibodies]were determined. RESULTS:Of the 14 patients with severe gastricatrophy,as demonstrated by histology and serum markers,and no evidence for an ongoing H.pylori infection,eight showed H.pylori antibodies by immunoblotting.All eight had elevated PCA and 4/8 also had IF antibodies.Of the six immunoblot-negative patients with severe corpus atrophy,PCA and IF antibodies were detected in four.Among the patients with low to moderate grade atrophic gastritis(all except one with an ongoing H.pylori infection),serum markers for gastric atrophy and autoimmunity were seldom detected.However,one H.pylori negative patient with mild atrophic gastritis had PCA and IF antibodies suggestive of a pre-atrophic autoimmune gastritis. CONCLUSION:Signs of H.pylori infection in autoimmune gastritis,and positive autoimmune serum markers in H.pylori gastritis suggest an etiological role for H.pylori in autoimmune gastritis.

Helicobacter pylori infection and low serum pepsinogen I level as risk factors for gastric carcinoma

AIM: To study whether examination of CagA antibodies could increase the odds ratio for gastric cancer in a casecontrol study, and how often other serum markers of gastric cancer risk could be found in Helicobacter pylorinegative patients.METHODS: H pylori CagA and parietal cell antibodies(PCAs), and serum pepsinogen Ⅰ (SPGI) levels were compared between patients with gastric cancer and controls who received endoscopic examination due to reasons other than gastrointestinal malignancy.RESULTS: The odds ratio (OR) for gastric cancer was2.9 (95% CI 1.4-5.8) in H pylori+ patients, and 2.4 (95%CI 1.2-4.9) in CagA+ patients. When results of H pylori and CagA antibodies were combined, OR increased to5.0 (95% CI 2.5-10.0). Furthermore, if cardia cancer patients were excluded, the OR increased to 6.8 (95% CI3.1-14.8). Among patients with a low SPGI level, the OR was 12.0 (95% CI 4.1-35.3). However, the risk was significant only in the older age group. The number of patients with low SPGI was significantly higher in H pylori-/CagA+ patients as compared to other cancer patients.CONCLUSION: Examination of both H pylori and CagA antibodies increases the OR for gastric cancer in our casecontrol study. CagA antibodies are important in detecting previous H pylori infection in advanced atrophic gastritis or cancer when spontaneous decline of H pylori antibodies occurs. SPGI may be helpful in screening elderly gastric cancer patients.

Milk protein IgG and IgA:The association with milk-induced gastrointestinal symptoms in adults

AIM:To study the association between serum levels of milk protein IgG and IgA antibodies and milk-related gastrointestinal symptoms in adults.METHODS:Milk protein IgG and IgA antibodies were determined in serum samples of 400 subjects from five outpatient clinics in Southern Finland.Subjects were randomly selected from a total of 1900 adults undergoing laboratory investigations in primary care.All 400 participants had completed a questionnaire on abdominal symptoms and dairy consumption while waiting for the laboratory visit.The questionnaire covered the nature and frequency of gastrointestinal problems,the provoking food items,family history and allergies.Twelve serum samples were disqualifi ed due to insuff icient amount of sera.The levels of specif ic milk protein IgG and IgA were measured by using the ELISA technique.The association of the milk protein-specific antibody level was studied in relation to the milk-related gastrointestinal symptoms and dairy consumption.RESULTS:Subjects drinking milk(n=265) had higher levels of milk protein IgG in their sera than non-milk drinkers(n=123,P<0.001).Subjects with gastrointestinal problems related to milk drinking(n=119) consumed less milk but had higher milk protein IgG levels than those with no milk-related gastrointestinal symptoms(n=198,P=0.02).Among the symptomatic subjects,those reporting dyspeptic symptoms had lower milk protein IgG levels than non-dyspeptics(P<0.05).However,dyspepsia was not associated with milk drinking(P=0.5).The association of high milk protein IgG levels with constipation was close to the level of statistical signif icance.Diarrhea had no association with milk protein IgG level(P=0.5).With regard to minor symptoms,flatulence and bloating(P=0.8),were not associated with milk protein IgG level.Milk protein IgA levels did not show any association with milk drinking or abdominal symptoms.The levels of milk protein IgA and IgG declined as the age of the subjects increased(P<0.004).CONCLUSION:Milk protein IgG but not milk IgA seems to be associated with self-reported milk-induced gastrointestinal symptoms.

Tuberculosis Case Notification and Treatment Outcomes in West Gojjam Zone, Northwest Ethiopia: A Five-Year Retrospective Study

Introduction: Tuberculosis (TB) is a major public health concern in Ethiopia. Analysis of TB case notification and treatment outcomes is crucial to understand the TB control program performance. The current study was carried out to assess trends of TB case notifications, treatment success rate and factors associated with unsuccessful treatment outcome among TB patients in West Gojjam Zone of Amhara Region, Ethiopia. Method: A retrospective cohort study was conducted in West Gojjam Zone. Demographic and clinical data were reviewed for all TB patients registered between July 2007 and June 2012 at 30 randomly selected public health facilities of the study zone. In addition, annual case notification reports of the study zone were used to analyze trends in TB case notifications. Logistic regression analysis was used to assess the association between potential predictor variables and unsuccessful treatment outcomes. Results: Tuberculosis case notification for all forms of TB decreased from 203/100,000 population in 2007 to 155/100,000 population in 2012. Among patients whose treatment outcomes were evaluated, 94.4% were successfully treated, 0.3% had treatment failure, 1.5% defaulted and 3.7% died. In multivariate analysis, the odds of unsuccessful treatment outcome was higher among retreatment cases than new cases (adjusted OR, 3.44;95% CI: 1.92, 6.19). HIV co-infected cases were more likely to have unsuccessful treatment outcome compared to HIV negatives (adjusted OR, 2.68;95% CI: 1.92, 3.72). Conclusion: Tuberculosis case notification rates showed a decreasing trend between 2007 and 2012. The treatment success rate exceeded the 90% treatment success rate target as set by the WHO. Special attention is required for patients with high risk of unsuccessful treatment outcome. Therefore, retreatment cases, and HIV positive cases need strict follow up throughout their treatment period.

Fecal microbiota transplantation as novel therapy in gastroenterology:A systematic review

AIM:To study the clinical efficacy and safety of Fecal microbiota transplantation(FMT).We systematically reviewed FMT used as clinical therapy.METHODS:We searched MEDLINE,EMBASE,the Cochrane Library and Conference proceedings from inception to July,2013.Treatment effect of FMT was calculated as the percentage of patients who achieved clinical improvement per patient category,on an intention-to-treat basis.RESULTS:We included 45 studies;34 on Clostridium difficile-infection(CDI),7 on inflammatory bowel disease,1 on metabolic syndrome,1 on constipation,1 on pouchitis and 1 on irritable bowel syndrome(IBS).In CDI 90% resolution of diarrhea in 33 case series(n = 867) was reported,and 94% resolution of diarrhea after repeated FMT in a randomized controlled trial(RCT)(n = 16).In ulcerative colitis(UC) remission rates of 0% to 68% were found(n = 106).In Crohn's disease(CD)(n = 6),no benefit was observed.In IBS,70% improvement of symptoms was found(n = 13).100% Reversal of symptoms was observed in constipation(n = 3).In pouchitis,none of the patients(n = 8) achieved remission.One RCT showed significant improvement of insulin sensitivity in metabolic syndrome(n = 10).Serious adverse events were rare.CONCLUSION:FMT is highly effective in CDI,and holds promise in UC.As for CD,chronic constipation,pouchitis and IBS data are too limited to draw conclusions.FMT increases insulin sensitivity in metabolic syndrome.

Physiology of gangliosides and the role of antiganglioside antibodies in human diseases

Gangliosides are structurally and functionally polymorphic sialic acid containing glycosphingolipids that are widely distributed in the human body.They play important roles in protecting us against immune attacks,yet they can become targets for autoimmunity and act as receptors for microbes,like the influenza viruses,and toxins,such as the cholera toxin.The expression patterns of gangliosides vary in different tissues,during different life periods,as well as in different animals.Antibodies against gangliosides(AGA)can target immune attack e.g.,against neuronal cells and neutralize their complement inhibitory activity.AGAs are important especially in acquired demyelinating immune-mediated neuropathies,like Guillain–Barrésyndrome(GBS)and its variant,the Miller–Fisher syndrome(MFS).They can emerge in response to different microbial agents and immunological insults.Thereby,they can be involved in a variety of diseases.In addition,antibodies against GM3 were found in the sera of patients vaccinated with Pandemrix®,who developed secondary narcolepsy,strongly supporting the autoimmune etiology of the disease.

Interleukin 8 Gene Polymorphisms Are Not Associated with Tuberculosis Susceptibility in the Chinese Population

Interleukin 8 （IL8） is an important chemokine that elicits host immune response against tuberculosis （TB）. However, whether there is an association between IL8 gene polymorphism and TB susceptibility in the Chinese population is unknown. IL8 gene was amplified and sequenced to search for nucleotide polymorphisms among the Chinese population. Four single nucleotide polymorphisms （SNPs） were identified, selected, and analyzed in a cohort of 438 patients with TB and 536 healthy controls. Allelic, genotypic, and haplotypic analysis demonstrated that the distribution of the four IL8 SNPs between patients with TB and healthy controls was not significantly different （P〉0.05）. The four IL8 SNPs detected in this study were not associated with TB susceptibility in the Chinese population. Secretion of IL8 by peripheral blood cells was greatly stimulated upon exposure to Mycobacterium tuberculosis whole cell extract, but such enhanced secretion was not associated with the IL8 rs4073 alleles.

2000年中国结核病流行病学抽样调查菌株分子流行病学特征

目的:判定北京基因型结核分枝杆菌在中国结核分枝杆菌复合群中的群结构特征。设计:采用spoligotyping和IS6110限制性片段长度多态性(RFLP)方法对441株分枝杆菌分离株行基因分型鉴定。分离株取自2000年中国结核病流行病学抽样调查,耐药性和流行病学资料已知。结果:spoligotyping法共鉴定408株结核分枝杆菌,其中北京基因型结核分枝杆菌占64.9%(265/408)。卡方检验北京基因型结核分枝杆菌在性别、年龄、地区所占的比例没有统计学显著性差异,但是耐多药(MDR)有统计学显著性差异。北京基因型在耐多药和药物敏感性分离株中所占的比例分别为77.8%(42/54)和60.2%(213/354)。多元回归分析提示北京基因型与不同地区存在联系,但与耐多药无关。结论:北京基因型结核分枝杆菌在中国流行至少50年,并且与地区有关。

Prokaryotic expression, localization and function of the infectious laryngotracheitis virus glycoprotein G

The infectious laryngotracheitis virus (ILTV) glycoprotein G (gG) gene of E3 and Zhonghai strains was cloned, sequenced and compared with the gG gene of other Type I animal herpesviruses. To find the localization and the function of the gG in the infected cells, the 35 kD fusion pro-tein (His-GG) was expressed by inserting the coding region of gG except for the signal peptide into pET30a (+). After puri-fication of the His-GG fusion protein, the rats’ antibody to the His-GG was prepared and purified by using the protein G Sepharose. Results of laser scanning confocal microscopy (LSCM) detection showed that the ILTV gG was in the peri-nuclear region and membrane of chicken embryo liver (CEL) and kidney (CEK) cells, and that the gG accumulated more in the coalescent part than in the other parts of the adjacent CEL or CEK cells. The plaque size and the one-step growth curve tests suggested that the ILTV gG was required for viral growth by cell-to-cell direct infection in tissue-cultured CEL cells.

The putative NAD(P)H Nitroreductase,Rv3131,is the Probable Activating Enzyme for Metronidazole in Mycobacterium Tuberculosis

Tuberculosis(TB),which is caused by the intracellular pathogen,Mycobacterium tuberculosis(Mtb),remains one of the deadliest infectious diseases worldwide,accounting for 1.45 million deaths in 2018[1].Thus,there is an increasing need to develop new antibiotics or to repurpose old drugs that have the potential to shorten the current duration of therapy.Metronidazole(MTZ)is a 5-nitroimidazole antibiotic that is mainly used in the treatment of anaerobic and protozoal infections[2].The action of mechanism for metronidazole has not been fully established.As a prodrug,MTZ is inactive until taken up and reduced by anaerobic bacteria and protozoa.It is possible that the reduction of Mtz leads to the production of toxic metabolites,which bind to deoxyribonucleic acid and electron-transport proteins of microbial pathogens,thus blocking nucleic acid synthesis[3].The impressive potency of MTZ against anaerobic bacteria indicates the potential for killing Mtb under anaerobic conditions.A previous in vitro study demonstrated that MTZ has potent activity against tubercle bacilli under anaerobic conditions[4].In a non-human primate model,MTZ has been shown to prevent reactivation of latent Mtb infection,indicating the in vivo role in targeting Mtb within an anaerobic environment[5].Although there is strong evidence for the role of MTZ in affecting anaerobic bacilli,the mechanism of action has not been elucidated in Mtb considering that MTZ must be catalyzed to a nitro-free radical form with endogenous nitroreductase.To provide insight=into how MTZ is activated in anaerobic Mtb bacilli,we first characterized the nitroreductase gene,Rv3131,and identified the key residue conferring the activation of MTZ in Mtb.

Do multiparous women need to work or exercise extra hard to control gestational diabetes?

In the current issue of the Journal of Sport and Health Science,Wang et al.^(1)describe their prospective study,where they analyzed whether or not multiple pregnancies would influence glycemia and the glycemic response to physical exercise in gestational diabetes mellitus(GDM).2 Multiparous women are known to be at a higher risk for GDM than primiparas.GDM carries an increased risk for adverse perinatal outcomes,not only for the mothers but also for the newborn babies.Due to a contemporary environment that is conducive to a sedentary lifestyle and obesity,the incidence of GDM among pregnant women is increasing.

Diabetes mellitus is associated with increased mortality during tuberculosis treatment:a prospective cohort study among tuberculosis patients in SouthEastern Amahra Region,Ethiopia

Background:There is growing evidence suggesting that diabetes mellitus(DM)affects disease presentation and treatment outcome in tuberculosis(TB)patients.This study aimed at investigating the role of DM on clinical presentations and treatment outcomes among newly diagnosed TB patients.Methods:A prospective cohort study was conducted in South-Eastern Amhara Region,Ethiopia from September 2013 till March 2015.Study subjects were consecutively recruited from 44 randomly selected health facilities in the study area.Participants were categorized into two patient groups,namely,patients with TB and DM(TBDM)and TB patients without DM(TBNDM).Findings on clinical presentations and treatment outcomes were compared between the two patient groups.Cox proportional hazard regression analysis was applied to identify factors associated with death.Results:Out of 1314 TB patients enrolled in the study,109(8.3%)had coexisting DM.TBDM comorbidity[adjusted hazard ratio(AHR)3.96;95%confidence interval(C.I.)(1.76–8.89)],and TB coinfection with human immunodeficiency virus(HIV)[AHR 2.59;95%C.I.(1.21–5.59)]were associated with increased death.TBDM and TBNDM patients did not show significant difference in clinical symptoms at baseline and during anti-TB treatment period.However,at the 2nd month of treatment,TBDM patients were more symptomatic compared to patients in the TBNDM group.Conclusions:The study showed that DM is associated with increased death during TB treatment.DM has no association with clinical presentation of TB except at the end of the intensive phase treatment.Routine screening of TB patients for DM is recommended for early diagnosis and treatment of patients with TBDM comorbidity.

Applying tuberculosis management time to measure the tuberculosis infectious pool at a local level in Ethiopia

Background:Measuring the size of the infectious pool of tuberculosis(TB)is essential to understand the burden and monitor trends of TB control program performance.This study applied the concept of TB management time to estimate and compare the size of the TB infectious pool between 2009 and 2014 in West Gojjam Zone of Amhara Region,Ethiopia.Methods:New sputum smear-positive and smear-negative pulmonary TB(PTB)and retreatment cases who attended 30 randomly selected public health facilities in West Gojjam Zone from October 2013 to October 2014 were consecutively enrolled in the study.In order to determine the infectious period,the TB management time(number of days from the onset of cough until start of anti-TB treatment)was computed for each patient category.The number of undiagnosed TB cases was estimated and hence the TB management time for the undiagnosed category was calculated.The total size of the TB infectious pool during the study period for the study zone was estimated as the annual number of infectious person days.Results:New smear-positive and smear-negative PTB cases contributed 25,050 and 12,931 infectious person days per year to the TB infectious pool,respectively.The retreatment and presently undiagnosed cases contributed 8840 and 34,310 infectious person days per year,respectively.The total size of the TB infectious pool in West Gojjam Zone during the study period was estimated at 81,131 infectious person days per year or 3405 infectious person days per 100,000 population per year.Compared to a similar study done in 2009 in the study area,the current study showed reduction of the TB infectious pool by 244,279 infectious person days.Conclusions:TB management time is a simple and practical tool that may help to estimate and compare the changes in the size of the TB infectious pool at local level.It may also be used as an indicator to monitor the changes in TB control program performance.