The Antibody Response to Brucella: Immunoglobulin Response Measured by Enzyme-linked lmmunosorbent Assay and Conventional Tests

Abstract: The antibody response to Brucella: immunoglobulin response measured by enzymelinked immunosorbent assay and conventional tests. G. L. Gilbert and L. A. Hawes, Aust. N.Z. J. Med ., 1981, 11, pp 40–45.
An enzyme-linked immunosorbent assay was adapted to measure total and individual classes of brucella antibody. The results were compared with those of conventional tests for brucella antibody on the sera of a number of healthy seropositive abattoir workers and several patients with either acute or suspected chronic brucellosis. IgG was the class of brucella specific immunoglobulin most commonly detected in all groups. IgM was present in the sera of 40% of seropositive abattoir workers, all but one of the patients with recent acute brucellosis or seroconversion and none of those with suspected chronic brucellosis. Many of the abattoir workers' sera which contained brucella specific IgM gave negative results in the direct agglutination test. The presence of brucella specific lgM in the sera of these men was, in most cases, associated with no past history of acute brucellosis and a relatively short period of employment in the abattoir. It is suggested that the presence of brucella specific IgM in the serum of a person occupationally exposed to 6. abortus, probably indicates a relatively recent primary infection, either symptomatic or sub clinical and has no prognostic significance. Repeated or prolonged exposure is associated with IgG brucella antibodies, often in high titre, irrespective of symptoms. It was not possible, on the basis of any serological tests performed in this study to distinguish healthy people exposed to brucella from those with symptoms consistent with chronic brucellosis     

Unusual complication of breast implants: Brucella infection

Brucellosis is hyperendemic in Saudi Arabia. We report infection in a breast implant as an unusual complication of laboratory-acquired brucellosis. A 48-year-old female developed fever, rigors, headache, arthralgia and weight loss. A blood culture for Brucella was positive. A 6-week course of antibiotics effected a clinical and bacteriological cure but the resolution was short-lived. Six weeks later a relapse of her febrile symptoms occurred together with the appearance of a breast abscess. Cultures of the abscess and blood yielded brucella. A second course of antibiotics together with surgical drainage and subsequent reconstruction resulted in a cure. Received: August 30, 2000 · Revision accepted: July 10, 2001     

布氏菌胶体金免疫层析快速检测板使用效果评价

目的对布氏菌胶体金免疫层析快速检测板(CG IRST)的使用效果进行评价,为布氏菌病防治提供一种新的、快速、简易、准确的好方法。方法我们用布氏菌纯蛋白衍化物做为该检测板的包被抗原,用金标记prote in A,利用间接法检测人和不同动物血液样品中的抗布氏菌抗体。待测样品中的抗布氏菌抗体与检测板上的金标记prote in A结合后沿着特殊纤维素膜移动,与膜上的固相抗原结合形成肉眼可见的红色线条。结果本检测板只与有抗布氏菌抗体的样品有特异性反应,与正常人、兔和小鼠的样品以及有抗结核抗体的样品无交叉反应。布氏菌CG IRST与布氏菌SAT比较,从被检测的部份已知样品看,两法的阳性符合率为100%。结论用布氏菌CG IRST检测血样中的布氏菌抗体,灵敏、快速、简便、特异性好,无需仪器设备,对基层单位开展布病监测,不失为一种有效的好方法,应有良好的应用前景。     

布鲁氏菌BP26蛋白的表达纯化及其抗原性的研究

目的获得布鲁氏菌感染血清学诊断优势抗原BP26重组蛋白。方法应用PCR技术从布鲁氏菌减毒株(104M)中扩增出bp26目的基因片段,将其克隆于表达载体PET30a中,并转化入大肠杆菌BL21(DE3)中,IPTG诱导表达,用亲和层析纯化BP26重组蛋白,然后分别用Western-blot,间接ELISA检测其抗原性。结果DNA测序及酶切鉴定证实PET-30a/bp26原核表达载体构建成功,并在大肠杆菌中高效表达,免疫印迹和间接ELISA实验证明BP26重组蛋白可与布鲁氏菌阳性血清产生特异性结合。结论成功获得了布鲁氏菌中序列全长为696bp,编码232个氨基酸的BP26蛋白,通过血清学反应证实,该蛋白为人和动物布鲁氏菌病临床诊断试剂盒的研发奠定了基础。     

<Emphasis Type="Italic">cagA</Emphasis> Gene and Protein Status Among Iranian <Emphasis Type="Italic">Helicobacter pylori</Emphasis> Strains

The wide geographic genetic diversity of Helicobacter pylori (Hp) and, in particular, the varying prevalence of cagA in different countries has been documented repeatedly. This study was designed to determine the frequency of cagA in Iranian Hp strains by means of genotyping and assessment of host antibodies. Helicobacter pylori strains from 235 patients, including 174 non-ulcer dyspepsia, 25 peptic ulcer and 36 gastric cancer patients, were studied. The frequencies of the 5′, middle and 3′ terminal regions of the cagA gene were 90.6, 57.6, 89%, respectively, with no correlation to the clinical outcomes. Antibodies against the CagA protein were present in 90.7% of patients. Multiple biopsy sampling in 97 cases revealed multiple infection in 16.5% of the patients. Sequencing of the seven variants of the 3′ end of the cagA gene revealed no clustering and the distribution of the Iranian strains among those of other countries. Our results from the genotyping and serology analyses confirm that the majority of Iranian Hp strains are cagA-positive.     

布鲁杆菌外膜蛋白OMP31和bp26基因的克隆、表达及鉴定

目的克隆并测序中国布鲁杆菌外膜蛋白基因OMP31和bp26,重组表达OMP31和bp26蛋白, 并对其进行初步的血清学鉴定。方法从布鲁杆菌基因组中获得OMP31和bp26基因,连接入PMD18-T克隆质粒并测序。测序后分别克隆入融合表达载体PGEX-4T-1,在大肠埃希菌中表达融合蛋白。用Western-blot技术分析重组表达的GST-OMP31和GST-bp26的免疫学特性。结果 OMP31和bp26基因的测序结果与 GeneBank中已报道的布鲁杆菌株完全一致,并在大肠埃希菌中成功表达了GST-OMP31和GST-bp26融合蛋白。布鲁杆菌免疫兔血清能特异性识别所表达的蛋白而不与载体蛋白发生反应。结论成功表达了布鲁杆菌外膜蛋白OMP31和bp26,它可以被布鲁杆菌免疫血清特异性识别,表现出良好的抗原性,为之后的实验室诊断和亚单位疫苗的研究打下良好的物质基础。     

CD34-related coexpression of <Emphasis Type="Italic">MDR1 and BCRP</Emphasis> indicates a clinically resistant phenotype in patients with acute myeloid leukemia (AML) of older age

Clinical resistance to chemotherapy in acute myeloid leukemia (AML) is associated with the expression of the multidrug resistance (MDR) proteins P-glycoprotein, encoded by the MDR1/ABCB1 gene, multidrug resistant-related protein (MRP/ABCC1), the lung resistance-related protein (LRP), or major vault protein (MVP), and the breast cancer resistance protein (BCRP/ABCG2). The clinical value of MDR1, MRP1, LRP/MVP, and BCRP messenger RNA (mRNA) expression was prospectively studied in 154 newly diagnosed AML patients ≥60 years who were treated in a multicenter, randomized phase 3 trial. Expression of MDR1 and BCRP showed a negative whereas MRP1 and LRP showed a positive correlation with high white blood cell count (respectively, p < 0.05, p < 0.001, p < 0.001 and p < 0.001). Higher BCRP mRNA was associated with secondary AML (p < 0.05). MDR1 and BCRP mRNA were highly significantly associated (p < 0.001), as were MRP1 and LRP mRNA (p < 0.001) expression. Univariate regression analyses revealed that CD34 expression, increasing MDR1 mRNA as well as MDR1/BCRP coexpression, were associated with a lower complete response (CR) rate and with worse event-free survival and overall survival. When adjusted for other prognostic actors, only CD34-related MDR1/BCRP coexpression remained significantly associated with a lower CR rate (p = 0.03), thereby identifying a clinically resistant subgroup of elderly AML patients.     

布鲁杆菌M16株pgm基因突变活菌苗株毒力初步评价

目的通过减毒性实验评价布鲁杆菌M16株pgm基因突变活菌苗株毒力减弱情况,为新型减毒活疫苗的研制和布鲁杆菌感染免疫防御的研究奠定基础。方法进行布鲁杆菌侵袭实验等动物和细胞实验评价突变菌株的毒力减弱情况。结果突变菌株在小鼠体内能生存,但较强毒株存活数量有很大减少（P〈0.001）;突变菌株较强毒株对多粘菌素B更为敏感,存活率与强毒株相比有明显下降（P〈0.001）;突变菌株LPS较强毒株LPS明显缺失;突变菌株在体外He-La细胞中能存活但复制减弱。结论布鲁杆菌M16株pgm基因突变菌株毒力减弱。可作为未来布鲁杆菌新型减毒活疫苗的候选者,有进一步研究的价值。     

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.     

<Emphasis Type="Italic">p16</Emphasis><Superscript><Emphasis Type="Italic">INK4A</Emphasis></Superscript> Alterations are accompanied by aberrant protein immunostaining in endometrial carcinomas

Purpose To date, the significance of p16^INK4A tumor suppressor gene inactivation in sporadic endometrial cancer (EC) has only rarely been described. In this study, we examined the alteration type and frequency of gene alterations [point mutations, aberrant promoter methylation and loss of heterozygosity (LOH)] in 50 sporadic ECs, and correlated the genetic findings with the immunohistochemical expression of the p16^INK4A protein and the classical clinicopathological features.Methods Gene mutations were detected by PCR-SSCP-sequencing analysis, promoter hypermethylation by methylation-specific PCR (MSP), and LOH by PCR of the STS-marker c5.1.Results In total, p16^INK4A alterations were found in 14 of 50 (28%) sporadic ECs. In six (12%) cases, two alterations occurred simultaneously. Partial p16^INK4A deletions were found in four of 50 (8%) samples. There was one missense mutation (codon 70; CCCGCC) and one frameshift mutation (1-bp deletion in exon 2). Only 2 of 47 (4.2%) tumors exhibited aberrant promoter methylation. An allelic loss was detected in 12 of 50 (24%) carcinomas with a higher incidence in advanced endometrial carcinomas than in early-stage uterine tumors. p16^INK4A alterations were generally accompanied by gene silencing, confirmed by aberrant protein immunostaining (r=-0.442; P=0.001). There was a significant difference in the frequency of p16^INK4A alterations between early (stage I; 18%) and advanced (stages II–IV; 58%) ECs (P=0.022). One case showed complete protein loss, but absence of genetic alterations.Conclusions Our data indicate that p16^INK4A inactivation plays a role in the tumorigenesis of the subset of sporadic ECs, particularly in cases exhibiting an aggressive clinical behavior. We demonstrate that p16^INK4A methylation can act efficiently and similarly to other genetic alterations as one of the two necessary hits according to the Knudson two-hit hypothesis of tumor suppressor gene inactivation.     

Occurrence of the root-knot nematode <Emphasis Type="Italic">Meloidogyne arenaria</Emphasis> on balm and in a mixed population with <Emphasis Type="Italic">M. javanica</Emphasis> on grapevine in Greece

Summary The root-knot nematode Meloidogyne arenaria was found in Greece infecting balm (Melissa officinalis L.) and grapevine (Vitis vinifera L.). The isolate from balm was M. arenaria while the one from grapevine was a mixture of M. arenaria and M. javanica (prevalent species). This is the first report of the M. arenaria species in the country in which identification was based on biochemical methods and its occurrence on balm is a new host record.     

Manifestations cliniques des infections à Chlamydia pneumoniae

Chlamydia pneumoniae is a newly described and ubiquitous bacterium. Most infections are asymptomatic as shown by a high worldwide séroprévalence (> 50% of cases). It is a common cause of acute respiratory infections, mainly pneumonia (> 50% of cases) and other acute respiratory tract infections (25% of acute bronchitis, < 5% of sinusitis, otitis and pharyngitis). About 10% of the community acquired pneumonia cases have been associated with Chlamydia pneumoniae infection. This incidence depends on a cyclic epidemiology with a high incidence for 2 to 3 years-followed by a low prevalence for 3 to 4 years. Most chlamydial infections are mild but occasionnaly severe with death especially in old people. Mostly acute infections are reccurent infections. The seroprevalence is higher in asthmatic patients, its role in acute exacerbation of chronic bronchitis is not definitely established. Extra-respiratory acute infections are less frequent, either fever alone, or cardiovascular diseases (acute myocarditis, pericarditis and endocarditis) or neurological (encephalitis, meningitis or Gudlain-Barré syndrome). In addition, seroepidemiology studies have shown an association with coronary artery disease, Chlamydia pneumoniae was detected in coronary atheroma by immunochemistry, polymerase chain reaction and by electron microscopy. Chlamydia pneumoniae may be involved in the atherosclerotc process. To define the clinical spectrum of infection requires precise laboratory diagnosis, the most efficient tests (PCR, direct immunofluorescence and culture) are done in specialized laboratories, serological tests are less reliable. Macrolides, cyclines and fluoroquinolones are the most potent antibiotics but with differences in vitro within and between these families of antibiotics. Bacteriological failures are described despite the in vitro activity. A lot of questions on clinical aspects, epidemiology and treatement are unanswered, we need more studies.     

Occurence of <Emphasis Type="Italic">Pomphorhynchus laevis</Emphasis> (Acanthocephala) in the Marsh Frog (<Emphasis Type="Italic">Rana ridibunda</Emphasis> Pallas, 1771), from Turkey

Summary   Pomphorhynchus laevis (Müller, 1776) was recovered from Marsh frogs (Rana ridibunda), collected in three localities, Yesilirmak River (Amasya Province), Civril Isikli Lake and Bagbasi district (Denizli Province) from Turkey. This is the first report of Pomphorhynchus laevis from a frog in Turkey.     

Occurrence of <Emphasis Type="Italic">Xiphinema</Emphasis> species in grapevine areas of Tyrnavos with comments on the distribution of <Emphasis Type="Italic">X. italiae</Emphasis> in Greece

Summary A survey for presence of Xiphinema species was undertaken in the viticulture areas of Tyrnavos, Thessally, Greece. Soil samples were collected from existing vineyards and from fields where grapevines had been uprooted and were currently under fallow or a cereal crop. The species X. index, X. italiae and X. pachtaicum were present in 37% of the samples with a prevalence of the last two species in uprooted fields. The occurrence of X. italiae in association with light sandy soils is discussed and compared with similar findings from other Greek grapevine areas.     

Mutation Analysis of <Emphasis Type="Italic">SPINK1</Emphasis> and <Emphasis Type="Italic">CFTR</Emphasis> Gene in Korean Patients with Alcoholic Chronic Pancreatitis

Several genetic mutations have been reported to increase susceptibility to chronic pancreatitis. However, their roles in alcoholic chronic pancreatitis are controversial. We investigated the prevalence of SPINK1 N34S and new CFTR Q1352H mutations in alcoholic chronic pancreatitis in Korea. Forty-three patients with alcoholic chronic pancreatitis were enrolled and 35 healthy individuals served as controls. The SPINK1 N34S mutation was detected by the PCR-RFLP technique. The CFTR Q1352H mutation was examined with PCR direct sequencing. Mean age of chronic pancreatitis and control groups was 53.2 and 51.3 years, respectively. A SPINK1 N34S was detected as a heterozygote in one (2.4%) patient with alcoholic chronic pancreatitis and a heterozygote CFTR Q1352H was detected in one other patient. In the control population, neither SPINK1 nor CFTR mutation was detected. This study shows that SPINK1 N34S and CFTR Q1352H mutations are uncommon and do not play an important role in chronic alcoholic pancreatitis in Korea.     

Joint effects of HLA, <Emphasis Type="Italic">INS</Emphasis>, <Emphasis Type="Italic">PTPN22</Emphasis> and <Emphasis Type="Italic">CTLA4</Emphasis> genes on the risk of type 1 diabetes

Background/hypothesis HLA, INS, PTPN22 and CTLA4 are considered to be confirmed type 1 diabetes susceptibility genes. HLA, PTPN22 and CTLA4 are known to be involved in immune regulation. Few studies have systematically investigated the joint effect of multiple genetic variants. We evaluated joint effects of the four established genes on the risk of childhood-onset type 1 diabetes. Methods We genotyped 421 nuclear families, 1,331 patients and 1,625 controls for polymorphisms of HLA-DRB1, −DQA1 and −DQB1, the insulin gene (INS, −23 HphI), CTLA4 (JO27_1) and PTPN22 (Arg620Trp). Results The joint effect of HLA and PTPN22 on type 1 diabetes risk was significantly less than multiplicative in the case-control data, but a multiplicative model could not be rejected in the trio data. All other two-way gene–gene interactions fitted multiplicative models. The high-risk HLA genotype conferred a very high risk of type 1 diabetes (OR 20.6, using the neutral-risk HLA genotype as reference). When including also intermediate-risk HLA genotypes together with risk genotypes at the three non-HLA loci, the joint odds ratio was 61 (using non-risk genotypes at all loci as reference). Conclusion Most established susceptibility genes seem to act approximately multiplicatively with other loci on the risk of disease except for the joint effect of HLA and PTPN22. The joint effect of multiple susceptibility loci conferred a very high risk of type 1 diabetes, but applies to a very small proportion of the general population. Using multiple susceptibility genotypes compared with HLA genotype alone seemed to influence the prediction of disease only marginally. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorised users.     

The first record of <Emphasis Type="Italic">Dracunculus mulbus</Emphasis> (Nematoda: Dracunculidae) in the Papuan olive python <Emphasis Type="Italic">Apodora papuana</Emphasis> (Ophidia: Boidae)

Summary A male specimen of Dracunculus (Nematoda: Dracunculidae), collected in 1973 from the lung of the Papuan olive python Apodora papuana (Peters et Doria) and now deposited in the helminthological collection of the Natural History Museum in London, is identified as Dracunculus mulbus Jones et Mulder, 2007, a species recently described from the water python Liasis fuscus Peters in northern Australia. Apodora papuana is a new host record for D. mulbus and Papua New Guinea is apparently a new geographical record.     

<Emphasis Type="Italic">Rhodococcus equi</Emphasis> infection during treatment of a C-ANCA positive vasculitis: a case report

Rhodococcus equi is a rare form of opportunistic infection in humans, more common in the immunocompromised. We present a case of pulmonary infection and subsequent cerebral abscess secondary to R. equi in a patient receiving immunosuppression for a c-ANCA positive vasculitis. Heightened awareness of R. equi infection is important when considering immunocompromised patients presenting with sepsis, especially those with cavitating pulmonary lesions and normal respiratory commensals on culture. Delays in diagnosis and treatment can cause increased patient morbidity and mortality.     

First evidence of <Emphasis Type="Italic">Paraheligmonina gracilis</Emphasis> and <Emphasis Type="Italic">Hymenolepis sulcata</Emphasis> among fat dormice (<Emphasis Type="Italic">Glis glis</Emphasis> L.) from Croatia

Summary The present study reports for the first time on the helminth species occurring in the gastro-intestinal system of fat dormice (Glis glis) in Croatia. Out of 55 dormice, 63.7 % harboured helminths belonging to two species, the nematode Paraheligmonina gracilis (syn. Longistriata elpatievskii) (Heligmonellidae, Trichostrogyloidea) in the prevalence of 52.7 %, and the cestode Hymenolepis sulcata (Hymenolepididae, Cyclophyllidea) in the prevalence of 32.7 %. Concurrent infections of both parasites were found in 12 fat dormice, P. gracilis alone was detected in 17 hosts and H. sulcata alone in 6 samples. No influence of parasitic infestation on animal weight was observed. Glirid helminths do not represent zoonotic pathogens despite the fact that dormice occasionally inhabit cottages and village houses, and are used in human nutrition.     

Clinical Relevance of cagE Gene from Helicobacter pylori Strains in Japan

It has been reported that H. pylori-containing cagE was associated with duodenal ulcer. The aims of the present study were to clarify the association between the cagE gene and clinical outcome and to analyze the relationship between the cagE gene and two other virulence factors—cagA and vacA—in two areas in Japan (Fukui and Okinawa) where the prevalence of duodenal ulcer and gastric cancer risk are quite different. Eighty of 81 isolates possessed the cagE gene, and all isolates possessed the cagA gene. The vacA genotype s1c/m1 was a major genotype in both areas in Japan. There was no significant association between cagE, cagA status, or vacA genotype and clinical outcome. Phylogenetic analysis of the cagE gene indicated that most Japanese isolates formed a different cluster from strains isolated in the West with an association with the vacA genotype. In conclusion, the strains with cagE, cagA, and the s1c/m1 genotype of vacA are predominant in Japan regardless of clinical outcome and construct a different phylogenetic cluster from those in the West.