Immunological study of a population of hemophiliacs treated with non-commercial fractions

We studied lymphocyte subpopulations, serum beta 2 microglobulin, and viral markers in 115 hemophiliacs who are followed and treated at our center. The only clinical or biological anomaly observed is hypergammaglobulinemia. This frequent finding is independent of hemophilia type and of the intensity of substitutive treatment. This group is characterized by the fact that the patients are treated only by locally prepared coagulation fractions, essentially cryoprecipitates for hemophiliacs A, and not containing any derivative of commercial origin. This series is compared to other recent reports, some of which present a high incidence of immunologic anomalies.     

LFA-1 and CD2 Synergize for the Erk1/2 Activation in the Natural Killer (NK) Cell Immunological Synapse

Natural killer (NK) cell recognition and formation of a conjugate with target cells, followed by intracellular signal pathway activation and degradation of cytolytic granules, are essential for NK cell cytotoxicity. In this study, NK92 cells were used to investigate synapse formation and subsequent signaling after binding to the target cell. The binding rate of the NK92-target cell was associated with NK92 cell cytotoxicity. Confocal results showed that adhesion molecules, LFA-1 (CD11a) and CD2, accumulated at the interface of the NK92-K562 contact. Ligation with K562 cells activated the Erk1/2 signal pathway of NK92 cells. The blocking of the NK-target conjugate by EDTA or anti-CD11a or/and anti-CD2 antibody decreased the phosphorylation of Erk1/2 and NK cell cytotoxicity. Inhibition of Erk1/2 phosphorylation by the chemical inhibitor U0126 suppressed the cytolytic activity of NK92 cells, but had no effect on NK-target conjugate formation. Thus, conjugate formation of the NK92-target cell was prerequisite to NK cell activation, and subsequent signal transduction was also required for NK cell cytotoxicity.Natural killer (NK)³ cells are a population of granular lymphocytes that play an essential role in cellular immune defense against a variety of tumor cells, virus-infected cells, or allogeneic cells (1–3). NK cells are critical for host immunity for their ability for a quick cytotoxic response and to produce a wide variety of cytokines and chemokines to modulate other cellular components of the immune system (4, 5). NK cells express two functional types of receptors: activating and inhibitory receptors (6–8). The effector function of NK cells is regulated by a balance between opposite signals delivered by the MHC class I-specific inhibitory receptors and the activating receptors responsible for NK cell triggering to permit elimination of pathogens (6).NK cell recognition and binding to target cells, as well as formation of conjugates, are essential for NK cell cytotoxicity (9). Conjugate formation by the NK cell with a target cell is a process mediated by integrins and immunoglobulin superfamily molecules including CD2, CD11a (LFA-1), CD11b, CD11c, and CD28, which also participate in the promotion of NK cell function (10–12). They participate in adhesion between the NK cell and the target cell, and blocking antibodies suppress the adhesion. In addition to possessing an adhesive role, ligation of CD2 induces kinase function and lipid raft polarization (11), whereas ligation of CD11a, CD11b, and CD11c induces phosphorylation-dependent NK cell activation (13, 14). The interaction of specific cell surface receptors with their ligands on a target cell at their interface forms specific activating NK cell immunological synapses and leads to the activation of a cascade of intracellular signals, resulting in Ca²⁺ flux, polarization of granules, and subsequent release of lytic molecules (13, 15, 16). The Erk1/2 (p44/42 mitogen-activated protein kinase) pathway plays an important role in NK cell cytotoxicity (17–21). Inhibition of Erk1/2 might block NK cell cytolytic activity by compromising the release of perforin (22). In this study, the roles of adhesion molecules in NK92-target cell conjugate formation of immunological synapse, and subsequent Erk1/2 activation in NK92 cells was investigated.     

NMR protocol for determination of oxidation susceptibility of serum lipids and application of the protocol to a chocolate study

A protocol for determination of oxidation susceptibility of serum lipids based on proton nuclear magnetic resonance ((1)H NMR) spectroscopy is presented and compared to the commonly used spectrophotometric method. Even though there are methodological differences between these two methods, the NMR-based oxidation susceptibility correlates well (r(2)?=?0.73) with the lag time determined spectrophotometrically. In addition to the oxidizability of serum lipids, the NMR method provides also information about the lipid profile. The NMR oxidation assay was applied to the chocolate study including fasting serum samples (n?=?45) from subjects who had consumed white (WC), dark (DC) or high-polyphenol chocolate (HPC) daily for 3?weeks. The oxidation susceptibility of serum lipids decreased in the HPC group, and there was a significant difference between the WC and HPC groups (P?=?0.031). According to the random forest analysis, the consumption of the HPC chocolate induced changes to the amounts of HDL, phosphatidylcholine, sphingomyelin, and nervonic, docosahexaenoic and myristic acids. Furthermore, arachidonic, docosahexaenoic, docosapentaenoic and palmitic acids, gamma-glutamyl transferase, hemoglobin, HDL, phosphatidylcholine and choline containing phospholipids explained about 60% of the oxidation susceptibility values. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s11306-011-0323-2) contains supplementary material, which is available to authorized users.     

Immunological effectiveness of a 2d revaccination against diphtheria performed at intervals extended up to 7-10 years

The present article deals with the data on the duration of antidiphtheria immunity, induced by the primary cycle of immunization with adsorbed DPT vaccine, with the aim of considering the problems of optimizing the immunization schedule for children. The prolongation of the interval before the second booster immunization to 7-10 years produces no negative influence on the effectiveness of immunization. Besides, the study has shown that after the main complex of immunizations with adsorbed DPT vaccine protective immunity to diphtheria is retained in 90.3-96.8% of children covered by this study for 9-10 years. These data point to high immunological effectiveness of adsorbed DPT vaccine and to the possibility of prolonging the intervals between booster immunizations to 10 years, as well as decreasing the number of booster immunizations in the immunization schedule for children.     

The 49,XYYYY syndrome: apropos of a case detected at birth and followed for 2 1/2 years

A male child, mentally and physically retarded shows a facial dysmorphy, fingers' abnormalities and a radio-ulnar synostosis. These features are common in the 49,XYYYY syndrom, in which external genitalia, normal at birth, remain undevelopped at the time of puberty. Four others publications through the literature report tetrasomic cells for Y chromosome, but only two of them are real 49,XYYYY (with 88% to 100% of affected cells). Our case has 96,7% tetrasomic cells.     

Immunological evidence for the capability of free-living Rhizobium japonicum to synthesize a portion of a nitrogenase component

Immunodiffusion tests conducted under aerobic conditions demonstrated that cross-reactive material to antiserum prepared against the MoFe protein component of nitrogenase from soybean nodule bacteroids was detectable in extracts of free-living Rhizobium japonicum cells cultured in a standard medium under: aerobic conditions; aerobic conditions with nitrate; aerobic conditions with ammonia; anaerobic conditions with nitrate; and anaerobic conditions with nitrate and ammonia. The most intense precipitin bands resulted from cross-section of the antiserum with extracts of cells cultured anaerobically with nitrate or anaerobically with ammonia and nitrate. Immunodiffusion experiments with crude bacteroid extract and purified MoFe protein revealed a greater number of precipitin bands in tests conducted under aerobic conditions than those conducted under anaerobic conditions. These results indicate that some of the cross-reactive material observed under aerobic conditions resulted from breakdown of the MoFe protein. Bacteroid extracts of nodules from plants supplied with ammonia exhibited only a trace of nitrogenase activity. The addition of an excess of the Fe protein component of nitrogenase, however, resulted in a 270-fold enhancement of activity indicating the presence of active MoFe protein in these extracts.Our experiments together with results published elsewhere provide evidence that the genetic information for synthesis of a part of the MoFe component of nitrogenase is carried by Rhizobium.     

Assessment of radiographic progression in the spines of patients with ankylosing spondylitis treated with adalimumab for up to 2 years

Introduction  

Ankylosing spondylitis (AS) is a chronic rheumatic disease associated with spinal inflammation that subsequently leads to progression of structural damage and loss of function. The fully human anti-tumor necrosis factor (anti-TNF) antibody adalimumab reduces the signs and symptoms and improves overall quality of life in patients with active AS; these benefits have been maintained through 2 years of treatment. Our objective was to compare the progression of structural damage in the spine in patients with AS treated with adalimumab for up to 2 years versus patients who had not received TNF antagonist therapy.     

National validation study of a swab protocol for the recovery of Bacillus anthracis spores from surfaces

Twelve Laboratory Response Network (LRN) affiliated laboratories participated in a validation study of a macrofoam swab protocol for the recovery, detection, and quantification of viable B. anthracis (BA) Sterne spores from steel surfaces. CDC personnel inoculated steel coupons (26 cm²) with 1-4 log₁₀ BA spores and recovered them by sampling with pre-moistened macrofoam swabs. Phase 1 (P1) of the study evaluated swabs containing BA only, while dust and background organisms were added to swabs in Phase 2 (P2) to mimic environmental conditions. Laboratories processed swabs and enumerated spores by culturing eluted swab suspensions and counting colonies with morphology consistent with BA. Processed swabs were placed in enrichment broth, incubated 24 h, and cultured by streaking for isolation. Real-time PCR was performed on selected colonies from P2 samples to confirm the identity of BA. Mean percent recovery (%R) of spores from the surface ranged from 15.8 to 31.0% (P1) and from 27.9 to 55.0% (P2). The highest mean percent recovery was 31.0% (sd 10.9%) for P1 (4 log₁₀ inoculum) and 55.0% (sd 27.6%) for P2 (1 log₁₀ inoculum). The overall %R was higher for P2 (44.6%) than P1 (24.1%), but the overall reproducibility (between-lab variability) was lower in P2 than in P1 (25.0 vs 16.5%CV, respectively). The overall precision (within-lab variability) was close to identical for P1 and P2 (44.0 and 44.1, respectively), but varied greatly between inoculum levels. The protocol demonstrated linearity in %R over the three inoculum levels and is able to detect between 26 and 5 × 10⁶ spores/26 cm². Sensitivity as determined by culture was > 98.3% for both phases and all inocula, suggesting that the culture method maintains sensitivity in the presence of contaminants. The enrichment broth method alone was less sensitive for sampled swabs (66.4%) during P2, suggesting that the presence of background organisms inhibited growth or isolation of BA from the broth. The addition of real-time PCR testing to the assay increased specificity from > 85.4% to > 95.0% in P2. Although the precision was low at the 1 log₁₀ inoculum level in both phases (59.0 and 50.2%), this swab processing protocol, was sensitive, specific, precise, and reproducible at 2-4 log₁₀/26 cm² spore concentrations.     

A study to determine the underlying reason for abnormal glandular cytology and the formulation of a management protocol

A retrospective review is presented of 89 patients with glandular dyskaryosis in order to formulate a management protocol. Fifteen patients had cervical intraepithelial neoplasia (CIN) without glandular abnormality (17%). One patient had adenocarcinoma in situ of the cervix and one patient had vaginal intraepithelial neoplasia (VAIN) grade III. Twenty‐two patients had endometrial carcinoma (24.5%) and 11 patients had cervical carcinoma (12.5%). Of the patients presenting with post‐menopausal bleeding as well as having glandular dyskaryosis, 69% had a gynaecological malignancy. In conclusion, colposcopy and out‐patient endometrial sampling are recommended in all cases. Patients with abnormal endometrial sampling require hysteroscopy. Cone biopsy is necessary to exclude occult glandular disease if cytology remains abnormal despite negative colposcopy and sampling.     

Acupuncture for chronic prostatitis/chronic pelvic pain syndrome: study protocol for a randomized controlled trial

Background

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common condition affecting men of all ages. Acupuncture may be an effective treatment option for CP/CPPS, but evidence is limited. We propose to evaluate the effectiveness of acupuncture in a rigorously conducted trial.

Methods

Ten hospitals will recruit 440 participants with CP/CPPS in China from October 2017 to December 2019. Participants will be randomly allocated to acupuncture or sham acupuncture with a 1:1 ratio using computerized simple random sampling. The whole study consists of 2-week baseline, 8-week treatment, and 24-week follow up. Twenty 30-mintute sessions of acupuncture or sham acupuncture treatment will be provided between week 1 and 8. The two co-primary outcomes are the proportion of responders at week 8 and week 32. Secondary outcomes include proportion of responders in the two groups at different time points; change in the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) total score; change in the NIH-CPSI subscales; change in the International Prostate Symptom Score; change in the Hospital Anxiety and Depression Scale; expectation assessments; proportions of participants in each response category of the Global Response Assessment; change in the International Index of Erectile Function 5; change in the five-level EuroQol five-dimensional questionnaire and a visual analogue scale; and changes in peak and average urinary flow rate.

Discussion

This study will provide robust evidence on whether acupuncture is effective for relieving symptoms of CP/CPPS.

Trials registration

ClinicalTrials.gov, NCT03213938. Registered on 5 July 2017.

     

Microbial contamination of cell cultures: a 2 years study.

Cell line contamination is a major drawback of main cell banks of the world and it has cost of losing important biological products or valuable research. The causative agents are different chemicals, invertebrates, bacteria, fungi, parasites, viral species and even other cell lines. In this retrospective study, cell lines from various species such as human, fish, insect, animals either offered or accessed through usual official accession in CGBRI were studied during 2 years (2002-2004) to detect their microbial contaminations and the causative organisms. Samples were taken for sterility test upon cell lines receipt and upon each cell line sub-culture. Samples were examined for bacterial (including mycoplasmas) and fungal contamination using conventional microbiological techniques. The study excluded parasites, viruses and other contaminating agents. This study revealed 39% of specimens were contaminated. The major contaminating agents were mycoplasmas (19%) followed by mixed infection (8%), fungi (8%) and bacteria (4%). Among various bacterial species (except mycoplasmas) Bacillus sp., Enterococcus sp. and Staphylococcus sp. are main bacterial agents and among various fungi Aspergillus sp. followed by Penicillium sp., Sepedonium sp. and Botrytis sp. were main fungal causative agents of CGBRI cell line contamination. Our study also delineates each cell line contamination rate and its causative agents. This is the first report of cell culture contamination from cell banks of Middle-East countries like Iran.     

Immunological study of antisera to to artificial O-antigen of Pseudomonas aeruginosa

O-sera were obtained by immunizing rabbits with artificial antigens: polysaccharide extracted from the lipopolysaccharide of P. aeruginosa (strain No. 868; 03a, 3d, 3e), or the high-molecular-weight or low-molecular-weight fraction of this polysaccharide, complexed with a natural protein (human IgG or rabbit globulin). The antisera to these antigenic complexes were highly O-specific. Antisera to the complexes of polysaccharide-protein and high-molecular-weight fraction-protein were more active in the passive haemagglutination reaction, slide agglutination test and immunodiffusion test in agar gel than was antiserum to the low-molecular-weight fraction-protein complex. The artificial antigens prepared and employed in the study are apt to be used for the preparation of monospecific immune sera.     

Rearrangement of VHa1-encoding Ig gene segment to the a2 chromosome in an a1/a2 heterozygous rabbit. Evidence for trans recombination

VDJ genes were cloned from leukemic B cells of an a1/a2 heterozygous Emu-cmyc transgenic rabbit. Restriction mapping and nucleotide sequence analysis indicated that one clone, 5C3, had a VHa1-encoding gene segment functionally rearranged to a JH gene segment from the a2 chromosome. This VDJ gene may be the result of a trans recombination between a VH gene on the a1 chromosome and a JH gene segment on the a2 chromosome or, it may be the result of a cis recombination if the a2 chromosome contains VHa1-encoding gene segments.