Phage-displayed mimotopes recognizing a biologically active anti-HIV-1 gp120 murine monoclonal antibody

Antibody-dependent cellular cytotoxicity (ADCC) is a host defense mechanism in which Fc receptor-bearing effector cells in combination with antigen-specific antibodies recognize and kill antigen-expressing target cells. The authors previously described a murine monoclonal antibody (MAb-ID6) that mediated ADCC activity against HIV-infected cells. It was demonstrated that the specificity of MAb-ID6 maps to the first 204 amino acids of gp120; however, the exact epitope was not identified. In the present work, by screening phage display libraries with MAb-ID6, the authors have mapped the corresponding epitope to amino acids 86-100 (HIV-1 gp120 sequence). This epitope lies within the C1 region of gp120 and is highly conserved among all subtypes and circulating recombinant forms of HIV-1. Thus, these phage mimotopes of C1 may serve as components of a vaccine for the induction of gp120-specific antibodies mimicking MAb-ID6.     

A TaqI polymorphism in the 3'UTR of the IL-12 p40 gene correlates with increased IL-12 secretion

Interleukin-12 (IL-12) is a key cytokine for the induction of Th1 immune responses. We evaluated whether a TaqI polymorphism in the 3'UTR of the IL-12 p40 gene affects secretion of IL-12 in vitro, and whether this polymorphism is associated with susceptibility to Crohn's disease (CD). IL-12 p40 and p70 secretion by monocytes in relation to genotype was determined in 63 healthy donors. Genotype and allele frequencies of the TaqI polymorphism in 150 CD patients were compared with 145 ethnically matched healthy controls (HC). No significant association was found between genotype and IL-12 p40 secretion after stimulation of monocytes with SAC+IFNgamma. In contrast, increasing IL-12 p70 secretion was found across the categories of non-carriers, heterozygotes and homozygotes for the variant allele (median values+/-SEM: 147+/-27, 282+/-51 and 482+/-34 pg/ml, respectively; P<0.005). The allele and genotype frequencies of this polymorphism in patients with CD did not differ statistically significantly from HC. The presence of a TaqI polymorphism in the IL12 p40 3'UTR correlates with increased in vitro IL-12 p70, but not p40 secretion. While this polymorphism does not appear to be correlated with susceptibility to CD in the limited population of patients tested here, it could influence the occurrence of the disease in certain subsets of patients.     

Cellular distribution of bovine leukemia virus proteins gp51SU, Pr72(env), and Pr66(gag-pro) in persistently infected cells

Monoclonal antibodies (mAbs) against bovine leukemia virus (BLV) mature proteins and precursors were used to map the localization of these proteins in persistently infected non-lymphocytic cell lines using immunofluorescence assay (IFA) and immuno-electron microscopy. IFA staining was observed in the basolateral surface of live FLK-BLV cells. When using a mAb against Pr66(gag-pro), mottled pinpoint fluorescence was seen in the cell surface of polarized cells, but no reaction was observed in cells undergoing mitosis. However, a mAb against Pr72(env) stained only mitotic cells and cellular fragments. Additionally, in these dividing cells, this envelope (Env) precursor polyprotein was not evenly distributed but concentrated predominantly in only one daughter cell. To the best of our knowledge, this observation has not been reported previously, either for BLV or for other retroviruses. The results of immunogold electron microscopy confirmed the specificity of the mAbs in the intracellular level. In infected cells, Pr72(env) and gp51SU were seen in proximity at the plasma membrane in incipient budding sites. Additionally, the mAb against Pr72(env) also reacted with Env precursor polyproteins in the mitochondria of BLV-bat(2) ultrathin sections. These mAbs may be used as a tool for mapping virus excretion sites in the cell surface of naturally or in vitro infected cells in the different stages of the cell cycle.     

Expression of the proteoglycans versican and mel-CSPG in dysplastic nevi

Nevi with architectural disorder and cytologic atypia of melanocytes (NAD) (also called dysplastic nevi) have been controversial with regard to their relationship with melanoma risk and to their gradation in 3 degrees of atypia. Versican and the melanoma-associated proteoglycan (mel-CSPG) are 2 major proteoglycans expressed by malignant melanoma, and they have a role in the regulation of cell adhesion, migration, and differentiation. We evaluated the differences in versican and mel-CSPG expression in nevi, NAD with several degrees of atypia, and primary malignant melanoma. Immunoreactivity for versican was negative in benign melanocytic nevi, positive in NAD (ranging from weakly to intensely positive), and intensely positive in malignant melanoma. Immunostaining for mel-CSPG was negative in benign melanocytic nevi and mild to moderately positive in NAD and melanoma. Our results suggest that versican expression may be of value for distinguishing NAD from benign melanocytic nevi and for distinguishing severe NAD from mild and moderate NAD.     

Identification of 26 new constitutional RB1 gene mutations in Spanish, Colombian, and Cuban retinoblastoma patients

Constitutional mutations in the RB1 gene predispose to retinoblastoma development. Hence genetic screening of retinoblastoma patients and relatives is important for genetic counseling purposes. In addition, RB1 gene mutation studies may help decipher the molecular mechanisms leading to tumors with different degrees of penetrance or expressivity. In the course of genetically screening of 107 hereditary and non-hereditary retinoblastoma patients (11 familiar bilateral, 4 familiar unilateral, 49 sporadic bilateral and 43 sporadic unilateral) and kindred from Spain, Colombia and Cuba, using direct PCR sequencing, we observed 45 distinct mutations and four RB1 deletions in 53 patients (9 familiar bilateral, 2 familiar unilateral, 31 sporadic bilateral and 11 sporadic unilateral). Most of these mutations (26/45, 57%) have not been reported before. In 32 patients, the predisposing mutations correspond to nonsense (mainly CpG transitions) and small insertions or deletions whose expected outcome is a truncated Rb protein that lacks the functional pockets and tail. Five single aminoacid replacements and seventeen mutations affecting splicing sites were also observed in retinoblastoma patients. Two of these sixteen mutations are of unclear pathogenic nature.     

HPLC analysis of 16 compounds from Artemisia ordosica

Objective: To establish a high-performance liquid chromatographic method (HPLC) for the simultaneous determination of sixteen compounds from Artemisia ordosica. Methods: HPLC was used to analyze 16 quality indicators of A. ordosica. The HPLC conditions were as follows: Agilent Eclipse Plus C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-water (B) as mobile phase, gradient elution: 0?10 min, 75%?65% B; 10?30 min, 65%?35 % B; and finally 30?40 min, 35%?15% B. The flow rate was 1.0 mL/min, the column temperature was 40 °C, the injection volume was 10 μL, and monitored by absorbance at 285 nm for compounds 1?10, 12 and 225 nm for compounds 11, 13?16. Results: Under the selected experimental chromatographic conditions, compounds 1?16 showed good linearity (r > 0.9993) in a wide concentration range. Their average recoveries were 99.50%, 95.38%, 97.75%, 96.00%, 98.20%, 97.50%, 95.50%, 99.33%, 96.75%, 96.50%, 98.50%, 97.83%, 99.20%, 95.33%, 97.33% and 96.30%, respectively, and the RSD were 1.99%, 1.81%, 1.63%, 1.98%, 1.67%, 1.92%, 1.74%, 1.67%, 1.90%, 1.72%, 1.88%, 1.83%, 1.79%, 1.76%, 1.81% and 1.96%, respectively. Conclusion: Based on the results of the HPLC analysis, it was concluded that p-hydroxycinnamic acid (1), O-hydroxycinnamic acid (2), coniferyl alcohol (5), 5,4''-dihydroxy-7,3''-dimethoxyflavanone (8), 5,4''-dihydroxy-7-methoxyflavanone (9), 5-hydroxy-7,4''-dimethoxyflavanone (12), dehydrofalcarindiol (13), arteordoyn A (14), dehydrofalcarinol (15) and capillarin (16) are best suited for the role of quality indicators of A. ordosica grown in different ecological environments.     

ALPPS与采用不同栓塞材料PVE对肝再生及手术切除率影响研究

目的 比较不同栓塞材料的门静脉栓塞术（PVE）与联合肝脏分隔和门静脉结扎的二步肝切除术（ALPPS）对剩余肝体积（FLR）增长速率的影响,比较各组FLR的增长速率,二期手术切除率、术中数据和术后并发症。方法 采用单中心、前瞻性、非随机对照的对比研究。2014年11月至2019年12月,海军军医大学第三附属医院共126例因FLR不足导致无法切除的肝细胞癌（HCC）或肝内胆管癌（ICC）病人,将其分为4组：ALPPS组及分别采用氰基丙烯酸正丁酯（NBCA）、微球、明胶海绵作为栓塞材料的PVE组。主要终点为FLR增长速率和二期手术切除率。结果 各组的手术切除例数及二期手术切除率分别为：ALPPS组38例（99.4%）,NBCA组32例（76.2%）,明胶海绵组20例（60.6%）,微球组10例（83.3%）。ALPPS组、NBCA组、微球组的FLR增长速率分别为15.1 mL/d,10.0 mL/d和 8.5 mL/d,均高于明胶海绵组（3.7 mL/d）。结论 采用NBCA及微球作为栓塞材料的PVE导致FLR增长速率低于ALPPS,两种栓塞材料的PVE二期手术切除率相当。使用NBCA作为栓塞材料的PVE其FLR增长速率高于微球,且这两种栓塞材料的栓塞效果均优于明胶海绵。     

Alterations in iron metabolism in chronic anti-HCV positive hepatitis. The relationship with response to interferon treatment

A prospective study in which the existence of iron metabolism alterations (increase in the serum levels of iron and ferritin and the presence of iron in liver tissue) in a group of 53 patients diagnosed with chronic anti-HCV positive hepatitis was performed. The aim of the study was to determine whether these parameters influence the response to interferon treatment. Elevations were observed in the serum levels of iron and/or ferritin in 17 (32%) of the patients. Higher than normal values of serum iron or ferritin in pretreatment analyses were associated with worse therapeutic response. The basal serum levels of iron and ferritin were significantly higher in non responding patients. No relationship was found between the presence of iron in the hepatic parenchyma and response to interferon treatment.     

Underreporting of percutaneous exposure accidents in a teaching hospital in Spain

The study's objectives were to determine the frequency of biological-risk accidents involving percutaneous exposure and to identify factors associated with underreporting. Two hundred fifty healthcare professionals from inpatient services at high risk for exposure at the Puerta del Mar University Hospital of Cádiz, Spain, participated in the study. A questionnaire was used to measure personal and work variables, the number of accidents suffered and reported in the last year, and the circumstances motivating the reporting or nonreporting. Two hundred thirty-two persons (92.8%) completed the questionnaire. The accident rate was 12 per 100,000 hours worked. Physicians were the most frequent accident victims (rate 22/100,000 hours). The general surgery and emergency services had higher rates than other services (rates 19.82 and 14.17, respectively). Sixty-six percent of the accidents were not reported to the register. The main predictors of the underreporting were length of professional service greater than 19 years, working in the surgery service, and the perception that the accidents did not involve health risk. The true accident rate was higher than that reflected in the Accident Register. Underreporting was high. The main variables associated with underreporting were length of professional service, work area or department, and perception of risk from the accident.     

Minimum basic set of data of hospital discharges as a source of information for a study of congenital abnormalities

BACKGROUND: The purpose of this study is that of assessing the validity of the computerized diagnoses of hospital discharges of congenital defects by comparing them with the information included in the medical history. METHODS: Based on the discharge records generated over a one-year period at 7 hospitals in the Autonomous Region of Valencia, 100 children were selected at random from each hospital. As a standard, the diagnoses stated in the medical histories were indexed and coded. Solely those discharges having taken place during the first year of life were considered. A study was also made of the type, seriousness and individual or combinations of congenital defects. A calculation was made of the sensitivity, specificity, predictive values and the 95% confidence intervals thereof by the exact binomial method for the case studies (children) and the positive predictive value and sensitivity for the study of diagnoses. RESULTS: 126 children were detected as having congenital defects, and 201 diagnoses in medical records, and 83 children with congenital defects and 108 diagnoses on record. For the detection of cases, the records showed a 64% sensitivity, a 99.1% specificity and some positive and negative predictive values of over 90%. With regard to the detection of diagnoses, the sensitivity was 46% and the positive predictive value 83%. The sensitivity varied a great deal depending upon the diagnoses. CONCLUSIONS: The hospital discharge records revealed a high degree of specificity and high predictive values, but a low degree of sensitivity. These facts must be considered when these records are used as a source of cases for the epidemiological studies of congenital defects.