Novel germline BRCA1 and BRCA2 mutations in breast and breast/ovarian cancer families from the Czech Republic

Germline mutations in breast cancer susceptibility genes, BRCA1 and BRCA2, are responsible for a substantial proportion of high‐risk breast and breast/ovarian cancer families. To characterize the spectrum of BRCA1 and BRCA2 mutations, we screened Czech families with breast/ovarian cancer using the non‐radioactive protein truncation test, heteroduplex analysis and direct sequencing. In a group of 100 high‐risk breast and breast/ovarian cancer families, four novel frame shift mutations were identified in BRCA1 and BRCA2 genes. In BRCA1, two novel frame shift mutations were identified as 3761‐3762delGA and 2616‐2617ins10; in BRCA2, two novel frame shift mutations were identified as 5073‐5074delCT and 6866delC. Furthermore, a novel missense substitution M18K in BRCA1 gene in a breast/ovarian cancer family was identified which lies adjacent just upstream of the most highly conserved C3HC4 RING zinc finger motif. To examine the tertiary structure of the RING zinc finger domain and possible effects of M18K substitution on its stability, we used threading techniques according to the crystal structure of RAG1 dimerization domain of the DNA‐binding protein. © 2000 Wiley‐Liss, Inc.     

Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of maedi-visna virus

A single-chain antibody fragments (scFv) was developed directed against transmembrane envelope glycoprotein gp46 of the virus maedi-visna, by the application of the antibody phage display library. To get specific scFv binders, the library was panned against the biotinylated peptide of 20 amino acids corresponding to the principal immunodominant domain of gp46 protein. The number of positively binding scFvs was evaluated by scFv-phage ELISA, BstN1 fingerprinting and DNA sequencing. The scFvs were expressed in soluble form and purified by immobilized metal affinity chromatography (IMAC) with a yield of 2-2.5 mg/l. Two scFvs have shown to recognize gp46 and gp150 proteins in Western blot analysis. The scFvs also recognized the virus in infected cells as shown by immunofluorescence assay. The affinity of the obtained antibody fragments to gp46 peptide was measured by surface plasmon resonance, and the resulting K(A) was in the 10(6)-10(7)lmol(-1) range. The application of characterized scFvs for expression as intrabodies in intracellular immunization against virus maedi-visna infection and for the diagnosis of this virus is discussed.     

Fully synthetic carbohydrate-based vaccines in biochemically relapsed prostate cancer: clinical trial results with alpha-N-acetylgalactosamine-O-serine/threonine conjugate vaccine.

PURPOSE: We report the synthesis of a mucin-related O-linked glycopeptide, alpha-N-acetylgalactosamine-O-serine/threonine (Tn), which is highly simplistic in its structure and can induce a relevant humoral response when given in a trimer or clustered (c) formation. We tested for an antitumor effect, in the form of a change in the posttreatment versus pretreatment prostate-specific antigen (PSA) slopes, that might serve as a surrogate for effectiveness of vaccines in delaying the time to radiographic progression. METHODS: We compared the antibody response to immunization with two conjugates, Tn(c)-keyhole limpet hemocyanin (KLH) and Tn(c)-palmitic acid (PAM) with the saponin immunologic adjuvant QS21, in a phase I clinical trial in patients with biochemically relapsed prostate cancer. Patients received Tn(c)-KLH vaccine containing either 3, 7, or 15 microg of Tn(c) per vaccination. Ten patients received 100 microg of Tn(c)-PAM. QS21 was included in all vaccines. Five vaccinations were administered subcutaneously during 26 weeks with an additional booster vaccine at week 50. RESULTS: Tn(c), when given with the carrier molecule KLH and QS21, stimulated the production of high-titer immunoglobulin M (IgM) and IgG antibodies. Inferior antibody responses were seen with T(c)-PAM. There was no evidence of enhanced immunogenicity with increasing doses of vaccine. An antitumor effect in the form of a decline in posttreatment versus pretreatment PSA slopes was also observed. CONCLUSION: A safe synthetic conjugate vaccine in a trimer formation was developed that can break immunologic tolerance by inducing specific humoral responses. It seemed to affect the biochemical progression of the disease as determined by a change in PSA log slope.     

Novel α + β Zr Alloys with Enhanced Strength

Low-alloyed zirconium alloys are widely used in nuclear applications due to their low neutron absorption cross-section. These alloys, however, suffer from limited strength. Well-established guidelines for the development of Ti alloys were applied to design new two-phase ternary Zr alloys with improved mechanical properties. Zr-4Sn-4Nb and Zr-8Sn-4Nb alloys have been manufactured by vacuum arc melting, thermo-mechanically processed by annealing, forging, and aging to various microstructural conditions and thoroughly characterized. Detailed Scanning electron microscopy (SEM) analysis showed that the microstructural response of the alloys is rather similar to alpha + beta Ti alloys. Duplex microstructure containing primary alpha phase particles surrounded by lamellar alpha + beta microstructure can be achieved by thermal processing. Mechanical properties strongly depend on the previous treatment. Ultimate tensile strength exceeding 700 MPa was achieved exceeding the strength of commercial Zr alloys for nuclear applications by more than 50%. Such an improvement in strength more than compensates for the increased neutron absorption cross-section. This study aims to exploit the potential of alpha + beta Zr alloys for nuclear applications.     

Predictors of mortality and cost among surgical patients requiring rapid response team activation

BackgroundPrior studies of rapid response team (RRT) implementation for surgical patients have demonstrated mixed results with respect to reductions in poor outcomes. The aim of this study was to identify predictors of in-hospital mortality and hospital costs among surgical inpatients requiring RRT activation.MethodsWe analyzed data prospectively collected from May 2012 to May 2016 at The Ottawa Hospital. We included patients who were at least 18 years of age, who were admitted to hospital, who received either preoperative or postoperative care, and and who required RRT activation. We created a multivariable logistic regression model to describe mortality predictors and a multivariable generalized linear model to describe cost predictors.ResultsWe included 1507 patients. The in-hospital mortality rate was 15.9%. The patient-related factors most strongly associated with mortality included an Elixhauser Comorbidity Index score of 20 or higher (odds ratio [OR] 3.60, 95% confidence interval [CI] 1.96–6.60) and care designations excluding admission to the intensive care unit and cardiopulmonary resuscitation (OR 3.52, 95% CI 2.25–5.52). The strongest surgical predictors included neurosurgical admission (OR 2.09, 95% CI 1.17–3.75), emergent surgery (OR 2.04, 95% CI 1.37–3.03) and occurrence of 2 or more operations (OR 1.73, 95% CI 1.21–2.46). Among RRT factors, occurrence of 2 or more RRT assessments (OR 2.01, 95% CI 1.44–2.80) conferred the highest mortality. Increased cost was strongly associated with admitting service, multiple surgeries, multiple RRT assessments and medical comorbidity.ConclusionRRT activation among surgical inpatients identifies a population at high risk of death. We identified several predictors of mortality and cost, which represent opportunities for future quality improvement and patient safety initiatives.     

Phase Composition of Al-Si Coating from the Initial State to the Hot-Stamped Condition

The chemical and phase composition of the coating and the coating/substrate interface of an Al-Si-coated 22MnB5 hot stamped steel was investigated by means of SEM-EDS, XRD, micro-XRD and electron diffraction. Moreover, the surface profile was analyzed by XPS and roughness measurements. The XPS measurements showed that the thickness of the Si and Al oxide layers increased from 14 to 76 nm after die-quenching, and that the surface roughness increased as well as a result of volume changes caused by phase transformations. In addition to the FeAl(Si) and Fe₂Al₅ phases and the interdiffusion layer forming complex structures in the coating, electron diffraction confirmed the presence of an Fe₂Al₅ phase, and also revealed very thin layers of Fe₃(Al,Si)C, Fe₂(Al,Si)₅ and Al-bearing rod-shaped particles in the immediate vicinity of the steel interface. Moreover, the scattered nonuniform layer of the Fe₂Al₈Si phase was identified in the outermost layer of the coating. Despite numerous studies devoted to researching the phase composition of the Al-Si coating applied to hot stamped steel, electron diffraction revealed very thin layers and particles on the coating/substrate interface and outermost layer, which have not been analyzed in detail.     

Albendazolesulphoxide concentrations in plasma and hydatid cyst and prediction of parasitological and clinical outcomes in patients with liver hydatidosis caused by Echinococcus granulosus

Aim

To investigate the relationship between plasma and cyst concentrations of albendazolesulphoxide (ASO) and their effects on parasitological findings and disease recurrence in patients with liver hydatidosis.

Methods

The study was conducted at the University Hospital for Infectious Diseases “Dr. Fran Mihaljević,” Zagreb, Croatia, between August 2006 and January 2011. Consecutive patients (N = 48, age 6-77 years) were treated with albendazole (3 × 5 mg/kg/d) over 28 days before surgical cyst removal (n = 34) or percutaneous evacuation (PAIR) (n = 14). Plasma ASO was determined on days 10 and 28 of treatment and cyst concentrations at surgery/PAIR.

Results

Disease recurred in 3 surgically treated patients. Variability of ASO concentrations was substantial. Plasma concentrations on day 10 were higher than on day 28 (geometric means ratio [GMR] 2.00; 95%CI 1.38-2.91, P < 0.001) and higher than cyst concentrations at the time of treatment (GMR = 1.58, 1.01-2.34, P = 0.045). Higher cyst (but not plasma) concentrations were independently associated with lower odds of protoscolex motility (OR = 0.23, 0.01-0.70, P < 0.001) and higher odds of protoscolex destruction (OR = 1.17, 1.04-1.46, P < 0.001). With adjustment for age and protoscolex motility, higher day 10 plasma concentrations (but not cyst concentrations) were associated with lower odds of disease recurrence (OR = 0.49, 0.09-0.97, P = 0.035). Plasma concentrations did not predict cyst concentrations.

Conclusion

Viability of protoscolices progressively decreased with increasing ASO concentrations in the cyst. Data strongly suggested that higher plasma concentrations reduced the risk of disease recurrence.Echinococcosis or hydatid cyst disease is an anthropozoonosis caused by the larvae of E. granulosus, E. multilocularis, E. vogeli, and E. oligarthus (1). In transient hosts (eg, sheep, cattle, pigs, humans), the parasite develops in the form of hydatid cyst(s) of different sizes. The cyst consists of the outer layer (ectocyst), made of dense fibrous tissue; the middle layer (endocyst), which is elastic and lamellar in structure; and the inner (germinative) layer, which gives rise to buds that develop into scolices. The cyst content is hydatid fluid, produced by the germinative layer. In humans, the most commonly invaded organs are the liver and the lungs, but practically all organs can be affected (1-3). In Croatia, the disease is caused exclusively by E. granulosus. According to the European Hospital Morbidity Database (4) for the period 2010-2012, age-adjusted annual hospital admission rates due to echinococcosis (ICD-B67) in Croatia varied between 0.011 and 0.0167/1000 population (ie, between 51 and 86 cases/y), indicating that the diseases is relatively rare but still stably present.Treatments for hydatid cyst disease include surgical removal of the cyst(s) (still most commonly used method); percutaneous aspiration of the cyst with instillation of a scolicidal agent (95% ethanol or hypertonic saline [15%-20%] or albendazole), ie, the PAIR (puncture, aspiration, instillation, and re-aspiration) procedure, which seems to have greater clinical efficacy and lower rate of complications than the surgical procedure; and treatment with benzimidazole anthelmintic drugs. The latter, pharmacological option might be used as a monotreatment in the case of smaller cysts or when invasive approaches are not feasible, but it is typically adjunctive treatment to surgery or PAIR used to prevent dissemination of scolices from ruptured cysts. In this setting, benzoimidazoles are used over at least 4 weeks before the definitive treatment (5-8). Among benzoimidazoles, albendazole is considered a cornerstone pharmacological treatment of echinococcosis. Although some controversies still exist regarding optimal dosing, the most widely accepted regimen implies administration of 10-15 mg/kg/d (5-8). Upon oral ingestion (with a fatty meal to increase bioavailability), albendazole is rapidly metabolized (first-pass metabolism in the liver) into the active form albendazolesulphoxide (ASO), which inhibits tubuline polymerization in the parasite microtubules and inactivates cell division (9). Greater systemic bioavailability is considered an important advantage of albendazole over mebendazole, the other member of the group (9). Although ASO has been shown to penetrate both the hepatic and non-hepatic cysts (9,10), the prognostic value of plasma and/or cyst concentrations has not been elucidated. Therefore, we aimed to relate ASO concentrations in the plasma and in the cysts, and to investigate their relationship with the parasitological and clinical outcomes in patients with liver hydatidosis treated with albendazole over one month prior to surgical treatment or PAIR.     

Tuberculosis Disease Diagnosis Using Artificial Immune Recognition System

Background: There is a high risk of tuberculosis (TB) disease diagnosis among conventional methods.Objectives:This study is aimed at diagnosing TB using hybrid machine learning approaches.Materials and Methods: Patient epicrisis reports obtained from the Pasteur Laboratory in the north of Iran were used. All 175 samples have twenty features. The features are classified based on incorporating a fuzzy logic controller and artificial immune recognition system. The features are normalized through a fuzzy rule based on a labeling system. The labeled features are categorized into normal and tuberculosis classes using the Artificial Immune Recognition Algorithm.Results:Overall, the highest classification accuracy reached was for the 0.8 learning rate (α) values. The artificial immune recognition system (AIRS) classification approaches using fuzzy logic also yielded better diagnosis results in terms of detection accuracy compared to other empirical methods. Classification accuracy was 99.14%, sensitivity 87.00%, and specificity 86.12%.     

Intracardiac shrapnel in a polytraumatized child

Penetrating cardiac trauma is a life-threatening condition and presents a therapeutic challenge for the surgeon. Additional multiple organ-system injuries, as are common in the setting of war, further complicate the management of such patients. We present the case of a 9-year-old girl who sustained multiple injuries from an unexploded artillery shell, resulting in a retained intracardiac shrapnel. Her cardiac pathology consisted of a shrapnel located in the interventricular septum accompanied by a pneumopericardium and a right-sided hemopneumothorax. The presentation and management of this patient are the subjects of this report.