FISHing in the dark: How the combination of FISH and conventional karyotyping improves the diagnostic yield in CpG‐stimulated chronic lymphocytic leukemia

Despite significant advances in molecular genetic approaches, fluorescence in situ hybridization (FISH) remains the gold standard for the diagnostic evaluation of genomic aberrations in patients with chronic lymphocytic leukemia (CLL). Efforts to improve the diagnostic utility of molecular cytogenetic testing have led to the expansion of the traditional 4‐probe CLL FISH panel. Not only do these efforts increase the cost of testing, they remain hindered by the inherent limitations of FISH studies ‐ namely the inability to evaluate genomic changes outside of the targeted loci. While array‐based profiling and next generation sequencing (NGS) have critically expanded our understanding of the molecular pathogenesis of CLL, these methodologies are not routinely used by diagnostic laboratories to evaluate copy number changes or the mutational profile of this disease. Mitogenic stimulation of CLL specimens with CpG‐oligonucleotide (CpG‐ODN) has been identified as a reliable and reproducible means of obtaining a karyotype, facilitating a low‐resolution genome‐wide analysis. Across a cohort of 1255 CpG‐ODN‐stimulated CLL specimens, we describe the clinical utility associated with the combinatorial use of FISH and karyotyping. Our testing algorithm achieves a higher diagnostic yield (～10%) through the detection of complex karyotypes, well‐characterized chromosomal aberrations not covered by the traditional CLL FISH panel and through the detection of concurrent secondary malignancies. Moreover, the single cell nature of this approach permits the evaluation of emerging new clinical concepts including clonal dynamics and clonal evolution. This approach can be broadly applied by diagnostic laboratories to improve the utility of traditional and molecular cytogenetic studies of CLL. Am. J. Hematol. 91:978–983, 2016. © 2016 Wiley Periodicals, Inc.     

Application of Saccharomyces cerevisiae/Calcium Alginate Composite Beads for Cephalexin Antibiotic Biosorption from Aqueous Solutions

Cephalexin (CPX) is recognized as a water pollutant, and it has been listed in a number of countries with a risk factor greater than one. Herein, the present work focused on the synthesis, characterization and biosorption capacity evaluation of Saccharomyces cerevisiae immobilized in calcium alginate as a biosorbent to remove CPX from aqueous solutions. Biosorbent was characterized by SEM and FTIR techniques. Batch biosorption experiments were conducted in order to evaluate the effect of the initial pH, biosorbent dose and CPX initial concentration. The removal efficiency, in considered optimal conditions (pH = 4, CPX initial concentration = 30 mg/L, biosorbent dose = 1 g/L) was 86.23%. CPX biosorption was found to follow the pseudo–second-order kinetics. The equilibrium biosorption data were a good fit for the Langmuir model with correlation coefficient of 0.9814 and maximum biosorption capacity was 94.34 mg/g. This study showed that the synthesized biosorbent by immobilization technique is a low-cost one, easy to obtain and handle, eco-friendly, with high feasibility to remove CPX antibiotic from aqueous solution. The findings of this study indicate that the biosorbents based on microorganisms immobilized on natural polymers have the potential to be applied in the treatment of wastewater.     

Respiratory viruses are continuously detected in children with chronic tonsillitis throughout the year

Objective

To evaluate the oscillations on the viral detection in adenotonsillar tissues from patients with chronic adenotonsillar diseases as an indicia of the presence of persistent viral infections or acute subclinical infections.

Study design

Cross-sectional prospective study.

Setting

Tertiary hospital.

Methods

The fluctuations of respiratory virus detection were compared to the major climatic variables during a two-year period using adenoids and palatine tonsils from 172 children with adenotonsillar hypertrophy and clinical evidence of obstructive sleep apnoea syndrome or recurrent adenotonsillitis, without symptoms of acute respiratory infection (ARI), by TaqMan real-time PCR.

Results

The rate of detection of at least one respiratory virus in adenotonsillar tissue was 87%. The most frequently detected viruses were human adenovirus in 52.8%, human enterovirus in 47.2%, human rhinovirus in 33.8%, human bocavirus in 31.1%, human metapneumovirus in 18.3% and human respiratory syncytial virus in 17.2%. Although increased detection of human enterovirus occurred in summer/autumn months, and there were summer nadirs of human respiratory syncytial virus in both years of the study, there was no obvious viral seasonality in contrast to reports with ARI patients in many regions of the world.

Conclusion

Respiratory viruses are continuously highly detected during whole year, and without any clinical symptomatology, indicating that viral genome of some virus can persist in lymphoepithelial tissues of the upper respiratory tract.     

Outcomes for pregnant women infected with the influenza A (H1N1) virus during the 2009 pandemic in Porto Alegre, Brazil

Objective

To study the epidemiologic characteristics and underlying conditions that place pregnant women infected with H1N1 virus at increased risk for being admitted to the intensive care unit (ICU).

Methods

In this cross-sectional study conducted in Porto Alegre, Brazil, with 57 pregnant women hospitalized with the H1N1 influenza during the 2009 pandemic, we collected epidemiologic characteristics and assessed the rates of ICU admission according to pregnancy duration and the presence or absence of comorbidities.

Results

The median (range) of maternal age was 26 years (15-41 years), the pregnancy duration at the time of infection was 29 weeks (8-41 weeks), and the birth weight was 3180 g (740-3900 g). Five patients (8.8%) were in the first, 22 (38.6%) in the second, and 30 (52.6%) in the third trimester, and (22.8%) had comorbidities. Antiviral drugs were administered to all, and 46 (80.7%) patients received an early treatment. There were no maternal, fetal, or neonatal deaths. Eight patients (14%) required ICU admission and 15 (50%) of the patients who gave birth during their hospitalization underwent a cesarean delivery. The risk of being treated at the ICU did not increase for patients with comorbidities (P = 0.22) or an advanced pregnancy (P = 0.31). The study revealed a relationship between early initiation of an antiviral treatment and a lower mortality rate.

Conclusion

Neither an advanced pregnancy nor comorbidities increased the risk of being admitted to the ICU but, compared with the results of other studies, a prompt treatment lowered mortality.     

Design and Evaluation of Orally Dispersible Tablets Containing Amlodipine Inclusion Complexes in Hydroxypropyl-β-cyclodextrin and Methyl-β-cyclodextrin

The development of new orally dispersible tablets containing amlodipine (AML) inclusion complexes in hydroxypropyl-β-cyclodextrin (HP-β-CD) and in methyl-β-cyclodextrin (Me-β-CD) was studied. The methods of obtaining amlodipine and the physical and chemical properties of the inclusion complexes using the two cyclodextrins was investigated separately. Solid inclusion complexes were obtained by three methods: kneading, coprecipitation, and lyophilization, at a molar ratio of 1:1. For comparison, a physical mixture in the same molar ratio was prepared. The aim of the complexation process was to improve the drug solubility. As the lyophilization method leads to a complete inclusion of the drug in the guest molecule cavity, for both used cyclodextrins, these types of compounds were selected as active ingredients for the design of orally dispersible tablets. Subsequently, the formulation of the orodispersible tablets containing AML-HP-β-CD and AML-Me-β-CD inclusion complexes and quality parameters of the final formulation were evaluated. The results prove that F1 and F4 formulations, based on silicified microcrystalline cellulose, which contains insignificant proportions of very small or very large particles, had the lowest moisture degree (3.52% for F1 and 4.03% for F4). All of these demonstrate their porous structure, which led to good flowability and compressibility performances. F1 and F4 formulations were found to be better to manufacture orally dispersible tablets.     

Is PostureScreen® Mobile app an accurate tool for dentists to evaluate the correlation between malocclusion and posture?

ABSTRACT

Objective

To determine whether PostureScreen® Mobile app is a useful, simple tool that is accessible for dentists to evaluate the correlation between malocclusion and posture.     

Contents and distributions of the proteoglycans decorin and biglycan in normal and osteoarthritic human articular cartilage

The study was designed to determine the contents and distributions of the proteoglycans decorin and biglycan in adult human femoral condylar cartilage and whether these may change in osteoarthritis. New radioimmunoassays were established using peptides representing the amino-terminal 21 amino acid sequence of each proteoglycan (to which a tyrosine was added for radioiodination) and antibodies in a rabbit antiserum raised to both these molecules. Cartilage was extracted with 4 M guanidine HCl to determine total content, and extracts were analyzed by chromatography to determine molecular sizes. Frozen sections were cut parallel to the articular surface and were extracted to determine distribution within the tissue. Gel chromatography on Sepharose CL-2B under dissociative conditions revealed molecules with a partition coefficient of 0.7–0.75 in both normal and osteoarthritic cartilage. In normal adult cartilage, the mean contents of the core proteins of biglycan and decorin were calculated to be approximately 0.34 and 0.48 mg per gram wet weight, respectively. These represented molar contents similar to that of aggrecan. In osteoarthritic cartilage, there were no overall significant changes in the content and distribution of these molecules. There was, however, considerable individual variation in both distribution and content. Analyses indicated that there was a trend in osteoarthritic cartilage toward a loss of biglycan and decorin from the more superficial layers of intact cartilage, where both these molecules are normally more concentrated. This was accompanied by maintenance of proteoglycan content deeper in the cartilage, regardless of the degree of degeneration.