Extracellular β-1,6-glucanase activity during growth of seven strains of Saccharomyces cerevisiae

Two standard strains and five local isolates of Saccharomyces cerevisiae all produced extracellular -1,6-glucanases in a growth-associated manner. Glucose at 2% (w/v) enhanced enzyme production up to 0.8 U ml^-1. Optimum activity was at pH 5.0.The authors are with the Department of Microbiology. University of Dhaka, Dhaka 1000, Bangladesh     

Design and Optimization of Anti-amyloid Domain Antibodies Specific for β-Amyloid and Islet Amyloid Polypeptide

Antibodies with conformational specificity are important for detecting and interfering with polypeptide aggregation linked to several human disorders. We are developing a motif-grafting approach for designing lead antibody candidates specific for amyloid-forming polypeptides such as the Alzheimer peptide (Aβ). This approach involves grafting amyloidogenic peptide segments into the complementarity-determining regions (CDRs) of single-domain (V_H) antibodies. Here we have investigated the impact of polar mutations inserted at the edges of a large hydrophobic Aβ42 peptide segment (Aβ residues 17–42) in CDR3 on the solubility and conformational specificity of the corresponding V_H domains. We find that V_H expression and solubility are strongly enhanced by introducing multiple negatively charged or asparagine residues at the edges of CDR3, whereas other polar mutations are less effective (glutamine and serine) or ineffective (threonine, lysine, and arginine). Moreover, Aβ V_H domains with negatively charged CDR3 mutations show significant preference for recognizing Aβ fibrils relative to Aβ monomers, whereas the same V_H domains with other polar CDR3 mutations recognize both Aβ conformers. We observe similar behavior for a V_H domain grafted with a large hydrophobic peptide from islet amyloid polypeptide (residues 8–37) that contains negatively charged mutations at the edges of CDR3. These findings highlight the sensitivity of antibody binding and solubility to residues at the edges of CDRs, and provide guidelines for designing other grafted antibody fragments with hydrophobic binding loops.     

Effect of sea water concentration on hyphal growth and antimicrobial metabolite production in marine fungi

We studied the effect of sea water concentration in a culture medium on fungal growth and the production of antimicrobial metabolites. Most of the marine fungal isolates were identified as members of the same genera as terrestrial isolates, such asAspergillus andTrichoderma. Many of the marine fungi isolated grew more abundantly as the sea water concentration increased. The production of antimicrobial materials was improved as the sea water concentration increased. Even though the marine fungi were considered to be similar to fungi from terrestrial environments, from a mycological perspective, the two types have different physiological characteristics. The fungi from marine samples are useful microbial resources in the search for new bioactive compounds.     

Asymptomatic Plasmodium falciparum Malaria in Pregnant Women in the Chittagong Hill Districts of Bangladesh

Background

Pregnancy is a known risk factor for malaria which is associated with increased maternal and infant mortality and morbidity in areas of moderate-high malaria transmission intensity where Plasmodium falciparum predominates. The nature and impact of malaria, however, is not well understood in pregnant women residing in areas of low, unstable malaria transmission where P. falciparum and P. vivax co-exist.

Methods

A large longitudinal active surveillance study of malaria was conducted in the Chittagong Hill Districts of Bangladesh. Over 32 months in 2010–2013, the period prevalence of asymptomatic P. falciparum infections was assessed by rapid diagnostic test and blood smear and compared among men, non-pregnant women and pregnant women. A subset of samples was tested for infection by PCR. Hemoglobin was assessed. Independent risk factors for malaria infection were determined using a multivariate logistic regression model.

Results

Total of 34 asymptomatic P. falciparum infections were detected by RDT/smear from 3,110 tests. The period prevalence of asymptomatic P. falciparum infection in pregnant women was 2.3%, compared to 0.5% in non-pregnant women and 0.9% in men. All RDT/smear positive samples that were tested by PCR were PCR-positive, and PCR detected additional 35 infections that were RDT/smear negative. In a multivariate logistic regression analysis, pregnant women had 5.4-fold higher odds of infection as compared to non-pregnant women. Malaria-positive pregnant women, though asymptomatic, had statistically lower hemoglobin than those without malaria or pregnancy. Asymptomatic malaria was found to be evenly distributed across space and time, in contrast to symptomatic infections which tend to cluster.

Conclusion

Pregnancy is a risk factor for asymptomatic P. falciparum infection in the Chittagong Hill Districts of Bangladesh, and pregnancy and malaria interact to heighten the effect of each on hemoglobin. The even distribution of asymptomatic malaria, without temporal and spatial clustering, may have critical implications for malaria elimination strategies.     

Is Serum Zinc Level Associated with Prediabetes and Diabetes?: A Cross-Sectional Study from Bangladesh

Aims

To determine serum zinc level and other relevant biological markers in normal, prediabetic and diabetic individuals and their association with Homeostasis Model Assessment (HOMA) parameters.

Methods

This cross-sectional study was conducted between March and December 2009. Any patient aged ≥30 years attending the medicine outpatient department of a medical university hospital in Dhaka, Bangladesh and who had a blood glucose level ordered by a physician was eligible to participate.

Results

A total of 280 participants were analysed. On fasting blood sugar results, 51% were normal, 13% had prediabetes and 36% had diabetes. Mean serum zinc level was lowest in prediabetic compared to normal and diabetic participants (mean differences were approximately 65 ppb/L and 33 ppb/L, respectively). In multiple linear regression, serum zinc level was found to be significantly lower in prediabetes than in those with normoglycemia. Beta cell function was significantly lower in prediabetes than normal participants. Adjusted linear regression for HOMA parameters did not show a statistically significant association between serum zinc level, beta cell function (P = 0.07) and insulin resistance (P = 0.08). Low serum zinc accentuated the increase in insulin resistance seen with increasing BMI.

Conclusion

Participants with prediabetes have lower zinc levels than controls and zinc is significantly associated with beta cell function and insulin resistance. Further longitudinal population based studies are warranted and controlled trials would be valuable for establishing whether zinc supplementation in prediabetes could be a useful strategy in preventing progression to Type 2 diabetes.     

Serological Cross-Reaction Between O-Antigens of Shigella dysenteriae Type 4 and an Environmental Escherichia albertii Isolate

An environmental freshwater bacterial isolate, DM104, appearing as Shigella-like colonies on selective agar plates was found to show strong and specific serological cross-reactivity with Shigella dysenteriae type 4. Biochemical identification according to the analytical profile index, molecular serotyping by restriction of the amplified O-antigen gene cluster (rfb-RFLP), together with phylogenetic analysis of the 16S rRNA gene and multi-locus sequence analysis, identified the isolate as Escherichia albertii. rfb-RFLP of DM104, revealed a profile different from that of S. dysenteriae type 4. However, western blot analysis of extracted lipopolysaccharides demonstrated strong cross-reactivity with S. dysenteriae type 4 using specific monovalent antisera and a lipopolysaccharide gel banding profile similar to that of S. dysenteriae type 4. The observed O-antigen cross-reaction between an E. albertii isolate and S. dysenteriae extends our knowledge of the extent of O-antigen cross-reaction within the Escherichia/Shigella group of organisms, and offers the possibility of using DM104 and similar cross-reacting strains as shigellosis vaccine candidates.     

Drought Tolerance in Mung Bean is Associated with the Genotypic Divergence,Regulation of Proline,Photosynthetic Pigment and Water Relation

Drought is one of the critical conditions for the growth and productivity of many crops including mung bean (Vigna radiata L. Wilczek). Screening of genotypes for variations is one of the suitable strategies for evaluating crop adaptability and global food security. In this context, the study investigated the physiological and biochemical responses of four drought tolerant (BARI Mung-8, BMX-08010-2, BMX-010015, BMX-08009-7), and four drought sensitive (BARI Mung-1, BARI Mung-3, BU Mung-4, BMX-05001) mung bean genotypes under wellwatered (WW) and water deficit (WD) conditions. The WW treatment maintained sufficient soil moisture (22% ± 0.5%, i.e., 30% deficit of available water) by regularly supplying water. Whereas, the WD treatment was maintained throughout the growing period, and water was applied when the wilting symptom appeared. The drought tolerant (DT) genotypes BARI Mung-8, BMX-08010-2, BMX-010015, BMX-08009-7 showed a high level of proline accumulation (2.52–5.99 mg g⁻¹ FW), photosynthetic pigment (total chlorophyll 2.96–3.27 mg g⁻¹ FW at flowering stage, and 1.62–2.38 mg g⁻¹ FW at pod developing stage), plant water relation attributes including relative water content (RWC) (82%–84%), water retention capacity (WRC) (12–14) as well as lower water saturation deficit (WSD) (19%–23%), and water uptake capacity (WUC) (2.58–2.89) under WD condition, which provided consequently higher relative seed yield. These indicate that the tolerant genotypes gained better physiobiochemical attributes and adaptability in response to drought conditions. Furthermore, the genotype BMX- 08010-2 showed superiority in terms of those physio-biochemical traits, susceptibility index (SSI) and stress tolerance index (STI) to other genotypes. Based on the physiological and biochemical responses, the BMX-08010-2 was found to be a suitable genotype for sustaining yield under drought stress, and subsequently, it could be recommended for crop improvement through hybridization programs. In addition, the identified traits can be used as markers to identify tolerant genotypes for drought-prone areas.     

Comparative Analysis of Serum Zinc, Copper, Manganese, Iron, Calcium, and Magnesium Level and Complexity of Interelement Relations in Generalized Anxiety Disorder Patients

The purpose of the study was to determine the concentration of serum trace and other essential elements of generalized anxiety disorder patients and to find out the relationship between element levels and nutritional status or socioeconomic factors. The study was conducted among 50 generalized anxiety disorder patients and 51 healthy volunteers. Patients were selected and recruited in the study with the help of a clinical psychologist by random sampling. The concentrations of serum trace elements (Zn, Cu, Mn, and Fe) and other two essential elements (Ca and Mg) were determined by graphite furnace and flame atomic absorption spectroscopy. Data were analyzed by independent t test, Pearson’s correlation analysis, regression analysis, and analysis of variance. The serum concentrations of Zn, Cu, Mn, Fe, Ca, and Mg in generalized anxiety disorder patients were 1.069?±?0.40, 1.738?±?0.544, 1.374?±?0.750, 3.203?±?2.065, 108.65?±?54.455, and 21?±?4.055 mg/L, while those were 1.292?±?0.621, 0.972?±?0.427, 0.704?±?0.527, 1.605?±?1.1855, 101.849?±?17.713, and 21.521?±?3.659 mg/L in control subjects. Significantly decreased (p?<?0.05) serum Zn concentration was found in the patient group compared to the control group while serum level of Cu, Mn, and Fe was significantly (p?<?0.05) higher, but the differences of the concentration of Ca and Mg between the patient and control groups were not significant (p?>?0.05). Socioeconomic data revealed that most of the patients were in the lower middle class group and middle-aged. Mean BMI of the control group (23.63?±?3.91 kg/m²) and the patient group (23.62?±?3.77 kg/m²) was within the normal range (18.5–25.0 kg/m²). The data obtained from different interelement relations in the generalized anxiety disorder patients and control group strongly suggest that there is a disturbance in the element homeostasis. So changes in the serum trace element level in generalized anxiety disorder patients occur independently and they may provide a prognostic tool for the diagnosis and treatment of this disease.     

The synergistic effects of NGF and IGF-1 on neurite growth in adult sensory neurons: convergence on the PI 3-kinase signaling pathway

Nerve growth factor (NGF) and insulin-like growth factor-1 (IGF-1) play an important role in promoting axonal growth from dorsal root ganglion (DRG) neurons. Adult DRG neurons exhibit neurotrophin-independent survival, providing an excellent system with which to study trophic factor effects on neurite growth in the absence of significant survival effects. Using young adult rat DRG neurons we have demonstrated a synergistic effect of NGF plus IGF (N + I), compared with either factor alone, in promoting neurite growth. Not only does the presence of NGF and IGF-1 enhance neurite initiation, it also significantly augments the extent of neurite branching and elongation. We have also examined potential mechanism(s) underlying this synergistic effect. Immunoblotting experiments of classical growth factor intermediary signalling pathways (PI 3-K-Akt-GSK-3 and Ras-Raf-MAPK) were performed using phospho-specific antibodies to assess activation state. We found that activation of Akt and MAPK correlated with neurite elongation and branching. However, using pharmacological inhibitors, we observed that a PI 3-K pathway involving both Akt and GSK-3 appeared to be more important for neurite extension and branching than MAPK-dependent signalling. In fact, inhibition of activation of MAPK with U0126 resulted in increased neuritic branching, possibly as a result of the concomitant increase observed in phospho-Akt. Furthermore, inhibition of GSK3 (which is negatively regulated by phosphorylation on S9/S21) also resulted in increased growth. Our data point to signalling convergence upon the PI 3-K-Akt-GSK-3 pathway that underlies the NGF plus IGF synergism. In addition, to our knowledge, this is the first report in primary neurons that inhibition of GSK3 results in an enhanced neurite growth.     

Pre-clinical tumor models of primary brain tumors: Challenges and opportunities

Primary brain tumors are a heterogeneous group of malignancies that originate in cells of the central nervous system. A variety of models tractable for preclinical studies have been developed to recapitulate human brain tumors, allowing us to understand the underlying pathobiology and explore potential treatments. However, many promising therapeutic strategies identified using preclinical models have shown limited efficacy or failed at the clinical trial stage. The inability to develop therapeutic strategies that significantly improve survival rates in patients highlight the compelling need to revisit the design of currently available animal models and explore the use of new models that allow us to bridge the gap between promising preclinical findings and clinical translation. In this review, we discuss current strategies used to model glioblastoma, the most malignant brain tumor in adults and highlight the shortcomings of specific models that must be circumvented for the development of innovative therapeutic strategies.