Casson fluid model for pulsatile flow of blood under periodic body acceleration

Pulsatile flow of a Casson fluid under the influence of a periodic body acceleration has been studied in this paper. An implicit finite difference numerical procedure has been used to analyze the flow. Applicability of this method has been checked by comparing the obtained results with the analytical solution for Newtonian flow and explicit scheme solution. The agreement between the implicit and explicit scheme solutions and the analytical solution is good (error less than 1%). Flow variables have been computed at three locations in cardiovascular system (wide (femoral) and narrow (arteriole and coronary) tubes). Effects of yield stress, tube radius and pressure gradient combined, body acceleration amplitude and frequency etc., on flow have been studied. The following observations have been made: (i) Initial transient time It changes with yield stress in narrow tubes are insignificant, whereas in wide tubes It decreases with yield stress; (ii) The axial velocity and fluid acceleration variations with yield stress are uniform (changes only quantitatively, profiles shape remain same) in narrow tubes, whereas in wide tubes these variations are non-uniform (profiles change qualitatively as well as quantitatively); (iii) Yield stress effects on wall shear amplitude are insignificant in narrow tubes (congruent to 0.3% in arteriole and congruent to 6% in femoral); and (iv) For Newtonian fluid, mean flow rate does not change with body acceleration amplitude a0 and frequency fb but it increases (decreases) with a0(fb) for Casson fluid.     

Capsular serotype and antibiotic resistance of Streptococcus pneumoniae isolates in Malaysia

Background

Streptococcus pneumoniae is a major causative agent of severe infections, including sepsis, pneumonia, meningitis, and otitis media, that has since become a major public health concern. In this study, the serotypes distribution of pneumococcal isolates was investigated to predict the efficacy of the 7-valent pneumococcal conjugate vaccine (PCV7) among the Malaysian populations.

Methodology/Principal Findings

A total of 151 clinical isolates were serotyped using multiplex PCR assays. Out of them, there were 21.2% penicillin-resistant, 29.1% penicillin-intermediate, and 49.7% penicillin-susceptible S.pneumoniae strains. Serotypes detected among the Malaysian isolates were 1, 3, 10A, 11A/11D, 12F/12A, 14, 15A, 15B/15C, 16F, 18C/18B/18A/18F, 19A, 19F, 23F, 35B, 35F/47F, 6A/6B, 7C/7B/40, 7F/7A, 9V/9A, and 34. Serotype 19F and 23F were the two most prevalent serotypes detected. Serotypes are highly associated with invasiveness of isolates (p = 0.001) and penicillin susceptibility (p<0.001). Serotype 19F was observed to have increased resistance against penicillin while serotype 19A has high invasive tendency. Age of patients was an important factor underlying the pneumococcal serotypes (p = 0.03) and clinical sites of infections (p<0.001). High prevalence of pneumococcal isolates were detected among children <5 years old at nasopharyngeal sites while elderly adults ≥60 years old were at increased risk for pneumococcal bacteremia.

Conclusion/Significance

Current study revealed that a number of serotypes, especially those associated with high penicillin resistance, have been formulated in the PCV7. Therefore, the protections expected from the routine use of PCV7 would be encouraging for the Malaysian. However, it is not possible to predict serotypes that might become predominant in the future and hence continued surveillance of circulating serotypes will be needed.     

High synergistic antibacterial,antibiofilm, antidiabetic and antimetabolic activity of Withania somnifera leaf extract-assisted zinc oxide nanoparticle

Nanotechnology is currently gaining immense attention to combat food borne bacteria, and biofilm. Diabetes is a common metabolic disease affecting majority of people. A better therapy relies on phytomediated nanoparticle synthesis. In this study, W. somnifera leaf extract-assisted ZnO NPs (Ws-ZnO NPs) was synthesized and characterized. From HR-TEM analysis, it has been found that the hexagonal wurtzite particle is 15.6 nm in size and − 12.14 mV of zeta potential. A greater antibacterial effect of Ws-ZnO NPs was noticed against E. faecalis and S. aureus at 100 µg mL⁻¹. Also, the biofilm of E. faecalis and S. aureus was greatly inhibited at 100 µg mL⁻¹ compared to E. coli and P. aeruginosa. The activity of α-amylase and α-glucosidase enzyme was inhibited at 100 µg mL⁻¹ demonstrating its antidiabetic potential. The larval and pupal development was delayed at 25 µg mL⁻¹ of Ws-ZnO NPs. A complete mortality (100%) was recorded at 25 µg mL⁻¹. Ws-ZnO NPs showed least LC₅₀ value (9.65 µg mL⁻¹) compared to the uncoated ZnO NPs (38.8 µg mL⁻¹) and leaf extract (13.06 µg mL⁻¹). Therefore, it is concluded that Ws-ZnO NPs are promising to be used as effective antimicrobials, antidiabetic and insecticides to combat storage pests.

     

Efficient Method of Genotyping Ob/Ob Mice Using High Resolution Melting Analysis

Objective

Direct health care costs of obesity continue to grow throughout the world and research on obesity disease models are on the rise. The ob/ob mouse is a well-characterized model of obesity and associated risk factors. Successful breeding and backcrossing onto different backgrounds are essential to create knockout models. Ob/ob mice are sterile and heterozygotes must be identified by genotyping to maintain breeding colonies. Several methods are employed to detect the ob mutant allele, a single nucleotide polymorphism (SNP). Gel based methods are time consuming and inconsistent, and non-gel based assays rely upon expensive and complex reagents or instruments. A fast, high-throughput, cost effective, and consistent method to identify Lep^ob mutation is much needed.

Design and Methods

Primers to produce an amplicon for High Resolution Melting Analysis (HRM) of the Lep^ob SNP were designed and validated.

Results

Fluorescence normalized high resolution melting curve plots delineated ob/+, ob/ob, and WT genotypes. Genotypes were also confirmed phenotypically.

Conclusions

HRM of the Lep^ob SNP allows closed-tube identification of the Lep^ob mutation using a real-time PCR machine now common to most labs/departments. Advantages of this method include assay sensitivity/accuracy, low cost dyes, less optimization, and cost effectiveness as compared to other genotyping techniques.     

Candidate Gene Identification for Systemic Lupus Erythematosus Using Network Centrality Measures and Gene Ontology

Systemic lupus erythematosus (SLE) commonly accredited as “the great imitator” is a highly complex disease involving multiple gene susceptibility with non-specific symptoms. Many experimental and computational approaches have been used to investigate the disease related candidate genes. But the limited knowledge of gene function and disease correlation and also lack of complete functional details about the majority of genes in susceptible locus, encumbrances the identification of SLE related candidate genes. In this paper, we have studied the human immunome network (undirected) using various graph theoretical centrality measures integrated with the gene ontology terms to predict the new candidate genes. As a result, we have identified 8 candidate genes, which may act as potential targets for SLE disease. We have also carried out the same analysis by replacing the human immunome network with human immunome signaling network (directed) and as an outcome we have obtained 5 candidate genes as potential targets for SLE disease. From the comparison study, we have found these two approaches are complementary in nature.     

Development of SCAR marker in Casuarina equisetifolia for species authentication

Casuarina equisetifolia is one of the most extensively introduced tree species outside its natural range and is a true multipurpose species, providing a range of services and products for industrial and local end users. Natural hybrids of C. equisetifolia have been reported and a putative hybrid of C. junghuhniana and C. equisetifolia is commercially cultivated in Thailand and India. In Taiwan, studies have also revealed that most plants of Casuarina are the result of introgressive hybridization involving C. equisetifolia, C. glauca, and C. cunninghamiana causing difficulty in species identification. The present study was aimed at developing DNA markers for the identification of C. equisetifolia. Seven ISSR primers were amplified in 120 individuals belonging to three species of Casuarina (C. equisetifolia, C. glauca and C. junghuhniana) and two Allocasuarina species (A. littoralis and A. huegeliana). One species-specific amplicon at 650 bp amplified in all individuals of C. equisetifolia was cloned, sequenced and primer pairs were developed and designated as IFGTBCE01. The sequence characterized amplified region (SCAR) marker was multiplexed with internal SSR primer pair (positive control) and amplified in ten randomly selected individuals of C. equisetifolia, C. junghuhniana, C. glauca, A. littoralis and A. huegeliana. The species-specific band amplified only in C. equisetifolia at 500 bp. Subsequently, the SCAR marker was validated in 30 individuals collected from 10 provenances belonging to 9 countries and in 10 locally selected clones. The marker was validated in the natural male hybrid of C. equisetifolia × C. junghuhniana which is widely planted in India for fast growth, wide adaptability and good form. Hence, this marker can be used for accurate and rapid identification of the species during certification and determination of putative hybrids in breeding programs.     

Specific microtubule-depolymerizing agents augment efficacy of dendritic cell-based cancer vaccines

Background

Damage-associated molecular patterns (DAMPs) are associated with immunogenic cell death and have the ability to enhance maturation and antigen presentation of dendritic cells (DCs). Specific microtubule-depolymerizing agents (MDAs) such as colchicine have been shown to confer anti-cancer activity and also trigger activation of DCs.

Methods

In this study, we evaluated the ability of three MDAs (colchicine and two 2-phenyl-4-quinolone analogues) to induce immunogenic cell death in test tumor cells, activate DCs, and augment T-cell proliferation activity. These MDAs were further evaluated for use as an adjuvant in a tumor cell lysate-pulsed DC vaccine.

Results

The three test phytochemicals considerably increased the expression of DAMPs including HSP70, HSP90 and HMGB1, but had no effect on expression of calreticulin (CRT). DC vaccines pulsed with MDA-treated tumor cell lysates had a significant effect on tumor growth, showed cytotoxic T-lymphocyte activity against tumors, and increased the survival rate of test mice. In vivo antibody depletion experiments suggested that CD8⁺ and NK cells, but not CD4⁺ cells, were the main effector cells responsible for the observed anti-tumor activity. In addition, culture of DCs with GM-CSF and IL-4 during the pulsing and stimulation period significantly increased the production of IL-12 and decreased production of IL-10. MDAs also induced phenotypic maturation of DCs and augmented CD4⁺ and CD8⁺ T-cell proliferation when co-cultured with DCs.

Conclusions

Specific MDAs including the clinical drug, colchicine, can induce immunogenic cell death in tumor cells, and DCs pulsed with MDA-treated tumor cell lysates (TCLs) can generate potent anti-tumor immunity in mice. This approach may warrant future clinical evaluation as a cancer vaccine.     

Application of autologous bone marrow mononuclear cells in six patients with advanced chronic critical limb ischemia as a result of diabetes: our experience

Background aimsPrevious clinical studies have reported that the injection of bone marrow (BM)-derived mononuclear cells (MNC) results in improvement in symptoms and healing of ulcers in patients with critical limb ischemia (CLI) up to stage IV of Fontaine's classification. However, most patients with Fontaine stage IV CLI limbs had to undergo amputation even after stem cell therapy. We report on six patients, who had poorly controlled diabetes with extensive ulceration and gangrene of limbs because of Fontaine stage IV CLI and had been advised amputation elsewhere, who underwent injection of autologous BM MNC.MethodsIn all six patients, BM was aspirated and the isolated MNC from the BM were injected intralesionally at various sites of the ulcer and its surroundings after necessary debridement. The patients were followed up at regular intervals for at least 6 months.ResultsAt the end of the 6-month follow-up, the lower limb pain and ulcers had improved significantly in all patients. The mean toe–brachial index had increased from 0.26 to 0.36. One patient died a month after therapy because of causes unrelated to the procedure. Limb salvage was possible in the remaining five patients and they had a pain-free walking distance of 100 m within 6 months.ConclusionsLimb salvage was possible in all six diabetic patients with Fontaine stage IV CLI following autologous BM MNC injection. The procedure was safe without any adverse outcomes.     

Cellulose-Based Matrix Microspheres of Prednisolone Inclusion Complex: Preparation and Characterization

The purpose of the present investigation was to encapsulate pure prednisolone (PRD) and PRD–hydroxypropyl-β-cyclodextrin (HPβCD) complex in cellulose-based matrix microspheres. The system simultaneously exploits complexation technique to enhance the solubility of low-solubility drug (pure PRD) and subsequent modulation of drug release from microspheres (MIC) at a predetermined time. The microspheres of various compositions were prepared by an oil-in-oil emulsion–solvent evaporation method. The effect of complexation and presence of cellulose polymers on entrapment efficiency, particle size, and drug release had been investigated. The solid-state characterization was performed by Fourier transform infrared spectroscopy, thermogravimetry, differential scanning calorimetry, and powder X-ray diffractometry. The morphology of MIC was examined by scanning electron microscopy. The in vitro drug release profiles from these microspheres showed the desired biphasic release behavior. After enhancing the solubility of prednisolone by inclusion into HPβCD, the drug release was easily modified in the microsphere formulation. It was also demonstrated that the CDs in these microspheres were able to modulate several properties such as morphology, drug loading, and release properties. The release kinetics of prednisolone from microspheres followed quasi-Fickian and first-order release mechanisms. In addition to this, the f ₂-metric technique was used to check the equivalency of dissolution profiles of the optimized formulation before and after stability studies, and it was found to be similar. A good outcome, matrix microspheres (coded as MIC5) containing PRD–HPβCD complex, showed sustained release of drug (95.81%) over a period of 24 h.     

A simple electrochemical approach to fabricate a glucose biosensor based on graphene-glucose oxidase biocomposite

We report a simple electrochemical approach for the immobilization of glucose oxidase (GOx) on reduced graphene oxide (RGO). The immobilization of GOx was achieved in a single step without any cross linking agents or modifiers. A simple solution phase approach was used to prepare exfoliated graphene oxide (GO), followed by electrochemical reduction to get RGO-GOx biocomposite. The direct electrochemistry of GOx was revealed at the RGO-GOx modified glassy carbon electrode (GCE). The electrocatalytic and electroanalytical applications of the proposed film were studied by cyclic voltammetry (CV) and amperometry. It is notable that the glucose determination has been achieved in mediator-free conditions. RGO-GOx film showed very good stability, reproducibility and high selectivity. The developed biosensor exhibits excellent catalytic activity towards glucose over a wide linear range of 0.1-27mM with a sensitivity of 1.85μAmM(-1)cm(-2). The facile and easy electrochemical approach used for the preparation of RGO-GOx may open up new horizons in the production of cost-effective biosensors and biofuel cells.