Chimeric antigen receptor-natural killer cell therapy: current advancements and strategies to overcome challenges.

Chimeric antigen receptor-natural killer (CAR-NK) cell therapy is a novel immunotherapy targeting cancer cells via the generation of chimeric antigen receptors on NK cells which recognize specific cancer antigens. CAR-NK cell therapy is gaining attention nowadays owing to the ability of CAR-NK cells to release potent cytotoxicity against cancer cells without side effects such as cytokine release syndrome (CRS), neurotoxicity and graft-versus-host disease (GvHD). CAR-NK cells do not require antigen priming, thus enabling them to be used as "off-the-shelf" therapy. Nonetheless, CAR-NK cell therapy still possesses several challenges in eliminating cancer cells which reside in hypoxic and immunosuppressive tumor microenvironment. Therefore, this review is envisioned to explore the current advancements and limitations of CAR-NK cell therapy as well as discuss strategies to overcome the challenges faced by CAR-NK cell therapy. This review also aims to dissect the current status of clinical trials on CAR-NK cells and future recommendations for improving the effectiveness and safety of CAR-NK cell therapy.

A case series on safety and tolerability of human umbilical cord-derived mesenchymal stem cells on patients in Malaysia.

Umbilical cord-derived mesenchymal stem cells for regenerative therapy are a promising treatment option for chronic illnesses. Umbilical cord-derived mesenchymal stem cells offer several advantages over other sources, which makes them an attractive option in tissue repair and regeneration. This clinical study describes a 1-year follow-up on the safety and tolerance of umbilical cord-derived mesenchymal stem cell therapy on nine patients in Malaysia. Patients were assessed for adverse effects, and liver function tests were carried out on both pre- and post-treatments. Umbilical cord-derived mesenchymal stem cells' effectiveness and safety were assessed by follow-up evaluations. All nine patients responded positively towards umbilical cord-derived mesenchymal stem cell therapy, without any adverse effects. After umbilical cord-derived mesenchymal stem cell therapy, a significant improvement was observed in liver functioning test outcomes, as haematological parameters and tumour markers were stable. The present study concludes that umbilical cord-derived mesenchymal stem cell therapy is well tolerated by Malaysian patients; however, further clinical screening must be done over a large number of patients population.

Comprehensive review on ideas, designs and current techniques in solar dryer for food applications.

Due to the expansion of residents, the consumption of non-renewable energy increased enormously, thus indirectly increasing pollution and affecting the surroundings. To reduce pollutions in the surroundings, it is recommended to choose non-conventional energy sources. By satisfying this, we can probably decrease the non-renewable sources of energy, by consuming the solar power in day-to-day life in the application of food drying process. In this review article, we have discussed the classification of solar dryer and the impact of design modifications performed in the components of solar dryer and assessed the various types of solar dryer performance, cost estimations and designs performed in solar dryer of food applications which were not discussed in the earlier research. The primary and critical task in designing the solar dryer is to achieve higher efficiency at minimum cost. Hence, proper analysis of drying application, selection of suitable components and suitable design must be carried out to attain efficient dryer. Considering these characteristics, this paper primarily focuses on the effective design parameters incorporated with various efficiency enhancement processes of the solar dryer in the applications of food drying techniques. Thus, this review paper delivers the various classifications, design parameters, performance enhancement methods, properties and valuable assets of solar dryer, which helps to develop the sustainable green eco-friendly environment most primarily, in the application of food drying process. This review article concreted the way for upcoming considerations and provided the techniques for the studies to convey the work for promoting method enhancements.

Synthesis, Characterization, Cytotoxicity Analysis and Evaluation of Novel Heterocyclic Derivatives of Benzamidine against Periodontal Disease Triggering Bacteria.

Periodontal disease (PD) is multifactorial oral disease that damages tooth-supporting tissue. PD treatment includes proper oral hygiene, deep cleaning, antibiotics therapy, and surgery. Despite the availability of basic treatments, some of these are rendered undesirable in PD treatment due to side effects and expense. Therefore, the aim of the present study is to develop novel molecules to combat the PD triggering pathogens. The study involved the synthesis of 4-((5-(substituted-phenyl)-1,3,4-oxadiazol-2-yl)methoxy)benzamidine (5a-e), by condensation of 2-(4-carbamimidoylphenoxy)acetohydrazide (3) with different aromatic acids; and synthesis of 4-((4-(substituted benzylideneamino)-4H-1,2,4-triazol-3-yl)methoxy)benzamidine (6a-b) by treatment of compound 3 with CS2 followed by hydrazination and a Schiff reaction with different aromatic aldehydes. Synthesized compounds were characterized based on the NMR, FTIR, and mass spectrometric data. To assess the effectiveness of the newly synthesized compound in PD, new compounds were subjected to antimicrobial evaluation against P. gingivalis and E. coli using the micro-broth dilution method. Synthesized compounds were also subjected to cytotoxicity evaluation against HEK-293 cells using an MTT assay. The present study revealed the successful synthesis of heterocyclic derivatives of benzamidine with significant inhibitory potential against P. gingivalis and E. coli. Synthesized compounds exhibited minimal to the absence of cytotoxicity. Significant antimicrobial potential and least/no cytotoxicity of new heterocyclic analogs of benzamidine against PD-triggering bacteria supports their potential application in PD treatment.

Removing microplastics from wastewater using leading-edge treatment technologies: a solution to microplastic pollution-a review.

Microplastics (MPs) in environmental studies have revealed that public sewage treatment plants are a common pathway for microplastics to reach local surroundings. Microplastics are becoming more of a worry, posing a danger to both marine wildlife and humans. These plastic items not only contribute to the macrocosmic proliferation of plastics but also the scattering of microplastics and the concentration of other micropollutant-containing objects, increasing the number of pollutants identified. Microplastics' behavior, movement, transformation, and persistence mechanisms, as well as their mode of action in various wastewater effluent treatment procedures, are still unknown. They are making microplastics made from wastewater a big deal. We know that microplastics enter wastewater treatment facilities (WWTPs), that wastewater is released into the atmosphere, and that this wastewater has been considered to represent a threat to habitats and ground character based on our literature assessment. The basic methods of wastewater and sewage sludge, as well as the treatment procedure and early characterization, are covered throughout the dissection of the problematic scientific conceptualization.

Dual-Use Vaccine for Diarrhoeal Diseases: Cross-Protective Immunogenicity of a Cold-Chain-Free, Live-Attenuated, Oral Cholera Vaccine against Enterotoxigenic Escherichia coli (ETEC) Challenge in BALB/c Mice.

In low- and middle-income countries, diarrhoeal diseases are the second most common cause of mortality in children, mainly caused by enterotoxin-producing bacteria, such as Shigella, Vibrio, Salmonella, and Escherichia coli. Cholera and traveller's diarrhoea are caused by Vibrio cholerae (O1 and O139 serogroups) and enterotoxigenic Escherichia coli (ETEC), respectively. The cholera toxin (CT) produced by V. cholerae and the heat-labile enterotoxin (LT) of ETEC are closely related by structure, function, and the immunological response to them. There is no exclusive vaccine for ETEC; however, cholera vaccines based on the CT-B component elicit a short-term cross-protection against ETEC infection. In this context, the cross-protective efficacy of MyCholTM, a prototype cold-chain-free, live-attenuated, oral cholera vaccine against V. cholerae O139 was evaluated in BALB/c mice. The 100% lethal dose (LD100) of 109 CFU/mL of the ETEC H10407 strain was used for the challenge studies. The mice immunised with MyChol™ survived the challenge by producing anti-CT antibodies, which cross-neutralised the LT toxin with no body weight loss and no sign of diarrhoea. Compared to unimmunised mice, the immunised mice elicited the neutralising antitoxin that markedly decreased ETEC colonisation and fluid accumulation caused by ETEC H10407 in the intestines. The immunised mice recorded higher antibody titres, including anti-CT IgG, anti-LT IgG, anti-CT-B IgG, and anti-LTB IgG. Only a two-fold rise in anti-CT/CT-B/LT/LT-B IgA was recorded in serum samples from immunised mice. No bactericidal antibodies against ETEC H10407 were detected. This investigation demonstrates the safety, immunogenicity, and cross-protective efficacy of MyCholTM against the ETEC H10407 challenge in BALB/c mice.

Live, Genetically Attenuated, Cold-Chain-Free Cholera Vaccine-A Research and Development Journey: Light at the End of a Long Tunnel.

Cholera, a diarrheal disease caused by Vibrio cholerae (V. cholerae) O139 and O1 strains, remains a public health problem. The existing World Health Organization (WHO)-licenced, killed, multiple-dose oral cholera vaccines demand 'cold-chain supply' at 2 °C-8 °C. Therefore, a live, single-dose, cold-chain-free vaccine would relieve significant bottlenecks and costs of cholera vaccination campaigns. Our cholera vaccine development journey started in 2000 at Universiti Sains Malaysia with isolation of the hemA gene from V. cholerae, followed by development of a gene mutant vaccine candidate VCUSM2 against V. cholerae O139 in 2006. In 2010, VCUSM2 reactogenicity was reduced by replacing its two wild-type ctxA gene copies with mutated ctxA to produce strain VCUSM14. Introducing the hemA gene into VCUSM14 created VCUSM14P, a strain with the 5-aminolaevulinic acid (ALA) prototrophic trait and excellent colonisation and immunological properties (100% protection to wild-type challenged rabbits). It was further refined in Asian Institute of Medicine, Science and Technology (AIMST University), with completion of single- and repeated-dose toxicity evaluations in 2019 in Sprague Dawley (SD) rats, followed by development of a novel cold-chain-free VCUSM14P formulation in 2020. VCUSM14P is unique for its intact cholera toxin B, a known mucosal adjuvant. The built-in adjuvant makes VCUSM14P an ideal vaccine delivery platform for emerging diseases (e.g. severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2] and tuberculosis). Our vaccine formulation mimics natural infection, remains non-reactogenic and immunogenic in vivo, and protects against infection and disease. It will also cost less and be less cumbersome to distribute due to its stability at room temperature. These features could revolutionise the outreach of this and other vaccines to meet global immunisation programmes, particularly in low-resourced areas. The next stage of our journey will be meeting the requisite regulatory requirements to produce the vaccine for rollout to countries where it is most needed.

Synthesis, Characterization and Biological Evaluation of Novel Benzamidine Derivatives: Newer Antibiotics for Periodontitis Treatment.

Periodontal disease (PD) is complex polymicrobial disease which destroys tooth-supporting tissue. Although various synthetic inhibitors of periodontitis-triggering pathogens have been recognized, their undesirable side effects limit their application. Hence, the present study intended to perform the synthesis, characterization, antimicrobial evaluation, and cytotoxicity analysis of novel benzamidine analogues (NBA). This study involved the synthesis of novel imino bases of benzamidine (4a-c), by reacting different aromatic aldehydes with 2-(4-carbamimidoylphenoxy) acetohydrazide (3), which was synthesized by the hydrazination of ethyl 2-(4-carbamimidoylphenoxy) acetate (2), the derivative of 4-hydroxybenzene carboximidamide (1). This was followed by characterization using FTIR, 1H, 13C NMR and mass spectrometry. All synthesized compounds were further tested for antimicrobial potential against PD-triggering pathogens by the micro broth dilution method. The cytotoxicity analysis of the NBA against HEK 293 cells was conducted using an MTT assay. The present study resulted in a successful synthesis of NBA and elucidated their structures. The synthesized NBA exhibited significant antimicrobial activity values between 31.25 and 125 µg/mL against tested pathogens. All NBA exhibited weak cytotoxicity against HEK 293 cells at 7.81 µg, equally to chlorhexidine at 0.2%. The significant antimicrobial activity of NBA against PD-triggering pathogens supports their potential application in periodontitis treatment.

Review on phase change materials for solar energy storage applications.

The energy storage application plays a vital role in the utilization of the solar energy technologies. There are various types of the energy storage applications are available in the todays world. Phase change materials (PCMs) are suitable for various solar energy systems for prolonged heat energy retaining, as solar radiation is sporadic. This literature review presents the application of the PCM in solar thermal power plants, solar desalination, solar cooker, solar air heater, and solar water heater. Even though the availability and cost of PCMs are complex and high, the PCMs are used in most solar energy methods due to their significant technical parameters improvisation. This review's detailed findings paved the way for future recommendations and methods for the investigators to carry work for further system developments.

Mechanical and Abrasive Wear Performance of Titanium Di-Oxide Filled Woven Glass Fibre Reinforced Polymer Composites by Using Taguchi and EDAS Approach.

Two-body abrasive wear behavior of glass fabric reinforced (GC) epoxy and titanium dioxide (TiO2) filled composites have been conducted out by using a tribo test machine. GC and TiO2 filled GC composites were produced by the hand layup technique. The mechanical performances of the fabricated composites were calculated as per ASTM standards. Three different weight percentages were mixed with the polymer to develop the mechanical and abrasive wear features of the composites. Evaluation Based on Distance from Average Solution (EDAS), a multi-criteria decision technique is applied to find the best filler content. Based on the output, 2wt% TiO2 filler gave the best result. Abrasive wear tests were used to compare GC and TiO2 filled GC composites. The abrasion wear mechanisms of the unfilled and TiO2 filled composites have also been studied by scanning electron microscopy. The outcome of the paper suggests the correct proportion of filler required for the resin in order to improve the wear resistance of the filled composites. Taguchi combined with Multi-Criteria Decision Method (MCDM) is used to identify the better performance of the TiO2 filled epoxy composites.

Microstructure, Mechanical Properties, and Corrosion Behavior of Boron Carbide Reinforced Aluminum Alloy (Al-Fe-Si-Zn-Cu) Matrix Composites Produced via Powder Metallurgy Route.

In this paper, Al-Fe-Si-Zn-Cu (AA8079) matrix composites with several weight percentages of B4C (0, 5, 10, and 15) were synthesized by powder metallurgy (PM). The essential amount of powders was milled to yield different compositions such as AA8079, AA8079-5 wt.%B4C, AA8079-10 wt.%B4C, and AA8079-15 wt.%B4C. The influence of powder metallurgy parameters on properties' density, hardness, and compressive strength was examined. The green compacts were produced at three various pressures: 300 MPa, 400 MPa, and 500 MPa. The fabricated green compacts were sintered at 375 °C, 475 °C, and 575 °C for the time period of 1, 2 and 3 h, respectively. Furthermore, the sintered samples were subjected to X-ray diffraction (XRD) analysis, Energy Dispersive Analysis (EDAX), and Scanning Electron Microscope (SEM) examinations. The SEM examination confirmed the uniform dispersal of B4C reinforcement with AA8079 matrix. Corrosion behavior of the composites samples was explored. From the studies, it is witnessed that the rise in PM process parameters enhances the density, hardness, compressive strength, and corrosion resistance.

Mechanical and Microstructural Characterization of Friction Stir Welded SiC and B4C Reinforced Aluminium Alloy AA6061 Metal Matrix Composites.

This study focuses on the properties and process parameters dictating behavioural aspects of friction stir welded Aluminium Alloy AA6061 metal matrix composites reinforced with varying percentages of SiC and B4C. The joint properties in terms of mechanical strength, microstructural integrity and quality were examined. The weld reveals grain refinement and uniform distribution of reinforced particles in the joint region leading to improved strength compared to other joints of varying base material compositions. The tensile properties of the friction stir welded Al-MMCs improved after reinforcement with SiC and B4C. The maximum ultimate tensile stress was around 172.8 ± 1.9 MPa for composite with 10% SiC and 3% B4C reinforcement. The percentage elongation decreased as the percentage of SiC decreases and B4C increases. The hardness of the Al-MMCs improved considerably by adding reinforcement and subsequent thermal action during the FSW process, indicating an optimal increase as it eliminates brittleness. It was seen that higher SiC content contributes to higher strength, improved wear properties and hardness. The wear rate was as high as 12 ± 0.9 g/s for 10% SiC reinforcement and 30 N load. The wear rate reduced for lower values of load and increased with B4C reinforcement. The microstructural examination at the joints reveals the flow of plasticized metal from advancing to the retreating side. The formation of onion rings in the weld zone was due to the cylindrical FSW rotating tool material impression during the stirring action. Alterations in chemical properties are negligible, thereby retaining the original characteristics of the materials post welding. No major cracks or pores were observed during the non-destructive testing process that established good quality of the weld. The results are indicated improvement in mechanical and microstructural properties of the weld.

Comprehensive review on various parameters that influence the performance of parabolic trough collector.

Among the different kinds of renewable energy sources, solar energy plays a major role because it is safe and inexpensive at all times. Several techniques are developed for steam and electricity generation by solar energy, in which the parabolic trough collector is an advantageous method for generating steam and electricity. Different types of collectors for various temperatures, in which PTCs are used to produce medium temperature ranges using the readily available solar energy, were developed, produced, and tests. Many theoretical and experimental studies have been carried out to improvise parabolic trough collectors' optical and thermal characteristics. The modifications are reviewed in this paper to enhance the design modification, optical and thermal properties utilized in the collector. This analysis paper also elucidates the use of PTC desalination, various integrated parabolic trough collector methods for power generation, and the economic aspects of parabolic trough collector.

Immunogenicity and protective efficacy of a live, oral cholera vaccine formulation stored outside-the-cold-chain for 140 days.

BACKGROUND: Cholera, an acute watery diarrhoeal disease caused by Vibrio cholerae serogroup O1 and O139 across the continents. Replacing the existing WHO licensed killed multiple-dose oral cholera vaccines that demand 'cold chain supply' at 2-8 °C with a live, single-dose and cold chain-free vaccine would relieve the significant bottlenecks and cost determinants in cholera vaccination campaigns. In this direction, a prototype cold chain-free live attenuated cholera vaccine formulation (LACV) was developed against the toxigenic wild-type (WT) V. cholerae O139 serogroup. LACV was found stable and retained its viability (5 × 106 CFU/mL), purity and potency at room temperature (25 °C ± 2 °C, and 60% ± 5% relative humidity) for 140 days in contrast to all the existing WHO licensed cold-chain supply (2-8 °C) dependent killed oral cholera vaccines. RESULTS: The LACV was evaluated for its colonization potential, reactogenicity, immunogenicity and protective efficacy in animal models after its storage at room temperature for 140 days. In suckling mice colonization assay, the LACV recorded the highest recovery of (7.2 × 107 CFU/mL) compared to those of unformulated VCUSM14P (5.6 × 107 CFU/mL) and the WT O139 strain (3.5 × 107 CFU/mL). The LACV showed no reactogenicity even at an inoculation dose of 104-106 CFU/mL in a rabbit ileal loop model. The rabbits vaccinated with the LACV or unformulated VCUSM14P survived a challenge with WT O139 and showed no signs of diarrhoea or death in the reversible intestinal tie adult rabbit diarrhoea (RITARD) model. Vaccinated rabbits recorded a 275-fold increase in anti-CT IgG and a 15-fold increase in anti-CT IgA antibodies compared to those of rabbits vaccinated with unformulated VCUSM14P. Vibriocidal antibodies were increased by 31-fold with the LACV and 14-fold with unformulated VCUSM14P. CONCLUSION: The vaccine formulation mimics a natural infection, is non-reactogenic and highly immunogenic in vivo and protects animals from lethal wild-type V. cholerae O139 challenge. The single dose LACV formulation was found to be stable at room temperature (25 ± 2 °C) for 140 days and it would result in significant cost savings during mass cholera vaccination campaigns.

Design and synthesis of newer 1,3,4-oxadiazole and 1,2,4-triazole based Topsentin analogues as anti-proliferative agent targeting tubulin.

A set of two series of 1,3,4-oxadiazole (11a-n) and 1,2,4-Triazole (12a, c, e, g, h, j-n) based topsentin analogues were prepared by replacing imizadole moiety of topsentin through a multistep synthesis starting from indole. All the compounds synthesized were submitted for single dose (10 µM) screening against a NCI panel of 60-human cancer cell lines. Among all cancer cell lines, colon (HCC-2998) and Breast (MCF-7, T-47D) cancer cell lines were found to be more susceptible for this class of compounds. Among the compounds tested, compounds 11a, 11d, 11f, 12e and 12h, were exhibited good anti-proliferative activity against various cancer cell lines. Compounds 11d, 12e and 12h demonstrated better activity with IC50 2.42 µM, 3.06 µM, and 3.30 µM respectively against MCF-7 human cancer cell line than that of the standard drug doxorubicin IC50 6.31 µM. Furthermore, 11d induced cell cycle arrest at G0/G1 phase and also disrupted mitochondrial membrane potential with reducing cell migration potential of MCF-7 cells in dose dependent manner. In vitro microtubule polymerization assays found that compound 11d disrupt tubulin dynamics by inhibiting tubulin polymerization with IC50 3.89 µM compared with standard nocodazole (IC50 2.49 µM). In silico docking studies represented that 11d was binding at colchicine binding site of ß-tubulin. Compound 11d emerged as lead molecule from the library of compounds tested and this may serve as a template for further drug discovery.

Novel 9-(2-(1-arylethylidene)hydrazinyl)acridine derivatives: Target Topoisomerase 1 and growth inhibition of HeLa cancer cells.

A series of 9-(2-(1-arylethylidene)hydrazinyl)acridine and its analogs were designed, synthesized and evaluated for biological activities. Various biochemical assays were performed to determine the free radical scavenging capacity of synthesized compounds (4a-4j). Anticancer activity of these compounds was assessed against two different human cancer cell lines viz cervical cancer cells (HeLa) and liver cancer cells (HepG2) as well as normal human embryonic kidney cell line (HEK 293). Compounds 4b, 4d and 4e showed potential anti-proliferative effects on HeLa cells. Based on results obtained from antioxidant and cytotoxicity studies, 4b, 4d and 4e were further studied in detail for different biological activities. 4b, 4d and 4e reduced the cell growth, inhibited metastatic activity and declined the potential of cell migration in HeLa cell lines. Topoisomerase1 (Top1) treated with compounds 4b, 4d and 4e exhibited inhibition of Top1 and prevented DNA replication. Molecular docking results validate that interaction of compounds 4b, 4d and 4e with Top1-DNA complex, which might be accountable for their inhibitory effects. Further it was concluded that compounds 4b, 4d and 4e arrests the cells at S phase and consequently induces cell death through DNA damage in HeLa cells.

Deciduous DPSCs Ameliorate MPTP-Mediated Neurotoxicity, Sensorimotor Coordination and Olfactory Function in Parkinsonian Mice.

Parkinson's disease (PD) is a neurodegenerative disorder defined by progressive deterioration of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Dental pulp stem cells (DPSCs) have been proposed to replace the degenerated dopaminergic neurons due to its inherent neurogenic and regenerative potential. However, the effective delivery and homing of DPSCs within the lesioned brain has been one of the many obstacles faced in cell-based therapy of neurodegenerative disorders. We hypothesized that DPSCs, delivered intranasally, could circumvent these challenges. In the present study, we investigated the therapeutic efficacy of intranasally administered DPSCs in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of PD. Human deciduous DPSCs were cultured, pre-labelled with PKH 26, and intranasally delivered into PD mice following MPTP treatment. Behavioural analyses were performed to measure olfactory function and sensorimotor coordination, while tyrosine hydroxylase (TH) immunofluorescence was used to evaluate MPTP neurotoxicity in SNpc neurons. Upon intranasal delivery, degenerated TH-positive neurons were ameliorated, while deterioration in behavioural performances was significantly enhanced. Thus, the intranasal approach enriched cell delivery to the brain, optimizing its therapeutic potential through its efficacious delivery and protection against dopaminergic neuron degeneration.

Repeated dose toxicity evaluation of a cold chain-free, live, attenuated oral cholera vaccine in Sprague Dawley rats.

The repeated dose toxicity of a prototype cold chain-free, live, attenuated oral cholera vaccine containing 5 × 106 CFU/mL of the VCUSM14P strain was evaluated in Sprague Dawley (SD) rats (single dose administered daily for 30 days) to ascertain its safety for clinical use. Repeated dose toxicity studies for cholera vaccines in the literature have administered 2 or 3 fixed doses at 7, 14, 21 or 69 day intervals. The present study reports an evaluation of 30 repeated doses of cholera vaccine administered at three different concentrations (Group II (1.25 × 106 CFU), Group III (2.5 × 106 CFU) and Group IV (5 × 106 CFU)) in SD rats. The liquid vaccine was administered orally to the rats with the respective dose every day, and normal saline was administered to the control group (Group I). No significant difference (P > 0.05) was observed in the body weights and biochemical parameters of the rats after 15 and 30 repeated doses compared to those of the control group. However, compared to those of Group I, a significant increase (P < 0.05) in the organ to body weight ratios of the lungs, ureter, liver, kidney, heart and spleen was found in G-II, G-III and G-IV. In the haematological analysis, a significant increase in the WBC was observed in G-II and G-IV compared to that in G-I. The histopathological findings indicated mild to moderate degeneration in the liver, kidney, heart and spleen in the treated rats. Mild to moderate lymphocytic infiltration in the lungs was observed in the G-II and G-III rats, and severe infiltration was observed in the G-IV rats. These histopathological findings may be attributed to the 30 doses of vaccine given in daily succession without an interval. In the acute toxicity study, a single dose of vaccine up to 10 × 106 CFU did not cause any adverse effects and lethality in SD rats.

Complete Genome Sequence of Lytic Bacteriophage VPUSM 8 against O1 El Tor Inaba Vibrio cholerae.

The complete genome sequence of bacteriophage VPUSM 8 against O1 El Tor Inaba Vibrio cholerae is reported here. The isolated VPUSM 8 has potential use in future phage therapy or as a biocontrol agent for the prevention and treatment of cholera.

Alteration in apyrase enzyme attenuated virulence of Shigella flexneri.

Virulence of Shigella is attributed to the genes presence in chromosome or in the megaplasmid. The apy gene which is located in the megaplasmid of Shigella species encodes for apyrase enzyme, a pathogenesis-associated enzyme causing mitochondrial damage and host cell death. In this study we constructed an apy mutant of Shigella flexneri by insertional activation using a kanamycin resistant gene cassette. The wild type apy gene of S. flexneri 2a was PCR amplified, cloned and mutated with insertion of kanamycin resistant gene cassette (aphA). The mutated construct (apy: aphA) was subcloned into a conjugative suicidal vector (pWM91) at the unique Sma1 and Sac1 sites. The mutation of the wild apy gene in the construct was confirmed by DNA sequencing. The mutated construct was introduced into wild type S. flexneri 2a by conjugation with Escherichia coli. After undergoing homologous recombination, the wild apy gene was deleted from the construct using the sucrose selection method. Non-functional activity of the apyrase enzyme in the constructed strain by colorimetric test indicated the successful mutation of the apyrase enzyme. This strain with mutated apy gene was evaluated for its protective efficacy using the guinea pig keratoconjunctivitis model. The strain was Sereny negative and it elicited a significant protection following challenge with wild S. flexneri strain. This apy mutant strain will form a base for the development of a vaccine target for shigellosis.