Hepatic endogenous defense potential of propolis after mercury intoxication

Exposure to mercuric chloride (HgCl₂; 5 mg kg^?1 body weight; i.p.) induced oxidative stress in mice and substantially increased lipid peroxidation (LPO) and oxidized glutathione (GSSG) levels, decreased the level of reduced glutathione (GSH) and various antioxidant enzymes in liver and also increased the activities of liver marker enzymes in serum. Therapy with propolis extract, a resinous wax‐like beehive product (200 mg kg^?1 orally, after mercury administration), for 3 days inhibited LPO and the formation of GSSG and increased the level of GSH in the liver. Release of serum transaminases, alkaline phosphatase, lactate dehydrogenase and γ‐glutamyl transpeptidase were significantly restored after propolis treatment. The activities of antioxidant enzymes, that is, superoxide dismutase, catalase, glutathione‐S‐transferase and glucose‐6‐phosphate dehydrogenase, were also concomitantly restored towards normal levels after propolis administration. These observations clearly demonstrate that propolis treatment augments antioxidant defense against mercury‐induced toxicity and provide evidence that propolis has therapeutic potential as a hepatoprotective agent.     

Comparative Analysis of CD4+ and CD8+ T Cells in Tumor Tissues,Lymph Nodes and the Peripheral Blood from Patients with Breast Cancer

Background: CD4+ and CD8+ T cells are the main types of lymphocytes in cell-mediated immunity and play a central role in the induction of efficient immune responses against tumors. The frequencies of T cell subtypes in the peripheral blood and tumor tissues, and draining lymph nodes (dLN) can be considered as useful markers for evaluation of the immune system in cancers. Methods: In this study, the frequencies of CD4+ and CD8+ T cells in blood, tumor tissues, and dLN samples of breast cancer patients were compared with each other and with similar tissues from normal individuals. Immunophenotyping was carried out by flow cytometry and the expression levels of CXCL10, granzyme B, and mammaglobin were evaluated by real-time PCR. Results: In the peripheral blood, there were no differences in the T cell subsets between the patients and the normal individuals. The frequency of CD8+ T cells was significantly higher in tumor tissue than normal breast tissues while granzyme B expression was similar. Based on mammaglobin expression levels, dLN have been classified into micro- and macro-metastatic dLN. We found significantly lower frequency of CD4+ in macro-metastatic dLN than micro-metastatic dLN. CD8+ frequency was similar in both dLN; however, granzyme B expression was higher in micro-metastatic ones. There was not any significant difference in CXCL10 expression between the two types of dLN. Conclusion: Based on our results, although the tumor does not affect the systemic immunity, tumoral cells affect the local immune system in the tumoral tissues and the metastatic dLN. Key Words: Breast neoplasms, CD4+ T lymphocyte, CD8+ T lymphocytes, CXCL10, Granzymes     

Spontaneous ordering of oxide nanostructures

We report the spontaneous formation of uniformly distributed arrays of "tips" (tall conical hillocks) upon oxidation of palladium (Pd) thin films. The formation of the palladium oxide tips depended on the thickness and granularity of the Pd film and on annealing and oxidation conditions. As the Pd film thickness increased from 40 to 200 nanometers, the average height of the tips increased from 0.5 to 1.2 micrometers, their height distribution became broader, and their density decreased from 55 x 10(6) to 12 x 10(6) per square centimeter. Enhanced photoelectron emission from locations corresponding to the tips suggests their possible use in field emission applications.     

Brucella abortus RB51: enhancing vaccine efficacy and developing multivalent vaccines

Brucella abortus vaccine strain RB51 is an attenuated, stable rough mutant that is being used in many countries to control bovine brucellosis. Our earlier study demonstrated that the protective efficacy of strain RB51 can be significantly enhanced by overexpressing Cu–Zn superoxide dismutase (SOD), a homologous protective antigen. We have also previously demonstrated that strain RB51 can be engineered to express heterologous proteins and mice vaccinated with such recombinant RB51 strains develop a strong Th1 type of immune response to the foreign proteins. The present study is aimed at combining these two characteristics to generate new recombinant RB51 vaccines with enhanced abilities to protect against brucellosis and simultaneously able to protect against infections by Mycobacterium spp. We constructed two recombinant RB51 strains, RB51SOD/85A which overexpresses SOD with simultaneous expression of the 85A, a protective protein of Mycobacterium spp., and RB51ESAT which expresses ESAT-6, another protective protein of M. bovis, as a fusion protein with the signal sequence and few additional amino terminal amino acids of SOD. Mice vaccinated with these recombinant strains developed specific immune responses to the mycobacterial proteins and significantly enhanced protection against Brucella challenge compared to the mice vaccinated with strain RB51 alone.     

Whitebacked planthopper Sogatella furcifera (Horváth) (Homoptera: Delphacidae) resistance in rice variety Sinna Sivappu

Whitebacked planthopper (WBPH) along with brown planthopper (BPH) has emerged as a major pest of rice in several Asian countries. Development and cultivation of varieties resistant to both planthoppers is an ecologically acceptable strategy to manage these pests. Sinna Sivappu, a Sri Lankan landrace, was reported to be resistant to both planthoppers. While inheritance of BPH resistance has been reported, the genetics of WBPH resistance in this variety is not known. Using a mapping population of 255 F_2:3 families from Taichung Native (TN)1/Sinna Sivappu cross and 128 polymorphic simple sequence repeat (SSR) markers, genes or quantitative trait loci (QTLs) for WBPH resistance quantified in ten phenotypic tests were identified, adopting classical Mendelian segregation, correlation and QTL analyses. The inheritance pattern suggested that a single recessive gene controlled regulation of seedling damage score. Antixenosis or nymphal preference was influenced by two complementary recessive genes, whereas tolerance in terms of days to wilt was under the influence of a single dominant gene. Several of these phenotypic tests recorded high degree of positive or negative correlation between them, suggesting dependence or redundancy of the tests. QTL analysis revealed 13 loci associated with nine traits. Five major-effect QTLs were detected for damage score (chromosome 6), nymphal survival (chromosome 12), and days to wilt (three QTLs on chromosome 4). We suggest involvement of four WBPH resistance genes in Sinna Sivappu, designated as wbph9(t), wbph10(t), wbph11(t), and Wbph12(t). One of the recessive genes could be allelic to any of the recessive genes reported in cluster C on chromosome 6 which might confer resistance to both BPH and WBPH.     

Identification of a Mycoplasma ovis-like organism in a herd of farmed white-tailed deer (Odocoileus virginianus) in rural Indiana

Mycoplasma ovis is a hemoplasma parasite of sheep, goats, and reindeer; however, natural hemoplasma infection in white-tailed deer has not previously been reported. Subsequent to finding many coccoid, bacillary, and ring-shaped organisms, consistent with hemotropic mycoplasmas, on RBCs from a 72-day-old female white-tailed fawn, we sought to (1) identify the putative hemoplasma observed in blood from the fawn, (2) evaluate others in the herd for hemoplasma infection, and (3) identify clinicopathologic characteristics of hemoplasma-infected white-tailed deer. EDTA-anticoagulated whole blood was collected from the fawn and 8 apparently healthy does in the same herd. CBCs were performed on 7 nonclotted samples from the fawn and 6 does. DNA was extracted from all samples, followed by PCR amplification of bacterial (16S rDNA) and protozoal (18S rDNA) genes. The nearly complete 16S rDNA product from the fawn's sample was directly sequenced and compared with known sequences in the GenBank database. Samples from the fawn and 7 of 8 does were PCR-positive using hemoplasma-specific and M ovis-specific protocols. The fawn was PCR-negative for Anaplasma spp., Babesia spp., and Theileria spp. The 16S rDNA sequence from the fawn (GenBank accession number, FJ824847) was most closely related to M ovis (AF338268), having 98.5% sequence identity. The fawn had a mild nonregenerative anemia, a neutrophilic left-shift with toxic change, aspiration bronchopneumonia, and gastrointestinal disease. Hematologic values, including blood film evaluation, in infected does were unremarkable. The M ovis-like organism may have acted as either an opportunistic or primary pathogen in the fawn. The high occurrence of subclinical infections in the does suggests that white-tailed deer may act as wildlife reservoirs for M ovis.     

Effect of ammonia on the electrolyte status of an Indian major carp Catla catla

Ammonia is present in the aquatic environment due to agricultural runoff, sewage effluents and decomposition of biological waste. High level of ammonium in the aquatic environment or in aquaculture farms is a serious problem for aquatic organisms. In the present investigation, the sublethal effects of total ammonia nitrogen (TAN) at 9.98 mg L^?1 on plasma electrolytes (Na⁺, K⁺ and Cl^?) level of Indian major carp Catla catla fingerlings was studied for a period of 35 days under semistatic conditions. Exposure of fish to sublethal concentration of TAN at 9.98 mg L^?1 caused changes in the levels of plasma electrolytes (Na⁺, K⁺ and Cl^?) compared with control fish. No mortality was observed in any group during the treatment period. The present study reveals that the levels of plasma sodium and potassium (except on 7th day) electrolytes were significantly (P < 0.05) decreased throughout the study period. However, the decrease in plasma chloride level was found to be not significant when compared with control group. The present study concludes that the measurements of plasma electrolytes levels can be used as potential biomarkers for monitoring nitrogenous chemicals in aquatic environment.     

Polyclonal antibody‐based farmer‐friendly flow‐through test for the detection of acute hepatopancreatic necrosis disease in shrimp

Early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) is currently the most significant disease of shrimp in farms of Vietnam, Thailand, Malaysia, China and Mexico, and there is a great risk that it may spread to other shrimp farming countries. Although, an array of sophisticated detection tools for AHPND available, there is a need for a sensitive, simple and rapid detection method. In this study, a simple, sensitive, rapid and polyclonal antibody‐based farmer‐friendly flow‐through assay (FTA) test has been developed for the detection of AHPND pathogen. The recombinant Photorhabdus insect‐related (Pir) A toxin‐like protein of AHPND pathogen was used to immunize rabbits at 21‐day interval observed for highest antibody titre after third booster by ELISA. The raised rabbit antiserum was purified by affinity chromatography and characterized by Western blot. The antiserum showed no cross‐reactivity with AHPND‐free Vibrio parahaemolyticus, V. anguillarum, White Spot Virus (WSV), Aeromonashydrophila and Aphanomycesinvadans. This polyclonal rabbit antiserum was used to develop a farmer‐friendly FTA test for the detection of AHPND pathogen. This simple FTA testis is more sensitive and could detect PirA^VP toxin up to 0.121 µg/ml, compared with 0.242 µg/ml by immunodot assay. Furthermore, FTA test requires only 8–10 min for completion, compared with 3 hr by immunodot thus found to be more sensitive, specific and cost‐effective. Collectively, sensitive FTA test would help shrimp farmers to take real‐time management decisions, especially emergency harvest and finally be a better hope for the prevention of AHPND.     

Alteration in certain enzymological parameters of an Indian major carp, Cirrhinus mrigala exposed to short- and long-term exposure of clofibric acid and diclofenac

The extensive use of pharmaceuticals in human and veterinary medicine may enter the aquatic environment and pose a serious threat to non-target aquatic organisms like fish. In this study, Indian major carp Cirrhinus mrigala was exposed to different concentrations (1, 10 and 100 μg L^?l) of most commonly used pharmaceutical drugs clofibric acid (CA) and diclofenac (DCF) to evaluate its impacts on certain enzymological parameters during short- and long-term exposures. During short-term (96 h) exposure period, plasma glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and gill Na⁺/K⁺-ATPase activity were significantly altered at all concentrations of both the CA- and DCF-treated fish. In long-term exposure (35 days), gill Na⁺/K⁺-ATPase activity was found to be significantly increased at all concentration of CA and DCF exposures throughout the study period (except at the end of 7th day in 10 and 100 µg L-1) . However, a biphasic trend was observed in plasma GOT and GPT activity when compared to the control groups. In both short- and long-term exposure, a significant (P < 0.01 and P < 0.05) changes were observed in all enzymological parameters of fish C. mrigala exposed to different concentrations of CA and DCF. The alterations of these enzymological parameters can be effectively used as potential biomarkers in monitoring of pharmaceutical toxicity in aquatic environment and organisms.     

Development and characterization of five novel cell lines from snubnose pompano,Trachinotus blochii (Lacepede, 1801), and their application in gene expression and virological studies

Five novel permanent cell lines have been established from gill, heart, kidney, eye and fin of snubnose pompano, Trachinotus blochii. They were designated as snubnose pompano gill (SPG), snubnose pompano heart (SPH), snubnose pompano kidney (SPK), snubnose pompano eye (SPE) and snubnose pompano fin (SPF), respectively. All these cell lines were characterized and cryopreserved successfully at different passage levels. Cell lines were passaged every alternate day; SPG, SPH, SPK, SPE and SPF cell lines attained passage levels of 68, 74, 82, 79 and 106, respectively, since the initiation of their development in 2019. The cell lines grew well in Leibovitz's 15 medium containing 15% foetal bovine serum at 28°C. Immunophenotyping of the cell lines revealed the presence of fibronectin and pancytokeratin. No mycoplasma contamination was found. The transfection study revealed the gene expression efficiency of these cell lines by expressing the green fluorescent protein (GFP). The authentication on origin of cell lines from T. blochii was confirmed by amplification of species-specific mitochondrial cytochrome oxidase I gene. The results showed the susceptibility of these cell lines to fish nodavirus (FNV) and tilapia lake virus (TiLV) and resistance to cyprinid herpesvirus 2 (CyHV-2). The FNV infection in the cell lines was confirmed by RT-PCR, Western blot, ELISA and immunocytochemistry, while TiLV infection was confirmed by RT-PCR assay. These results revealed that these cell lines are suitable for virological and foreign gene expression studies.