RelCoVax®, a two antigen subunit protein vaccine candidate against SARS-CoV-2 induces strong immune responses in mice.

The COVID-19 pandemic has spurred an unprecedented movement to develop safe and effective vaccines against the SARS-CoV-2 virus to immunize the global population. The first set of vaccine candidates that received emergency use authorization targeted the spike (S) glycoprotein of the SARS-CoV-2 virus that enables virus entry into cells via the receptor binding domain (RBD). Recently, multiple variants of SARS-CoV-2 have emerged with mutations in S protein and the ability to evade neutralizing antibodies in vaccinated individuals. We have developed a dual RBD and nucleocapsid (N) subunit protein vaccine candidate named RelCoVax® through heterologous expression in mammalian cells (RBD) and E. coli (N). The RelCoVax® formulation containing a combination of aluminum hydroxide (alum) and a synthetic CpG oligonucleotide as adjuvants elicited high antibody titers against RBD and N proteins in mice after a prime and boost dose regimen administered 2 weeks apart. The vaccine also stimulated cellular immune responses with a potential Th1 bias as evidenced by increased IFN-γ release by splenocytes from immunized mice upon antigen exposure particularly N protein. Finally, the serum of mice immunized with RelCoVax® demonstrated the ability to neutralize two different SARS-CoV-2 viral strains in vitro including the Delta strain that has become dominant in many regions of the world and can evade vaccine induced neutralizing antibodies. These results warrant further evaluation of RelCoVax® through advanced studies and contribute towards enhancing our understanding of multicomponent subunit vaccine candidates against SARS-CoV-2.

Arsenic aggravated reproductive toxicity in male rats exposed to lead during the perinatal period.

The aim of this study was to assess the reproductive toxic effects of arsenic on adult Wistar rats exposed to lead during the perinatal period. The pregnant rats were allowed ad libitum access to tap water containing 819 mg of lead (Pb) per L or without Pb from conception until weaning. Litter size, survival rate and developmental milestones of the pups delivered by Pb exposed dams were comparable to those of the control rats. Conversely, the pups exposed to Pb during the perinatal period exhibited significant delay in cliff avoidance, negative geotaxis, surface righting reflex, ascending wire mesh and testis descent. The control and perinatal Pb-exposed male rats were maintained on tap water containing 2.3 mg of arsenite (As) per L or without arsenite from the pubertal period (post-natal day 55) to adulthood (post-natal day 115) and assessed for reproductive end points. The results revealed that the (1) relative weights of the testis, epididymis, seminiferous tubules and ventral prostate; (2) daily sperm production; (3) epididymal sperm density and (4) numbers of motile, viable, and HOS tail swelled sperm declined significantly in the rats exposed to either Pb or As. The activity levels of testicular 3ß- and 17ß-hydroxysteroid dehydrogenases were also significantly decreased in the experimental rats. Significant elevation in the levels of reactive oxygen species and lipid peroxidation in association with reduced activities of antioxidant enzymes in the testis and different epididymal regions was recorded in the experimental rats. In the fertility study, although each male in the control and experimental groups produced a copulatory plug and impregnated a female, the mean conception time significantly increased in the experimental groups. The mean number of implantations decreased significantly in the females mated with the experimental males. Moreover, the results of the present study also indicate that reproductive alterations were more deteriorated in the Pb-exposed rats treated with arsenic when compared to individual exposures. In conclusion, the data clearly suggest that reproductive toxicity in male rats exposed to Pb during the perinatal period is exacerbated by As treatment during the pubertal period.

Behavioral and neurochemical consequences of perinatal exposure to lead in adult male Wistar rats: protective effect by Centella asiatica.

The present study evaluated the protective effects of Centella asiatica (CA) leaf extract on behavioral deficits and neurotoxicity in adult rat exposed to lead during perinatal period. Adult Wistar rats were exposed to 0.15% lead acetate (Pb) from gestation day 6 through drinking water and the pups were exposed lactationally to Pb till weaning. Significant perturbations in locomotor activity and exploratory behavior were observed in rats exposed to Pb during perinatal period. The levels of lipid peroxidation increased significantly with a reduction in levels of glutathione and activity levels of acetylcholinesterase and antioxidant enzymes in hippocampus, cerebrum, cerebellum, and medulla of brains excised from Pb-exposed rats. Oral supplementation of CA during postweaning period provided significant protection against Pb-induced behavioral impairments and neurotoxicity, without chelating tissue Pb levels. The possible neuroprotective efficacy of CA may be due to its antioxidant potential but not by lowering effects of brain Pb content.

Altered spermatogenesis, steroidogenesis and suppressed fertility in adult male rats exposed to genistein, a non-steroidal phytoestrogen during embryonic development.

This article focuses on the effects of prenatal exposure to genistein on the mother, her pregnancy and reproductive functions of the male progeny, since these issues have ethological relevance in both animals and humans. Pregnant Wistar rats received i.p. injections of genistein at a dose level of 2, 20 or 100 mg/kg body weight daily from 12th to 19th day of gestation. Male pups from control and genistein exposed animals were weaned and allowed to develop until 100 days of age; however, when they were 90 days old, twelve males from each group were cohabited with untreated 90-day old females for 8 days. Results revealed a significant decrease in indices of reproductive organs in adult male rats exposed to genistein during embryonic development. Dose dependent reduction was observed in daily sperm production and epididymal sperm density and quality in genistein treated rats. Significant decrease was observed in the activity levels of 3ß- and 17ß-hydroxysteroid dehydrogenases in testis of experimental rats with a decline in plasma testosterone levels. Histological examination of testis of genistein treated rats indicated deterioration in testicular architecture. In the fertility study, the mean number of implantations and live fetuses per dam mated with 100 mg genistein exposed males was reduced.

Effects of maternal exposure to aflatoxin B1 during pregnancy on fertility output of dams and developmental, behavioral and reproductive consequences in female offspring using a rat model.

A suboptimal in utero environment can have detrimental effects on the pregnancy and long-term adverse "programing" effects on the offspring. Aflatoxin B1 is one of the potent reproductive toxicants and currently detected in both milk and tissues. This article focuses on the effects of prenatal exposure to graded doses of aflatoxin B1 on the pregnancy outcomes of dams and postnatal developments of the female offspring, since these issues have ethological relevance in both animals and humans. Pregnant Wistar rats were injected intramuscularly with vehicle or aflatoxin B1 (10, 20, 50 or 100 µg/kg body weight/day) on days 12-19 of gestation. At parturition, newborns were observed for clinical signs of toxicity and survival. The female offspring were examined through a battery of tests in order to evaluate their developmental, behavioral and reproductive end points. All animals were born alive. The litter size of the aflatoxin B1 treated rats was comparable to the controls. However, the birth weight of the pups in the experimental group was significantly lower when compared to controls. Significant and persistent lags in cliff avoidance, negative geotaxis, surface rightening activity and ascending wire mesh, with a delay in elapsed time for vaginal opening were detected in the female progeny exposed to aflatoxin B1 during embryonic development. The locomotor activity and exploratory behavior in experimental females were significantly decreased than that of controls. Embryonic exposure to aflatoxin B1 also resulted in prolonged stress response, irregular estrus and suppressed fertility output in the progeny at their adulthood. These results indicate that in utero exposure to aflatoxin B1 severely compromised postnatal development of neonatal rats and caused irregular estrus that was accompanied by suppressed fertility output.