Functional annotation of Candida albicans hypothetical proteins: a bioinformatics approach.

Candida albicans is a member of the ascomycetes class of fungi and it is an opportunistic pathogen species responsible for a wide range of fungal infections in humans. Bioinformatics and sequencing analysis of Candida proteomics has disclosed that around 69% proteome is still uncharacterized which needs to be annotated with functions. The NCBI-Genome has termed them as hypothetical proteins (HPs) in the whole proteome of Candida. Interpretation of this substantial portion of the proteome can reveal novel pharmacological targets for markers, drug development, and other therapeutics and so on. In this article, we have assigned functional annotation to these hypothetical proteins using bioinformatics methodologies. The advanced and robust computational models have been used to assign the preliminary functions to these putative HPs with high level of confidence. The findings of this study unveil some novel pharmacological targets for drug therapy and vaccines and it would help to identify novel molecular mechanisms underlying the fungal pathogenesis.

Acute and Sub-acute Oral Toxicity Assessment of Ferrum phosphoricum in Rats.

BACKGROUND: Ferrum phosphoricum (FP) has been used by traditional medicine practitioners for various ailments since ancient times. However, scientific evidence on the safety of FP is still unavailable. Thus, the current study aimed to investigate the acute and sub-acute oral toxicity of homeopathic FP in experimental rats. METHODS: In an acute toxicity investigation, a single dose of 2,000 µL/kg of FP 6c, 30c and 200c was administered to female Wistar rats, which were monitored for up to 14 days according to the Organization for Economic Cooperation and Development (OECD) guideline 423. For a sub-acute toxicity study, FP 6c, 30c and 200c (200 µL/kg) were administered to male and female rats for 28 days as per the OECD guideline 407. All the animals were observed for mortality, clinical signs and body weight during the study. At the end of the experiment, hematological, biochemical and histopathological assessments were performed. RESULTS: During the acute toxicity study, no mortality was observed in rats administered with FP, and thus the median lethal dose (LD50) was identified as >2,000 µL/kg. In the sub-acute study, no mortality or adverse clinical signs were noticed with FP treatment. Moreover, weekly body weight gain was normal. Hematological and biochemical investigations revealed no abnormalities. Furthermore, histological analysis of FP-treated rats' vital organs revealed no pathological changes. CONCLUSION: Overall, our findings imply that FP 6c, 30c and 200c potencies are safe and do not cause toxicity when given orally to Wistar albino rats for an extended period at a dose of 200 µL/kg.

Impact of green synthesized WcAgNPs on in-vitro plant regeneration and withanolides production by inducing key biosynthetic genes in Withania coagulans.

KEY MESSAGE: Withania coagulans (L.) Dunal bio-synthesized silver nanoparticles (WcAgNPs) worked as an abiotic elicitor or auto-catalyst that enhanced root regeneration and withanolides production in in-vitro regenerated W. coagulans. Rapid development in the production / consumption of silver nanoparticles (AgNPs) raised serious concern over its effects on the growth of natural plant community. The knowledge related to impact of AgNPs on plant growth and biocompatibility is increasing day by day, but comprehensive mechanism and gaps regarding their impacts on plant health have yet to be addressed. In the present study, we investigated the impact of Withania coagulans biosynthesized AgNPs (WcAgNPs) on in-vitro plant growth and withanolides production. Obtained results showed that the low concentrations of WcAgNPs significantly induced the plant growth by regulating oxidative stress via anti-oxidative defense system. Physiological, morphology and anatomical features also reflected healthy plant growth under low WcAgNPs exposure. While higher concentrations of WcAgNPs have a negative impact on W. coagulans plant growth due to induced lipid peroxidation, ROS accumulation, and root cell death. At lower concentrations, WcAgNPs have shown a positive effect on in-planta withanolides biosynthesis stimulating withanolide A and withaferin A up to 11.15-22.8-fold, respectively. Furthermore, the expression of withanolides biosynthetic genes were also quantified upon WcAgNPs exposure and terpenes biosynthetic genes showed over-expression. Thus, the present study concludes that the lower concentrations of WcAgNPs positively induced plant growth via improved root organogenesis and also have potential to act as an elicitor for withanolides production.

Short term UV-B radiation mediated modulation of physiological traits and withanolides production in Withania coagulans (L.) Dunal under in-vitro condition.

Accumulation of secondary metabolites is a key process in the growth and development of plants under different biotic/abiotic constraints. Many studies highlighted the regulatory potential of UV-B treatment towards the secondary metabolism of plants. In the present study, we examined the impact of UV-B on the physiology and secondary metabolism of Withania coagulans, which is an important ayurvedic plant with high anti-diabetic potential. Results showed that in-vitro UV-B exposure negatively influenced chlorophyll content and photosynthetic machinery. However, Fv/Fm ratio was found non-significantly altered up to 3 h UV-B exposure. The maximum lipid peroxidation level was recorded with 46.8% higher malondialdehyde content in the plants supplemented with 5 h UV-B radiation, that was indicated the oxidative stress in W. coagulans. Conversely, UV-B treatment significantly increased the plant's stress protective compounds like carotenoids, anthocyanin, phenol and proline, in W. coagulans. Free radical scavenging activity was also significantly increased ~ 18% than the control with 3 h UV-B treatment. The maximum antioxidative enzymes activities were observed with the short-term (up to 3 h) UV-B treatment. Specifically, UV-B radiation exposure significantly increased the content of withaferin A and withanolide A in W. coagulans with maximum 1.38 and 3.42-folds, respectively. Additionally, withanolides biosynthesis related genes transcript levels were found over-expressed under the response of UV-B elicitation. The acquired results suggested that short-term UV-B supplementation triggers secondary metabolism along with combating oxidative stress via improving the antioxidative defense system in W. coagulans. Also, UV-B can be used as an efficient abiotic elicitor to increase pharmaceutical compounds (withanolides) production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01046-7.

Epigenetic control of UV-B-induced flavonoid accumulation in Artemisia annua L.

MAIN CONCLUSION: UV-B-induced flavonoid biosynthesis is epigenetically regulated by site-specific demethylation of AaMYB1, AaMYC, and AaWRKY TF-binding sites inAaPAL1promoter-causing overexpression ofAaPALgene inArtemisia annua. The present study was undertaken to understand the epigenetic regulation of flavonoid biosynthesis under the influence of ultraviolet-B radiation using Artemisia annua L. as an experimental model. In-vitro propagated and acclimatized plantlets were treated with UV-B radiation (2.8 W m-2; 3 h), which resulted in enhanced accumulation of total flavonoid and phenolics content as well as eleven individual flavonoids measured through HPLC-DAC. Expression of eight genes (phenylanaline ammonia lyase, cinnamate-4-hydroxylase, 4-coumarate: CoA ligase; chalcone synthase, chalcone isomerase, cinnamoyl reductase, flavonoid-3'-hydroxylase, and flavones synthase) from upstream and downstream flavonoid biosynthetic pathways was measured through RT-PCR and RT-Q-PCR and all were variably induced under UV-B irradiation. Among them, AaPAL1 transcript and its protein were most significantly upregulated. Global DNA methylation analysis revealed hypomethylation of genomic DNA in A. annua. Further epigenetic characterization of promoter region of AaPAL1 revealed cytosine demethylation at five sites, which in turn caused epigenetic activation of six transcription factor-binding sites including QELEMENT, EBOXBNNAPA/MYCCONSENSUSAT, MYBCORE, MYBCOREATCYCB1, and GCCCORE. MYB transcription factors are positive regulators of flavonoid biosynthesis. Epigenetic activation of transcription-enhancing cis-regulatory elements in AaPAL1 promoter and subsequent overexpression of AaMYB1 and AaMYC and AaWRKY transcription factors under UV-B irradiation may probably be the reason for higher AaPAL1 expression and hence greater biosynthesis of flavonoids in A. annua L. The present study is the first report that provides mechanistic evidence of epigenetic regulation of flavonoid biosynthesis under UV-B radiation in A. annua L.

Ultraviolet Radiations: Skin Defense-Damage Mechanism.

UV-radiations are the invisible part of light spectra having a wavelength between visible rays and X-rays. Based on wavelength, UV rays are subdivided into UV-A (320-400 nm), UV-B (280-320 nm) and UV-C (200-280 nm). Ultraviolet rays can have both harmful and beneficial effects. UV-C has the property of ionization thus acting as a strong mutagen, which can cause immune-mediated disease and cancer in adverse cases. Numbers of genetic factors have been identified in human involved in inducing skin cancer from UV-radiations. Certain heredity diseases have been found susceptible to UV-induced skin cancer. UV radiations activate the cutaneous immune system, which led to an inflammatory response by different mechanisms. The first line of defense mechanism against UV radiation is melanin (an epidermal pigment), and UV absorbing pigment of skin, which dissipate UV radiation as heat. Cell surface death receptor (e.g. Fas) of keratinocytes responds to UV-induced injury and elicits apoptosis to avoid malignant transformation. In addition to the formation of photo-dimers in the genome, UV also can induce mutation by generating ROS and nucleotides are highly susceptible to these free radical injuries. Melanocortin 1 receptor (MC1R) has been known to be implicated in different UV-induced damages such as pigmentation, adaptive tanning, and skin cancer. UV-B induces the formation of pre-vitamin D3 in the epidermal layer of skin. UV-induced tans act as a photoprotection by providing a sun protection factor (SPF) of 3-4 and epidermal hyperplasia. There is a need to prevent the harmful effects and harness the useful effects of UV radiations.

Investigation of The Effect of Metal-based Medicine Arsenicum album on Humoral Immune Response in SRBC-immunized Mice.

BACKGROUND: In complementary and alternative medicinal systems, the Arsenicum album in ultra-high dilution was used in various therapeutic conditions, considering its effects on the body's immune system, including the COVID-19 pandemic. However, scientific evidence regarding its immunomodulatory effects is insufficient. OBJECTIVE: The current study aimed to investigate the immunomodulatory effects of Arsenicum album in an experimental mouse model. MATERIALS AND METHODS: Immunomodulatory activity of potentized dilutions of Arsenicum album i.e., 6C, 30C, 200C in BALB/c mice was evaluated by humoral antibody titer and delayed- type hypersensitivity assays wherein a fixed concentration (0.5 ml of 1× 109 cells/ml) of freshly prepared sheep RBC was administered as a foreign antigen to generate primary and secondary antibodies. RESULTS: Arsenicum album showed significant immunomodulatory activity by increasing primary antibody titer evaluated on day 21 of the treatment in all the dilutions as compared to SRBC and vehicle control group in humoral immune response assay without showing any effect on delayed-type hypersensitivity. CONCLUSION: The results of this preliminary study indicate that oral administration of Arsenicum album has the potential to augment primary humoral response at all dilutions. Hence, the possibility of using the Arsenicum album could be explored to treat immunological conditions, infections, etc., as an alternative therapy alongwith modern medicines.

Neuromodulatory potential of Asparagus racemosus and its bioactive molecule Shatavarin IV by enhancing synaptic acetylcholine level and nAChR activity.

Cholinergic dysfunction has been commonly known to be associated with plethora of neurodegenerative disorders and also serves as a biomarker. Recently, cholinergic system demonstrated that acetylcholine has major role in regulation of its function therefore the main therapeutic regimens towards disease management have been focused on increasing acetylcholine levels. The current study explores the potential of Asparagus racemosus extract (ARE) and its bioactive molecule Shatavarin IV (SIV) in improving cholinergic transmission via utilizing Caenorhabditis elegans considering as a model system. Observations and results obtained through this study have clearly showed significant modulation in cholinergic function by increasing acetylcholine (ACh) levels and the nicotinic acetylcholine receptors (nAChRs) activity. Further exploration on mechanistic facet pointed towards ARE and SIV modulatory potential through increased synaptic ACh level by blocking acetyl cholinesterase at enzyme level and by regulating increment in transcript level of cha-1, and cho-1 that are directly responsible for the synthesis of ACh. Further, the up-regulation of unc-38 and unc-50 transcripts could be the reason for enhanced nAChR activity and investigation on stress modulator activity showed excellent efficiency of ARE and SIV in diminishing ROS thereby lowering the oxidative damage.

Interactive role of salicylic acid and nitric oxide on transcriptional reprogramming for high temperature tolerance in lablab purpureus L.: Structural and functional insights using computational approaches.

Salicylic acid (SA) and nitric oxide (NO) are considered as putative plant growth regulators that are involved in the regulation of an array of plant's growth and developmental functions under environmental fluctuations when applied at lower concentrations. The possible involvement of NO in SA induced attenuation of high temperature (HT) induced oxidative stress in plants is however, still vague and need to be explored. Therefore, the present study aimed to investigates the biochemical and physiological changes induced by foliar spray of SA and NO combinations to ameliorate HT induced oxidative stress in Lablab purpureus L. Foliar application of combined SA and NO significantly improved relative water content (27.8 %), photosynthetic pigment content (67.2 %), membrane stability (45 %), proline content (1.0 %), expression of enzymatic antioxidants (7.1-18 %) along with pod yield (1.0 %). Heat Shock Factors (HSFs) play crucial roles in plants abiotic stress tolerance, however there structural and functional classifications in L. purpureus L. is still unknown. So, In-silico approach was also used for functional characterization and homology modelling of HSFs in L. purpureus. The experimental findings depicted that combine effect of SA and NO enhances tolerance in HT stressed L. purpureus L. plants by regulating physiological functions, antioxidants, expression and regulation of stress-responsive genes via transcriptional regulation of heat shock factor.

Effect of Vanadium Supplementation on Production Performance, Nutrient Utilization, Plasma Mineral Concentration, and Mineral Balance in Lactating Goats.

Vanadium (V) has not been elucidated as an essential mineral in ruminants, though in lower organisms and rat model, its role is well known as insulin-a mimetic agent for catalyzing enzymatic activities. The objective of this study was to determine the effect of V supplementation on production performance, milk composition, and mineral profile in lactating goats. Twenty-four crossbred goats (body weight 34.83 ± 0.25 kg) were blocked in four groups and randomly assigned to four treatment groups (n = 6) on body weight and milk yield basis. All the animals were kept on similar feeding regimen except that different treatment groups were supplemented with 0, 2, 4, and 6 ppm inorganic V/kg DM. Feed intake, milk yield, milk composition, nutrient utilization, minerals in plasma and milk, and their balance studies post vanadium supplementation were observed during the 150-day experimental period. V supplementation did not change dry matter intake (DMI), milk yield, and composition during the experimental period. Calcium levels in plasma were improved (P < 0.05) on vanadium supplementation. Nutrient digestibility remained similar among goats fed on basal or V-supplemented diets. Blood and milk V concentration showed a positive correlation with supplemental V levels but no difference was observed in levels of other minerals. The results of present findings indicated that up to the supplemented level, dietary inorganic V does not affect the production and mineral profile in milk and plasma.

Green and cost effective synthesis of silver nanoparticles from endangered medicinal plant Withania coagulans and their potential biomedical properties.

Silver nanoparticles (AgNPs) have great potential for their mechanistic role in biomedical researches. Recently, green biosynthetic approaches have been received much attention in plant science for nanoparticles production. Therefore, in the present study AgNPs have been synthesized utilizing in-vitro grown leaf extract of anti-diabetic medicinal plant Withania coagulans Dunal by the reduction of silver nitrate solution. W. coagulans synthesized silver nanoparticles (WcAgNPs) were characterized by UV-visible spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, X-ray powder diffraction and Fourier transform Infra-Red spectroscopy. All cumulative results showed that WcAgNPs were ~14 nm in size having spherical shape with face centered cubic structure. High performance liquid chromatography confirmed the involvement of withanolides in AgNPs synthesis as a reducing/capping agent. Synthesized WcAgNPs showed greater antioxidative potential when compared with W. coagulans leaf extract. WcAgNPs have efficient antimicrobial potential and suppresses the growth of both gram positive and gram negative bacteria. In our finding we also observed cytotoxicity of WcAgNPs against SiHa (cervical cancerous, hyper-triploid) cell lines and apoptosis in SiHa cells after 48 hour incubation with 13.74 µg ml-1 (IC50) concentration of WcAgNPs. As results suggested, this is the first report which explain that W. coagulans leaf extract have potential as bio-reducing agent for synthesis of silver nanoparticles, which can be exploited as anti-oxidant, antimicrobial and anti-cancerous agent and depicting an effective way for utilizing bioactive resources in restoration of medicinal properties of this plant with high efficacy.

Vanadium in Biosphere and Its Role in Biological Processes.

Ultra-trace elements or occasionally beneficial elements (OBE) are the new categories of minerals including vanadium (V). The importance of V is attributed due to its multifaceted biological roles, i.e., glucose and lipid metabolism as an insulin-mimetic, antilipemic and a potent stress alleviating agent in diabetes when vanadium is administered at lower doses. It competes with iron for transferrin (binding site for transportation) and with lactoferrin as it is secreted in milk also. The intracellular enzyme protein tyrosine phosphatase, causing the dephosphorylation at beta subunit of the insulin receptor, is inhibited by vanadium, thus facilitating the uptake of glucose inside the cell but only in the presence of insulin. Vanadium could be useful as a potential immune-stimulating agent and also as an antiinflammatory therapeutic metallodrug targeting various diseases. Physiological state and dose of vanadium compounds hold importance in causing toxicity also. Research has been carried out mostly on laboratory animals but evidence for vanadium importance as a therapeutic agent are available in humans and large animals also. This review examines the potential biochemical and molecular role, possible kinetics and distribution, essentiality, immunity, and toxicity-related study of vanadium in a biological system.

Inorganic Vanadium Supplementation in Crossbred Calves: Effects on Antioxidant Status, Immune Response and Haemato-Biochemical Attributes.

The aim of the study was to assess the effect of inorganic vanadium (V) supplementation on antioxidant enzymes, immune status, and haemato-biochemical attributes of growing crossbred calves. Twenty-four male Karan Fries calves (Tharparkar × Holstein Friesian) (initial body mass 72.83 ± 2.5 kg; age 3-9 month) were randomly allocated to four groups: the control (received basal diet devoid of supplemental V), the 3 ppm (received basal diet with 3 mg/kg V), the 6 ppm (received basal diet with 6 mg/kg V) and the 9 ppm group (received basal diet with 9 mg/kg V). All the calves were fed for 150 days as per ICAR (2013) feeding standards to meet their nutrient requirements for 500 g growth rate/day. Peripheral blood samples were collected at the start of experiment and subsequently at 30, 60, 90, 120 and 150 days post-V supplementation for determination of antioxidant enzyme activity, immunological parameters and haemato-biochemical attributes. Results indicated that dietary supplementation of V did not affect daily gain, feed intake and haematological parameters. Crossbred calves fed with 9 mg V/kg diet showed reduced (P < 0.05) plasma total cholesterol concentration; however, plasma total protein and glucose concentration remained unaltered. Glutathione peroxidase (GPx) activity as well as immunoglobulin G (IgG) concentration was significantly (P < 0.05) higher in group supplemented with 9 mg V/kg DM; however, superoxide dismutase (SOD), catalase activity and total plasma immunoglobulin (Ig) concentration were similar in all experimental group. Dietary V supplementation showed a negative relation with plasma thiobarbituric acid reactive substances (TBARS) concentration, whereas non-esterified fatty acid (NEFA) concentration remained unaltered among all groups. Plasma V level increased (P < 0.05) with increasing dietary V levels without affecting levels of Ca, Mg, Fe, Cu and Zn. In conclusion, a dietary addition of 9 mg V/kg DM reduced cholesterol content and improved antioxidant and immune response in growing crossbred calves.

Influence of Feeding Inorganic Vanadium on Growth Performance, Endocrine Variables and Biomarkers of Bone Health in Crossbred Calves.

The nutritional essentialities of transition element vanadium (V) as micro-nutrient in farm animals have not yet been established, though in rat model, vanadium as vanadate has been reported to exert insulin-mimetic effect and shown to be needed for proper development of bones. The objective of this study was to determine the effect of V supplementation on growth performance, plasma hormones and bone health status in calves. Twenty-four crossbred calves (body weight 72.83 ± 2.5 kg; age 3-9 months) were blocked in four groups and randomly assigned to four treatment groups (n = 6) on body weight and age basis. Experimental animals were kept on similar feeding regimen except that different groups were supplemented with either 0, 3, 6 or 9 ppm inorganic V/kg DM. Effect of supplementation during 150-day experimental period was observed on feed intake, body weight gain, feed efficiency, body measures, endocrine variables, plasma glucose and biomarkers of bone health status. Supplementation of V did not change average daily gain (ADG), dry matter intake (DMI), feed efficiency and body measures during the experimental period. During the post-V supplementation period plasma insulin-like growth factor-1 (IGF-1), triiodothyronine (T3) and thyroxin (T4) concentrations were increased and observed highest in 9 mg V/kg DM fed calves; however, levels of insulin, glucose, parathyroid hormone (PTH) and calcitonin hormones remained similar among calves fed on basal or V-supplemented diets. Bone alkaline phosphatase (Bone-ALP) concentration was increased (P < 0.05); however, plasma protein tyrosine phosphatase (PTP) level decreased (P < 0.05) in 6 and 9 mg V/kg DM supplemented groups. Plasma hydroxyproline (Hyp) and tartrate-resistant acid phosphatase (TRAP) concentration were unchanged by V supplementation. Blood V concentration showed positive correlation with supplemental V levels. These results suggest that V may play a role in modulation of the action of certain endocrine variables and biomarkers of bone health status in growing crossbred calves.

Silicon bioavailability in exocarp of Cucumis sativus Linn.

Scanning electron microscopy (SEM) and electron probe micro-analyzer (EPMA) techniques have been used to detect the silicon bioavailability in the exocarp of warty cucumber surface. Warts appear at the time of anthesis and are remnant part of spines/trichomes which on further fruit maturation abscised from the exocarp. Results of EPMA and phytolith analysis clearly revealed that the surface of exocarp (fruit) of Cucumis sativus Linn. containing warts has greater quantity of silicon as compared to the other part of the fruit. Besides silicon, some other elements were also found, on the fruit exocarp and its surrounding area. The other elements are magnesium (Mg), aluminum (Al), phosphorus (P), sulfur (S), potassium (K), calcium (Ca), iron (Fe), nickel (Ni), copper (Cu), and sodium. The percentage of silica is highest followed by Ni, Ca, Al, P, Mg, Fe, S, Cu, K, and Cl. Thus, this study clearly demonstrates that Cucumis sativus Linn. fruits which are used as salads and appetizers on daily basis are loaded with silicon and other useful elements and possess numerous health benefits.