Natural Bioactive Polypeptides: A Study towards the Clinical Application in The Bahamas and Trinidad & Tobago

Synthetic medication is being overprescribed, resulting in adverse complications linked to major public health issues such as antimicrobial resistance, drug dependency, and the current opioid crisis. According to the United Nations Office on Drugs and Crime, this global burden of overuse of synthetic accounts for the heaviest burden of disease attributable to drug use disorders. In 2015, with almost 12 million disability-adjusted life year (DALYs), or 70% of the global burden of disease attributable to opioid addiction. With suitable alternatives available, peptide-based drugs will help to reduce the global burden, appease the patient preference of naturally derived medication and ensure safer patient usage.

Traditional Medicines and Dietary Therapy in COVID-19: Exclusive Analysis and Prospects

• The international committee on taxonomy of viruses declared the "severe acute respiratory syndrome coronavirus 2" (SARS-CoV-2) as the new virus which has quickly transmitted from human to human and speed expeditiously across the globe causing Coronavirus Disease 2019 (COVID-19) pandemic. • At present, COVID-19 is treated using chloroquine, hydroxychloroquine, remdesivir, etc,; however, these drugs have demonstrated inadequate clinical response while causing serious adverse effects. This facilitates the study of inhibition of COVID-19 protease by traditional herbal plants and dietary therapy. • India has always been a rich reservoir of medicinal plants because of several agro-climatic zones. Indian herbal plants such as harsingar (night jasmine or parijat), giloy (moonseed plant or guduchi) and aloe vera (ghrit kumari) are particularly interesting in terms of their inhibition potentials. Chinese Herbal medicines (CHM) has been recommended and included in the interim guidelines for the treatment of COVID-19 as Chinese medicine is on the mainstream health care system in China. • The key functional food plants with immunomodulatory and antiviral; antibacterial; antifungal; anti-inflammatory; antioxidant; anticancer activity included liquorice (Glycyrrhiza glabra L.), garlic (Allium sativum L.), tea (Camellia sinensis [L.] Kuntez), ginger (Zingiber officinale Roscoe), turmeric (Curcuma longa L.) pomegranate (Punica granatum L.), black pepper (Pipper nigram L.) etc. • The present study explores the scientific data to provide exclusive analysis and establish the prospects of traditional medicine (TM) and dietary therapy (DT) in treatment and prevention of Coronavirus Disease 2019 (COVID-19) patients.

Methane production and estimation from livestock husbandry: A mechanistic understanding and emerging mitigation options.

Globally, livestock is an important contributor to methane (CH4) emissions. This paper reviewed the various CH4 measurement and estimation techniques and mitigation approaches for the livestock sector. Two approaches for enteric livestock CH4 emission estimation are the top-down and bottom-up. The combination of both could further improve our understanding of enteric CH4 emission and possible mitigation measures. We discuss three mitigation approaches: reducing emissions, avoiding emissions, and enhancing the removal of emissions from livestock. Dietary management, livestock management, and breeding management are viable reducing emissions pathways. Dietary manipulation is easily applicable and can bring an immediate response. Economic incentive policies can help the livestock farmers to opt for diet, breeding, and livestock management mitigation approaches. Carbon pricing creates a better option to achieve reduction targets in a given period. A combination of carbon pricing, feeding management, breeding management, and livestock management is more feasible and sustainable CH4 emissions mitigation strategy rather than a single approach.

Synthetic and antimicrobial studies on natural cyclopolypeptide from Caribbean marine-sponge and its analogs

Objective: A natural cyclic peptide, rolloamide, previously isolated from marine-sponge, was synthesized by coupling of tri and tetrapeptide units Boc­Phe-Pro- Val­OMe and Boc­Pro-Leu-Pro-Ile­OMe after proper deprotection at carboxyl and amino terminals followed by cyclization of linear heptapeptide segment. Design and Methodology: Solution phase technique was adopted for the synthesis of cycloheptapeptide. Required tri and tetrapeptide units were prepared by coupling of Boc-protected dipeptides viz. Boc­Phe-Pro­OH and Boc­Pro-Leu­OH with respective amino acid methyl ester hydrochloride Val-OMe.HCl and dipeptide methyl ester Pro-Ile-OMe. Cyclization of linear heptapeptide unit was done by p-nitrophenyl ester method. Similarly, two analogs of rolloamide were prepared by modification of tripeptide unit. The structures of synthesized cyclopeptide and its analogs were elucidated by spectral and elemental analysis. The newly synthesized peptide was subjected to antimicrobial screening and compared with biopotential of analogs. Results: Synthesis of cyclopeptide was accomplished with >84% yield utilizing diisopropylcarbodiimide (DIPC) as coupling agent. Newly synthesized peptide possessed promising activity against C. albicans and P. aeruginosa, K. pneumonia as compared to standard drugs, in addition to moderate activity against dermatophytes. Synthesized peptide analogs showed better antimicrobial potential against C. albicans and dermatophytes. Conclusions: Solution phase technique employing N,Ndiisopropylcarbodiimide (DIPC) and triethylamine (TEA) proved to be effective for the synthesis of natural cycloheptapeptide. N-methyl morpholine (NMM) was found to be a better base for cyclization of linear heptapeptide unit in comparison to TEA and pyridine. Promising antimicrobial potential was seen for newly synthesized cyclic peptide and its analogs.

Propagation of amorphous oxide nanowires via the VLS mechanism: growth kinetics.

This work reports the growth kinetics of amorphous nanowires (NWs) developed by the vapour-liquid-solid (VLS) mechanism. The model presented here incorporates all atomistic processes contributing to the growth of amorphous oxide NWs having diameters in the 5-100 nm range. The steady state growth condition has been described by balancing the key atomistic process steps. It is found that the 2D nano-catalyst liquid and NW solid (L-S) interface plays a central role in the kinetic analysis. The balance between the 2D Si layer crystallization and oxidation rate is quantitatively examined and compared with experimental values. The atomistic process dependencies of the NW growth rate, supersaturation (C/C 0), desolvation energy (Q D) barrier and NW diameter have been analyzed in detail. The model successfully predicts the reported NW growth rate to be in the range of 1-10 µm s-1. A novel seed/catalyst metal-based synthesis strategy for the preparation of amorphous silica NWs is reported. A nickel thin film on Si is used as a seed metal for the Au assisted VLS growth of silica NWs. The experimental results provide evidence of the creation of SiO under the given conditions followed by Si injection in the Au-Si nano-catalyst solution. The usage of seed metal was observed to reduce the growth temperature compared to the methods reported in the literature and obtain similar growth rates. The technique presented here holds promise for the synthesis of sub-100 nm diameter NWs.

Correction: Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells.

The authors report that there is a mistake in the representative picture of Fig. 4D (top row: PC3-miR1260b inh-0h) in the original version. The correct version of Fig. 4 with the original pictures for both PC3 miR-NC inh-0h and PC3-miR1260b inh-0h are provided below.

Climate change impact of livestock CH4 emission in India: Global temperature change potential (GTP) and surface temperature response.

Two climate metrics, Global surface Temperature Change Potential (GTP) and the Absolute GTP (AGTP) are used for studying the global surface temperature impact of CH4 emission from livestock in India. The impact on global surface temperature is estimated for 20 and 100 year time frames due to CH4 emission. The results show that the CH4 emission from livestock, worked out to 15.3 Tg in 2012. In terms of climate metrics GTP of livestock-related CH4 emission in India in 2012 were 1030 Tg CO2e (GTP20) and 62 Tg CO2e (GTP100) at the 20 and 100 year time horizon, respectively. The study also illustrates that livestock-related CH4 emissions in India can cause a surface temperature increase of up to 0.7mK and 0.036mK over the 20 and 100 year time periods, respectively. The surface temperature response to a year of Indian livestock emission peaks at 0.9mK in the year 2021 (9 years after the time of emission). The AGTP gives important information in terms of temperature change due to annual CH4 emissions, which is useful when comparing policies that address multiple gases.

MicroRNA-383 located in frequently deleted chromosomal locus 8p22 regulates CD44 in prostate cancer.

A major genomic alteration in prostate cancer (PCa) is frequent loss of chromosome (chr) 8p with a common region of loss of heterozygosity (LOH) at chr8p22 locus. Genomic studies implicate this locus in the initiation of clinically significant PCa and with progression to metastatic disease. However, the genes within this region have not been fully characterized to date. Here we demonstrate for the first time that a microRNA component of this region-miR-383-is frequently downregulated in prostate cancer, has a critical role in determining tumor-initiating potential and is involved in prostate cancer metastasis via direct regulation of CD44, a ubiquitous marker of PCa tumor-initiating cells (TICs)/stem cells. Expression analyses of miR-383 in PCa clinical tissues established that low miR-383 expression is associated with poor prognosis. Functional data suggest that miR-383 regulates PCa tumor-initiating/stem-like cells via CD44 regulation. Ectopic expression of miR-383 inhibited tumor-initiating capacity of CD44+ PCa cells. Also, 'anti-metastatic' effects of ectopic miR-383 expression were observed in a PCa experimental metastasis model. In view of our results, we propose that frequent loss of miR-383 at chr8p22 region leads to tumor initiation and prostate cancer metastasis. Thus, we have identified a novel finding that associates a long observed genomic alteration to PCa stemness and metastasis. Our data suggest that restoration of miR-383 expression may be an effective therapeutic modality against PCa. Importantly, we identified miR-383 as a novel PCa tissue diagnostic biomarker with a potential that outperforms that of serum PSA.

In vitro growth inhibitory efficacy of some target specific novel drug molecules against Theileria equi.

The in vitro growth inhibitory efficacies of five drug molecules against Theileria equi were evaluated in in vitro cultured parasites. A continuous microaerophilic stationary-phase culture (MASP) system was established for propagation of T. equi parasites. This in vitro culture system was used to assess the growth inhibitory effect of harmaline hydrochloride dihydrate (HHD), hexadecyltrimethylammonium bromide (HDTAB), hesparidin methyl chalcone (HMC), andrographolide and imidocarb dipropionate against T. equi. The 50% inhibitory concentration value of HHD, HDTAB, HMC, and imidocarb dipropionate for T. equi growth were 17.42 µM, 14.00 µM, 246.34 µM and 0.279 µM (equivalent to 0.139 µg/ml), respectively (P<0.05). The andrographolide was not effective in inhibiting in vitro growth of T. equi in the present study. Furthermore, the in vitro cytotoxicity of these five drugs was evaluated on horse PBMC. At 2000 µM concentration of HHD, HDTAB, HMC, andrographolide and imidocarb dipropionate were 8.34, 46.44, 58.53, 31.06, 15.14% cytotoxic on PBMC, respectively. Out of our four tested drug molecules, HHD was having low IC50 value along with least cytotoxicity, as compared to reference drug imidocarb dipropionate. The difference in IC50 value of HDTAB and HHD was significant, but HDTAB was moderately more cytotoxic on PBMC cell lines. HHD and HDTAB are selective inhibitor for heat shock protein 90 (Hsp90) and choline kinase pathway. It can be concluded that HHD and HDTAB are potential drug molecules against T. equi parasite by acting on Hsp90 and choline kinase pathway.

Genistein downregulates onco-miR-1260b and upregulates sFRP1 and Smad4 via demethylation and histone modification in prostate cancer cells.

BACKGROUND: Recently several microRNAs (miRNAs) have been found to be regulated by genistein in cancer cells. In this study, we focused on the gene regulatory effect of genistein on microRNA and its target genes in prostate cancer (PC). METHODS: Initially, we investigated the effect of genistein on prostate cancer cells and identified that the expression of miRNA-1260b was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA-1260b expression and prostate cancer patient outcomes. Two target genes (sFRP1 and Smad4) of miR-1260b were identified based on computer algorithm and 3'UTR luciferase assay was carried out to determine direct miRNA regulation of the genes. RESULTS: Genistein promoted apoptosis while inhibiting prostate cancer cell proliferation, invasion and TCF reporter activity in PC cells. MiR-1260b was highly expressed in prostate cancer tissues and significantly downregulated by genistein in PC cells. After knocking down miR-1260b, cell proliferation, invasion, migration and TCF reporter activity were decreased in PC cells. Western analysis and 3'UTR luciferase assay showed that the two target genes (sFRP1 and Smad4) were directly regulated by miR-1260b. The expression of sFRP1 and Smad4 was significantly decreased in prostate cancer tissues. Genistein also increased expression of these two genes via DNA demethylation and histone modifications. CONCLUSIONS: Our data suggest that genistein exerts its anti-tumour effect via downregulation of miR-1260b that targeted sRRP1 and Smad4 genes in prostate cancer cells. The expression of sFRP1 and Smad4 was also modulated by genistein via DNA methylation or histone modifications in PC cell lines.

Comparative evaluation of serum fluoride levels in patients with and without chronic abdominal pain.

BACKGROUND: Fluorosis ranks high among the major environmental health problems in India. Non-ulcer dyspeptic complaints are common in humans and it is a known fact that fluoride in drinking water, food and other items can cause these symptoms. METHODS: Fifty adult outpatients (mean age: 35.2±12.7 y) with chronic abdominal pain of unexplained origin were tested for their serum, urinary, and drinking water fluoride (F) concentrations. These concentrations were compared with those of 50 asymptomatic outpatients (mean age: 37.4±11.5 y) and analysed statistically. RESULTS: Serum F concentrations were higher than normal in 62% of the study group I and in 42% of the control group II with a mean of 0.065±0.03 ppm (range: 0.010-0.421) in the former and 0.023±0.028 ppm in the latter. Statistical analysis of the data by Student's t-test (unpaired) revealed a significant correlation (p<0.05) between chronic abdominal pain and elevated serum F. Urinary fluoride concentrations in group I were 0.87±1.67 (0.01-3.7) ppm. Seventy-three percent of the patients examined for urinary fluoride concentrations were having higher values than normal, whereas 27% patients had normal range urinary fluoride concentrations despite raised serum fluoride concentrations. CONCLUSIONS: In the cases of chronic pain abdomen, chronic fluoride ingestion from drinking water and other sources can be the cause and should be evaluated in patients in which other parameters are normal.

Genistein downregulates onco-miR-1260b and inhibits Wnt-signalling in renal cancer cells.

BACKGROUND: Wnt-signalling has an important role in renal cancer and it is modulated by genistein in other cancers. Recently, microRNAs (miRNAs) have emerged as new regulators of gene expression. Thus, we focused on miRNAs to examine the regulatory mechanism of genistein on the Wnt-signalling pathway in renal cell carcinoma (RCC). METHODS: Initially, we investigated the effect of genistein on Wnt-signalling (TOPflash reporter assay (TCF reporter assays)) in renal cancer cells, and using microarray identified candidate miRNAs whose expression was decreased by genistein. We performed functional analyses and investigated the relationship between miRNA expression and renal cancer patient outcomes. We also did 3'UTR luciferase assays to look at direct miRNA regulation of Wnt-signalling-related genes. RESULTS: Genistein promoted apoptosis while inhibiting RCC cell proliferation and invasion. Genistein also decreased TCF reporter activity in RCC cells. We found that miR-1260b was highly expressed and significantly downregulated by genistein in RCC cells. The expression of miR-1260b was significantly higher in renal cancer tissues compared with normal, and significantly related to overall shorter survival. In addition, miR-1260b promoted renal cancer cell proliferation and invasion in RCC cells. The 3'UTR luciferase activity of target genes (sFRP1, Dkk2, Smad4) was significantly decreased and their protein expression significantly upregulated in miR-1260b inhibitor-transfected renal cancer cells. CONCLUSION: Our data suggest that genistein inhibited Wnt-signalling by regulating miR-1260b expression in renal cancer cells.

microRNA-183 is an oncogene targeting Dkk-3 and SMAD4 in prostate cancer.

BACKGROUND: The purpose of this study was to identify prostate cancer (PC) oncogenic microRNAs (miRs) based on miR microarray and to investigate whether these oncogenic miRs may be useful as PC biomarkers. METHODS: Initially, we carried out miR microarray and real-time PCR using RWPE-1, PC-3, DU-145 and LNCaP cells. To investigate the function of miR-183, we used a miR-183 knockdown inhibitor in cell growth and wound-healing assays. We used several algorithms and confirmed that they are directly regulated by miR-183. RESULTS: We identified three potential oncogenic miRs (miR-146a, miR-183 and miR-767-5P). The expression of miR-183 in PC cells (PC-3, DU-145 and LNCaP) was upregulated compared with RWPE-1 cells. MiR-183 expression was also significantly higher in PC tissues compared with that in matched normal prostate tissues. Additionally, miR-183 expression was correlated with higher prostate-specific antigen, higher pT and shorter overall survival. MiR-183 knockdown decreased cell growth and motility in PC cells and significantly decreased prostate tumour growth in in vivo nude mice experiments. We identified Dkk-3 and SMAD4 as potential target genes of miR-183. CONCLUSION: Our data suggest that oncogenic miR-183 may be useful as a new PC biomarker and that inhibition of miR-183 expression may be therapeutically beneficial as a PC treatment.

Tumour suppressor microRNA-584 directly targets oncogene Rock-1 and decreases invasion ability in human clear cell renal cell carcinoma.

BACKGROUND: The purpose of this study was to identify new tumour suppressor microRNAs (miRs) in clear cell renal cell carcinoma (ccRCC), carry out functional analysis of their suppressive role and identify their specific target genes. METHODS: To explore suppressor miRs in RCC, miR microarray and real-time PCR were performed using HK-2 and A-498 cells. Cell viability, invasion and wound healing assays were carried out for functional analysis after miR transfection. To determine target genes of miR, we used messenger RNA (mRNA) microarray and target scan algorithms to identify target oncogenes. A 3'UTR luciferase assay was also performed. Protein expression of target genes in ccRCC tissues was confirmed by immunohistochemistry and was compared with miR-584 expression in ccRCC tissues. RESULTS: Expression of miR-584 in RCC (A-498 and 769-P) cells was downregulated compared with HK-2 cells. Transfection of miR-584 dramatically decreased cell motility. The ROCK-1 mRNA was inhibited by miR-584 and predicted to be target gene. The miR-584 decreased 3'UTR luciferase activity of ROCK-1 and ROCK-1 protein expression. Low expression of miR-584 in ccRCC tissues was correlated with high expression of ROCK-1 protein. The knockdown of ROCK-1 by siRNA inhibited cell motility. CONCLUSION: miR-584 is a new tumour suppressor miR in ccRCC and inhibits cell motility through downregulation of ROCK-1.

The functional significance of microRNA-145 in prostate cancer.

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that have important roles in numerous cellular processes. Recent studies have shown aberrant expression of miRNAs in prostate cancer tissues and cell lines. On the basis of miRNA microarray data, we found that miR-145 is significantly downregulated in prostate cancer. METHODS AND RESULTS: We investigated the expression and functional significance of miR-145 in prostate cancer. The expression of miR-145 was low in all the prostate cell lines tested (PC3, LNCaP and DU145) compared with the normal cell line, PWR-1E, and in cancerous regions of human prostate tissue when compared with the matched adjacent normal. Overexpression of miR-145 in PC3-transfected cells resulted in increased apoptosis and an increase in cells in the G2/M phase, as detected by flow cytometry. Investigation of the mechanisms of inactivation of miR-145 through epigenetic pathways revealed significant DNA methylation of the miR-145 promoter region in prostate cancer cell lines. Microarray analyses of miR-145-overexpressing PC3 cells showed upregulation of the pro-apoptotic gene TNFSF10, which was confirmed by real-time PCR and western analysis. CONCLUSION: One of the genes significantly upregulated by miR-145 overexpression is the proapoptotic gene TNFSF10. Therefore, modulation of miR-145 may be an important therapeutic approach for the management of prostate cancer.

Down-regulation of frizzled-7 expression decreases survival, invasion and metastatic capabilities of colon cancer cells.

BACKGROUND: The canonical Wnt signalling pathway is activated in most sporadic colorectal cancers (CRCs). We previously reported that FZD7 functions as a receptor for the canonical Wnt signalling pathway in colon cancer cells. METHODS AND RESULTS: In this study, we examined the function of FZD7 in survival, invasion and metastatic capabilities of colon cancer cells. FZD7_siRNA transfection decreased cell viability of HT-29 and HCT-116 colon cancer cells. Expression of c-Jun, phosphorylation of JNK and c-Jun, and activation of RhoA were suppressed after FZD7_siRNA transfection into HCT-116 cells. In vitro invasion activity and Wnt target gene expression were also reduced in HCT-116 cells transfected with FZD7_siRNA. Liver metastasis of stable FZD7_siRNA HCT-116 cell transfectants in scid mice was decreased to 40-50% compared to controls. The mRNA levels of FZD7 in 135 primary CRC tissues were examined by real-time PCR. FZD7 mRNA levels were significantly higher in stage II, III or IV tumours than in non-tumour tissues (P<0.005), and overall survival was shorter in those patients with higher FZD7 expression (P<0.001). CONCLUSION: These data suggest that FZD7 may be involved in enhancement of survival, invasion and metastatic capabilities of colon cancer cells through non-canonical Wnt signalling pathways as well as the canonical pathway.

Superoxide dismutase analog (Tempol: 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine 1-oxyl) treatment restores erectile function in diabetes-induced impotence.

We hypothesized that the administration of the superoxide dismutase (SOD) mimetic Tempol (4-hydroxy-2, 2, 6, 6-tetramethylpiperidine 1-oxyl) may reverse diabetes-induced erectile dysfunction. To test this hypothesis, reactive oxygen species-related genes (SOD1, SOD2, GP x 1, CAT, NOS2, NOS3) were tested, erectile functional studies and immunohistochemical analysis were carried out in diabetic rats treated with or without Tempol. Thirty Sprague-Dawley (3-4 months old) rats were divided into three groups (n=10 each), 20 with diabetes (diabetic control and Tempol treatment) and 10 healthy controls. At 12 weeks after the induction of diabetes by streptozotocin and Tempol treatment, all groups underwent in vivo cavernous nerve stimulation. Rat crura were harvested and the expression of antioxidative defense enzymes were examined by semi-quantitative reverse transcriptase PCR (RT-PCR). To confirm the RT-PCR results, we carried out immunohistochemistry (IHC) for catalase (CAT) and iNOS (NOS2). Nitration of tyrosine groups in proteins was also examined by IHC. Mean intracavernous pressure in the diabetic group was significantly lower than in the healthy controls (P <0.001) and was reversed by Tempol treatment (P <0.0108). NOS2 protein expression was significantly increased in diabetic animals compared with healthy controls and Tempol restored NOS2 protein level. Nitrotyrosine was also higher in diabetic animals and although Tempol treatment decreased its formation, it remained higher than that found in healthy controls. This study suggests that Tempol treatment increased erectile function through modulating oxidative stress-related genes in diabetic rats. This is the first report about the relationship between diabetes-induced erectile dysfunction and oxidative stress, and antioxidative therapy using the superoxide dismutase mimetic, Tempol, to restore erectile function.

miR-449a targets HDAC-1 and induces growth arrest in prostate cancer.

Histone deacetylases (HDACs) are frequently overexpressed in broad range of cancer types, where they alter cellular epigenetic programming to promote cell proliferation and survival. However, the mechanism by which HDACs become overexpressed in human cancers remains somewhat of a mystery. In this study, we investigated the expression and functional significance of miR-449a in prostate cancer cells. Using real-time PCR, we found that miR-449a is downregulated in prostate cancer tissues relative to patient-matched control tissue. Introduction of miR-449a into PC-3 prostate cancer cells resulted in cell-cycle arrest, apoptosis and a senescent-like phenotype. In silico analysis of 3'-UTR regions identified a number of genes involved in cell-cycle regulation as putative targets of miR-449a. Using a luciferase 3'-UTR reporter system, we established that HDAC-1 (histone deacetylase 1), a gene that is frequently overexpressed in many types of cancer, is a direct target of miR-449a. Further, our data indicate that miR-449a regulates cell growth and viability in part by repressing the expression of HDAC-1 in prostate cancer cells. Our findings provide new insight into the function of miRNA in regulating HDAC expression in normal versus cancerous tissue.