Vaginal Infections during Pregnancy Increase Breast Milk Microbiome Alpha Diversity and Alter Taxonomic Composition.

We previously reported that breast milk from women with (W) or without (WO) vaginal yeast infection during pregnancy differs in its immunological and antimicrobial properties, especially against pathogenic vaginal Candida sp.. Here, we investigated the differences in microbiota profiles of breast milk from these groups. Seventy-two breast milk samples were collected from lactating mothers (W, n=37; WO, n=35). The DNA of bacteria was extracted from each breast milk sample for microbiota profiling by 16S rRNA gene sequencing. Breast milk from the W-group exhibited higher alpha diversity than that from the WO-group across different taxonomic levels of class (P=0.015), order (P=0.011), family (P=0.020), and genus (P=0.030). Compositional differences between groups as determined via beta diversity showed marginal differences at taxonomic levels of phylum (P=0.087), family (P=0.064), and genus (P=0.067). The W-group showed higher abundances of families Moraxellaceae (P=0.010) and Xanthomonadaceae (P=0.008), and their genera Acinetobacter (P=0.015), Enhydrobacter (P=0.015), and Stenotrophomonas (P=0.007). Meanwhile, the WO-group showed higher abundances of genus Staphylococcus (P=0.046) and species Streptococcus infantis (P=0.025). This study shows that, although breast milk composition is affected by vaginal infection during pregnancy, this may not pose a threat to infant growth and development.

Breast milk from healthy women has higher anti-Candida properties than women with vaginal infections during pregnancy.

The aim of this study was to investigate the different immunological and antimicrobial properties of breast milk from women with (W) or without (WO) vaginal yeast infections during pregnancy in 85 lactating women (W, n = 43; WO, n = 42). Concentrations of IL-10, IgA, IgM, IgG, EGF, and TGF-α were similar in both groups. However, breast milk of women aged below 31 years old from the W-group showed higher concentration of EGF than the WO-group (p = 0.031). Breast milk from WO-group exhibited higher anti-Candida properties than W-group, both via growth inhibition and aggregation of yeast cells (p < 0.001). Correlation analysis showed that breast milk concentration of TGF-α positively correlated with concentrations of IL-10 (p = 0.001) and IgA (p = 0.021) in the W-group. Data from our present study shows that although breast milk from women with vaginal infections during pregnancy may not sufficiently hinder Candida growth, other immuno-modulatory bioactives may substitute for such a protective effect.

Sex-divergent expression of cytochrome P450 and SIRTUIN 1-7 proteins in toxicity evaluation of a benzimidazole-derived epigenetic modulator in mice.

Efforts in precision medicine to combat aberrant epigenome have led to the development of epigenetic targeting drugs. We have previously reported the capability of the BZD9L1 epigenetic modulator to impede colorectal tumour growth in vitro and in vivo through sirtuin (SIRT) inhibition. Although most benzimidazole derivatives are commonly less toxic, their effects on SIRTs and cytochrome P450 (CYP) regulations have not been explored alongside toxicity assessments. SIRTs are histone deacetylases that are crucial in maintaining metabolic homeostasis, whereas CYP is essential in drug metabolism. This study aims to determine the toxicology profile of BZD9L1 through oral acute and repeated dose toxicity evaluations, along with molecular analyses of SIRT, CYP and relevant toxicity markers through western blot and quantitative polymerase chain reaction (qPCR). BZD9L1 demonstrated no sign of acute toxicity at the limit dose (2000 mg/kg). The 28-day toxicity study highlighted the tolerability of repeated dose administration without adverse effects. BZD9L1 showed a sex-divergent regulation of hepatic SIRT1-7, CYP2A5 and CYP2D proteins. Furthermore, BZD9L1 did not induce the expression of organ injury proteins or alter the gene expression of cellular function indicators in mouse liver and kidneys, hence demonstrating, at least in part, the safety of BZD9L1 in short-term evaluations. The present study cautions for personalised strategies when employing benzimidazole-derived epigenetic therapeutics.

Probiotic Bifidobacterium lactis Probio-M8 treated and prevented acute RTI, reduced antibiotic use and hospital stay in hospitalized young children: a randomized, double-blind, placebo-controlled study.

PURPOSE: The development of probiotics has seen tremendous growth over the years, with health benefits ranging from gut health to respiratory. We thus aimed to investigate the effects of probiotic Bifidobacterium lactis Probio-M8 (2 × 1010 log CFU/day) against acute respiratory tract infections (RTI), use of antibiotics, hospitalization period and elucidate the possible mechanisms of action in hospitalized young children. METHOD: A prospective, randomized, double-blind and placebo-controlled study was performed in RTI-hospitalized children. Patients were randomized to either the probiotic (n = 60, mean age 13.81 ± 0.90 months) or placebo (n = 60, mean age 12.11 ± 0.73 months) which were administered upon admission, continued during hospitalization and 4-week post-discharged. RTI and gut health parameters were assessed at these time points using validated questionnaires while concentrations of inflammatory cytokines were assessed via oral swabs. RESULTS: Probio-M8 reduced the duration of nasal, pharyngeal and general flu-like symptoms compared to the placebo during the hospitalization period and 4-week post-discharged (P < 0.05) as compared to the placebo, with a more prevalent effect against lower respiratory tract infections (LRTI). Probio-M8 reduced prescription of antibiotic (P = 0.037), prevented new prescription of antibiotic in non-prescribed patients (P = 0.024) and reduced hospitalization period in antibiotic-prescribed patients (P = 0.004) as compared to the placebo. Oral cytokine levels of TNF-α decreased in the Probio-M8 group (P = 0.001) accompanied by increased in IL-10 (P = 0.018) over 4-week post-discharged, while the placebo group did not exhibit such an effect. Increased IL-10 in the Probio-M8 group was correlated with decreased body ache (r = - 0.296, P = 0.001), headache (r = - 0.295, P = 0.001) and pain during swallow (r = - 0.235, P = 0.010). CONCLUSION: Data from our present study show that B. lactis Probio-M8 could be a potential natural and non-drug strategy for the management of RTI in young children in a safe manner. CLINICAL TRIAL REGISTRATION: Clinical studies (Approval No. USM/JEPeM/19030177) were registered at ClinicalTrials.gov (Identifier No. NCT04122495) on September 30, 2019.

Methadone, Buprenorphine, and Clonidine Attenuate Mitragynine Withdrawal in Rats.

Background: Kratom or Mitragyna speciosa Korth has been widely used to relieve the severity of opioid withdrawal in natural settings. However, several studies have reported that kratom may by itself cause dependence following chronic consumption. Yet, there is currently no formal treatment for kratom dependence. Mitragynine, is the major psychoactive alkaloid in kratom. Chronic mitragynine treatment can cause addiction-like symptoms in rodent models including withdrawal behaviour. In this study we assessed whether the prescription drugs, methadone, buprenorphine and clonidine, could mitigate mitragynine withdrawal effects. In order to assess treatment safety, we also evaluated hematological, biochemical and histopathological treatment effects. Methods: We induced mitragynine withdrawal behaviour in a chronic treatment paradigm in rats. Methadone (1.0 mg/kg), buprenorphine (0.8 mg/kg) and clonidine (0.1 mg/kg) were i.p. administered over four days during mitragynine withdrawal. These treatments were stopped and withdrawal sign assessment continued. Thereafter, toxicological profiles of the treatments were evaluated in the blood and in organs. Results: Chronic mitragynine treatment caused significant withdrawal behaviour lasting at least 5 days. Methadone, buprenorphine, as well as clonidine treatments significantly attenuated these withdrawal signs. No major effects on blood or organ toxicity were observed. Conclusion: These data suggest that the already available prescription medications methadone, buprenorphine, and clonidine are capable to alleviate mitragynine withdrawal signs rats. This may suggest them as treatment options also for problematic mitragynine/kratom use in humans.

Mitragynine Attenuates Morphine Withdrawal Effects in Rats-A Comparison With Methadone and Buprenorphine.

BACKGROUND: Opiate addiction is a major health problem in many countries. A crucial component of the medical treatment is the management of highly aversive opiate withdrawal signs, which may otherwise lead to resumption of drug taking. In a medication-assisted treatment (MAT), methadone and buprenorphine have been implemented as substitution drugs. Despite MAT effectiveness, there are still limitations and side effects of using methadone and buprenorphine. Thus, other alternative therapies with less side effects, overdosing, and co-morbidities are desired. One of the potential pharmacotherapies may involve kratom's major indole alkaloid, mitragynine, since kratom (Mitragyna speciosa Korth.) preparations have been reported to alleviate opiate withdrawal signs in self-treatment in Malaysian opiate addicts. METHODS: Based on the morphine withdrawal model, rats were morphine treated with increasing doses from 10 to 50 mg/kg twice daily over a period of 6 days. The treatment was discontinued on day 7 in order to induce a spontaneous morphine abstinence. The withdrawal signs were measured daily after 24 h of the last morphine administration over a period of 28 abstinence days. In rats that developed withdrawal signs, a drug replacement treatment was given using mitragynine, methadone, or buprenorphine and the global withdrawal score was evaluated. RESULTS: The morphine withdrawal model induced profound withdrawal signs for 16 days. Mitragynine (5-30 mg/kg; i.p.) was able to attenuate acute withdrawal signs in morphine dependent rats. On the other hand, smaller doses of methadone (0.5-2 mg/kg; i.p.) and buprenorphine (0.4-1.6 mg/kg; i.p.) were necessary to mitigate these effects. CONCLUSIONS: These data suggest that mitragynine may be a potential drug candidate for opiate withdrawal treatment.

MicroRNA deregulation and cancer and medicinal plants as microRNA regulator

MircroRNAs (miRNAs) are short non-coding RNAs with a length of approximately 20-22 nucleotides, which interact with their target mRNAs at 3'-untranslated region by partial pairing. The miRNA- mRNA interaction leads to induction of mRNA degradation and eventually translational inhibition. Thus, miRNAs play an important role in virtually all cellular processes, especially differentiation, proliferation, migration, and apoptosis. The deregulation of miRNAs may lead to serious diseases including cancer. There is mounting evidence demonstrating the participation of miRNA regulation during carcinogenesis. In this review, we discuss an updated miRNA biogenesis, mechanisms involved in their deregulation, and their role in cancer development. This review also summarizes updated information on potential medicinal plants which regulate miRNA expression as a promising molecular miRNA therapeutic approach for cancers.

Methadone hydrochloride and leukemia cells: Effects on cell viability, DNA fragmentation and apoptotic proteins expression level.

Leukemia is a type of blood cancer where abnormal and immature leucocytes are produced in the bone marrow. Methadone hydrochloride is a man-made drug that is commonly used in the maintenance treatment for drug addiction. The objective of this research was to determine the cytotoxic activity and apoptotic effects of methadone hydrochloride treatment towards two leukemia cell lines which are CCRF-CEM and HL-60. CCRF-CEM and HL-60 cells were treated with methadone hydrochloride for 24 and 48 hours to determine the cytotoxic activity. IC50 at 24 hours obtained for CCRF-CEM was 121.6µmol/L while IC50 for HL-60 cells was 97.18µmol/L. Result obtained from DNA fragmentation assay showed no characteristic DNA ladder pattern in CCRF-CEM leukemia cells treated with methadone hydrochloride. Characteristics DNA ladder pattern was observed in methadone hydrochloride treated HL-60 cells. Formation of comets was seen in methadone hydrochloride treated CCRF-CEM and HL-60 cells with varying degree of DNA damage. The comets formed by methadone hydrochloride treated HL-60 cells were more prominent as compared to methadone-treated CCRF-CEM cells. The expression of apoptotic-related proteins in methadone-treated CCRF-CEM and HL-60 cells were checked by incubating the cell lysate with Raybio® Human Apoptosis Antibody Array. Significant alterations in expression level of apoptosis-related proteins in methadone hydrochloride treated CCRF-CEM cells were found involving upregulation of caspase-8 expression and downregulation of survivin expression. Methadone hydrochloride induced apoptosis in HL-60 cells involved upregulation of Bid and caspase-8 expression and downregulation of Bcl-2, p21 and survivin expression.

Lactobacillus plantarum DR7 Reduces Cholesterol via Phosphorylation of AMPK That Down-regulated the mRNA Expression of HMG-CoA Reductase.

Hypercholesterolemia is one of the primary risk factors for cardiovascular diseases. The use of lactobacilli probiotics to reduce blood cholesterol levels have been extensively reported. However, more information is needed to evaluate the possible mechanisms involved and to identify possible targets for further therapeutic development. In this study, strains of lactobacilli were screened based on the ability to assimilate cholesterol, and prevention of cholesterol accumulation in hepatic (HepG2) and intestinal (HT-29) cells. Cell free supernatant (CFS) from Lactobacillus plantarum DR7 showed a higher ability to assimilate cholesterol, reduction in cholesterol accumulation in both HepG2 and HT-29 cells, accompanied by reduced mRNA expression of HMG-CoA reductase (HMGCR) in HepG2 (p<0.05), compared to other lactobacilli. The reduction of HMGCR expression was also diminished in the presence of an AMPK inhibitor (Compound C), suggesting that L. plantarum DR7 exerted its effect via the AMPK pathway, typically via the phosphorylation of AMPK instead of the AMPK mRNA expression in HepG2 (p<0.05). Altogether, our present study illustrated that lactobacilli could exert cholesterol lowering properties along the AMPK pathway, specifically via phosphorylation of AMPK that led to reduced expression of HMGCR.

Antioxidant activity of methanol extracts of different parts of Lantana camara

<p><b>OBJECTIVE</b>To investigate the antioxidant activity of methanolic extracts of Lantana camara (L. camara) various parts and the determination of their total phenolics content.</p><p><b>METHODS</b>The extract was screened for possible antioxidant activities by free radical scavenging activity(DPPH), xanthine oxidase inhibition activity and Griess-Ilosvay method.</p><p><b>RESULTS</b>The results showed that all the plant parts possessed antioxidant properties including radical scavenging, xanthine oxidase inhibition and nitrites scavenging activities. The antioxidative activities were correlated with the total phenol. The leaves extract of L. camara was more effective than that of other parts.</p><p><b>CONCLUSIONS</b>This study suggests that L. camara extracts exhibit great potential for antioxidant activity and may be useful for their nutritional and medicinal functions.</p>

Pharmacological screening of methanolic extract of Ixora species

<p><b>OBJECTIVE</b>To investigate the antimicrobial activity of methanolic extracts of different parts of Ixora species.</p><p><b>METHODS</b>Antimicrobial activity was carried out using disc diffusion assay against fungi, gram-positive and gram-negative bacteria.</p><p><b>RESULTS</b>All methanolic extracts of different parts of Ixora species showed a broad-spectrum of antibacterial and antiyeast activities, which inhibited the growth of at least one bacterium or yeast. There was no remarkable difference between different Ixora species observed in this study.</p><p><b>CONCLUSIONS</b>The significant antimicrobial activity shown by this Ixora species suggests its potential against infections caused by pathogens. The extract may be developed as an antimicrobial agent.</p>

Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and Vero cell lines

<p><b>OBJECTIVE</b>To investigate the cytotoxic effect of Elaeis guineensis methanol extract on MCF-7 and Vero cell.</p><p><b>METHODS</b>In vitro cytotoxicity was evaluated in by MTT assay. Cell morphological changes were observed by using light microscope.</p><p><b>RESULTS</b>The MTT assay indicated that methanol extract of the plant exhibited significant cytotoxic effects on MCF-7. Morphological alteration of the cell lines after exposure with Elaeis guineensis extract were observed under phase contrast microscope in the dose dependent manner.</p><p><b>CONCLUSIONS</b>The results suggest the probable use of the Elaeis guineensis methanol extract in preparing recipes for cancer-related ailments. Further studies on isolation of metabolites and their in vivo cytotoxicity are under investigation.</p>

Antioxidant and antibacterial activity of different parts of Leucas aspera

<p><b>OBJECTIVE</b>To evaluate antioxidant, antimicrobial and cytotoxic activity of different parts (root, flower, leaf and stem) of Leucas aspera (L. aspera) (Labiatae).</p><p><b>METHODS</b>Different parts of L. aspera were extracted with 80% (v/v) methanol. The methanol extracts were subjected to antioxidant, antimicrobial and brine shrimp lethality assay.</p><p><b>RESULTS</b>All the extracts showed moderate to potent antioxidant activity, among which the root extract demonstrated the strongest antioxidant activity with the IC50 value of 6.552 µg/mL. Methanol extract of root possessed antioxidant activity near the range of vitamin E and thus could be a potential rich source of natural antioxidant. In case of antimicrobial screening, crude extracts of root, flower, leaf and stem showed notable antibacterial activity against tested microorganisms. The root extract showed the highest mean zone of inhibition ranging from 9.0-11.0 mm against tested microorganisms, at a concentration of 100 mg/mL. In the brine shrimp lethality bioassay, it was evident that the methanol root extract did not show significant toxicity. The LC50 value for 12 h and 24 h observation was 2.890 mg/mL and 1.417 mg/mL, respectively.</p><p><b>CONCLUSIONS</b>The present finding suggests that the methanol root extract of L. aspera could be developed as pharmaceutical products.</p>

Cytotoxicity of methanol extracts of Elaeis guineensis on MCF-7 and Vero cell lines

To investigate the cytotoxic effect of Elaeis guineensis methanol extract on MCF-7 and Vero cell. Methods: In vitro cytotoxicity was evaluated in by MTT assay. Cell morphological changes were observed by using light microscope. Results: The MTT assay indicated that methanol extract of the plant exhibited significant cytotoxic effects on MCF-7. Morphological alteration of the cell lines after exposure with Elaeis guineensis extract were observed under phase contrast microscope in the dose dependent manner. Conclusions: The results suggest the probable use of the Elaeis guineensis methanol extract in preparing recipes for cancer-related ailments. Further studies on isolation of metabolites and their in vivo cytotoxicity are under investigation.

Isolation and identification of salmonella from curry samples and its sensitivity to commercial antibiotics and aqueous extracts of Camelia sinensis (L.) and Trachyspermum ammi (L.).

OBJECTIVE: To isolate Salmonella from curry samples and to evaluate the drug sensitivity of the food-borne Salmonella and its susceptibility to specific plant extracts. METHODS: Salmonella was isolated from the curry samples by standard microbiological methods and was confirmed by biochemical tests. The antibiotic susceptibility test was conducted by disc diffusion method using commercially available antibiotics such as ampicillin, tetracycline, chloramphenicol, kanamycin, and penicillin. In addition, the susceptibility of the food-borne Salmonella was also evaluated against the aqueous extracts of Camelia sinensis (L.) Theaceae (tea leaves) and the Trachyspermum ammi (L.) Apiaceae ( ajwain or omum seeds). RESULTS: Out of fifty curry samples, only seven samples were identified to have Salmonella contamination. The Salmonella isolates showed a significant drug resistance pattern except for kanamycin. The plant extracts showed a considerable antibacterial activity against the isolates, indicating the presence of antimicrobial principle which can be exploited after complete pharmacological investigations. CONCLUSIONS: The present study demonstrates the occurrence of Salmonella in the curry samples, and shows significant drug resistance against most of the commercially available antibiotics, except kanamycin. Antimicrobial effect of the plant extracts against the food-bone Salmonella suggests that dietary including medicinal herbs would be one strategy to manage food borne pathogens.

Epidemiology of Helicobacter pylori among multiracial community in Northern Peninsular, Malaysia: effect of age across race and gender

OBJECTIVE@#To study the epidemiology of Helicobacter pylori (H. pylori) infection according to age group.@*METHODS@#H. pylori infection data among 1 965 consecutive patients referred to the Endoscopy Unit collected at Sungai Petani Hospital for oesophagogastro-duodenoscopy (OGD). The patients were divided into 9 age groups (10-19, 20-29, 30-39, 40-49, 50-59, 60-69, 70-79, 80-89 and 90-99 years). In addition these groups were further divided into three minor group namely young adults (10-39), older adults (40-69) and geriatric groups (70-99).@*RESULTS@#Overall prevalence of infection of H. pylori was analyzed and found that the prevalence increase with age (P<0.05). When the patients divided by ethnic and gender group with age, prevalence rate among young adults and older adults significantly higher (P<0.05) compared to geriatric groups across all races and gender (P<0.05). Furthermore, significantly higher number of males were infected compared to female (P<0.05) but such trend was only observed among older adult groups. In addition, there is a significant differences in H. pylori infection prevalence rates among ethnic groups (highest in Indians adults, followed Chinese and low in Malays, P<0.05).@*CONCLUSIONS@#The overall prevalence of H. pylori did increase with age group across ethnicity and gender, in Northern Peninsular Malaysia.

Antioxidant activity and phytochemical screening of the methanol extracts of Euphorbia hirta L

OBJECTIVE@#To assess antioxidant activities of different parts of Euphorbia hirta (E. hirta), and to search for new sources of safe and inexpensive antioxidants.@*METHODS@#Samples of leaves, stems, flowers and roots from E. hirta were tested for total phenolic content, and flavonoids content and in vitro antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH) assay and reducing power was measured using cyanoferrate method.@*RESULTS@#The leaves extract exhibited a maximum DPPH scavenging activity of (72.96±0.78)% followed by the flowers, roots and stems whose scavenging activities were (52.45±0.66)%, (48.59±0.97)%, and (44.42±0.94)%, respectively. The standard butylated hydroxytoluene (BHT) was (75.13±0.75)%. The IC(50) for leaves, flowers, roots, stems and BHT were 0.803, 0.972, 0.989, 1.358 and 0.794 mg/mL, respectively. The reducing power of the leaves extract was comparable with that of ascorbic acid and found to be dose dependent. Leaves extract had the highest total phenolic content [(206.17±1.95) mg GAE/g], followed by flowers, roots and stems extracts which were (117.08±3.10) mg GAE/g, (83.15±1.19) mg GAE/g, and (65.70±1.72) mg GAE/g, respectively. On the other hand, total flavonoids content also from leave had the highest value [(37.970±0.003) mg CEQ/g], followed by flowers, roots and stems extracts which were (35.200±0.002) mg CEQ/g, (24.350±0.006) mg CEQ/g, and (24.120±0.004) mg CEQ/g, respectively. HPTLC bioautography analysis of phenolic and antioxidant substance revealed phenolic compounds. Phytochemical screening of E. hirta leaf extract revealed the presence of reducing sugars, terpenoids, alkaloids, steroids, tannins, flavanoids and phenolic compounds.@*CONCLUSIONS@#These results suggeste that E. hirta have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidant agents, which can be used to treat various oxidative stress-related diseases.

A transmission electron microscopy study of the diversity of Candida albicans cells induced by Euphorbia hirta L. leaf extract in vitro

<p><b>OBJECTIVE</b>To determine the major changes in the microstructure of Candida albicans (C. albicans) after treatment with Euphorbia hirta (E. hirta) L. leaf extract.</p><p><b>METHODS</b>Transmission electron microscopy was used to study the ultrastructural changes caused by E. hirta extract on C. albicans cells at various exposure time.</p><p><b>RESULTS</b>It was found that the main abnormalities were the alterations in morphology, lysis and complete collapse of the yeast cells after 36 h of exposure to the extract. Whereas the control cultures showed a typical morphology of Candida with a uniform central density, typically structured nucleus, and a cytoplasm with several elements of endomembrane system and enveloped by a regular, intact cell wall.</p><p><b>CONCLUSIONS</b>The significant antifungal activity shown by this methanol extract of E. hirta L. suggests its potential against infections caused by C. albicans. The extract may be developed as an anticandidal agent.</p>

In vivo toxicity study of Lantana camara

<p><b>OBJECTIVE</b>To investigate the toxicity of methanol extract of various parts (Root, Stem, Leaf, Flower and Fruit) of Lantana camara (L. Camara) in Artemia salina.</p><p><b>METHODS</b>The methanol extracts of L. camara were tested for in vivo brine shrimp lethality assay.</p><p><b>RESULTS</b>All the tested extract exhibited very low toxicity on brine shrimp larva. The results showed that the root extract was the most toxic part of L. camara and may have potential as anticancer agent.</p><p><b>CONCLUSIONS</b>Methanolic extract of L. camara is relatively safe on short-term exposure.</p>