A Scoping Review on Hepatoprotective Mechanism of Herbal Preparations through Gut Microbiota Modulation.

Liver diseases cause millions of deaths globally. Current treatments are often limited in effectiveness and availability, driving the search for alternatives. Herbal preparations offer potential hepatoprotective properties. Disrupted gut microbiota is linked to liver disorders. This scoping review aims to explore the effects of herbal preparations on hepatoprotective mechanisms, particularly in the context of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), and hepatic steatosis, with a focus on gut microbiota modulation. A systematic search was performed using predetermined keywords in four electronic databases (PubMed, Scopus, EMBASE, and Web of Science). A total of 55 studies were included for descriptive analysis, covering study characteristics such as disease model, dietary model, animal model, intervention details, comparators, and study outcomes. The findings of this review suggest that the hepatoprotective effects of herbal preparations are closely related to their interactions with the gut microbiota. The hepatoprotective mechanisms of herbal preparations are shown through their effects on the gut microbiota composition, intestinal barrier, and microbial metabolites, which resulted in decreased serum levels of liver enzymes and lipids, improved liver pathology, inhibition of hepatic fatty acid accumulation, suppression of inflammation and oxidative stress, reduced insulin resistance, and altered bile acid metabolism.

Oxidative Stress and DNA Damage Effect of Dioscorea hispida Dennst. on Placental Tissues of Rats.

Dioscorea hispida Dennst. locally known as "ubi gadung" has been used as a traditional remedy and source of carbohydrate among Malaysians. To assess the effect of Dioscorea hispida aqueous extract (DHAE) on the production of reactive oxygen species (ROS) and their effects on DNA damage in Sprague Dawley rat's placental tissues, pregnant rats were randomly divided into four groups. The animals were orally treated with distilled water (negative control) and three different concentrations of DHAE (250, 500 and 1000 mg/kg body weight (BW)) from gestation day 6 until 20. The oxidative stress in placental tissues was evaluated at day 21 by measuring the level of ROS, superoxide dismutase (SOD) and lipid peroxidation biomarker, malondialdehyde (MDA) while comet assay was used for DNA damage. There was no significant production of ROS and SOD activities in all groups. Significant changes were observed in the MDA level at 1000 mg/kg BW DHAE. Comet assay revealed a significant increase (p < 0.05) of DNA damage on animals treated with 250 and 500 mg/kg BW DHAE but not at the highest concentration. It was postulated that the placental cells could have undergone necrosis which destroys all components including DNA. This occurrence simultaneously reduces the levels of DNA damage which can be represented by lower level of tail moments. This finding correlates with our histopathological examination where necrotic cells of spongiotrophoblast were observed in the basal zone of placental tissue. The high amount of hydrogen cyanide and other compounds in 1000 mg/kg BW DHAE could elevate the lipid peroxidation and directly induce cell necrosis which requires further investigation.

Natural dead sea salt and retrospective dosimetry.

Accidents resulting in widespread dispersal of radioactive materials have given rise to a need for materials that are convenient in allowing individual dose assessment. The present study examines natural Dead Sea salt adopted as a model thermoluminescence dosimetry system. Samples were prepared in two different forms, loose-raw and loose-ground, subsequently exposed to 60Co gamma-rays, delivering doses in the range 2-10 Gy. Key thermoluminescence (TL) properties were examined, including glow curves, dose response, sensitivity, reproducibility and fading. Glow curves shapes were found to be independent of given dose, prominent TL peaks for the raw and ground samples appearing in the temperature ranges 361-385 ºC and 366-401 ºC, respectively. The deconvolution of glow curves has been undertaken using GlowFit, resulting in ten overlapping first-order kinetic glow peaks. For both sample forms, the integrated TL yield displays linearity of response with dose, the loose-raw salt showing some 2.5 × the sensitivity of the ground salt. The samples showed similar degrees of fading, with respective residual signals 28 days post-irradiation of 66% and 62% for the ground and raw forms respectively; conversely, confronted by light-induced fading the respective signal losses were 62% and 80%. The effective atomic number of the Dead Sea salt of 16.3 is comparable to that of TLD-200 (Zeff 16.3), suitable as an environmental radiation monitor in accident situations but requiring careful calibration in the reconstruction of soft tissue dose (soft tissue Zeff 7.2). Sample luminescence studies were carried out via Raman and Photoluminescence spectroscopy as well as X-ray diffraction, ionizing radiation dependent variation in lattice structure being found to influence TL response.

Ameliorative Effects of Aquilaria malaccensis Leaves Aqueous Extract on Reproductive Toxicity Induced by Cyclophosphamide in Male Rats.

BACKGROUND: Cyclophosphamide (CP) is a widely used anti-neoplastic and immunosuppressive agent that is associated with adverse side effects including reproductive toxicity. Aquilaria malaccensis (AM) is a traditional medicinal plant which was reported to exhibit high anti-oxidant and free radical scavenging properties. The present study was aimed to evaluate the protective effects of AM leaves extract on sperm quality following toxic exposure to CP. METHODS: Forty-eight male Sprague Dawley rats were allocated into eight groups of six rats (n = 6): control, CP only (200 mg kg-1), AM only (100 mg kg-1, 300 mg kg-1 and 500 mg kg-1) and CP + AM (100 mg kg-1, 300 mg kg-1 and 500 mg kg-1). Animals were sacrificed after 63 days of treatment and the sperm from the caudal epididymis was taken for sperm analysis. RESULTS: The body and the reproductive organs weight, sperm count and motility did not differ between CP and other groups (P > 0.05). A significant increase (P < 0.05) in percentage of the dead and abnormal sperm were seen in the CP alone treated group compared to the control group. Co-administration of AM to the CP exposed rats significantly reduced the (P < 0.05) percentage of abnormal sperm as compared to the CP only group. CONCLUSION: Overall, the present results represent the potential of AM to protect against CP induced reproductive toxicity.

The effect of freeze-dried Carica papaya leaf juice treatment on NS1 and viremia levels in dengue fever mice model.

BACKGROUND: Carica papaya leaf juice (CPLJ) was well known for its thrombocytosis activity in rodents and dengue patients. However, the effect of CPLJ treatment on other parameters that could contribute to dengue pathogenesis such as nonstructural protein 1 (NS1) production and viremia level have never been highlighted in any clinical and in vivo studies. The aim of this study is to investigate the effect of freeze-dried CPLJ treatment on NS1 and viremia levels of dengue fever mouse model. METHODS: The dengue infection in mouse model was established by inoculation of non-mouse adapted New Guinea C strain dengue virus (DEN-2) in AG129 mice. The freeze-dried CPLJ compounds were identified by Ultra-High Performance Liquid Chromatography High Resolution Accurate Mass Spectrometry analysis. The infected AG129 mice were orally treated with 500 mg/kg/day and 1000 mg/kg/day of freeze-dried CPLJ, starting on day 1 post infection for 3 consecutive days. The blood samples were collected from submandibular vein for plasma NS1 assay and quantitation of viral RNA level by quantitative reverse transcription PCR. RESULTS: The AG129 mice infected with dengue virus showed marked increase in the production of plasma NS1, which was detectable on day 1 post infection, peaked on day 3 post-infection and started to decline from day 5 post infection. The infection also caused splenomegaly. Twenty-four compounds were identified in the freeze-dried CPLJ. Oral treatment with 500 mg/kg/day and 1000 mg/kg/day of freeze-dried CPLJ did not affect the plasma NS1 and dengue viral RNA levels. However, the morbidity level of infected AG129 mice were slightly decreased when treated with freeze-dried CPLJ. CONCLUSION: Oral treatment of 500 mg/kg/day and 1000 mg/kg/day of freeze-dried CPLJ at the onset of viremia did not affect the plasma NS1 and viral RNA levels in AG129 mice infected with non-mouse adapted New Guinea C strain dengue virus.

The solar end game: bibliometric analysis, research and development evolution, and patent activity of hybrid photovoltaic/thermal-phase change material.

Solar photovoltaic-thermal hybrid with phase change material (PVT-PCM) emerges as an intelligent game changer to stimulate the clean, reliable, and affordable renewable energy technology. This PVT-PCM technology can be manipulated into generating both electricity and thermal energy that feature its practicality for residential and industrial applications. Hybridized of PCM into PVT design adds value to existing architecture with its capability to store excess heat that can be used during insufficient solar irradiation. Present work gives overview of the PVT-PCM system on technology innovation toward commercialization (viz, solar end game) subjected to bibliometric analysis, research and development evolution, and patent activity. A consolidation of these review articles was decluttered to focus on the performance and efficiency of PVT-PCM technology based on the fact that commercialization is ready once the technology is completed and qualified (at technology readiness level, TRL: 8). Economic review was conducted to understand the feasibility of the existing solar technologies and how it affects the PVT-PCM market price. Based on the contemporary findings, promising performance of PVT-PCM technology has underpinned its feasibility and technology readiness. China has predominant local and international framework and expected to be the PVT-PCM technology trendsetter in the next years through its strong international collaborative projects and pioneer in PVT-PCM patent filing. This present work underscores the solar end-game strategy and recommendation to create a path forward to achieve clean energy transition. Though, as to the date of submission of this article, no industry  has found to manufacture/sell this hybrid technology in the market.

Repeated dose 28-days oral toxicity study of Carica papaya L. leaf extract in Sprague Dawley rats.

Carica papaya L. leaves have been used in ethnomedicine for the treatment of fevers and cancers. Despite its benefits, very few studies on their potential toxicity have been described. The aim of the present study was to characterize the chemical composition of the leaf extract from 'Sekaki' C. papaya cultivar by UPLC-TripleTOF-ESI-MS and to investigate the sub-acute oral toxicity in Sprague Dawley rats at doses of 0.01, 0.14 and 2 g/kg by examining the general behavior, clinical signs, hematological parameters, serum biochemistry and histopathology changes. A total of twelve compounds consisting of one piperidine alkaloid, two organic acids, six malic acid derivatives, and four flavonol glycosides were characterized or tentatively identified in the C. papaya leaf extract. In the sub-acute study, the C. papaya extract did not cause mortality nor were treatment-related changes in body weight, food intake, water level, and hematological parameters observed between treatment and control groups. Some biochemical parameters such as the total protein, HDL-cholesterol, AST, ALT and ALP were elevated in a non-dose dependent manner. Histopathological examination of all organs including liver did not reveal morphological alteration. Other parameters showed non-significant differences between treatment and control groups. The present results suggest that C. papaya leaf extract at a dose up to fourteen times the levels employed in practical use in traditional medicine in Malaysia could be considered safe as a medicinal agent.

Phytochemical and In Vitro Genotoxicity Studies of Standardized Ficus deltoidea var. kunstleri Aqueous Extract.

The present study was carried out to assess the genotoxicity potential of Ficus deltoidea var. kunstleri aqueous extract (FDAE) using standard in vitro assays. The DNA damage of V79B cells was measured using the alkaline comet assay treated at 0.1 mg/mL (IC10) and 0.3 mg/mL (IC25) of FDAE together with positive and negative controls. For in vitro micronucleus assay, the V79B cells were treated with FDAE at five different concentrations (5, 2.5, 1.25, 0.625, and 0.3125 mg/mL) with and without S9 mixture. The bacteria reverse mutation assay of FDAE was performed on Salmonella typhimurium strains TA98, 100, 1535, 1537, and Escherichia coli strain WP2uvrA using pre-incubation method in the presence or in the absence of an extrinsic metabolic system (S9 mixture). FDAE at 0.1 and 0.3 mg/mL significantly increased DNA damage in both comet tail and tail moment (p < 0.05). No significant changes were detected in the number of micronucleated cell when compared to control. Tested at the doses up to 5000 µg/plate, the FDAE did not increase the number of revertant colonies for all strains. In conclusion, further investigation needs to be conducted in animal model to confirm the non-genotoxicity activities of FDAE.

Longitudinal Vitamin D Deficiency Among Malaysian Pregnant Women and Its Correlation With Neonatal Serum 25-Hydroxyvitamin D Levels.

Objective: This study aimed to investigate the longitudinal relationship between maternal vitamin D concentrations during pregnancy and neonatal vitamin D concentrations at birth. Materials and Methods: A prospective cohort of 236 healthy pregnant women from various ethnicity in early pregnancy (≤20 weeks of pregnancy) was followed at late pregnancy (28-40 weeks of pregnancy) and birth. Maternal serum 25-hydroxyvitamin D (25(OH)D) was assessed at early pregnancy (baseline) and late pregnancy, while neonatal cord serum 25(OH)D at birth. General estimating equations (GEE) were used to analyze the longitudinal association of maternal serum 25(OH)D levels during pregnancy and neonatal cord serum 25(OH)D levels at birth with adjusting for the time exposure, maternal weight gain, ethnicity, and skin type. Results: The results showed that the prevalence of vitamin D deficiency (25(OH)D <50 nmol/L) was at 89.9, 92.2, and 96.1% in early, late pregnancy and in neonatal cord serum, respectively. The GEE analysis showed a trend that longitudinal vitamin D deficiency during pregnancy leads to lower vitamin D concentrations in neonatal cord blood (RR = 1.17; 95% CI (1.05-1.36); p = 0.04). Conclusion: Longitudinal vitamin D deficiency during pregnancy leads to vitamin D deficiency in neonates at birth. A further trial is needed to affirm this association.

Herbal-Based Formulation Containing Eurycoma longifolia and Labisia pumila Aqueous Extracts: Safe for Consumption?

A combined polyherbal formulation containing tongkat ali (Eurycoma longifolia) and kacip fatimah (Labisia pumila) aqueous extracts was evaluated for its safety aspect. A repeated dose 28-day toxicity study using Wistar rats was conducted where the polyherbal formulation was administered at doses 125, 500 and 2000 mg/kg body weight to male and female treatment groups daily via oral gavage, with rats receiving only water as the control group. In-life parameters measured include monitoring of food and water consumption and clinical and functional observations. On day 29, blood was collected for haematological and biochemical analysis. The rats were necropsied and the organs were collected for histopathological examination. This study showed that the combined formulation did not induce any significant toxicity effect at any dose level in terms of morbidity, mortality, behaviour, functional observation, body weight, food and water consumption, whole blood haematology and serum biochemistry. However, there were some microscopic changes in the histopathological examinations of some organs given 2000 mg/kg body weight, which may suggest an early response to the polyherbal formulation. From this study, the no observed adverse effect level is estimated to be more than 500 mg/kg body weight but not exceeding 2000 mg/kg body weight. The observed effects at the highest dose indicate the need for further study of longer dosing duration.

Eurycoma longifolia-Infused Coffee-An Oral Toxicity Study.

Coffee infused with the additive Eurycoma longifolia, also known as Tongkat ali (TA), has become widely available in the Malaysian market. Safety evaluations for consumption of the products have been called for due to the herbal addition. This study investigates the acute, subacute and chronic effects of a commercial TA coffee in Sprague Dawley rats when given in a single, repeated and prolonged dosage. The dosages of 0.005, 0.05, 0.30 and 2 g/kg body weight (BW) were used in the acute study and 0.14, 0.29 and 1 g/kg BW were used in the repeated dose studies. The in-life parameters measured were food and water intake, body weight and clinical observations. Blood were collected for hematology and clinical biochemistry analyses. All animals were subjected to full necropsies. Non-toxicity-related changes were observed in the food and water consumption parameters. Body weight showed normal increments and none of the animals had any clinical signs of toxicity. Microscopically assessed organ tissues did not reveal any abnormalities. There was significant decrease of platelet count in all the chronic study male treated groups. Significant elevation of renal profile parameters in both gender groups given 0.29 g/kg BW, along with liver and lipid profile elevation in some female groups of the chronic study were noted. No dose-dependent relationship was apparent in the dosage range tested, though these changes may suggest an initial safety indication to the TA coffee. The study concludes that the no observed adverse effect level (NOAEL) for this commercial TA coffee was 1 g/kg BW.

Clinacanthus nutans Standardized Fraction Arrested SiHa Cells at G1/S and Induced Apoptosis via Upregulation of p53.

INTRODUCTION: Cervical cancer is a leading cause of death in women. Current cancer treatment comes with side effects. Clinacanthus nutans has been known traditionally to treat cancer. This study was aimed to characterize C. nutans standardized fraction (SF1) and to investigate its anticancer mechanism against SiHa cells. MATERIALS AND METHODS: SF1 was produced by optimized methodology for bioassay-guided fractionation. Fourier transform infrared (FTIR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) were carried out to characterize the SF1. SF1 was screened for cytotoxicity activity toward HeLa, SiHa, and normal cells (NIH) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The anticancer mechanism of SF1 was evaluated toward SiHa cells, which showed highest cytotoxicity toward SF1 treatment. The mechanism includes cell cycle progression and protein expression, which was detected using specific antibody-conjugated fluorescent dye, p53-FITC, by flow cytometry. RESULTS: Major constituents of SF1 were alkaloids with amines as functional group. SF1 showed highest cytotoxic activity against SiHa (half-maximal inhibitory concentration [IC50] < 10 µg/mL) compared to HeLa cells. Cytoselectivity of SF1 was observed with no IC50 detected on normal NIH cells. On flow cytometry analysis, SF1 was able to induce apoptosis on SiHa cells by arresting cell cycle at G1/S and upregulation of p53 protein. CONCLUSION: SF1 showed anticancer activity by inducing apoptosis through arrested G1/S cell cycle checkpoint-mediated mitochondrial pathway.

Orthosiphon stamineus as a potential antidiabetic drug in maternal hyperglycemia in streptozotocin-induced diabetic rats.

BACKGROUND: Maternal hyperglycemia is associated with increased risk of adverse outcomes for both mother and offspring. Insulin is the standard treatment of hyperglycemia with the aim to reduce risks of complications, however, due to several restrictions, the search for more effective drugs from traditional medicinal plants continues. METHODS: The antidiabetic effects of Orthosiphon stamineus (O . stamineus ) in non-pregnant and pregnant streptozotocin-induced Sprague Dawley rats were identified. The effect of different concentrations of O. stamineouson insulin level using isolated pancreatic islets in response to low and high concentrations of glucose was identified. Oral glucose tolerance test was performed in both pregnant and non-pregnant rats prior to and after treatment with O. stamineus (0.1 g/100 g of body weight). O. stamineus was given orally daily for 2 weeks in non-pregnant and 10 days in pregnant rats. RESULTS: Oral glucose tolerance test indicated that treatment with O. stamineus in non-pregnant and pregnant rats significantly reduced blood glucose level and stimulated glucose-induced insulin secretion. No mortality was recorded throughout the study and no signs of toxicity during the experimental period including in both mother and foetus. For plasma analysis, the interactions of peptides such as GLP-1 and ghrelin level might contribute to the glucose lowering effect by O. stamineus via stimulation of insulin. The incubation of islets showed that O . stamineussignificantly stimulated insulin release in response to high glucose. CONCLUSION: O. stamineus could be a potential source of a specific oral hypoglycaemic agent to treat glucose intolerance in pregnancy.

Histopathological changes in placenta and liver of pregnant rats administered with aqueous extract of Dioscorea hispida var. daemona (Roxb) Prain & Burkill.

Dioscorea hispida var. daemona (Roxb) Prain & Burkill (DH), also known a tropical yam or intoxicating yam is a bitter wild tuber which is consumed as a staple food and traditionally used as a remedy in Malaysia. However, DH is also notorious for its intoxicating effects and there is currently a dearth of study of possible effects of DH on liver and placental tissues and hence its safe consumption warrants in-depth investigation. This study was therefore designed to investigate into the effect of DH on liver and placenta of pregnant rat via histopathological examination. Thirty pregnant Sprague-Dawley rats were randomly divided into five groups consisting of a control (distilled water) and four DH aqueous extract groups (250, 500, 1000 and 2000 mg/kg body weight). The extracts were administered via oral gavage daily throughout the study and animals were sacrificed on day 21. Paraffin-embedded, hematoxylin and eosin stained sections of placenta and liver were examined. Significant changes (p < 0.05) were observed on relative liver and placental weights of animals treated with 2000 mg/kg body weight DH extract. The placental numbers were decreased with the increased of DH extract concentration. Liver histological examination in all treated groups showed that tissues underwent degeneration characterized by hepatocyte swelling, cytoplasmic vacuolation, cytolysis, margination and clumping of nucleus chromatin. Changes of the basal and labyrinth zone were observed in placental tissues in all treated groups. Glycogen cells were reduced with fibrin deposition in the basal zone, while irregular vessel formation was demonstrated in the labyrinth zone. UHPLC-ESI-MS analysis showed the presence four steroidal saponins DH. In conclusion, DH aqueous extract exert hepatotoxicity and adverse effects on the placenta of rats. However, the underlying mechanism and phytochemicals inducing the observed toxicity require further investigation.

Gene Expression Profiling associated with Hepatoxicity in Pregnant Rats treated with Ubi Gadong (Dioscorea hispida) Extract.

Dioscorea hispida (D.hispida) is the most well-known starchy tuber in Malaysia and called 'ubi gadong'. Despite concerns over toxicity effects, the tuber is known to possess therapeutic values due to the presence of bioactive compounds such as saponins. This study was performed to identify the changes in gene expression profiles associated with hepatoxicity in pregnant rat treated with D.hispida using RT² Profiler PCR Array. The identification of steroidal saponins from D.hispida was carried out by UHPLC/MS method. Treatment of D.hispida caused mortality when dosage above 2000 mg/kg b.w. was given to pregnant rats. The PCR array showed that several genes were significantly up and down-regulated upon treatment with D.hispida. Treatment of D.hispida at 2000 mg/kg b.w leads to significant upregulation of several genes such as Btg2, Gsr, L2hgdn, S100a8, Slc17a3, Bhmt, Cd68, Cyp1a2 whereas several genes were downregulated such as Abcb1a, Aldoa, Cdc14b, Icam1, Krt18, Hpn and Maob. The consumption of D.hispida extract when taken at lower dosage of 2000 mg/kg may not be harmful to rats. D.hispida extract given at the highest dosage to pregnant rats caused alterations of several genes categorized in different hepatotoxic group functions such as necrosis, cholestasis and phospholipodisis.

Study on the developmental toxicity of a standardized extract of Orthosiphon stamineus in rats

Infusions of Orthosiphon stamineus Benth., Lamiaceae, leaves are widely used in Southeastern Asia to treat different illnesses. Nonetheless, no data is available on the safety of O. stamineus for pregnant women and their babies. This study was undertaken to evaluate the developmental toxicity of O. stamineus standardized aqueous extract in female Sprague Dawley rats (n=21) at 0, 250, 500, 1000 and 2000 mg/kg/day, by gavage on gestation days 6-20. Clinical signs of maternal toxicity, body weight gain, and food and water consumption were recorded. Caesarean sections were performed on gestation day 21; resorptions and living and dead fetuses were counted. Fetuses were weighed and examined for external abnormalities. Half of the fetuses from each litter were cleared and stained with Alizarin red S for skeleton evaluation. O. stamineus standardized aqueous extract did not alter pregnancy body weight gain and food and water consumption and caused no other sign of maternal toxicity. Embryolethality and prenatal growth retardation were not observed either. O. stamineus standardized aqueous extract increased a few skeleton variations and a skull bone malformation (hyoid bone absent) in a non-dose dependent manner. Anogenital distance was increased in male and female fetuses exposed to the highest O. stamineus standardized aqueous extract dose, an indication that the extract could possibly contain androgenic compounds.

The in vivo rodent micronucleus assay of Kacip Fatimah (Labisia pumila) extract.

Kacip Fatimah also known as Labisia pumila (Myrsinaceae), is a traditional herbal medicine with a long history in the Malay community. It has been used by many generations of Malay women to induce and facilitate childbirth as well as a post-partum medicine. We tested the genotoxic potential of Kacip Fatimah in bone marrow cells obtained from Sprague-Dawley rats using micronuclei formation as the toxicological endpoints. Five groups of five male rats each were administered orally for two consecutive days with doses of 100, 700 and 2000 mg/kg body weight of Kacip Fatimah extract dissolved in distilled water. Micronucleus preparation was obtained from bone marrow cells of the animals following standard protocols. No statistically significant increase in micronucleated polychromatic erythrocytes (MNPCEs) was observed at any dose level and sacrifice/harvest time point (24, 48 and 72h). However, a significant decrease in polychromatic erythrocytes/normochromatic erythrocytes (PCE:NCE) ratio was observed from the highest dose level (2000 mg/kg of body weight) at 48h harvest time point. In this study, we investigated the effect of Kacip Fatimah on mammalian bone marrow cells using micronuclei formation to assess the genotoxicity of the herb.