Attenuation of CFA-induced arthritis through regulation of inflammatory cytokines and antioxidant mechanisms by Solanum nigrum L. leaves extracts.

Solanum nigrum L. is a popular traditional medicine for various inflammatory conditions including rheumatism and joint pain. The current study aimed to evaluate the anti-arthritic mechanism of Solanum nigrum L. Four extracts were prepared using n-hexane, methanol, chloroform, and water. The anti-nociceptive and anti-inflammatory activity was carried out with 100, 200, and 300 mg/kg body wt. PO of each extract by the hot plate and carrageenan-induced paw oedema methods, respectively. The anti-arthritic study was performed with chloroform and aqueous extracts (300 mg/kg) in complete Freund's adjuvant (CFA)-induced arthritis. Paw size (mm), ankle joint diameter (mm), and latency time (sec) were recorded on day 0 and every 4th day till 28 days. The hematological, inflammatory, and oxidative biomarkers were estimated. Results showed that significant analgesia (p < 0.05) and reduction in paw inflammation were achieved with all extracts. The highest percent inhibition in Carrageenan-induced inflammation was achieved with 300 mg/kg of chloroform (72.19%) and aqueous (71.30%) extracts, respectively. In the CFA model, both extracts showed a significant reduction in paw size and ankle joint diameter (p < 0.05). The RT-qPCR analysis revealed the upregulation of interleukin-4 and interleukin-10, and down-expression of interleukin-1ß, interleukin-6, tumor necrosis factor-α, cycloxygenase-2, nuclear factor-κB, prostaglandin E synthase 2, and interferon-γ. A significant increase in superoxide dismutase, catalase, and glutathione levels was observed. Hence, it is concluded that Solanum nigrum L. leaf extracts regulate the expression of inflammatory markers and improve oxidative stress resulting in the attenuation of CFA-induced arthritis.

Anticonvulsant potential of Grewia tiliaefolia in pentylenetetrazole induced epilepsy: insights from in vivo and in silico studies.

Epilepsy, a chronic neurological condition, impacts millions of individuals globally and remains a significant contributor to both illness and mortality. Available antiepileptic drugs have serious side effects which warrants to explore different medicinal plants used for the management of epilepsy reported in Traditional Indian Medicinal System (TIMS). Therefore, we explored the antiepileptic potential of the Grewia tiliaefolia (Tiliaeceae) which is known for its neuroprotective properties. Aerial parts of G. tiliaefolia were subjected to extraction with increasing order of polarity viz. hexane, chloroform and methanol. Antioxidant potential of hexane, chloroform and methanol extracts of G. tiliaefolia was evaluated by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) assay, total antioxidant capacity (TAC) assay, reducing power assay (RPA) and DNA nicking assay. Additionally, quantitative antioxidant assays were also conducted to quantify total phenolic (TPC) and total flavonoid content (TFC). As revealed by in vitro assays, methanol extract was found to contain more phenolic content. Hence, the methanol extract was further explored for its anticonvulsant potential in pentylenetetrazole (PTZ) induced acute seizures in mice. The methanol extract (400 mg/kg) significantly increased the latency to occurrence of myoclonic jerks and generalized tonic clonic seizures (GTCS). Additionally, it also reduced duration and seizure severity score associated with GTCS. The Grewia tiliaefolia methanol extract was further screened by Ultra High-Performance Liquid Chromatography (UHPLC) for presence of polyphenolic compounds, among which gallic acid and kaempferol were present in higher amount and were further analysed by in silico study to predict their possible binding sites and type of interactions these compounds show with gamma amino butyric acid (GABA) receptor and glutamate α amino-3- hydroxyl-5-methyl-4-isoxazolepropionic acid (Glu-AMPA) receptor. It was revealed that gallic acid and kaempferol had shown agonistic interaction for GABA receptor and antagonistic interaction for Glu-AMPA receptor. We concluded that G. tiliaefolia showed anticonvulsant potential possibly because of gallic acid and kaempferol possibly mediated through GABA and Glu-AMPA receptor.

Polyphenol-enriched Desmodium elegans DC. ameliorate scopolamine-induced amnesia in animal model of Alzheimer's disease: In Vitro, In Vivo and In Silico approaches.

The current study aims to quantify HPLC-DAD polyphenolics in the crude extracts of Desmodium elegans, evaluating its cholinesterase inhibitory, antioxidant, molecular docking and protective effects against scopolamine-induced amnesia in mice. A total of 16 compounds were identified which include gallic acid (239 mg g-1), p-hydroxybenzoic acid (11.2 mg g-1), coumaric acid (10.0 mg g-1), chlorogenic acid (10.88 mg g-1), caffeic acid (13.9 mg g-1), p-coumaroylhexose (41.2 mg g-1), 3-O-caffeoylquinic acid (22.4 mg g-1), 4-O-caffeoylquinic acid (6.16 mg g-1), (+)-catechin (71.34 mg g-1), (-)-catechin (211.79 mg g-1), quercetin-3-O-glucuronide (17.9 mg g-1), kaempferol-7-O-glucuronide (13.2 mg g-1), kaempferol-7-O-rutinoside (53.67 mg g-1), quercetin-3-rutinoside (12.4 mg g-1), isorhamnetin-7-O-glucuronide (17.6 mg g-1) and isorhamnetin-3-O-rutinoside (15.0 mg g-1). In a DPPH free radical scavenging assay, the chloroform fraction showed the highest antioxidant activity, with an IC50 value of 31.43 µg mL-1. In an AChE inhibitory assay, the methanolic and chloroform fractions showed high inhibitory activities causing 89% and 86.5% inhibitions with IC50 values of 62.34 and 47.32 µg mL-1 respectively. In a BChE inhibition assay, the chloroform fraction exhibited 84.36% inhibition with IC50 values of 45.98 µg mL-1. Furthermore, molecular docking studies revealed that quercetin-3-rutinoside and quercetin-3-O-glucuronide fit perfectly in the active sites of AChE and BChE respectively. Overall, the polyphenols identified exhibited good efficacy, which is likely as a result of the compounds' electron-donating hydroxyl groups (-OH) and electron cloud density. The administration of methanolic extract improved cognitive performance and demonstrated anxiolytic behavior among tested animals.

Challenges in estimating the encapsulation efficiency of proteins in polymersomes - Which is the best method?

Abstract Polymersomes are nanometric vesicles that can encapsulate large and hydrophilic biomolecules, such as proteins, in the aqueous core. Data in literature show large variation in encapsulation efficiency (%EE) values depending on the method used for calculation. We investigated different approaches (direct and indirect) to quantify the %EE of different proteins (catalase, bovine serum albumin-BSA, L-asparaginase and lysozyme) in Pluronic L-121 polymersomes. Direct methods allow quantification of the actual payload of the polymersomes and indirect methods are based on the quantification of the remaining non-encapsulated protein. The protein-loaded polymersomes produced presented approximately 152 nm of diameter (PDI ~ 0.4). Higher %EE values were obtained with the indirect method (up to 25%), attributed to partial entanglement of free protein in the polymersomes poly(Ethylene Glycol) corona. For the direct methods, vesicles were disrupted with chloroform or proteins precipitated with solvents. Reasonable agreement was found between the two protocols, with values up to 8%, 6%, 17.6% and 0.9% for catalase, BSA, L-asparaginase and lysozyme, respectively. We believe direct determination is the best alternative to quantify the %EE and the combination of both protocols would make results more reliable. Finally, no clear correlation was observed between protein size and encapsulation efficiency.

The Assessment of Trihalomethanes Concentrations in Drinking Water of Hamadan and Tuyserkan Cities, Western Iran and Its Health Risk on the Exposed Population.

BACKGROUND: The aim of the present study was to calculate and to assess the potential lifetime cancer risks for trihalomethanes from consuming chlorinated drinking water in Hamadan and Tuyserkan cities, western Iran in 2016-2017. STUDY DESIGN: A cross-sectional study. METHODS: Seventy-two water samples were collected from the distribution systems and from the outlet of water treatment plants (WTPs) and the experiments were carried out to determine the desired parameters. All the sampling and measurement methods were according to Standard Methods. The obtained data were analyzed using SPSS software. RESULTS: The mean concentration of total THMs in the summer and winter was 42.75 and 17.75 µg/L, respectively, below the WHO and Iranian standard. The positive correlation was observed between temperature and THMs levels. Moreover, THMs concentration in Shahid Beheshti's WTP was several times lower than in Ekbatan's WTP. Chloroform, the dominant species of THMs, was identified at different sampling points. The highest cancer risk in Hamadan was 1.4×10-5 and 4.8×10-5 for male and female, respectively; and the cancer risk was obtained to be 5.6×10-7-2.26×10-6 in Tuyserkan. CONCLUSION: The drinking water obtained from the studied area is safe in terms of THMs concentration. Nevertheless, the highest cancer risk was higher than the EPA's acceptable level of 10-6.

A general dose-response relationship for chronic chemical and other health stressors and mixtures based on an emergent illness severity model.

Current efforts to assess human health response to chemicals based on high-throughput in vitro assay data on intra-cellular changes have been hindered for some illnesses by lack of information on higher-level extracellular, inter-organ, and organism-level interactions. However, a dose-response function (DRF), informed by various levels of information including apical health response, can represent a template for convergent top-down, bottom-up analysis. In this paper, a general DRF for chronic chemical and other health stressors and mixtures is derived based on a general first-order model previously derived and demonstrated for illness progression. The derivation accounts for essential autocorrelation among initiating event magnitudes along a toxicological mode of action, typical of complex processes in general, and reveals the inverse relationship between the minimum illness-inducing dose, and the illness severity per unit dose (both variable across a population). The resulting emergent DRF is theoretically scale-inclusive and amenable to low-dose extrapolation. The two-parameter single-toxicant version can be monotonic or sigmoidal, and is demonstrated preferable to traditional models (multistage, lognormal, generalized linear) for the published cancer and non-cancer datasets analyzed: chloroform (induced liver necrosis in female mice); bromate (induced dysplastic focia in male inbred rats); and 2-acetylaminofluorene (induced liver neoplasms and bladder carcinomas in 20,328 female mice). Common- and dissimilar-mode mixture models are demonstrated versus orthogonal data on toluene/benzene mixtures (mortality in Japanese medaka, Oryzias latipes, following embryonic exposure). Findings support previous empirical demonstration, and also reveal how a chemical with a typical monotonically-increasing DRF can display a J-shaped DRF when a second, antagonistic common-mode chemical is present. Overall, the general DRF derived here based on an autocorrelated first-order model appears to provide both a strong theoretical/biological basis for, as well as an accurate statistical description of, a diverse, albeit small, sample of observed dose-response data. The further generalizability of this conclusion can be tested in future analyses comparing with traditional modeling approaches across a broader range of datasets.

3-methyadenine attenuates chloroform-induced hepatotoxicity via autophagy activation.

Chloroform is a common contaminant in the drinking water. Exposure of human to chloroform leads to severe hepatotoxicity. In the present study, chloroform-induced acute liver injury was investigated in mice using 3-methyadenine (3-MA), a common autophagy inhibitor. At 24 h after intraperitoneal injection of 0.5 mL/kg chloroform, serum alanine aminotransferase (ALT) levels were increased significantly; extensive necrosis and inflammation occurred as identified by histological examinations. Moreover, chloroform induced an increase in lipid peroxidation as demonstrated by increased formation of malondialdehyde (MDA) in the liver tissues. Hepatic antioxidants including glutathione (GSH) and superoxide dismutase (SOD) were decreased by chloroform treatment. All these changes were significantly inhibited by 3-MA treatment. Further mechanistic insights demonstrated that chloroform up-regulated pro-inflammatory cytokine, IL-1ß, in the livers and blood, which was suppressed by 3-MA. Surprisingly, Western blots results showed that after 24-hours of chloroform treatment 3-MA activated autophagy as indicated by decreased levels of LC3B II and p62 protein. Co-treatment of chloroquine with 3-MA to inhibit autophagy would abrogate the hepatoprotection of 3-MA in chloroform hepatotoxicity. Taken together, findings in the present study suggested that a widely-used autophagy inhibitor, 3-MA, significantly reduced chloroform hepatotoxicity in mice via autophagy activation. Findings in this study also suggested that caution should be exercised when using 3-MA to modulate autophagy in vivo.

Risk of leukemia in relation to exposure to ambient air toxics in pregnancy and early childhood.

There are few established causes of leukemia, the most common type of cancer in children. Studies in adults suggest a role for specific environmental agents, but little is known about any effect from exposures in pregnancy to toxics in ambient air. In our case-control study, we ascertained 69 cases of acute lymphoblastic leukemia (ALL) and 46 cases of acute myeloid leukemia (AML) from California Cancer Registry records of children

Keratocystic odontogenic tumors and Carnoy's solution: results and complications assessment.

OBJECTIVE: Keratocystic odontogenic tumors (KOTs) can be treated with Carnoy's solution, although this treatment modality is not free from complications. It is important to verify the incidence of complications after the use of Carnoy's solution and compare these with the literature. MATERIALS AND METHODS: This study verified the effects of a complementary treatment for KOTs and assessed the incidence of such complications as recurrence, infection, sequestrum formation, mandibular fracture, dehiscence, and neuropathy. RESULTS: Twenty-two KOTs treated with Carnoy's solution combined with peripheral ostectomy were included, and the follow-up period varied from 12 to 78months with a mean of 42.9months. Complications included recurrence (4.5%), dehiscence (22.7%), infection (4.5%), and paresthesia (18.2%). No difference was found among lesions associated (9.1%) or not (0%) with nevoid basal cell carcinoma syndrome (P>0.05). Dehiscence was influenced by marsupialization (P<0.05), and paresthesia was observed exclusively in cases of mandibular canal fenestration (P<0.01). CONCLUSIONS: Complementary treatment with Carnoy's solution and peripheral ostectomy appear to provide efficient treatment for KOTs. Complications originating from the use of the solution are less frequent and less serious than complications associated with cryotherapy. Neuropathy seems to be related to direct contact between the solution and the epineurium.

Exposure to trihalomethanes in drinking water and small-for-gestational-age births.

BACKGROUND: Trihalomethanes in drinking water have been associated with higher occurrence of small-for-gestational-age (SGA) births, although results have been inconsistent. METHOD: We geocoded residential address for mother of live, singleton, term births to 33 water distribution systems in a large metropolitan area of New South Wales, Australia (314,982 births between 1998 and 2004) and classified births into <10th percentile and ≥ 10 percentile of weight for gestational age. Mean trihalomethane exposure was estimated by trimester and for the entire pregnancy based on monthly sampling in each of the 33 water distribution systems. We estimated the relative risk (RR) of SGA for exposure to trihalomethanes using log-binomial regression adjusting for confounding. RESULTS: SGA births increased with mother's third-trimester exposure to chloroform (RR = 1.04 [95% confidence interval = 1.02-1.06], across an interquartile range [IQR] = 25 µg/L) and bromodichloromethane (1.02 [1.01-1.04], 5 µg/L). Larger associations were found for SGA less than third percentile. Smoking modified the effects of trihalomethane exposure, with generally larger associations in births to nonsmoking mother and weaker or protective associations in births to smoking mothers. CONCLUSIONS: : Mothers' exposures during pregnancy to total trihalomethane as well as to chloroform and bromodichloromethane were associated with SGA. These associations were modified by maternal smoking during pregnancy.

Human umbilical cord blood-derived mesenchymal stem cells improve glucose homeostasis in rats with liver cirrhosis.

Disturbance of glucose metabolism is a common feature in liver cirrhosis which is associated with insulin resistance and is an established risk factor for disease progression and survival in patients with chronic liver diseases. We investigated whether umbilical cord blood-derived mesenchymal stem cells (HMSCs) have an effect on the expression of molecules responsible for glucose utilization and hepatic gluconeogenesis, focusing on the insulin signaling pathway in rats with liver cirrhosis. Rats received a dose of CCl4 (100 µl/100 g 4:1 in corn oil) thrice-weekly. HMSCs were infused at 4 weeks after induction of liver cirrhosis by CCl4. Infusion of HMSCs improved insulin resistance which was associated with increased glucose levels and decreased insulin sensitivity in CCl4-induced cirrhotic rats. HMSCs increased activities in the proximal part of the insulin signaling cascade, as evidenced by increased expression of key enzymes such as phosphatidylinositol-3-kinase (PI 3-kinase), protein kinase B (PKB), protein kinase C-ζ (PKC-ζ), and the decrease of glycogen synthase kinase 3 (GSK-3) compared to CCl4-induced liver cirrhotic rats. We also observed that glucose-6-phosphatase (G-6-P) and phosphoenolpyruvate kinase (PEPCK), two hepatic enzymes involved in gluconeogenesis were strongly decreased over 40-50% after infusion of HMSCs. Taken together, our results showed that HMSCs could improve insulin resistance in CCl4-induced liver cirrhosis, thereby contributing to glucose homeostasis.

Development of a multi-pathway probabilistic health risk assessment model for swimmers exposed to chloroform in indoor swimming pools.

For swimmers, exposure to chloroform, a probable carcinogen, in indoor swimming pools can be through different pathways such as ingestion, dermal absorption, inhalation during swimming, and inhalation during resting. In order to evaluate health risk results from excessive exposure to chloroform, concentrations of chloroform in pool water were first collected and analyzed. Then, a two-layer model is used, which is capable of estimating the concentrations of chloroform in the boundary layer adjacent to the water surface and the concentrations of chloroform in indoor swimming pool air. The use of stratification model is important for estimating the risks for swimmers since they are exposed to these kinds of situations while performing swimming and resting in indoor swimming pools environment. The incremental lifetime cancer risk (ILCR) was then estimated using the multi-pathway exposure model. The results showed that the 95th percentile of ILCRs calculated for male and female swimmers were 2.80 × 10(-4) and 2.47 × 10(-4), respectively. The major exposure routes were found to be inhalation during swimming which contributes to more than 99% of the total health risk. Our study suggested that to protect swimmers from excessive exposure to chloroform, alternative methods or processes of disinfection should be considered for swimming pool managers.

Chloroform anesthesia and the Saville Kent murder in 1860.

The discovery of the anesthetic properties of chloroform by Sir James Simpson is one of the therapeutic triumphs of the 19th century. Queen Victoria requested chloroform anesthesia for the birth of her second son, Prince Leopold, and from then until the end of century chloroform was the most popular general anesthetic for obstetrics and surgery. Chloroform was so pleasant to inhale that it became a drug of abuse and was involved in all sorts of criminal activity. Despite its wide usage, chloroform was not thought to be part of the brutal murder of 3-year-old Saville Kent on the night of June 30, 1860, outside his manor house in southwest England. The events surrounding and after the murder have recently been documented by Kate Summerscale in a book entitled, "The Suspicions of Mr. Whicher," the Detective Inspector assigned to the case by Scotland Yard. According to Ms Summerscale's records, Mr Whicher ignored the possibility that chloroform was involved in Saville's death. However, evidence supports the view that chloroform played a critical role in the crime and indicates that the guilty plea by Samuel Kent's daughter, Constance, for which she spent 20 years to the day in prison, was as inaccurate as it was incomplete.

The effect of silver nitrate, chloroformic garlic extract, and normal saline in induction of sclerosing cholangitis in rabbits.

OBJECTIVE: To evaluate the effects of 0.5% silver nitrate, 20% chloroformic garlic extract, and 0.9% normal saline in induction of sclerosing cholangitis in the bile ducts of rabbits. METHODS: During a-6-months period from April to September 2006 in Shiraz University Laboratory Animal Research Center, we selected 3 equal groups of rabbits. We injected 0.5% silver nitrate, 20% chloroformic garlic extract, and 0.9% normal saline into the bile ducts of each group. The animals were euthanized, and autopsied after 4 months and the liver and bile ducts were removed and studied histopathologically. Cholangiography was undertaken to evaluate the presence and extent of any sclerosing cholangitis. RESULTS: Animals showed sclerosing cholangitis in silver nitrate group (7 [58%]), one (8%) in chloroformic garlic extract group and one (7%) in normal saline group. The difference between silver nitrate and chloroformic garlic extract groups were statistically significant and similar results were noticed between chloroformic garlic extract and normal saline groups. CONCLUSION: Twenty percent of chloroformic garlic extract had fewer complications such as sclerosing cholangitis, compared to other materials.

[Hygienic evaluation of the combined effect of portable water chloroform].

The paper considers the significance of complex entrance of chloroform from portable water into the human body (enterally, inhalationally, and through the intact skin). It shows it necessary to toughen the maximum permissible concentration (MPC) of chloroform in the portable drinking, by taking into account of the multiplicity of routes of its action on the population. The authors present the results of their own investigations of the levels of chloroform in the air of bath and shower rooms before and after taking a shower and filling the bath with water, as well as in the airspace layer above the water of an indoor swimming pool, by using chromatographic mass-spectrometry.