Intestinal Homogenates Research Articles

Influence of Rectal Ozone Application on the Intensity of Free Radical Destruction of Lipids and Intestinal Proteins in the Dynamics of Experimental Colitis.

We studied clinical status, content of products of LPO, and oxidative modification of proteins (OMP) in the lesion focus of the intestine in experimental colitis under conditions of rectal administration of ozone. Experimental colitis was simulated by two-stage administration of oxazolone; rectal insufflation of ozone in the ozone-oxygen mixture was performed daily. The disease activity index (DAI), the content of calprotectin in the feces, and LPO and OMP products in the intestinal homogenate were assessed. On days 2, 4, and 6 of the pathological process, DAI, concentration of calprotectin in feces, content of primary, secondary, and end-products of LPO in the heptane and isopropanol phases, and content of primary and secondary OMP products progressively increases. Under conditions of ozone application, DAI, concentration of calprotectin in feces, the levels of heptane- and isopropanol-soluble primary, secondary, and end-products of LPO, and the level of primary and secondary products of OMP decreased on days 4 and 6; the level of isopropanol-soluble primary, secondary, end-products of LPO increased on day 2 of experimental colitis. The severity of clinical manifestations weakens as the content of LPO and OMP products in the colon decreases on days 4 and 6 of observation.

Natural products-characterized moringa oleifera leaves methanolic extract and anti-diabetic properties mechanisms of its fractions in streptozotocin-induced diabetic rats

Background: The study characterized natural products and investigated the mechanisms of antidiabetic properties in Moringa oleifera leaves methanolic extract in Streptozotocin-induced diabetic male rats. Methods: the natural bioactive compounds in difffferent fractions were obtained by column chromatography on 60-120 silica gel mesh and assayed using gas chromatography-mass spectrometric (GC-MS) technique. Also, possible anti-diabetic properties mechanisms of the fractions of extracts investigated using standard methods. Results: Lethality study indicated safety of the extract at 5000mg/kgbwt. The GC- MS analysis of the methanolic extract revealed seven out of eleven common bioactive constituents with reported antidiabetic activity. Results of possible anti-diabetic properties mechanisms based on studied glucose homeostasis-related bio-indicators showed signifificant (P<0.05) increase in serum lipoprotein lipase, whole blood GLUT 1, liver homogenate G-6-P-DH, Intestine homogenate Na+ /K+ -ATPase, Muscle homogenate GLUT 4 and liver homogenate AMP-Kinase activities in rats administered with the drug (glibenclamide) and fractions 3,4,5 compared with the diabetic group. A signifificant (P<0.05) inhibition of serum α-amylase activity in rat administered drug and fractions 3,4,5 was observed when compared with diabetic control and fractions 1 and 2 . Histological outcomes of rats' pancreas corroborated these fifindings. Conclusion: Thus, characterized natural bioactive constituents of M. oleifera fractions exerted concerted potent anti-diabetic activity in streptozotocin-induced diabetic rats by probable mechanisms involving concerted α-amylase inhibition, AMP-kinase and Lipo-protein lipase activation and glucose transport Proteins (GLUT-1 and GLUT-4) and liver homogenate Glucose-6-phosphate dehydrogenase up-regulation.

Feeding-Induced Changes of Bacteriolytic Activity and the Pattern of Bacteriolytic Compounds in the Stomach and Small Intestine of the Haematophagous Bug Triatoma infestans (Klug, 1834) (Reduviidae, Triatominae)

Intestinal homeostasis mechanisms of the haematophagous triatomines regulate the development of mutualistic symbionts and other gut bacteria. Investigating antimicrobial compounds of these insects, we have determined spectrophotometrically that the bacteriolytic activity is between pH 3 and pH 9 using homogenates of fifth instar Triatoma infestans stomachs and small intestines from unfed bugs and up to 50 days after feeding. The activity against Gram-positive Micrococcus luteus was strongest at pH 4 and pH 7 and was higher in the stomach than in the small intestine. Symbiotic Rhodococcus triatomae were not lysed. Lysis of Gram-negative Escherichia coli showed a maximum at pH 7 in the stomach and at pH 5 in the small intestine. Bacteriolytic activity against both M. luteus and E. coli was reduced 24 h after feeding, then increased, and at 50 days after feeding was strongly reduced. In zymographs, the activity against M. luteus was mainly correlated to proteins of about 16 kDa. At different periods of time after feeding, seven bands of lysis appeared between 15 and 40 kDa and more bands using extracts of the small intestine than those of the stomach. This is the first proof for the synthesis of antibacterial proteins of 22–40 kDa in triatomines.

In vitro stability and ex vivo absorption of thymol monoglucosides in the porcine gut

Thymol α-D-glucopyranoside (TαG) and thymol β-D-glucopyranoside (TβG) are believed to have different kinetic behaviours in the porcine gut than its parent aglycon thymol. However, recently, it was shown that concentrations of both glucosides decreased rapidly in the stomach and proximal small intestine following oral supplementation to piglets as did thymol. Yet, the stability of thymol glucosides in gut contents and their absorption route remains obscure. Therefore, a series of in vitro incubations were performed, simulating the impact of pH, digestive enzymes, bacterial activity and mucosal extracts on stability of these glucosides. Their absorption mechanisms were investigated using the Ussing chamber model in the presence or the absence of inhibitors of sodium-dependent glucose linked transporter 1 and lactase phlorizin hydrolase. Both glucosides remained intact at physiological pH levels in the presence of digestive enzymes. Recoveries from TαG and TβG were below 90% when incubated with small intestinal homogenates from the distal jejunum or from all sampled sites, respectively. However, no aglycon could be detected in these samples. Bacterial inoculum of the small intestine, on the other hand, hydrolysed TβG quickly with up to 44% of free aglycon appearing. TαG proved more resistant to porcine gastro-intestinal bacterial glucosidases with only trace amounts (<1%) of free thymol at the end of the incubations. Electrophysiological measurements in Ussing chambers did not suggest active transport of the glucosides. Mucosal TαG and TβG concentrations were unchanged between start and end of the absorption measurements. Additionally, no TαG and only a very limited amount of TβG were retrieved from the serosal side. Tissue associated concentrations, although marginal (<1% of luminal concentration), were mainly as intact glucoside or as aglycon for TαG and TβG, respectively. Addition of both inhibitors significantly increased the amount of intact glucosides retrieved from the mucosal tissues as compared to controls. In conclusion, bacterial hydrolysis was identified as the most important source of TβG loss, whereas TαG seemed less prone to degradation or absorption in these in vitro and ex vivo models.

Comparing the Effect of Piperine and Ilepcimide on the Pharmacokinetics of Curcumin in SD Rats.

The poor bioavailability and rapid metabolism of curcumin (CUR) restrict its clinical application. Piperine (PIP), which was extracted from natural compounds, can increase the plasma concentration of curcumin in humanidad. As an artificial synthetic piperine analog, silepcimide (ILE) has significant advantages because of the low price and simple synthesis process. In this study, a simple and rapid HPLC-UV method was developed for determination of the plasma concentration of CUR, PIP,ILE and dihydrocurcumin (DHC, a metabolite of CUR) simultaneously. Meanwhile, the effects of PIP and ILE on the plasma concentration and pharmacokinetics of DHC in SD rats was studied to explore whether ILE could serve as a CUR bioavailability enhancer. The metabolic pathway of CUR was studied by comparing the differences of CUR plasma concentration between intravenous injection and oral administration over the same time period, and reacting with small intestine homogenate without microbes of SD rats. The results of drug-time curve showed that combined administration of ILE and CUR had significant effect on plasma concentrations of DHC. Repeated administration of PIP or ILE could significantly increase the plasma concentration of DHC. Plasma CUR could be detected in the samples of from intravenous injection of CUR rats, whereas, it couldn’t be detected in the plasma sample form oral administration rats. CUR incubated with intestinal homogenate without intestinal bacteria could not be transformed into DHC. In conclusion, our results show that ILE can improve the bioavailability of CUR. Additionally, it was inferred that most of the CUR was reduced to DHC by NADPH when it was absorbed from gastrointestinal tract, and our results demonstrated that this pathway might be mediated by gastrointestinal microorganisms.

Electromagnetic Fields Modify Redox Balance in the Rat Gastrointestinal Tract.

Objective: The aim of the study was to assess the influence of electromagnetic fields with divergent physical properties on the prooxidative and antioxidative balances in homogenates of the tongue, salivary glands, esophagus, stomach, and small and large intestines of rats.Material and Methods: Forty rats were randomly divided into four equal groups, namely, a control group, a group exposed to low-frequency electromagnetic fields (LF-EMFs; frequency: 50 Hz; intensity: 10 kV/m; magnetic induction: 4.3 pT), a group exposed to radiofrequency electromagnetic fields (RF-EMFs) emitted by mobile phones (frequency: 900 MHz), and a group exposed simultaneously to LF-EMFs and RF-EMFs emitted by mobile phones. After 28 consecutive days of the experiment, the following pro- and antioxidative markers were assessed in the gastrointestinal tract homogenates: superoxide dismutase (SOD) and its two isoenzymes (Mn-SOD, Cu,Zn-SOD) catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST), total antioxidative capacity (TAC), total oxidative status (TOS), and malondialdehyde (MDA).Results: In rats exposed to LF-EMFs, higher concentrations of the markers of prooxidant processes, MDA or TOS, were observed in the salivary glands, esophagus, and small intestine homogenates in comparison with the control group. Additionally, in the group of rats opposite to the control, antioxidant activity was observed. The main differences included a higher activity of Cu,Zn-SOD in homogenates of the tongue, salivary glands, and esophagus as well as decreased activity of CAT in homogenates of the tongue, esophagus, and small intestine. In animals exposed to RF-EMFs, the concentration of TOS was higher in the large intestine than in control rats. The main difference of antioxidant activity was presented by decreased Cu,Zn-SOD in homogenates of the salivary glands, stomach, small and large intestine as well as CAT in homogenates of the tongue, esophagus, stomach, and small and large intestine. Moreover, in rats exposed simultaneously to LF-EMFs and RF-EMFs, a lower concentration of TOS was observed. Antioxidant activity was presented by a decreased activity of CAT in homogenates of the tongue, esophagus, stomach, and small and large intestine in comparison to the control group.Conclusion: Among those applied in the study, electromagnetic fields of a low-frequency caused the most significant disturbances of oxidative stress in the rat gastrointestinal tract.

Spondias pinnata bark extract- an ameliorator of inflammatory derangement in etoposide induced mucositis: An experimental approach.

Background and Aim:Mucositis, one of the vulnerabilities of chemotherapy, affects the physiological activities and therapeutic strategies of patients because it can affect the normal cell population. Etoposide is a commonly used chemotherapeutic agent for cancers such as oral, lung, and gastrointestinal. In addition to the abnormal metabolic processes in the body caused by tumorigenesis, new metabolic alterations can occur, such as oxidative stress, antioxidant imbalance, and inflammatory reactions, all of which can contribute to existing patient vulnerability. Therapeutic adjuvants can help overcome these toxic effects. Spondias pinnata is a tropical tree omnipresent in the coastal and Western Ghat section of India that is used for culinary purposes and as a local analgesic. Therefore, we aimed to study the anti-inflammatory effects of S. pinnata in an etoposide-induced mucositis rat model.Materials and Methods:Small intestinal tissue homogenates from albino Wistar rats were used to estimate the levels of glutathione (GSH) and nitric oxide (NO), and activities of total antioxidant (TAO), myeloperoxidase (MPO) and Na+-K+ ATPase. The animals were grouped into: (1) normal control, (2) etoposide-induced mucositis (65 mg/kg bodyweight, single IP dose), (3) S. pinnata control group, and (4) etoposide followed by S. pinnata bark extract (200 mg/kg bodyweight, once in a day). Animals were sacrificed after 24, 48, 72, and 96 h and compared with that of the normal control group (n=6). Statistical analysis was performed using EZR software.Results:We observed a significant decrease in the TAO and GSH levels with a marked increase in NO, MPO, and Na+-K+ ATPase activity in the mucositis group. A tendency to recover from the decreased TAO and GSH levels existed in the treated group, showing the protective effects of S. pinnata bark extract against mucositis. In addition, this extract also showed anti-inflammatory effects as reflected by the recovery in MPO levels at the end of 96 h. Maintenance of Na+-K+ ATPase activity in the treated group demonstrates the protective effects of the extract against the increased levels observed in the etoposide-induced mucositis group.Conclusion:This study revealed the protective effects of S. pinnata bark extract against the oxidative and inflammatory changes that occurred during the development ofmucositis. This would decrease the pathological burden during chemotherapy and prevent any hurdles in therapeutic modalities.

Динамика хемилюминесценции в тонкой кишке на фоне анестетического прекондиционирования при геморрагической гипотензии

Introduction. The major pathogenetic factor of massive blood loss is hypoxia, which triggers activation of free-radical oxidation (FRO) processes in organs and tissues and the systemic inflammatory response. A universal factor of multiple organ dysfunction in blood loss is altered intestinal wall permeability with translocation of microflora and toxins into the systemic circulation during reperfusion. Recently, much of the attention has been focused on effects of anesthetic preconditioning, including during operations associated with hemorrhagic hypotension (HH). The aim of this study was to evaluate in experiment the dynamics of small intestinal FRO in HH during the use of the anesthetic sevoflurane, which has an effect of anesthetic preconditioning. Methods. Experiments were performed on 105 white male rats divided into two groups; groups 1 and 2 were exposed to HH with ether or sevoflurane as the anesthetic, respectively. Two groups of intact animals treated with ether or sevoflurane were used as the controls. Five animals died during the experiment. To evaluate FRO processes, samples of the duodenum, jejunum, and ileum were taken at 15 min, 30 min, 1 h, and 2 h of HH. The chemiluminescence (CL) study of small intestine homogenates was performed according to the Farukhutdinov method on a Fluorate ABLF-2T chemiluninometer. The following FRO indexes were recorded: spontaneous luminosity (SL), flash (F), and light sum (L∑). Significance of differences was determined with the Mann-Whitney test. Results. In the sevoflurane group 2 compared to the ether group after 15 min of HH, SL was increased in the jejunum by 33%; F was decreased in the duodenum by 50%, in the jejunum by 24%, and in the ileum by 36%; L∑ was decreased in the duodenum by 36%, in the jejunum by 45%, and in the ileum by 52%. At 30 min, SL in the jejunum was increased by 80%. In the HH+sevoflurane group, F was decreased in the duodenum by 38%, in the jejunum by 22%, and in the ileum by 27%; L∑ in the duodenum was decreased by 44%, in the jejunum by 45%, and in the ileum by 67%. After 1 h of HH+sevofluran, SL was increased in the jejunum twofold, in the duodenum by 38% and in the ileum by 15%; F was decreased in the duodenum by 67% and in the jejunum by 43%; L∑ in the duodenum was decreased by 62%, in the jejunum by 60%, and in the ileum by 70% compared to the ether group. After 2 h of HH+sevofluran, SL was increased in the jejunum and ileum by 80% and 67%, respectively, compared to the ether group. In this process, L∑ in the duodenum was decreased by 24% and in the jejunum by 15%. Conclusion. The HH+diethyl ether exposure was associated with activation of FRO processes in the small intestine. The sevoflurane preconditioning provided a significant restriction of oxidative stress in the rat small intestine due to activation of the antioxidant system in the duodenum, jejunum, and ileum at 1 h, 15 min, and 30 min of HH, respectively.

Peptidomics of enteroendocrine cells and characterisation of potential effects of a novel preprogastrin derived-peptide on glucose tolerance in lean mice

ObjectivesTo analyse the peptidomics of mouse enteroendocrine cells (EECs) and human gastrointestinal (GI) tissue and identify novel gut derived peptides. MethodsHigh resolution nano-flow liquid chromatography mass spectrometry (LC–MS/MS) was performed on (i) flow-cytometry purified NeuroD1 positive cells from mouse and homogenised human intestinal biopsies, (ii) supernatants from primary murine intestinal cultures, (iii) intestinal homogenates from mice fed high fat diet. Candidate bioactive peptides were selected on the basis of species conservation, high expression/biosynthesis in EECs and evidence of regulated secretionin vitro. Candidate novel gut-derived peptides were chronically administered to mice to assess effects on food intake and glucose tolerance. ResultsA large number of peptide fragments were identified from human and mouse, including known full-length gut hormones and enzymatic degradation products. EEC-specific peptides were largely from vesicular proteins, particularly prohormones, granins and processing enzymes, of which several exhibited regulated secretion in vitro. No regulated peptides were identified from previously unknown genes. High fat feeding particularly affected the distal colon, resulting in reduced peptide levels from GCG, PYY and INSL5. Of the two candidate novel peptides tested in vivo, a peptide from Chromogranin A (ChgA 435−462a) had no measurable effect, but a progastrin-derived peptide (Gast p59−79), modestly improved glucose tolerance in lean mice. ConclusionLC–MS/MS peptidomic analysis of murine EECs and human GI tissue identified the spectrum of peptides produced by EECs, including a potential novel gut hormone, Gast p59−79, with minor effects on glucose tolerance.

Effects of Cola anomala (K. Schum.) water/ethanol pods extract on the inflammation and intestinal secretion in lipopolysaccharide-induced diarrhea

Diarrhea is one of the leading causes of death among children in low and low-middle income countries and the management of this pathology is still a problem in these regions. The water/ethanol extract of the pods of Cola anomala (KEO) has been shown to possess antimicrobial and antidiarrheal effects in Shigella flexneri-induced diarrhea, but whether KEO is active on the toxemic part of this diarrhea is unknown. This study was undertaken to evaluate the effects of KEO on the intestinal secretion and inflammation induced by intraperitoneal administration of lipopolysaccharide (LPS). KEO obtained by maceration in water/ethanol (1:1) was administered orally (25, 50 and 100 mg/kg of body weight) against LPS-induced diarrhea in mice. The mass of feces, the intestinal nitric oxide (NO) and prostaglandin (PGE2) contents as well as myeloperoxidase (MPO) activity were assessed. KEO was also tested on LPS-induced enteropooling in rats. In this experiment, the intestinal fluid and its electrolytes (Na+, K+ and Cl-) contents were determined as well as NO, PGE2, TNF-α and IL-1β levels in the small intestine homogenate. Indomethacin (5 mg/kg) was used as reference drug. KEO significantly (p &lt; 0.001) reduced stools excretion, NO content and MPO activity in intestine but did not affect PGE2 in LPS-induced diarrhea. On the enteropooling model, KEO showed no effect on the intestinal fluid and electrolyte excretion, PGE2, TNF-α and IL-1β contents, but significantly (p &lt; 0.05) reduced the NO production. This study suggests that KEO does not have antisecretory effect, but has anti inflammatory activities. It can be concluded that the anti-toxemic effect of KEO contributes less to its antidiarrheal activity in infectious diarrhea.

Effect of dexpanthenol on colonic anastomosis healing in rats

The purpose of this study was to perform the clinical, histopathologic and biochemical evaluation of the effects of intraperitoneally (IP) administered dexpanthenol (dxp) on colonic anastomosis healing in rats. The study was conducted on a total of 28 rats divided into four groups comprising seven rats each. Group I was designated as the control group, group II as the dxp group, group III as the anastomosis group, and group IV as the anastomosis + dxp group. The groups were compared in terms of intestinal bursting pressure, adhesion formation, nitric oxide (NO) and malondialdehyde (MDA) levels, total antioxidant capacity (TAC) and total oxidant capacity (TOC) in blood and intestinal homogenates, as well as histopathologic findings. Dxp decreased adhesion formation (6 rats in group III and 4 rats in group IV). Mean bursting pressures were higher in the dxp groups than in the other groups (group II = 254.3 ± 42.1 mmHg, group IV = 109.3 ± 34.5 mmHg). Moreover, there was a remarkable decrease in the levels of NO and MDA and in blood oxidative stress parameters in the dxp groups. The results suggest that dxp increased intestinal bursting pressure by accelerating healing in the anastomosis line and decreased adhesion formation, positively affecting healing. Dexpanthenol, which was found to have positive effects in the experimental rat model, can be introduced into clinical practice.

Comparative In vitro metabolism of purified mogrosides derived from monk fruit extracts

Mogrosides are the primary components responsible for the sweet taste of Monk fruit which is derived from Siraitia grosvenorii (Swingle), a herbaceous plant native to southern China. Many mogrosides have been identified from Monk fruit extract, but the major sweetness component of Monk fruit by mass is mogroside V, comprising up to 0.5% of the dried fruit weight. Recent pharmacokinetic studies indicate that the parent mogrosides undergo minimal systemic absorption following ingestion and hydrolysis by digestive enzymes and/or intestinal flora and are excreted as mogrol (i.e., the aglycone) and its mono- and diglucosides. The objective of this study was to demonstrate whether individual mogrosides, are metabolized to a common and terminal deglycosylated metabolite, mogrol. An in vitro assay was conducted with pooled human male and female intestinal fecal homogenates (HFH) using mogrosides IIIe, mogroside V, siamenoside I, and isomogroside V at two concentrations over a 48 h period. The results show that various mogrosides that differ in the linkages and number of glucose units attached to a common cucurbitane backbone, share a common metabolic fate, and are metabolized within 24 h to mogrol. Aside from an apparent difference in the initial rate of deglycosylation between mogrosides at higher concentrations, no apparent difference in the rate of deglycosylation was observed between the male and female HFH. Given the similar structures of these mogrosides and a shared metabolic fate to mogrol, the study provides support for a reasonably conservative approach to assess safety based on bridging safety data from an individual mogroside (i.e., Mogroside V) to other mogrosides, and the establishment of a group Acceptable Daily Intake (ADI), rather than individual ADI's for mogrosides.

Bacillus subtilis Inhibits Viral Hemorrhagic Septicemia Virus Infection in Olive Flounder (Paralichthys olivaceus) Intestinal Epithelial Cells

Viral hemorrhagic septicemia virus (VHSV) is a highly pathogenic virus that infects a wide range of host fish species causing high economic losses in aquaculture. Epithelial cells in mucosal organs are target sites for VHSV entry into fish. To protect fish against VHSV infection, there is a need to develop antiviral compounds able to prevent establishment of infection at portals of virus entry into fish. Bacillus subtilis is a probiotic with excellent antiviral properties, of which one of its secretions, surfactin, has been shown to inhibit viral infections in mammals. Herein, we demonstrate its ability to prevent VHSV infection in olive flounder (Paralichthys olivaceus) intestinal epithelial cells (IECs) and infection in internal organs. Our findings show inhibition of VHSV infection in IECs by B. subtilis and surfactin. In addition, our findings showed inhibition of VHSV in Epithelioma Papulosum Cyprini (EPC) cells inoculated with intestinal homogenates from the fish pretreated with B. subtilis by oral exposure, while the untreated fish had cytopathic effects (CPE) caused by VHSV infection in the intestines at 48 h after the VHSV challenge. At 96 h post-challenge, samples from the untreated fish had CPE from head kidney and spleen homogenates and no CPE were observed in the intestinal homogenates, while the B. subtilis-pretreated fish had no CPE in all organs. These findings demonstrate that inhibition of VHSV infection at portals of virus entry in the intestines culminated in prevention of infection in internal organs. In summary, our results show that B. subtilis has the potential to prevent VHSV infection in fish and that its use as a probiotic in aquaculture has the potential to serve as an antiviral therapeutic agent against different viral infections.

Evaluation of crude adult Ascaris suum intestinal tract homogenate in inducing protective IgG production against A. suum larvae in BALB/c mice

Globally, ascariasis ranks as the second leading intestinal helminth infection. However, progress in developing better control strategies, such as vaccines, remains slow-paced. This study aims to measure antibody production and parasite load in male BALB/c mice immunized with crude Ascaris suum intestinal tract homogenate.Thirty-two (32) mice were randomized into: (1) unvaccinated, uninfected (UU); (2) unvaccinated, infected (UI); (3) vaccinated, uninfected (VU); and (4) vaccinated, infected (VI) groups. A 100-μL vaccine containing 50 μg of homogenized A. suum intestines and Complete Freund's Adjuvant (1:1) were introduced intraperitoneally. Immunizations were done on days 0, 10, and 20. Oral gavage with 1000 embryonated eggs was done on day 30. Blood was obtained at day 40. To measure serum IgG levels, indirect ELISA was done. Microtiter plates were coated with 100 μg larval homogenate, and HRP-conjugated anti-mouse IgG was used as secondary antibody. Parasite load was measured in lung and liver tissues.Tukey's HSD of signal to cut-off ratios of absorbance readings obtained in indirect ELISA procedure for the 1:200 serum dilution showed statistically significant difference between the UU and VI (p = 0.026) as well as between UI and VI (p = 0.003) groups. No statistically significant difference in parasite load was observed in the lungs (p = 0.074), liver (p = 0.130), and both lungs and liver (p = 0.101).Immunization elicited a significant larva-directed IgG production. However, there is no significant difference in parasite loads in either lung or liver tissues across all treatment groups as the larval counts obtained from the study were very low and may not be indicative of the actual parasite load in mice.

Oxidative stress mediated apoptotic potential of mefloquine on experimental trichinellosis

Conventional anthelmintics such as albendazole could not achieve complete cure of trichinellosis till now. The antimalarial mefloquine mediates oxidative stress and disrupts lysosomal functions leading to cell death. Therefore, the aim of this work was to investigate the effect of mefloquine on experimental acute and chronic trichinellosis and to clarify the possible mechanisms of such effects. Mice were divided into four groups; Group I: Uninfected untreated control (20 mice); Group II: Infected untreated control (40 mice); Group III: infected and treated with albendazole (400 mg/kg) (40 mice); Group IV: infected and treated with mefloquine (300 mg/kg) (40 mice). All infected treated groups were equally subdivided into 2 subgroups; (a) treated on the 2nd day post infection (dpi) for 3 days, (b) treated on the 35th dpi for 5 days. Parasitological adults and larvae counting besides immunohistopathological examination of intestines and muscles were done. Biochemical assay of oxidant/antioxidant status, apoptotic, cytoprotective and inflammatory biomarkers in intestinal and muscle homogenates were achieved. Results showed that both albendazole and mefloquine significantly reduced adults and larvae counts with higher efficacy of albendazole in the intestinal phase and superiority of mefloquine in the muscle phase. The superiority of mefloquine was indicated by increased inflammatory immune infiltration and decreased anti-apoptotic immunohistochemical markers expression in both jejunal and muscle tissues. Biochemically, mefloquine treatment showed highly significant oxidative, apoptotic and inflammatory effects. So, our results suggest that mefloquine might be a superior treatment for chronic trichinellosis.

High-Fat Diet Induces Disruption of the Tight Junction-Mediated Paracellular Barrier in the Proximal Small Intestine Before the Onset of Type 2 Diabetes and Endotoxemia.

A link between an impaired intestinal barrier, endotoxemia, and the pathogenesis of metabolic diseases, such as type 2 diabetes mellitus (T2DM), has been proposed. In previous work, we have demonstrated that the tight junction (TJ)-mediated intestinal barrier in ileum/colon was marginally changed in prediabetic mice; therefore, it does not seem to mainly contribute to the T2DM onset. In this study, the TJ-mediated epithelial barrier in the duodenum and jejunum was evaluated in mice during the development of type 2 prediabetes. HF diet induced prediabetes after 60days associated with a significant rise in intestinal permeability to the small-sized marker Lucifer yellow in these mice, with no histological signs of mucosal inflammation or rupture of the proximal intestine epithelium. As revealed by immunofluorescence, TJ proteins, such as claudins-1, -2, -3, and ZO-1, showed a significant decrease in junctional content in duodenum and jejunum epithelia, already after 15days of treatment, suggesting a rearrangement of the TJ structure. However, no significant change in total cell content of these proteins was observed in intestinal epithelium homogenates, as assessed by immunoblotting. Despite the changes in intestinal permeability and TJ structure, the prediabetic mice showed similar LPS, zonulin, and TNF-α levels in plasma or adipose tissue, and in intestinal segments as compared to the controls. Disruption of the TJ-mediated paracellular barrier in the duodenum and jejunum is an early event in prediabetes development, which occurs in the absence of detectable endotoxemia/inflammation and may contribute to the HF diet-induced increase in intestinal permeability.

Hepatoprotective Bile Acid Co-Drug of Isoniazid: Synthesis, Kinetics and Investigation of Antimycobacterial Potential

The long-course treatment of tuberculosis with isoniazid (INH) leads to hazardous side effects on liver and poor patient compliance. To overcome these toxic effects caused by INH, a unique hepatoprotective co-drug platform was developed by tethering INH with ursodeoxycholic acid (UDCA) – an antioxidant bile acid for possible synergistic outcome. INH and UDCA were linked into co-drug (UI ) through a bioreversible amide linkage by EDCI coupling. UI resisted hydrolysis in acidic (pH 1.2) and basic (pH 7.4) buffers and stomach homogenate of rat, whereas exhibited significant hydrolysis (83.38%) in intestinal homogenate over a period of 6 h. The effect of UI in attenuating oxidative stress and reinstating the normal physiology of liver was striking as it restored the levels of glutathione peroxidase and superoxide dismutase enzymes to normal. Result obtained for antimycobacterial activity assessment clearly demonstrated that UI was as potent as INH in lowering the mycobacterial load in mice. The outcomes of this exploration confirm that the proposed prodrug can add safety as well as efficiency to future medical procedures of tuberculosis management.

Phytochemical Characterization and In vitro Effects of Extracts Produced from Different Maytenus ilicifolia Matrices on the Activity of Intestinal Disaccharidases

Introduction: Maytenus ilicifolia Mart. Ex Reiss, Celastraceae, popularly known as “espinheira-santa” is traditionally used to treat gastrointestinal disorders and diabetes. However, studies proving efficacy for the treatment of diabetes are scarce. Furthermore, it is believed that the presence of chemical constituents responsible for pharmacological activity may be affected by environmental variations. Thus, the objective of this research was to evaluate the occurrence of variations in chemical composition, total polyphenol content, total tannin, antioxidant and antidiabetic activity in vitro for different matrices of M. ilicifolia.&#x0D; Methodology: Chemical characterization was determined by CG-MS. Total polyphenol and total tannin contents were determined by spectrophotometer readings using standard gallic acid and tannic acid curves, respectively. In vitro antioxidant potential was determined by reducing the DPPH radical. In vitro antidiabetic activity was determined by inhibiting intestinal disaccharidases (maltase, sucrase and lactase) from a commercial glucose measurement kit produced by incubating intestinal homogenates with their substrates.&#x0D; Results and Discussion: The results indicated the presence of variations in the chemical constituents and their concentrations, the total polyphenol content, total tannins and the in vitro antioxidant activity among the different tested extracts of M. ilicifolia. It is believed that these variations may be responsible for the differences found in inhibition of disaccharidases for the three intestinal enzymes.&#x0D; Conclusion: Extracts 116 and 122 showed the best results in disaccharidase inhibition, however further studies are needed to investigate the results and reproducibility in vivo.

Development and validation of an LC-MS/MS method for the quantitative analysis of milciclib in human and mouse plasma, mouse tissue homogenates and tissue culture medium

Milciclib is a promising cyclin-dependent kinase inhibitor currently in phase II clinical trials to treat several types of cancer. The first bioanalytical method for the quantitative analysis of milciclib in several biomatrices using liquid chromatography-tandem mass spectrometry is described here. This method was fully validated in human plasma according to FDA and EMA guidelines, and partially validated in mouse plasma, homogenates of mouse brain, kidney, liver, small intestine, spleen, and tissue culture medium. Palbociclib, an analog compound, was used as internal standard. A simple and fast sample pre-treatment by protein precipitation with acetonitrile was used, leading to efficient extraction of the analyte with recoveries between 95–100%. Chromatographic separation was achieved with a C18 analytical column and a gradient elution using 10 mM ammonium bicarbonate in water and 10 mM ammonium bicarbonate in water-methanol (1:9, v/v). This assay was selective, accurate, precise and linear in the concentration range of 1−1000 ng/mL. Moreover, samples above the upper limit of quantification can be integrally diluted up to 10-fold prior to analysis. The use of human plasma as a surrogate matrix to quantify milciclib in tissue culture medium and mouse matrices resulted in acceptable accuracy and precision, however tissue culture medium samples required a dilution with human plasma prior the pre-treatment. All performance parameters of the method complied with the acceptance criteria recommended by the guidelines, except for the carry-over, which was slightly above (22.9% of the lower limit of quantification) the recommended percentage (20%). Therefore, additional measures were taken to assure data integrity. Stability of milciclib in all matrices was evaluated, and in some matrices the analyte was unstable under the tested conditions. It is therefore recommended to keep these samples as briefly as possible at room temperature during the pre-treatment, and to store them at –70 °C to avoid analyte degradation. This method was successfully applied to support preclinical pharmacokinetic studies of milciclib.

Rat intestinal homogenate and pancreatic juice can induce the Z-isomerization of all-E-lycopene in vitro

Lycopene is one of the carotenoids often consumed by humans in their diet. Although lycopene exists mainly in the form of the all E-isomer in foods, the considerable quantity of its Z-isomers is found in the human plasma and liver. This observation suggested that the lycopene all-E-isomer was converted into Z-isomers in the human body. In this study, the Z-isomerization of the all-E-isomer was induced in vitro by the pancreatic juice and small intestinal homogenate of male rats under the conditions of 37 °C, pH = 7.5, nitrogen and darkness, as well as shaking. After 2 hours, the proportion of the all E-isomer decreased to 25% and Z-isomer amounts increased relatively. The converted products were identified as 5, 9, and 11 Z-isomers by electronic absorption spectroscopy and mass spectrometry (MS). The observations from this experiment suggested that the Z-isomerization site of the lycopene all E-isomer was located in the small intestinal wall of the rat.