Histopathological and biochemical changes of morphine sulphate administration on the cerebellum of albino rats

In this study the long-term effects of morphine sulphate treatment (MST) on histopathological and biochemical changes in the cerebellum was assessed in albino rats. Normal saline (5 ml) was given orally as placebo in the control group (n = 25). Morphine groups received morphine orally at a dose level of 5 mg/kg body weight day after day for 10, 20 and 30 days (n = 25/group). Light microscopy revealed that the molecular layer showed vacuolation. The Purkinje cells lost their specific shaped appearance, decreased in size and numbers. The granular cells highly degenerated. Electron microscopy revealed fragmentation of the cisterns of the both types of endoplasmic reticulum, resulted in a progressive depletion of total protein contents as well as general carbohydrates in all treated groups as supported by histochemical observation. Obvious destruction of mitochondrial inner membrane and cristae mediate cell death. Also, abnormal nucleus with deformed perforated nuclear membrane and deformation of the plasma membrane with degeneration of the synapses could interpreted as a sign of necrosis. Biochemical analysis revealed that dopamine (DA) and norepinephrine (NE) were significantly decreased in four brain areas (cortex striatum, thalamus/hypothalamus, and cerebellum). In contrast, serotonin (5-HT) level was increased in these brain regions; with an exception of 5-HT on day 10 and neurotransmitter levels in the pons were unaffected. The quantitative analysis showed a significant decrease (P < 0.05) in the diameter of Purkinje cells and in the thickness of both molecular and granular layers treated groups. Morphine sulphate induces may be a cell death or necrosis in the rat cerebellum and modulating neurotransmitter system. Our findings pointed out the risk of increased cerebellum damage due to long-term of morphine use.     

Spilanthes acmella ethanolic flower extract: LC-MS alkylamide profiling and its effects on sexual behavior in male rats

According to Indian Systems of Medicine, Spilanthes acmella (L.) Murr. (Family - Asteraceae), is considered effective in the treatment of sexual deficiencies especially due to ageing. In the present study, characterization of ethanolic extracts of the Spilanthes acmella flower and its effect on general mating pattern, penile erection and serum hormone levels of normal male Wistar albino rats were investigated and compared with sildenafil citrate. In vitro nitric oxide release was also investigated in human corpus cavernosum cell line. As N-alkylamides are a promising group, their profiling was performed using a gradient reversed phase high performance liquid chromatography/electrospray ionization ion trap mass spectrometry (HPLC/ESI-MS) method on an embedded polar column. MS¹ and MS² fragmentation data were used for identification purposes. For assessment of sexual behavior, animals were divided into five groups of eight male rats. The extracts (50, 100 and 150 mg/kg body weight/day) and sildenafil citrate (5 mg/kg body weight/day) (positive control) were administered orally for 28 days. The behavioral and sexual parameters were observed at days 0, 15, 28 and after a lapse of 7 and 14 days of discontinuance of drug treatment. Five N-isobutylamides, one 2-methylbutylamide and one 2-phenylethylamide were identified. The orally administered extract had a dose dependent positive effect on mounting frequency, intromission frequency and ejaculation frequency and the most significant effects (p < 0.05) were observed at 150 mg/kg treatment, even after a lapse of 7 and 14 days of discontinuance of drug treatment. A dose dependent effect was also observed on the FSH, LH and testosterone serum levels. With 150 mg/kg of ethanolic extract the values for FSH, LH and testosterone were 3.10 ± 0.25 mlU/ml, 6.87 ± 0.18 mlU/ml and 3.72 ± 0.12 ng/ml, respectively. In vitro nitric oxide release was 21.7 ± 2.9 μM, which was significantly higher compared to the control group (p < 0.01). Sildenafil citrate exhibited also a significant effect on NO release, but no effect on hormone levels of rats was observed. The aphrodisiac potential of an ethanolic Spilanthes acmella extract was demonstrated in vitro and in vivo. N-Alkylamides might attribute to the improved sexual potential. Study lends support to the traditional utilization of S. acmella as a sexual stimulating agent.     

Operculina turpethum attenuates N-nitrosodimethylamine induced toxic liver injury and clastogenicity in rats

The root extract of Operculina turpethum (OTE) has been used as an anti-inflammatory, purgative, and hepato-protective agent. N-Nitrosodimethylamine (NDMA) is a potent hepatotoxin that induces fibrosis of the liver. In the present study, we examined the therapeutic effects of OTE root extract against NDMA-induced hepatotoxicity and clastogenicity in rats. Hepatic fibrosis was induced in adult male albino rats through serial intraperitoneal administrations of NDMA at a concentration of 10 mg/kg body weight on three consecutive days of each week over a period of three weeks. A group of rats received OTE orally in doses of 75, 150 and 200 mg/kg body weight at 5 h after the administration of NDMA. The controls and treated animals were sacrificed on days-7, 14 and 21 after the start of the administration of NDMA. The progression of hepatic fibrosis as well as the amelioration effect of OTE was evaluated through histopathologically as well as by immunohistochemical staining for the activation of hepatic stellate cells. Alterations in serum and liver biochemical parameters and LDH isoenzymes were also studied. Serial administration of NDMA resulted in well formed fibrosis in the liver and induction of micronuclei in the bone marrow cells. Staining of α-SMA demonstrated activated stellate cells from day-7 onwards which was dramatically increased on day-21. An elevation of micronuclei count, liver function enzymes, serum hydroxyproline levels and LDH isoenzymes 4 and 5 were also observed. All these changes were remarkably reduced in OTE administered animals and fibrogenesis was completely absent. Our results suggest that OTE has hepatoprotective and anti-clastogenic effects against NDMA-induced hepatic fibrosis. Therefore OTE may be used as a hepatoprotective agent against various liver diseases including toxic liver injury.     

Hydrogen-rich saline reduces lung injury induced by intestinal ischemia/reperfusion in rats

Objective. Hydrogen has been reported to selectively reduce the hydroxyl radical, the most cytotoxic of reactive oxygen species. In this study we investigated the effects of hydrogen-rich saline on the prevention of lung injury induced by intestinal ischemia/reperfusion (I/R) in rats. Methods. Male Sprague-Dawley rats (n = 30, 200-220 g) were divided randomly into three experimental groups: sham operated, intestinal I/R plus saline treatment (5 ml/kg, i.v.), and intestinal I/R plus hydrogen-rich saline treatment (5 ml/kg, i.v.) groups. Intestinal I/R was produced by 90 min of intestinal ischemia followed by a 4 h of reperfusion. Results. Hydrogen-rich saline treatment decreased the neutrophil infiltration, the lipid membrane peroxidation, NF-κB activation and the pro-inflammatory cytokine interleukin IL-1β and TNF-α in the lung tissues compared with those in saline-treated rat. Conclusion. Hydrogen-rich saline attenuates lung injury induced by intestinal I/R.     

Accumulation and excretion of morphine by Calliphora stygia, an Australian blow fly species of forensic importance

This study examined the ability of the forensically important blow fly, Calliphora stygia to actively excrete morphine, thereby maintaining a low morphine level within its body when fed on a diet containing morphine at low (7 pmol g⁻¹) and high (17.5 pmol g⁻¹) concentrations. Morphine was accumulated within the bodies of maggots (≈70% within the tissues) at concentrations which were lower than that of the meat (3-24%). The morphine content of the initial developing stages (second and third instar maggots) maintained on the high morphine diet was higher than those on the low morphine diet. Morphine was cleared from the body with negatively exponential kinetics (High morphine group: Morphine (pmol g⁻¹ wet weight) = 8425e^−0.014t. Low morphine group: Morphine (pmol g⁻¹ wet weight) = 2180e^−0.010t). Clearance constants for morphine by animals in both groups were similar and thus both groups had a similar ability to excrete morphine. The Malpighian tubules of maggots were able to actively secrete morphine using a transport mechanism that transports small type II organic cations, such as morphine and quinine. The rate of morphine secretion by the Malpighian tubules could explain the clearance of the drug by the maggots. As the morphine was transported across the Malpighian tubules cells, a significant proportion was metabolised into a compound that is yet to be fully characterised.     

Effect of ceramide N-acyl chain and polar headgroup structure on the properties of ordered lipid domains (lipid rafts)

Ceramides are sphingolipids that greatly stabilize ordered membrane domains (lipid rafts), and displace cholesterol from them. Ceramide-rich rafts have been implicated in diverse biological processes. Because ceramide analogues have been useful for probing the biological function of ceramide, and may have biomedical applications, it is important to characterize how ceramide structure affects membrane properties, including lipid raft stability and composition. In this report, fluorescence quenching assays were used to evaluate the effect of analogues of ceramide with different N-acyl chains or different sphingoid backbones on raft stability and sterol content. The effect of replacing 18 mol% of sphingomyelin (SM) with ceramide in vesicles composed of a 1:1 (mol:mol) mixture of SM and dioleoylphosphatidylcholine (DOPC), with or without 25 mol% sterol, was examined. In the absence of sterol, the thermal stability of the SM-rich ordered domains increased with ceramide N-acyl chain length in the order C2:0 ∼ C6:0 ∼ C8:0 < no ceramide < C12:0 < C16:0. In vesicles containing 25 mol% cholesterol (1:1:0.66 sphingolipid:DOPC:cholesterol), the dependence of raft stability on ceramide N-acyl chain length increased in the order C8:0 ∼ C6:0 < C2:0 < C12:0 ∼ no ceramide < C16:0. We also studied the stability of lipid rafts in the presence of N-lauroyl- and N-palmitoylsphingosine analogues containing altered structures in or near the polar portion of the sphingoid base. In almost all cases, the analogues stabilized rafts to about the same degree as a normal ceramide containing the same acyl chain. The only exception was N-palmitoyl-4D-ribophytosphingosine, which was very strongly raft-stabilizing. We conclude that variations in sphingoid base structure induce only insignificant changes in raft properties. N-Lauroyl and N-palmitoylsphingosine and their analogues displaced sterol from rafts to a significant degree. Both C12:0 and C16:0 analogues of ceramide may be good mimics of natural ceramide, and useful for cellular studies in which maintenance of the normal physical properties of ceramide are important.     

Morphine glucuronidation increases its analgesic effect in guinea pigs

Aims

Morphine is extensively metabolized to neurotoxic morphine-3-glucuronide (M3G) and opioid agonist morphine-6-glucuronide (M6G). Due to these different roles, interindividual variability and co-administration of drugs that interfere with metabolism may affect analgesia. The aim of the study was to investigate the repercussions of administration of an inducer (2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD) and an inhibitor (ranitidine) of glucuronidation in morphine metabolism and consequent analgesia, using the Guinea pig as a suitable model.

Main methods

Thirty male Dunkin–Hartley guinea pigs were divided in six groups: control, morphine, ranitidine, ranitidine + morphine, TCDD and TCDD + morphine. After previous exposure to TCDD and ranitidine, morphine effect was assessed by an increasing temperature hotplate (35–52.5 °C), during 60 min after morphine administration. Then, blood was collected and plasma morphine and metabolites were quantified.

Key findings

Animals treated with TCDD presented faster analgesic effect and 75% reached the cut-off temperature of 52.5 °C, comparing with only 25% in morphine group. Animals treated with ranitidine presented a significantly lower analgesic effect, compared with morphine group (p < 0.05). Moreover, significant differences between groups were found in M3G levels and M3G/morphine ratio (p < 0.001 and p < 0.0001), with TCDD animals presenting the highest values for M3G, M6G, M3G/morphine and M6G/morphine, and the lowest value for morphine. The opposite was observed in the animals treated with ranitidine.

Significance

Our results indicate that modulation of morphine metabolism may result in variations in metabolite concentrations, leading to different analgesic responses to morphine, in an animal model that may be used to improve morphine effect in clinical practice.     

Liver and kidney toxicity in chronic use of opioids: An experimental long term treatment model

In this study, histopathological and biochemical changes due to chronic usage of morphine or tramadol in liver and kidney were assessed in rats. Thirty male Wistar rats (180–220 g) were included and divided into three groups. Normal saline (1 ml) was given intraperitoneally as placebo in the control group (n = 10). Morphine group (n = 10) received morphine intraperitoneally at a dose of 4, 8, 10 mg/kg/day in the first, second and the third ten days of the study, respectively. Tramadol group (n = 10), received the drug intraperitoneally at doses of 20, 40 and 80 mg/kg/day in the first, second and the third ten days of the study, respectively. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), creatinin, blood urea nitrogen (BUN) and malondialdehyde (MDA) levels were measured in the serum. Liver and kidney specimens were evaluated by light microscopy. Serum ALT, AST, LDH, BUN and creatinin levels were significantly higher in morphine group compared to the control group. Serum LDH, BUN and creatinin levels were significantly increased in the morphine group compared to the tramadol group. The mean MDA level was significantly higher in morphine group compared to the tramadol and control groups (P<0.05). Light microscopy revealed severe centrolobular congestion and focal necrosis in the liver of morphine and tramadol groups, but perivenular necrosis was present only in the morphine group. The main histopathologic finding was vacuolization in tubular cells in morphine and tramadol groups. Our findings pointed out the risk of increased lipid peroxidation, hepatic and renal damage due to long term use of opioids, especially morphine. Although opioids are reported to be effective in pain management, their toxic effects should be kept in mind during chronic usage Presented at the 10th XX Annual ESRA Congress, 6–9 April 2002, Warsaw, Poland.     

Toxoplasma gondii: A morphometric analysis of the wall and epithelial cells of pigs intestine

The aim of this study was to perform a morphometric analysis of the different layers of the jejunal wall and epithelial cells of pigs with toxoplasmosis. Experiments were conducted using 10, 88-day-old crossbred (Pietran × Wessex) pigs divided into two groups: control (n = 5) and experimental (n = 5). The experimental group consisted of animals inoculated orally with 5000 sporulated oocysts of a genotype III strain of Toxoplasma gondii. At 30 and 60 days following inoculation, the animals were anaesthetised for jejunal biopsy. The intestinal segments were processed routinely for histology. Transverse cuts (4 μm thick) were stained with haematoxylin and eosin (HE), Periodic Acid Schiff (PAS), Alcian Blue (AB), pH 2.5, and Alcian Blue (AB), pH 1.0. We observed hypertrophy of the jejunal wall, increased enterocyte height, and a decreased number of intraepithelial lymphocytes in the infected animals. There were no changes in the number of goblet cells.     

Beta-glucan from Saccharomyces cerevisiae reduces plasma lipid peroxidation induced by haloperidol

Since oxidative stress observed in schizophrenia may be caused partially by the treatment of patients with various antipsychotics, the aim of the study was to establish the effects of beta-d-glucan, polysaccharide derived from the yeast cell walls of species such as Saccharomyces cerevisiae, and the antipsychotics (the first generation antipsychotic (FGA) - haloperidol and the second generation antipsychotic (SGA) - amisulpride) action on plasma lipid peroxidation in vitro. Lipid peroxidation in human plasma was measured by the level of thiobarbituric acid reactive species (TBARS). The samples of plasma from healthy subjects were incubated with haloperidol or amisulpride in the presence of beta-glucan (4 μg/ml). The action of beta-d-glucan was also compared with the properties of a well characterized commercial monomeric polyphenol - resveratrol (3,4′,5-trihydroxystilbene, the final concentration - 4 μg/ml). The two-way analysis variance showed that the differences in TBARS levels were depended on the type of tested drugs (p = 7.9 × 10⁻⁶). We observed a statistically increase of the level of biomarker of lipid peroxidation such as TBARS after 1 and 24 h incubation of plasma with haloperidol compared to the control samples (p < 0.01, p < 0.02, respectively). Amisulpride, contrary to haloperidol (after 1 and 24 h) did not cause plasma lipid peroxidation (p > 0.05). We showed that in the presence of beta-glucan, lipid peroxidation in plasma samples treated with haloperidol was significantly decreased. Moreover, we did not observe the synergistic action of beta-glucan and amisulpride on the inhibition of plasma lipid peroxidation. However, the beta-d-glucan was found to be more effective antioxidant, than the solution of pure resveratrol. The presented results indicate that beta-glucan seems to have distinctly protective effects against the impairment of plasma lipid molecules induced by haloperidol.     

An automated microfluidic-based immunoassay cartridge for allergen screening and other multiplexed assays

A microfluidic cartridge and system for multiplexed immunoassays is described. The passive microfluidic cartridge was composed of three layers of injection molded plastic sealed together using a thermal staking technique. Using this platform technology, a specific immunoglobulin E (IgE) panel assay was constructed. Allergen extract targets, positive and negative controls, and IgE calibration standards were immobilized within the cartridge as a microarray. A computer-controlled solenoid array provided the necessary actuation force for pumping reagents within the cartridge to perform an automated, chemiluminescent indirect immunoassay. A 20-target allergen extract panel was demonstrated on the device with a total analysis time of 27 min. Allergen screening results showed 84% agreement for 3 house dust mites (N = 300) compared with a commercial test and 80% agreement overall (N = 978). Average coefficients of variation (N = 80) were measured as 20.5% for low/medium levels and 20.4% for medium/high levels. The average limit of detection (N = 160) was measured at 0.535 AU, and cutoff levels of 1.0 AU were estimated at less than 1 IU/ml (2.4 ng/ml). Such a system has potential applications in decentralized allergen screening as well as in other near-patient diagnostic immunoassays where multiplexed analysis, ease of use, and short analysis time are critical.     

An investigation of the inclusion complex of cyclomaltoheptaose (β-cyclodextrin) with N-methylanthranilic acid in the solid state

A 2:1 complex between cyclomaltoheptaose (β-cyclodextrin) and N-methylanthranilic acid has been studied in the solid state. The inclusion complex belongs to the triclinic system (space group P1) with unit cell dimensions a = 15.2773(15) Å, b = 15.4710(15) Å, c = 17.9627(18) Å, α = 99.632(5)°, β = 113.416(5)°, and γ = 102.818(5)°. The complex forms a head-to-head channel-type structure with the N-methylanthranilic acid lying between the β-cyclodextrin groups in a sandwich fashion, which is held in place by an extensive hydrogen-bonding network between the cyclodextrin molecules.     

Effect of prenatal androgens on click-evoked otoacoustic emissions in male and female sheep (Ovis aries)

Otoacoustic emissions (OAEs) were measured in male and female Suffolk sheep (Ovis aries). Some sheep had been administered androgens or estrogens during prenatal development, some were gonadectomized after birth, and some were allowed to develop normally. As previously reported for spotted hyenas, gonadectomy did not alter the OAEs for either sex; accordingly, the untreated/intact and the untreated/gonadectomized animals were pooled to form the control groups. The click-evoked otoacoustic emissions (CEOAEs) exhibited by the female control group (N = 12) were slightly stronger (effect size = 0.42) than those in the male control group (N = 15), which is the same direction of effect reported for humans and rhesus monkeys. Females administered testosterone prenatally (N = 16) had substantially weaker (masculinized) CEOAEs than control females (effect size = 1.15). Both of these outcomes are in accord with the idea that prenatal exposure to androgens weakens the cochlear mechanisms that underlie the production of OAEs. The CEOAEs of males administered testosterone prenatally (N = 5) were not different from those of control males, an outcome also seen in similarly treated rhesus monkeys. Males administered dihydrotestosterone (DHT) prenatally (N = 3) had slightly stronger (hypo-masculinized) CEOAEs than control males. No spontaneous otoacoustic emissions (SOAEs) were found in any ears, a common finding in non-human species. To our knowledge, this is the first ruminant species measured for OAEs.     

The end product of transglutaminase crosslinking: Simultaneous quantitation of [N-(γ-glutamyl) lysine] and lysine by HPLC-MS

Transglutaminases catalyze the formation of N^ε-(γ-glutamyl) isodipeptide crosslinks between proteins. These enzymes are thought to participate in a number of diseases, including neurological disease and cancer. A method associating liquid chromatography and multiple stage mass spectrometry has been developed for the simultaneous quantitation of [N^ε-(γ-glutamyl) lysine] isodipeptide and lysine on an ion trap mass spectrometer. Highly specific detection has been achieved in MS³ mode. The method includes a derivatization step consisting of butylation of carboxylic groups and acetylation of amide groups, a liquid-liquid extraction, and a 19-min separation on a 100 × 2.1-mm Beta-basic C₁₈ column with an acetonitrile gradient elution. ¹³C₆-¹⁵N₂ isotopes of the isodipeptide and the lysine serve as internal standards. The assay was linear in the range of 50 pmol/ml to 75 nmol/ml for the isodipeptide and the range of 10 nmol/ml to 3.5 μmol/ml for the lysine, with correlation coefficients greater than 0.99 for both ions. Intra- and inter-day coefficients of variation ranged from 3.5 to 15.9%. The method was successfully applied to human biological samples known to be crosslinked by transglutaminase such as cornified envelopes of epidermis, fibrin, and normal and Huntington disease brain.     

The hepatocyte phase of Gd-EOB-DTPA-enhanced MRI in the evaluation of hepatic fibrosis and early liver cirrhosis in a rat model: An experimental study

Aims

To evaluate the hepatocyte phase of Gd-EOB-DTPA-enhanced MRI in the early diagnosis of hepatic fibrosis and cirrhosis and assessment of liver function in a rat model.

Main methods

In 2 groups of SD rats, liver fibrosis was induced in experimental animals by repetitive carbon tetrachloride injections, while the control group received saline injections. Five experimental rats and 2 control rats were randomly selected at weeks 4, 8, 12. One week after carbon tetrachloride administration, MRI (FIRM T1WI) scan was performed. Gd-EOB-DTPA (0.08 mL) was injected into the rat's tail vein and hepatocyte phase images were obtained after 20 min. The pre-enhanced phase and hepatocyte phase signal intensities (SI) were measured, and the relative contrast enhancement index (RCEI) was calculated. ANOVA analysis (LSD) of RCEI values in controls (n = 6), hepatic fibrosis (n = 7), and histopathologically-determined early cirrhosis group (n = 6) was performed.

Key findings

RECI values showed a decreasing trend in the control group, hepatic fibrosis and early cirrhosis groups (1.11 ± 0.43, 0.96 ± 0.22, and 0.57 ± 0.33, respectively). While the difference between the control and early cirrhosis groups was statistically significant (p = 0.013), there was no significant difference in the hepatic fibrosis group vs the control (p = 0.416) and the hepatic fibrosis group vs the early cirrhosis group (p = 0.054).

Significance

Hepatocyte phase RCEI values obtained with Gd-EOB-DTPA-enhanced MRI scan indicate liver injury in hepatic fibrosis and early cirrhosis. RCEI values are helpful for early diagnosis of liver cirrhosis.     

Cytoarchitectural alterations in kidney of Wistar rat after oral exposure to cadmium chloride

Male Wistar rats were randomly divided into three groups - A, B and C. A dose of 5 mg and 10 mg of cadmium chloride/kg body weight/day was orally administered to groups B and C, respectively. Rats from group A served as control. Rats were sacrificed on 1st, 2nd, 4th, 6th, and 8th week after initiation of the experiment. Kidneys were removed immediately, fixed in Bouin's fixative, routinely processed and stained with hematoxylin and eosin. The present study showed that the histopathological changes were caused in kidney of rats by cadmium exposure. The changes noticed were mainly - the glomerular swelling (at initial stage), the shrinkage of glomerulus (at later stage), the tubular dilatation, hypertrophy of tubular epithelium, degeneration of glomerulus and renal tubules and deposition of eosin-positive substances in the glomerulus and renal tubules. However, lesions were depended upon the doses and duration of the treatment.     

PPAR’s and Diosgenin a chemico biological insight in NIDDM

Diosgenin a steroidal saponin found widely in nature is reported to contain several biological activities in recent years. The present work elaborates the modulation of the lipid and antioxidant profile by Diosgenin in diabetic condition. Type 2 diabetes was induced in experimental animals by feeding high fat diet (HFD) for 8 weeks followed by streptozotocin (STZ) injection (sub-diabetogenic dose; 35 mg/kg body weight). Diosgenin administered orally at two doses (40 and 80 mg/kg body weight) for 14 days reduced hyperglycemia, hypercholesterolemia and hypertriglyceridemia (p < 0.001). Oxidative stress a crucial marker of diabetes and obesity associated complications was analyzed and noteworthy changes were observed. Improved levels of the antioxidant enzymes SOD and GPx and a minimized level of lipid peroxidation were also observed in Diosgenin treated rats. Further, analyzing the lipid accumulation by Oil Red O staining in 3T3-L1 preadipocytes confirmed its adipogenic activity which was influenced by PPAR γ and PPAR α. This was also substantiated through docking studies of Diosgenin with the PPARs. Altogether, Diosgenin a phytochemical of natural origin is found to mitigate diabetes induced oxidative stress and dyslipidemia which is crucial in cardio-metabolic risks by modulating the PPARs.     

Regional variation in adipogenesis and IGF regulatory proteins in the fetal baboon

Intrauterine growth rate is associated with body distribution in adulthood suggesting differential response of fetal fat depots to nutritional modifications. We hypothesize that there is regional differences in fetal adipogenesis, in part, due to depot-specific regulation of the availability of insulin growth factors. In near-term baboon fetuses (n = 3-5), the subcutaneous abdominal vs. omental preadipocytes had (1) more extensive lipid accumulation as assessed by BODIPY (lipid staining) to DAPI (nuclei) absorbance ratios (mean ± SEM; 0.51 ± 0.21, 0.35 ± 0.09, p < 0.05), (2) lower (p < 0.05) secretion of IGF-binding protein 4 (9.6 ± 1.2 vs. 17.4 ± 2.8 ng/ml) and its protease pregnancy associated plasma protein A (24.6 ± 1.9 vs. 39.1 ± 6.3 μIU/ml), (3) lower protein expression of IGF2 “clearance” receptor in cell lysate (0.28 ± 0.03 vs. 0.53 ± 0.02 OD U/mm², p < 0.05); all variables were intermediate in femoral preadipocytes. The regional variation of the adipogenesis and the IGF regulatory pathway set the stage for differential responsiveness of fat depots to external signals.     

Olive (Olea europaea) leaf extract effective in patients with stage-1 hypertension: Comparison with Captopril

A double-blind, randomized, parallel and active-controlled clinical study was conducted to evaluate the anti-hypertensive effect as well as the tolerability of Olive leaf extract in comparison with Captopril in patients with stage-1 hypertension. Additionally, this study also investigated the hypolipidemic effects of Olive leaf extract in such patients. It consisted of a run-in period of 4 weeks continued subsequently by an 8-week treatment period. Olive (Olea europaea L.) leaf extract (EFLA^®943) was given orally at the dose of 500 mg twice daily in a flat-dose manner throughout the 8 weeks. Captopril was given at the dosage regimen of 12.5 mg twice daily at start. After 2 weeks, if necessary, the dose of Captopril would be titrated to 25 mg twice daily, based on subject's response to treatment. The primary efficacy endpoint was reduction in systolic blood pressure (SBP) from baseline to week-8 of treatment. The secondary efficacy endpoints were SBP as well as diastolic blood pressure (DBP) changes at every time-point evaluation and lipid profile improvement. Evaluation of BP was performed every week for 8 weeks of treatment; while of lipid profile at a 4-week interval. Mean SBP at baseline was 149.3 ± 5.58 mm Hg in Olive group and 148.4 ± 5.56 mm Hg in Captopril group; and mean DBPs were 93.9 ± 4.51 and 93.8 ± 4.88 mm Hg, respectively. After 8 weeks of treatment, both groups experienced a significant reduction of SBP as well as DBP from baseline; while such reductions were not significantly different between groups. Means of SBP reduction from baseline to the end of study were −11.5 ± 8.5 and −13.7 ± 7.6 mm Hg in Olive and Captopril groups, respectively; and those of DBP were −4.8 ± 5.5 and −6.4 ± 5.2 mm Hg, respectively. A significant reduction of triglyceride level was observed in Olive group, but not in Captopril group. In conclusion, Olive (Olea europaea) leaf extract, at the dosage regimen of 500 mg twice daily, was similarly effective in lowering systolic and diastolic blood pressures in subjects with stage-1 hypertension as Captopril, given at its effective dose of 12.5-25 mg twice daily.     

Schistosoma mansoni: Egg-induced downregulation of hepatic stellate cell activation and fibrogenesis

Eggs of Schistosoma mansoni trapped in human liver can lead to fibrosis. Since liver fibrosis requires activation of hepatic stellate cells (HSC) from a quiescent to a myofibroblastic phenotype, we investigated the effects of S. mansoni eggs on this process using in vitro co-cultures with human HSC and evaluated established biomarkers for activation and fibrosis. HSC demonstrate significantly reduced expression of α-smooth muscle actin (p < 0.001), connective tissue growth factor (p < 0.01) and type I collagen (p < 0.001) but significantly increased expression of peroxisome proliferator-activated receptor-γ (p < 0.01). Morphologically, HSC exhibited elongated fine cellular processes and reduced size, increased accumulation of lipid droplets and reduced expression and organization of α-smooth muscle actin and F-actin stress fibres. Additionally, schistosome eggs prevented the HSC fibrogenic response to exogenous transforming growth factor-β. In summary, schistosome eggs blocked fibrogenesis in HSC, a finding which may have implications for our understanding of the fibrotic pathology in S. mansoni infections.