Histological changes in placental rat apoptosis via FasL and cytochrome c by the nano-herbal Zanthoxylum acanthopodium

Zanthoxylum acanthopodium has several biological activities, such as antimicrobial, anti-inflammatory, and antioxidant properties. This strong antioxidant herb can be used as a drug for hypertension. FasL and cytochrome c expression play roles in the apoptotic pathway in the placenta. This study focused on the histological change in apoptosis via cytochrome c and Fas ligand expression by investigating whether Zanthoxylum acanthopodium (ZA) fruits affect apoptosis. The present study consisted of five treatments: Normal pregnant rats (C), Hypertension rats (C + ), hypertension rats + extra virgin olive oil (EVOO) (T1), Hypertension rats + ZA (T2), and hypertension rats + EVOO + ZA (T3). Hypertension was induced in rats by injecting 3 mlml of 6% NaCl. Nanoherbal of ZA (100 mg/kg BW) and EVOO (1 ml) were given on the 13th–19th days of pregnancy. Pregnant rats were dissected on the 20th day of pregnancy by cervical dislocation. ELISA assays were performed for the analysis of HSP-70 expression. Immunohistochemistry and TUNEL assays were used to analyse the histological changes in placental tissue. The results showed that cytochrome c and FasL protein exposure levels in the labyrinth, basal, and yolk sac zones were increased during hypertensive pregnancy (P < 0.0001) in rats. The administration of nanoherbal of ZA decreased the expression of cytochrome c and FasL. A significant difference was found in the combination of nanoherbal of ZA and EVOO.     

Sperm collection and cryopreservation for threatened newt species

The aims of this project were to transfer hormone-induced spermiation and sperm cryopreservation protocols developed in the model salamander species, Ambystoma tigrinum, to three threatened newt species. Additionally, we tested if supplementation with trehalose or thawing at different temperatures impacts post-thaw sperm parameters. Hormone stimulation protocols were applied to male Notophthalmus meridionalis (N = 10), Neurergus kaiseri (N = 5) and Tylototriton kweichowensis (N = 6) with sperm collected periodically up to 24–28 h post-spermiation dose. Samples of adequate sperm concentration (>70%) were cryopreserved in solutions of 10% Me₂SO + 1% BSA with or without a 10% trehalose cryodiluent. Frozen sperm samples were thawed at either 20 °C or 40 °C and examined for post-thaw motility parameters and abnormalities in head and tail structure. The spermiation response to exogenous hormone treatment was significantly different between newt species, with a success rate of 0% for N. kaiseri, 67% for T. kweichowensis, and 100% for N. meridionalis. Sperm concentration varied with time of collection after hormone administration in both T. kweichowensis and N. meridionalis. For N. meridionalis, structural abnormalities decreased in samples collected over the 24 h period (p < 0.0001) and a thaw temperature of 40 °C resulted in higher relative total sperm motility (p < 0.0001). This is the first study to describe the cryopreservation of sperm from two newt species and demonstrates the transferability of ART developed in a salamander to two newt species.     

Vascular mechanisms involved in angiotensin II-induced venoconstriction in hypertensive rats

To investigate the venoconstrictor effect of angiotensin II (Ang II) in spontaneously hypertensive rats (SHR), we used preparations of mesenteric venular beds and the circular muscle of the portal veins. Vessels were tested with Ang II in the presence or absence of losartan, PD 123319, HOE 140, L-NAME, indomethacin, or celecoxib. In the mesenteric venular bed of SHR, the effect of Ang II (0.1 nmol) was nearly abolished by losartan and enhanced by HOE 140, indomethacin, and celecoxib, while PD123319 and L-NAME had no effect. In portal vein preparations, cumulative-concentration response curves (CCRC) to Ang II (0.1–100 nmol/L) exhibited a lower maximal response (E_max) in SHR compared to Wistar rats. AT₁ receptor expression was similar in the two strains, while AT₂ receptor levels were lower in SHR portal veins when compared to Wistar. In SHR portal veins, losartan shifted the CCRC to Ang II to the right, while indomethacin and HOE 140 increased the E_max to Ang II. PD 123319, celecoxib, and L-NAME had no effect. Taken together, our results suggest that Ang II-induced venoconstriction in SHR is mediated by activation of AT₁ receptors and this effect may be counterbalanced by kinin B₂ receptor and COX metabolites. Furthermore, our data indicate that there are different cellular and molecular mechanisms involved in the regulation of venous tonus of normotensive and hypertensive rats. These differences probably reflect distinct factors that influence arterial and venous bed in hypertension.     

Effect of losartan on the blood-brain barrier permeability in diabetic hypertensive rats

Our previous publication has stressed the benefits of losartan, an angiotensin II receptor blocker, on the permeability of blood-brain barrier (BBB) and blood pressure during L-NAME-induced hypertension. This study reports the impacts of anti-hypertensive treatment by losartan on the brain endothelial barrier function and the arterial blood pressure, during acute hypertension episode, in experimentally diabetic hypertensive rats. Systolic blood pressure measurements were taken with tail cuff method before and during administration of L-NAME (0.5 mg/ml). We induced diabetes by using alloxan (50 mg/kg, i.p). Losartan (3 mg/kg, i.v) was given to rats following the L-NAME treatment. Acute hypertensive vascular injury was induced by epinephrine (40 microg/kg). The BBB disruption was quantified according to the extravasation of the Evans blue (EB) dye. L-NAME induced a significant increase in arterial blood pressure on day 14 in normoglycemic and hyperglycemic rats (p < 0.05). Losartan significantly reduced the increased blood pressure in hypertensive and diabetic hypertensive rats (p < 0.01). Epinephrine-induced acute hypertension in diabetic hypertensive rats increased the content of EB dye dramatically in cerebellum and diencephalon (p < 0.01) and slightly in both cerebral cortex (p < 0.05). Losartan treatment reduced the increased BBB permeability to EB dye in the brain regions of diabetic hypertensive rats treated with epinephrine (p < 0.05). This study indicates that, in diabetic hypertensive rats, epinephrine administration leads to an increase in microvascular-EB-albumin efflux to brain, however losartan treatment significantly attenuates this protein's transport to brain tissue.     

Antihypertensive and endothelium-dependent vasodilator effects of aqueous extract of Cistus ladaniferus

Cistus ladaniferus L. (Cistaceae) is a medicinal plant originated from the Mediterranean region which exerts different pharmacological effects. In the present study, our goal was to examine whether the plant possessed antihypertensive properties. Aqueous extract of Cistus leaves (AEC, 500 mg/kg/day) reduced systemic blood pressure (SBP) in two animal models of hypertension, the l-NAME and renovascular two kidney-one clip (2K-1C) hypertensive rats. In the former, AEC prevented the increase in SBP when co-administered with l-NAME during four weeks (164 ± 3 mm Hg in l-NAME vs. 146 ± 1 mm Hg in l-NAME + AEC, p < 0.001). In the latter, AEC reversed the increase in SBP when administered during four weeks after installation of the hypertension (146 ± 5 mm Hg with AEC vs. 179 ± 6 mm Hg without, p < 0.05). AEC treatment also reversed the endothelial dysfunction observed in both animal models of hypertension. A direct effect on cardiac and vascular tissue was also tested by examining the contractile effects of AEC in rat isolated aortic rings and Langendorff perfused hearts. AEC (10 mg/L) had no effect on left ventricular developed pressure and heart rate in isolated perfused heart. However, AEC produced a strong relaxation of pre-contracted rat aortic rings (80 ± 2% relaxation, n = 25). When the rings were denuded from endothelium or were incubated with 1 mM Nω-nitro-l-arginine (l-NNA), the relaxant effect of AEC was lost. We conclude that C. ladaniferus possesses antihypertensive properties which are mainly due to an endothelium-dependent vasodilatory action.     

Effect of estrogen on the expression of GnRH and kisspeptin in the hypothalamus of rats during puberty

The aim of this study was to assess whether changes in kisspeptin and GnRH levels could be attributed to sex steroids at puberty onset. We used the ovariectomy (OVX) model in rats treated with 17β-estradiol (E₂; OVX + E₂), or oil (OVX + oil), and in intact rats treated with E₂ (intact + E₂) or oil only (intact + oil) to determine gene expression changes of Kiss1 and Gnrh1 in the hypothalamus and protein expression of kisspeptin and GnRH in the different areas of the hypothalamus. In the intact + E₂ and OVX + E₂ rats on the day of the onset of puberty, GnRH-immunoreactive (ir) cell numbers decreased (P < 0.05) in the arcuate nucleus but were increased in the preoptic area; Kisspeptin-ir cells increased (P < 0.05) in the arcuate nucleus, periventricular nucleus, and preoptic area; no difference (P > 0.05) was found in the paraventricularis nucleus for GnRH-ir or kisspeptin-ir cells. Additionally, levels of Kiss1 and Gnrh1 messenger RNA in the hypothalamus were significantly higher (P < 0.05) in the OVX + E₂ or intact + E₂ rats than in the OVX + oil or intact + oil animals, respectively. In the OVX + oil rats, OVX significantly increased (P < 0.05) levels of Gnrh1 and Kiss1 messenger RNA and the expression of GnRH and kisspeptin in the hypothalamus compared to intact + oil animals. These results suggest that kisspeptin and GnRH play major roles in modulating the activity of estrogen circuits at the onset of puberty.     

Synergic effects of exercise training and octopamine on peroxisome proliferator-activated receptor-gamma coactivator -1a and uncoupling protein 1 mRNA in heart tissue of rat treated with deep frying oil

Octopamine (OCT) have an adverse effect on heart function. One of the positive effects of exercise training is improving cardiac function and cardiomyocytes signaling, which along with herbal supplements can have better effects on the heart tissue. Therefore, the aim of this study was to evaluate the effects of exercise training and OCT on changes of PGC1α and UCP1 expression in heart tissue of rat treated with deep frying oil (DFO). In this study, 45 male wistar rats were divided into 5 groups (n = 9 in each): I) control (Co), II) DFO, III) DFO + exercise, IV) DFO + OCT, and V) DFO + OCT + exercise. The quantification of apoptotic effects of DFO in heart tissue was assessed by TUNEL assay. Masson's trichrome stain applied to study cardiomyocytic fibers. Moreover, PGC1α and UCP1 genes and proteins expression in all groups were investigated using quantitative real-time PCR and immunohistochemical method. A significant increase in apoptotic cells was observed in the DFO-treated group (p < 0.05). In Masson's Trichrome stain study, more cardiomyocytic fibers were observed and some lymphocytic cells were present in some fibers. Also, the expression of PGC1α and UCP1 was significantly increase in DFO + exercise group, DFO + OCT group, and DFO + OCT + exercise group compare to DFO group (p < 0.05). Based on these findings, exercise and octopamine can be considered as factors affecting the expression of PGC1α genes and UCP1 as well as drug poisoning.     

Postnatal Nitric Oxide Inhibition Modifies Neurotensin Effect on ATPase Activity

We have previously showed that peptide neurotensin inhibits neuronal Na⁺, K⁺-ATPase activity, an effect which involves high affinity neurotensin receptor. Nitric oxide (NO) acts as a neurotransmitter or as a neuromodulator when it is synthesized by neuronal nitric oxide synthase. Neurotensin effect on Na⁺, K⁺-ATPase activity was evaluated in cortical synaptosomal membranes isolated from rats injected at 3, 4 and 5 postnatal days with saline (control) or N (ω)-nitro-l-arginine methyl esther (L-NAME), a nitric oxide synthase inhibitor. Assays were carried out at two stages: juvenile (35 days) and adult (56 days) ages. In an open field task, results recorded in juvenile rats markedly differed from those obtained in adult rats. The presence of neurotensin at 3.5 × 10⁻⁸–3.5 × 10⁻⁶ M concentration decreased 16–34% Na⁺, K⁺-ATPase activity in membranes purified from control animals. At variance, the peptide failed to alter this enzyme activity in membranes obtained after L-NAME treatment. After administration of L-NAME, [³H]-ouabain binding to membranes isolated from adult male rats decreased 64% in the presence of 1.0 × 10⁻⁶ M neurotensin, a peptide concentration which only slightly decreased binding to membranes isolated from juvenile rats. It is postulated that early postnatal NO dysfunction may exert a permanent change in neurotensin system that influence later Na⁺, K⁺-ATPase response to neurotensin.     

Earthworm compound and rock biofertilizer enriched in nitrogen by inoculation with free living diazotrophic bacteria

Free living diazotrophic bacteria are known to enrich nitrogen of organic matter sources. In this paper we report of experiments using rock biofertilizers mixed with two types of organic matter (earthworm compound and ice cream waste) inoculated with free living diazotrophic bacteria. The earthworm compound and P and K biofertilizers were mixed to form substrates S₁ (earthworm compound 3 dm³ + PK biofertilizer 1 dm³ and waste ice cream 1 dm³); S₂ (earthworm compound 2.5 dm³ + PK biofertilizer 1.5 dm³ and waste ice cream 1 dm³L) and S₃ earthworm compound 2.0 dm³ + PK biofertilizer 2.0 dm³ and waste ice cream 1 dm³), and subsequently inoculated (100 mL pot⁻¹) with 3 free living diazotrophic bacteria isolated from different Brazilian soils. The control was an uninoculated earthworm compound. Samples were collected at various incubation time (0; 15; 30 and 45 days) and analyzed for total N. Total N concentrations were highest in S₁, S₂ and S₃ substrates at 34, 27 and 29 days, respectively. The isolate NFB 1001 increased total N in all substrates and the best results were obtained at 34 days in S₁ substrate which contained the highest amount of earthworm compound. The isolates promoted a decline in N content after 30 days of growth, indicating the best time to produce the organic biofertilizer. The organic biofertilizer enriched in nitrogen by free living diazotrophic bacteria is of relevance to organic agriculture.     

Phytochemical analysis and toxicological evaluation of the ethanolic Leaves extract of Hypoestes rosea on the morphology and biochemical indices of the Kidneys of albino Wistar Rats

BackgroundHypoestes rosea (family: Acanthacea), has been harnessed and utilized for treatment of several ailments. However, there is the paucity of available data on nephrotoxicity associated with this herb. Here, we investigated the phytochemical profile and toxicological effect of H. rosea on Wistar Rats.MethodsTwenty rats (weight range: 75–100 g) were assigned into five study groups, viz; (a) control (without treatment) (b) treatment group 1, orally administered with 50 mg/kg (c) treatment group 2, orally administered with 100 mg/kg (d) treatment group 3, orally administered with 250 mg/kg, and (e) treatment group 4, orally administered with 300 mg/kg of H. rosea, respectively for 28 days of four rats per group. The rats were made unconscious by using oral administration of chloroform. Cardiac punctures were made, and blood samples collected into 10 ml labeled plain container, allowed to clot and spun to harvest serum for determination of sodium, potassium, chloride, bicarbonate, urea and creatinine using colorimetric, back-titrimetric, Urease-Berthelot and Jaffe’s reaction methods respectively. Kidneys of rats were harvested, weighed and immediately fixed in 10% neutral buffered formalin for histological analysis.ResultMean serum sodium (p = 0.049), potassium (p = 0.007), and urea (p < 0.001) levels were significantly higher among the treatment groups compared to controls. Histopathological findings of kidney sections revealed mild glomerular infiltration in treatment groups 2–4. Additionally, sclerosis was observed in groups 3–4. Phytochemical analysis of H. rosea revealed presence of alkaloids, flavonoids, saponins, tannins, terpenoids, steroids and reducing sugars.ConclusionFrom the findings in this study, H. rosea leaf extract causes significant damage to the kidneys of Wistar rats at higher doses. Of which, the damages were dose-dependent in direct proportionality manner. To better determine the safe dosage and ideal duration of consumption, there is the need for further studies on H. rosea.     

Effects of Canarium odontophyllum leaves on plasma glucose and T lymphocyte population in streptozotocin-induced diabetic rats

Type 1 diabetes mellitus is a chronic disease characterized by lack of insulin production. Immune mechanisms are implicated in the pathogenesis of Type 1 diabetes. Canarium odontophyllum (CO) fruits and leaves have been shown to possess high antioxidant activity. This study was conducted to evaluate the effects of CO leaves aqueous extract on the blood glucose and T lymphocyte population in the spleen of streptozotocin (STZ)-induced diabetic rats. Nineteen male Sprague–Dawley rats were randomly divided into three groups: normal, diabetic control and CO treated diabetic groups. Diabetes was induced by a single intraperitoneal injection of 65 mg STZ/kg body weight. The extract of CO leaves was administered orally by force feeding daily at the dose of 300 mg/kg for 28 days. The rats were sacrificed at the end of the study and the spleen was harvested for flow cytometry analysis. The results showed a significant decrease in body weight of diabetic and CO treated diabetic groups compared with the normal group (p < 0.05). The fasting blood glucose level of CO treated diabetic group was significantly lower than the diabetic group (p < 0.05). Diabetic and CO treated diabetic groups showed a significant increase in the percentage of spleen CD3⁺ CD4⁺ T lymphocytes (p < 0.05) when compared with the normal group. However, there was no significant difference in the percentage of spleen CD3⁺ CD8⁺ T lymphocytes among all experimental groups. The finding suggested that an aqueous extract of CO leaves has the ability to reduce blood glucose levels in diabetic rats.     

Resistance exercise acutely enhances mesenteric artery insulin-induced relaxation in healthy rats

Aims

We evaluated the mechanisms involved in insulin-induced vasodilatation after acute resistance exercise in healthy rats.

Main methods

Wistar rats were divided into 3 groups: control (CT), electrically stimulated (ES) and resistance exercise (RE). Immediately after acute RE (15 sets with 10 repetitions at 70% of maximal intensity), the animals were sacrificed and rings of mesenteric artery were mounted in an isometric system. After this, concentration–response curves to insulin were performed in control condition and in the presence of LY294002 (PI3K inhibitor), L-NAME (NOS inhibitor), L-NAME + TEA (K⁺ channels inhibitor), LY294002 + BQ123 (ET-A antagonist) or ouabain (Na⁺/K⁺ ATPase inhibitor).

Key findings

Acute RE increased insulin-induced vasorelaxation as compared to control (CT: R_max = 7.3 ± 0.4% and RE: R_max = 15.8 ± 0.8%; p < 0.001). NOS inhibition reduced (p < 0.001) this vasorelaxation from both groups (CT: R_max = 2.0 ± 0.3%, and RE: R_max = − 1.2 ± 0.1%), while PI3K inhibition abolished the vasorelaxation in CT (R_max = − 0.1 ± 0.3%, p < 0.001), and caused vasoconstriction in RE (R_max = − 6.5 ± 0.6%). That insulin-induced vasoconstriction on PI3K inhibition was abolished (p < 0.001) by the ET-A antagonist (R_max = 2.9 ± 0.4%). Additionally, acute RE enhanced (p < 0.001) the functional activity of the ouabain-sensitive Na⁺/K⁺ ATPase activity (R_max = 10.7 ± 0.4%) and of the K⁺ channels (R_max = − 6.1 ± 0.5%; p < 0.001) in the insulin-induced vasorelaxation as compared to CT.

Significance

Such results suggest that acute RE promotes enhanced insulin-induced vasodilatation, which could act as a fine tuning to vascular tone.     

Long-term enhanced nitrogen deposition increases ecosystem respiration and carbon loss from a Sphagnum bog in the Scottish Borders

Nitrogen (N) deposition has increased in the last few decades with implications for the functioning of Sphagnum mosses, the main peat forming genus in peatlands. However, there are few in situ measurements of the carbon balance, especially where the N additions have been realistically manipulated in the field, and none with respect to the effect of N form. The aim of this study was to look at the effects of experimental N additions as oxidized or reduced N, with and without phosphorus (P) and potassium (K), on CO₂ fluxes from Sphagnum capillifolium hummocks in a long-term N addition experiment, Whim bog, in the Scottish Borders. In situ static chamber measurements were made during 2008 on 20 plots (control, and N treatments receiving 56 kg N ha⁻¹ y⁻¹ of either nitrate (NO₃⁻) or ammonium (NH₄⁺) added, with and without PK) to assess N effects on CO₂ exchange. Almost all the measured fluxes were negative, i.e. Sphagnum hummocks lost CO₂ to the atmosphere, irrespective of the treatments applied. N treatment did not have a significant effect on ecosystem respiration (ER) or net ecosystem productivity (NEP) but adding PK with N increased gross photosynthesis (P_G) significantly, compared to the other treatments. Summed monthly averages of NEP for each treatment indicated that increasing N deposition increased CO₂ loss from the system. The form of N affected the response: compared to the control, adding nitrate increased the CO₂ loss more than ammonium, both with and without PK. Nitrogen (both forms) increased the ecosystem respiration fluxes at a certain temperature, adding PK with N further enhanced the response. The positive (increasing) temperature response of ecosystem respiration with N suggests that in high N deposition areas the potential increase in ecosystem respiration, CO₂ loss, will be exacerbated with climate change.     

Changes of sodium and ATP affinities of the cardiac (Na,K)-ATase during and after nitric oxide deficient hypertension

In the cardiovascular system, NO is involved in the regulation of a variety of functions. Inhibition of NO synthesis induces sustained hypertension. In several models of hypertension, elevation of intracellular sodium level was documented in cardiac tissue. To assess the molecular basis of disturbances in transmembraneous transport of Na⁺, we studied the response of cardiac (Na,K)-ATPase to NO-deficient hypertension induced in rats by NO-synthase inhibition with 40 mg/kg/day N^G-nitro-L-arginine methyl ester (L-NAME) for 4 four weeks. After 4-week administration of L-NAME, the systolic blood pressure (SBP) increased by 36%. Two weeks after terminating the treatment, the SBP recovered to control value. When activating the (Na,K)-ATPase with its substrate ATP, no changes in K_m and V_max values were observed in NO-deficient rats. During activation with Na⁺, the V_max remained unchanged, however the K_Na increased by 50%, indicating a profound decrease in the affinity of the Na⁺-binding site in NO-deficient rats. After recovery from hypertension, the activity of (Na,K)-ATPase increased, due to higher affinity of the ATP-binding site, as revealed from the lowered K_m value for ATP. The K_Na value for Na⁺ returned to control value. Inhibition of NO-synthase induced a reversible hypertension accompanied by depressed Na⁺-extrusion from cardiac cells as a consequence of deteriorated Na⁺-binding properties of the (Na,K)-ATPase. After recovery of blood pressure to control values, the extrusion of Na⁺ from cardiac cells was normalized, as revealed by restoration of the (Na,K)-ATPase activity. (Mol Cell Biochem 000: 000-000, 1999)     

Influence of the conformations of αA-crystallin peptides on the isomerization rates of aspartic acid residues

The isomerization rate of aspartic acid (Asp) residue is known to be affected by the three-dimensional structures of peptides and proteins. Although the isomerized Asp residues were experimentally observed, structural features which affect the isomerization cannot be elucidated sufficiently because of protein denaturation and aggregation. In this study, molecular dynamics (MD) simulations were conducted on three αA-crystallin peptides (T6, T10, and T18), each containing a single Asp residue with different isomerization rate (T18 > T6 > T10) to clarify the structural factors of Asp isomerization tendency. For MD trajectories, distances between side-chain carboxyl carbon of Asp and main-chain amide nitrogen of (n + 1) residue (C_γ–N distances), root mean square fluctuations (RMSFs), and polar surface areas for main-chain amide nitrogen of (n + 1) residues (PSA_N) were calculated, because these structural features are considered to relate to the formations of cyclic imide intermediates. RMSFs and PSA_N are indexes of peptide backbone flexibilities and solvent exposure of the amide nitrogen, respectively. The average C_γ–N distances of T10 was longer than those of the other two peptides. In addition, the peptide containing Asp residue with a higher isomerization rate showed higher flexibility of the peptide backbone around the Asp residue. PSA_N for amide nitrogen in T18 were much larger than those of other two peptides. The computational results suggest that Asp-residue isomerization rates are affected by these factors.     

Immunomodulation of tahneeq method in IL-12 and CD8+ T-Lymphocyte,an in-vivo study in neonatal rats

Stimulation of the neonatal immune system is quite important for the proliferation and differentiation of antigen-presenting cells (APCs) and T cells. Tahneeq is a traditional method to manually rub the palatal mucosa of newborn babies with premasticated Ajwa palm dates. The present study was to investigate the tahneeq effects on IL-12 expression of dendritic cells (DCs) and blood T lymphocytes expressing CD8⁺ in neonatal Wistar rats. The number of 90 healthy neonatal Wistar rats have randomly divided into three groups: control group received breastmilk only, treatment group (T1) receiving breast milk + mild-scratched intensity of tahneeq, and T2 group received breastmilk + strong-scratched intensity of tahneeq on the palatal and gingival mucosa immediately after birth.  Seven neonatal Wistar rats in all groups were then sacrificed in three hours after birth and days 1, 5, 7, 13, and 30 treatment. IL-12 expression in the palatal and gingival mucosa was determined using immunohistochemical staining, and blood CD8⁺ T-lymphocytes were quantified using a flow cytometer. One way ANOVA was used to analyze the percentage of IL-12 and CD8⁺ T-lymphocytes among neonatal Wistar rat groups.  The T1 and T2 newborn rat groups had significantly higher IL-12 expression than the control group (p<0.001). The increased IL-12 expression in T2 groups significantly increased (p<0.001) compared to the IL-12 expression in the T1 and control groups. The percentage of  CD8⁺ T lymphocytes in all neonatal rat groups increased on three hours after birth and day 30 treatment but remained constant on days 5 and 7 treatment and decreased on day 13 treatment. At 5, 13, and 30^th days treatment,  the percentage of CD8⁺ T lymphocytes in T1 and T2 neonatal rat groups was significantly higher (p<0.05) than that in the control group. In conclusion, the impact on systemic CD8⁺ T cells did not influence by the depth of the scratch. Both mild and strong tahneeq increased the systemic CD8⁺ T-lymphocytes in neonatal Wistar rats. The roles of anti-inflammatory cytokines and Treg cells should be further investigated to unravel those different results for the development of mucosal immunity in neonates.     

Contribution of genetic variant identified in HHEX gene in the overweight Saudi patients confirmed with type 2 diabetes mellitus

BackgroundThe rs7932837 polymorphism in the Hematopoietically expressed homeobox (HHEX) gene was discovered through genome-wide association studies and is a promising candidate for type 2 diabetes mellitus (T2DM), which is one of the risk factors for obesity and other complications. T2DM has been identified as a heterogeneous and multifactorial disease characterized by insulin resistance and secretion.AimThe aim of this study was to investigate the rs7932837 polymorphism in the HHEX gene in overweight patients diagnosed with T2DM in the Saudi Population.MethodsIn this case-control study, one hundred T2DM cases and 100 controls were selected based on inclusion and exclusion criteria. Genotyping was performed with polymerase chair reaction-restriction fragment length polymorphism analysis and statistical analysis was performed between T2DM cases and controls for clinical characteristics, genotype and allele frequencies and multiple logistic regression analysis.ResultsIn this study, T2DM cases were compared with healthy control subjects. Clinical characteristic analysis revealed the statistical analysis between age, weight, BMI, FBG, HDL-c, TC, TG and family history (p < 0.05). HWE analysis was in the accordance (p < 0.05). The rs7932837 polymorphism in the recessive model showed the positive association (AA + AG vs AA: 2.22 [1.25–3.96] & p = 0.006) and none of the genotypes or alleles were in the statistical association. Multiple logistic regression analysis revealed positive association with age, BMI and FBG (p < 0.05).ConclusionThis study concludes as rs7932837 polymorphism in the HHEX gene showed positive association with recessive model and future studies recommend to carry out with large number of sample size with additional polymorphisms in HHEX gene.     

Uptake and inhibition kinetics of nitrogen in Microcystis aeruginosa: Results from cultures and field assemblages collected in the San Francisco Bay Delta,CA

The nitrogen (N) uptake kinetic parameters for Microcystis field assemblages collected from the San Francisco Bay Delta (Delta) in 2012 and non-toxic and toxic laboratory culture strains of M. aeruginosa were assessed. The ¹⁵N tracer technique was used to investigate uptake of ammonium (NH₄⁺), nitrate (NO₃⁻), urea and glutamic acid over short-term incubations (0.5–1 h), and to study inhibition of NO₃⁻, NH₄⁺ and urea uptake by NH₄⁺, NO₃⁻ and NH₄⁺, respectively. This study demonstrates that Delta Microcystis can utilize different forms of inorganic and organic N, with the greatest capacity for NH₄⁺ uptake and the least for glutamic acid uptake, although N uptake did not always follow the classic Michaelis–Menten hyperbolic relationship at substrate concentrations up to 67 μmol N L⁻¹. Current ambient N concentrations in the Delta may be at sub-saturating levels for N uptake, indicating that if N loading (especially NH₄⁺) were to increase, Delta Microcystis assemblages have the potential for increased N uptake rates. Delta Microcystis had the highest specific affinity, α, for NH₄⁺ and the lowest for NO₃⁻. In culture, N uptake by non-toxic and toxic M. aeruginosa strains was much higher than from the field, but followed similar N utilization trends to those in the field. Neither strain showed severe inhibition of NO₃⁻ uptake by NH₄⁺ or inhibition of NH₄⁺ uptake on NO₃⁻, but both strains showed some inhibition of urea uptake by NH₄⁺.     

Losartan delays the progression of streptozotocin‐induced diabetic cataracts in albino rats

The ocular renin‐angiotensin system has become an interesting target for ocular diseases because it has been implicated in various ocular diseases such as diabetic retinopathy, glaucoma, age‐related macular degeneration, uveitis, and hypertensive cataracts. In the present study, we explored the effect of topically and orally administered losartan (an angiotensin receptor blocker) on streptozotocin‐induced diabetic cataract in albino rats. Topical treatment with losartan modulated neither the blood glucose level nor the polyol content but oral treatment with losartan decreased both. Topical and oral treatment with losartan significantly increased the antioxidants (glutathione, glutathione peroxidase, superoxide dismutase, and catalase), decreased the lipid peroxidant malondialdehyde, and restored soluble protein, and insoluble protein and various ions (Na⁺, K⁺, and Ca²⁺) in the lens; however, topical treatment had a better effect than oral treatment. These findings demonstrate that topical administration of losartan significantly reduces the risk of cataract formation without affecting either the blood glucose level or polyol contents.     

Independent case-control study in KCNJ11 gene polymorphism with Type 2 diabetes Mellitus

BackgroundType 2 Diabetes Mellitus (T2DM) is the most common form of diabetes in the aging population. This chronic metabolic disorder has discovered many candidate genes, and KCNJ11 was one of the genes associated with insulin secretion pathways mediated by potassium channels. There have been limited studies on the rs5210 polymorphism in T2DM patients, and none of them have been conducted in Saudi Arabia.AimThe aim of this study is to investigate at genotyping levels of rs5210 polymorphism in the KCNJ11 gene in older population with T2DM in the Saudi Population.MethodsBased on the sample size design, this case-control study included 102 T2DM cases and 102 controls. Using the PCR-RFLP assay, 204 patients extracted DNA was genotyped for the rs5210 polymorphism. SPSS software was used for statistical analysis, including t-tests, HWE, genotyping, and multiple logistic regression analysis.ResultsThe t-tests performed on T2DM cases and controls revealed a significant association in age, weight, BMI, FBG, Hb1Ac, SBP, DBP, HDLC, TC, and TG parameters (p < 0.05). HWE analysis found to be in consistent with rs5210 polymorphism. Allelic association was found in the rs5210 polymorphism (OR-1.64 [95 %CI: 1.08–2.49]; p = 0.01); however, no association (p > 0.05) was observed in the multivariate logistic regression assessment performed in this study.ConclusionThese results indicate that the rs5210 polymorphism was primarily associated with allele frequencies, which could be attributable to the small sample size. Large sample size studies will be required to determine whether KCNJ11 gene polymorphisms may be required as a risk marker for T2DM in the Saudi population.