Counts In Rats Research Articles

Using Waterless Alcohol-based Antiseptic for Skin Preparation and Active Thermal Support in Laboratory Rats.

Rodents are frequently used for models that require surgical procedures. At our institution, laboratory rats are increasingly preferred for investigations of neurologic disorders, cardiovascular interventions, and assessment and treatment of addictive and depressive behaviors. For these types of studies, surgical preparations of the head and neck areas are necessary for catheterization and instrumentation. Based upon our former work in laboratory mice, we sought to improve rat surgery outcomes and confirm the efficacy of a waterless alcohol-based (WAB) antiseptic for skin disinfection prior to incision. In addition, we wanted to investigate whether active warming efforts improved perioperative body temperatures for rats to aid in return to consciousness. Prior to cranial surgical incision and placement in stereotactic equipment, rats were assessed after skin preparation with WAB and after thermal interventions, including prewarming cages for 30 min before anesthesia and delivery of warmed fluid (NaCl) supplementation. Core temperatures were recorded and aerobic culture swabs collected from surgical sites at multiple time points. As previously shown in mice, bacterial counts in rats were effectively diminished by WAB agents. Assessment of intraoperative body temperature trajectories did not identify appreciable differences between control rats and rats that were exposed to prewarming or warmed fluid supplementation or both. However, heavier male rats recovered more rapidly from isoflurane anesthesia than did lighter male and female rats. Although these thermal support measures did not significantly improve anesthetic recovery times in rats, animals warmed for 30 min trended toward a faster return to righting reflex after exposure to isoflurane. These findings confirm that WAB antiseptic is an acceptable option for skin preparation in rats and suggest that continued evaluation of thermal interventions remains of interest for improved outcomes in rat surgery.

EFFECTS OF METABOLIC SYNDROME ON BLOOD CELLS OF RATS

Objective: The study is focused on the effects of metabolic syndrome induced by high-fat, high-carbohydrate diet on erythrocytes and platelets of rats. Design and method: The 6-week old Wistar rats (n = 23) were used for study. Rats were divided into control and experimental groups. The rats from the control group were fed standard rat chow. The rats from the experimental group had a high-fat, high-carbohydrate diet. Body weight, blood pressure (BP) were assessed. After 12 weeks of a diet load an oral glucose tolerance test was carried out. Hematological and biochemical parameters in plasma were analyzed. The conductivity of the KCa-channels of the erythrocyte membrane was measured by potentiometric method, and the platelet aggregation was determined by turbidimetric method. Results: High-fat, high-carbohydrate diet feeding in rats for 12 weeks led to obesity, BP elevation, hyperglycemia, impaired glucose tolerance, dyslipidemia with triglyceridemia. Elevation of number of leukocytes due to granulocytes and platelets were observed in the experimental group. The collagen-induced aggregation of platelets was elevated in rats with metabolic syndrome. Red blood cell count in rats of the experimental group did not significantly differ from the control group. The multidirectional changes in the membrane potential were observed in response to the stimulation of the KCa-channels of the erythrocyte membrane of the experimental group with the Ca2 + ionophore A23187 or the redox-system. Conclusions: The data obtained indicate that a high-fat, high-carbohydrate diet leads to metabolic and hemorheological disorders, that are characteristic of metabolic syndrome.

Toxicological evaluations of betulinic acid and ursolic acid; common constituents of Houttuynia cordata used as an anthelmintic by the Naga tribes in North-east India

BackgroundBetulinic acid (BA) and ursolic acid (UA) are two major phytoconstituents of Houttuynia cordata Thunb., (Saururaceae) which is used as an anthelmintic in the traditional medicine system of the Nagas in Nagaland, India. This study evaluates their toxic potentials using rodent models (Swiss albino mice and Wistar rats) according to the OECD (Organization for Economic Cooperation and Development) guidelines. Acute and 28-day sub-acute oral toxicity studies were conducted, and evaluations were made based on biochemical, hematological, and histopathological observations.ResultsAcute oral toxicity study revealed the oral LD50 of both the test compounds to be > 2000 mg/kg in mice. Sub-acute administration of BA at 10 mg/kg body weight (b.w.) revealed a significant increase in serum glutamic oxaloacetic transaminase (SGOT), alkaline phosphatase (ALP), urea concentrations and eosinophil and lymphocyte counts in rats. Animals administered with 10 mg/kg b.w. UA revealed elevated neutrophil count, SGOT, ALP, and urea concentrations, whereas white blood cells (WBC), lymphocyte, and platelet counts were found to be low. Histopathological examinations of body organs revealed alterations in the architecture of the liver, kidney, and spleen tissues. Notably, all these alterations were recoverable as evident in the satellite group, indicating a recovering pattern from the toxic effects caused by the oral administration of these phytocompounds.ConclusionAlthough UA and BA possess several therapeutic properties, their long-term usage can cause mild toxicity in their users. This study also paves way for evaluating the optimum effective and safe dose of these phytocompounds.

ПРОФИЛАКТИЧЕСКОЕ ВВЕДЕНИЕ БЕЛКА ТЕПЛОВОГО ШОКА HSP70 ОСЛАБЛЯЕТ ЛИХОРАДОЧНУЮ РЕАКЦИЮ ПРИ ЭНДОТОКСЕМИИ У ГОЛУБЕЙ ( COLUMBA LIVIA ) И КРЫС

The stress-inducible 70 kDa heat shock protein (Hsp70) can exert a protective effect on endotoxemia and sepsis due to its ability to interact with immune cells and modulate the immune response. However, it remains unknown whether Hsp70 is able to relieve endotoxemia-induced fever. We carried out a comparative study of the effects of preventive administration of the human recombinant Hsp70 (HSPA1A) on lipopolysaccharide (LPS)-induced endotoxemia in pigeons and rats with preliminarily implanted electrodes and thermistors for recording the thermoregulatory parameters (brain temperature, peripheral vasomotor reaction, muscular contractile activity). Additionally, we analyzed the dynamics of the white blood cell (WBC) count in rats under the same conditions. It was found that preventive administration of Hsp70 relieves the LPS-induced febrile reaction in pigeons and rats and accelerates the restoration of the WBC count in rats. These data suggest that these warm-blooded animals share the common physiological mechanism of the protective effect of Hsp70.

Human granulocyte-colony stimulating factor (G-CSF)/stem cell factor (SCF) fusion proteins: design, characterization and activity.

BackgroundStem cell factor (SCF) and granulocyte-colony stimulating factor (G-CSF) are well-characterized vital hematopoietic growth factors that regulate hematopoiesis. G-CSF and SCF synergistically exhibit a stimulatory effect on hematopoietic progenitors. The combination of G-CSF and SCF has been used for mobilization of peripheral blood progenitor cells in cancer and non-cancerous conditions. To overcome challenges connected with the administration of two cytokines, we developed two fusion proteins composed of human SCF and human G-CSF interspaced by an alpha-helix-forming peptide linker.MethodsThe recombinant proteins SCF-Lα-GCSF and GCSF-Lα-SCF were purified in three steps using an ion-exchange and mixed-mode chromatography. The purity and quantity of the proteins after each stage of purification was assessed using RP-HPLC, SDS-PAGE, and the Bradford assays. Purified proteins were identified using high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) and the Western blot analyses. The molecular weight was determined by size exclusion HPLC (SE-HPLC). The activity of heterodimers was assessed using cell proliferation assays in vitro. The capacity of recombinant fusion proteins to stimulate the increase of the absolute neutrophil count in rats was determined in vivo. The binding kinetics of the proteins to immobilized G-CSF and SCF receptors was measured using total internal reflection ellipsometry and evaluated by a standard Langmuir kinetics model.ResultsThe novel SCF-Lα-GCSF and GCSF-Lα-SCF proteins were synthesized in Escherichia coli. The purity of the heterodimers reached >90% as determined by RP-HPLC. The identity of the proteins was confirmed using the Western blot and HPLC/ESI-MS assays. An array of multimeric forms, non-covalently associated dimers or trimers were detected in the protein preparations by SE-HPLC. Each protein induced a dose-dependent proliferative response on the cell lines. At equimolar concentration, the heterodimers retain 70–140% of the SCF monomer activity (p ≤ 0.01) in promoting the M-07e cells proliferation. The G-CSF moiety in GCSF-Lα-SCF retained 15% (p ≤ 0.0001) and in SCF-Lα-GCSF retained 34% (p ≤ 0.01) of the monomeric G-CSF activity in stimulating the growth of G-NFS-60 cells. The obtained results were in good agreement with the binding data of each moiety in the fusion proteins to their respective receptors. The increase in the absolute neutrophil count in rats caused by the SCF-Lα-GCSF protein corresponded to the increase induced by a mixture of SCF and G-CSF.

Developmental and reproductive toxicity studies of BIA 10-2474

A series of regulatory studies were carried out to investigate the effects of the FAAH inhibitor BIA 10–2474 on fertility, embryo-fetal toxicity and pre- and post-natal development in rats and rabbits. Despite some reductions in sperm count in rats from 50 mg/kg, there were no major changes in male fertility up to 100 mg/kg. In female rats administered up to GD6, there were increases in pre-implantation loss at 50 and 100 mg/kg but neither post-implantation loss nor early embryonic development was affected. In contrast, when administered to female rats during pregnancy (GD6-GD17), BIA 10–2474 at 75 mg/kg/day reduced food consumption resulting in weight loss, increased post-implantation loss and reduced mean fetal body weight. In rabbits, the same maternal toxicity was seen but there were no effects in this species on post-implantation loss or fetal body weights. There were no teratological effects clearly due to BIA 10–2474 and developmental milestones and behavior of offspring were not affected. When administered during pregnancy and lactation (GD6-PND20), some post-implantation loss was seen from 20 mg/kg/day, but developmental milestones and behavior of the offspring were not affected, although males tended to have lower body weight. Based on these data the NOAEL for parental fertility was established as 50 mg/kg/day, the maternal NOAEL during pregnancy was 25 mg/kg/day in rats and developmental NOAEL was 25 and 75 mg/kg/day in rats and rabbits, respectively. When administered during post-natal development to rats the maternal NOAEL was 6 mg/kg/day. The parental reproductive NOAEL, the NOAEL for viability and growth of the F1 offspring, the F1 parental NOAEL and the F1 reproductive NOAEL were all considered to be 20 mg/kg/day.

Comparison between a novel tap water wound irrigation device with sterile saline device in an open traumatic wound animal model

Study objective Open traumatic wounds present to US emergency departments at a rate of approximately 12 million per year. Tap water wound irrigation usage has not been adopted by providers thus negating a potential cost saving of approximately US$65,600,000 per year when compared to saline irrigation. A novel tap water wound irrigation device can reduce bacterial counts in rats inoculated with MRSA as effectively as a commonly used sterile saline wound irrigation system. Methods Two groups of six Sprague–Dawley rats were used for this study. Two 3 cm incisions were made into the dorsum of each Sprague–Dawley rat and each incision was inoculated with 100 µl of MRSA. One wound served as the control, the other wound was either irrigated with the sterile saline wound irrigation system or a novel tap water wound irrigation device. The wounds were homogenized, plated, and bacterial colonies were enumerated to determine the final concentration of bacteria in each wound. Results There was no significant statistical difference in the reduction of MRSA in wounds treated with the sterile saline system relative to wounds treated with tap water (one-way ANOVA with post hoc Dunnet's test). Conclusion The novel tap water wound irrigation device is as effective as the sterile saline wound irrigation device at reducing bacterial counts in a rat laceration model when compared to non-irrigated controls. Adopting the use of a convenient, effective, and inexpensive wound irrigation system using tap water could help save the health care system millions of dollars in direct costs, shipping cost, and would not be affected by saline shortages.

The Role of Curcumin on Reduced Leukocyte and Platelet Counts in Rats After Exposure to Ultraviolet Light from Compact Fluorescent Lamps: A Morphological Study

Background: Compact fluorescent lamps (CFLs) could be a source of ultraviolet (UV) radiation with potential harmful effects. Objectives: We assessed the effects of ultraviolet radiation on leukocyte and platelet counts in rats and also investigated the preservative effects of curcumin on hematologic disorders caused by UV radiation. Methods: We randomly divided 48 male Sprague-Dawley rats into six groups (n = 8). Two control groups received ethyl oleate (0.2 mL, IP) for 10 (group I) and 30 days (group II), without exposure to fluorescent light. Two fluorescent groups treated daily with 12 hours fluorescent light exposure for 10 (group III) and 30 days (group IV). Group V and VI received curcumin (20 µmol, IP) and 12 hours fluorescent light exposure for 10 and 30 days, respectively. The blood samples were collected and the leukocyte indexes were measured by cell counter and blood smears were obtained. Results: The number of leukocytes was significantly lower in groups III (8.36 ± 0.23 × 103/µL) and IV (7.85 ± 0.18 × 103/µL) compared with groups I (8.78 ± 0.14 × 103/µL) and II (8.50 ± 0.20 × 103/µL). (P value III:I = 0.004, IV:II < 0.0001). The number of leukocytes insignificantly increased to normal levels of 8.85 ± 0.29 × 103/µL and 8.30 ± 0.16 × 103/µL in groups V and VI, respectively (P value V:I = 0.79, VI:II = 0.09). The number of platelets (PLTs) was significantly lower in the groups IV and VI in comparison with the group II (750 ± 6.52 × 103/µL, 808 ± 2.13 × 103/µL and 846 ± 2.73 × 103/µL, respectively (P value IV:II < 0.0001, VI:II < 0.0001). In the morphological pattern of leukocytes, the CFLs lead to atopic cells, bond cells, and cell lysis, however, administration of curcumin prevented this disturbance. Conclusions: Curcumin has a protective role in reducing complications caused by CFLs in reduced leukocyte and platelet counts, which is probably due to its antioxidant agent.

Goat Milk Whey Improves Nutritional Status, Fecal Microbial Composition and Intestinal Morphology in Female Rats Fed a Westernized Diet and Their Offspring

This study evaluated whether supplementation with goat milk whey (GMW) affects the nutritional status, intestinal histology and intestinal microbial composition of female rats fed a westernized diet from gestation to lactation, as well as whether changes can be replicated in the offspring at the end of weaning and at 45 days of life. Pregnant female rats were randomized into four groups: control group (CSAL), control group supplemented with GMW (CGOAT), westernized group (WSAL) or westernized group supplemented with GMW (WGOAT). After weaning, half of offspring were euthanized and the rest of the offspring were maintained under the same treatment applied to dams, up to 45 days of life. Body weight, food intake, intestinal histology and counting of fecal microbial groups were determined in both female rats and offspring. The offspring supplemented with GMW showed decreased body weight at weaning. After weaning, groups supplemented with GMW showed reduced body weight and visceral fat, increased fecal lactobacilli counts in rats and offspring and attenuation of damages induced by the westernized diet on intestinal epithelial cells. GMW supplementation caused a positive effect on fecal microbial composition, intestinal morphology and induced reduction in weight gain and visceral fat in female rats and offspring fed westernized diet. These effects appear to be dependent on the animal’s age and period time of GMW supplementation.

Prolonged Administration of Leaf Extract of Ocimum gratissimum Reduces RBC, PCV and Platelet Count but Increases Total WBC and Lymphocyte Counts in Rats

Background: Ocimum gratissimum is an ethnopharmacological plant with numerous healing potentials. But there is paucity in scientific literature on its impact on the blood cells.&#x0D; Aim and Objective: The present study was intended to evaluate the dose and time-dependent effect of aqueous leaf extract of Ocimum gratissimum on some haematological in albino Wistar rats.&#x0D; Methods: Twenty four rats were randomly assigned to four equal groups. Group 1 was control, groups 2, 3 and 4 were given 450 mg/kg, 800 mg/kg and 1800 mg/kg respectively of the extract daily for 30 days. All rats had free access to water and rat chow.&#x0D; Results: The result showed a significantly lower RBC count in groups 3 and 4 when compared with control (p&lt;0.05 and p&lt;0.01 respectively). PCV was significantly lower in group 4 than control (p&lt;0.05). Platelet count was significantly lower in group 4 than control (p&lt;0.05) and group 2 (p&lt;0.05). A significant increase in WBC count was observed in groups 2 and 3 when compared with control (p&lt;0.05 and p&lt;0.01 respectively). A significant reduction in neutrophil count was observed in groups 2, 3 and 4 compared with control (p&lt;0.05; p&lt; 0.05 and p&lt;0.01 respectively). There were significant increases in lymphocyte counts in groups 2, 3 and 4 when compared with control (p&lt;0.05, p&lt;0.01 and p&lt;0.01 respectively). There were no significant changes in MCV, MCH and MCHC in the groups.&#x0D; Conclusion: In conclusion, aqueous leaf extract of Ocimum gratissimum causes reduction in RBC count, PCV, platelet count and neutrophil count but causes increases in total WBC count and lymphocyte count.

Analysis of Morphology and Erythrocyte Count in Rat after Electrical Exposure

Electrical burns cause high mortality and morbidity and various complications, although the incidence is only 4% of all burns. Joule heating phenomenon occurs in electric burn patients which cause changes in erythrocyte morphology resulting hypercoagulability that contribute to the formation of thrombus. Changes in erythrocyte morphology may cause tissue oxygenation disorders and decrease in erythrocyte count leading to hypoxia. Until now there have been no reports of changes in erythrocyte morphology and reduction in the number of erythrocytes in burn patients due to electricity. This study aims to observe and analyze morphological and erythrocyte count in rats after electrical exposure of 140 V for 17 seconds. This is a true experimental laboratories study with post-test only control group design. The sample used were 24 rats consisting of one control group (K) and five treatment groups (P). Treatment groups were distinguished by days of blood sampling that is days 0 (P1), 3 (P2), 7 (P3), 10 (P4), and 14 (P5) post electrical exposure of 140 V for 17 seconds. Assessment in erythrocyte morphology was performed using giemsa staining that observed with 1000x magnification while erythrocyte count was performed using improved neubauer counting chamber. One Way Anova showed that there was no significant change in rats erythrocyte count after electrical exposure of 140 V for 17 seconds (p = 0.110). Descriptive analysis showed that there is morphological changes in erythrocyte morphology after electrical exposure of 140 V for 17 seconds, in terms of shape there are more fragmentosit, microsite, tear drops, stomatocytes, and cell target, in terms of size there are more microsite, and in terms of color there are more hypochromic erythrocytes.&#x0D; Keywords: Burn Injury, Erythrocyte Morphology, Erythrocyte Count

Effects of Aqueous Extract of Basella alba L. leaves on Blood Cell Count in Rats

The effect of aqueous extract of Basella alba (puishak) leaves on blood cell count of rats was studied. Forty rats of both sexes weighing between 100-150 gm were used. The rats were divided into four groups (7 rats in each group), with group A as the control group and experimental groups were denoted as B, C and D. With all aseptic precautions, aqueous extract of Basella alba (Puishak) leaves was administered into the three different doses (For group B: 60 mg/kg bw, group C: 80 mg/kg bw and group D: 100 mg/kg bw). Control group A also received distilled water as a placebo at the dose of 10 mg/kg bw for 14 days. At day 15, blood samples were collected and sent for haematological analysis. Data analysis of blood count profile of 28 rats revealed that there is an increased number of RBC, WBC and platelet count in experimental groups than in control group. ANOVA test revealed that increased blood cell counts following administration of aqueous leaves extract of keeves of B. alba were statistically significant (p value for each case was .0001&lt;.05) than control group. Paired samples t test was performed to compare between baseline hematological parameters and parameters after 14 days of intervention. Then comparison between Group A &amp; Group B, between Group A &amp; Group C and between Group A &amp; Group D were done. In all cases, probability (p) value &lt; 0.05 was considered as statistically significant.This it is clearly evident that aqueous extract of B.alba has positive stimulant effect on blood cells count of rats. Moreover, it was found that increment of doses also increases the cell count that is positively correlated with the hypothesis. Bangladesh Pharmaceutical Journal 22(1): 73-78, 2019

90-Day Oral Toxicity Assessment of Tropaeolum majus L. in Rodents and Lagomorphs.

Tropaeolum majus L., popularly known as nasturtium, is a species widely used in the form of infusions and salads. In the last years, the antihypertensive, diuretic, and calcium and potassium sparing activities of T. majus preparations were shown. Moreover, no preclinical 90-day oral toxicity studies were conducted. Thus, this study evaluated the toxicity of the hydroethanolic extract obtained from T. majus (HETM) leaves in female and male mice, rats, and rabbits. Swiss mice and Wistar rats were treated with HETM (75, 375, and 750 mg/kg). The doses of rabbits (30, 150, and 300 mg/kg) were calculated by allometric extrapolation. The control groups received vehicle. The animals were orally treated, daily, for 90 days. At the end, the animals were anesthetized, and body weight gain, relative weight of liver, kidney, and spleen, and histopathological changes were evaluated. Serum hematological and biochemical parameters were also analyzed. No alterations were found in body and organ weights or in histopathological and biochemical evaluation. Hematological analyses revealed small changes in lymphocytes and neutrophil counts in rats after administration of 750 mg/kg of HETM. These results showed that 90-day use of T. majus is safe in rodents and lagomorphs.

Guava leaf juice effect towards number of megakaryocytes in bone marrow of thrombocytopenic mice

BackgroundDengue virus infection that most frequently occur in tropical and subtropical regions can cause many symptoms, one of which is a decrease in thrombocyte count. Recent studies showed that guava leaf extract can increase the thrombocyte count in rats. The present study aimed to determine the effect of guava leaf juice in increasing the number of megakaryocytes in the bone marrow of thrombocytopenic mice.MethodsThis study was of experimental design. The study subjects were 24 mice (Mus musculus). The mice were randomly divided into four groups that were subjected to intervention for 14 days. Group 1 was given guava leaf juice (56 mg/kg) and quinine (14 mg/kg), group 2 guava leaf juice (56 mg/kg) only, group 3 was given quinine (14 mg/kg) and group 4 distilled water. After 14 days, from the bone marrow of the femoral bones of each of the mice, hematoxylin eosin stained histologic preparations were made. Anova test was used to analyze the data.ResultsThe mean megakaryocyte count per field of view in group 1 (2.83), group 2 (3.30), group 3 (2.24) and group 4 (2.93). Anova test results for all groups showed significant differences between groups (p&lt;0.05). The mean megakaryocyte count was increased in group 1 compared to group 3, but the difference was statistically not significant (p=0.206).Conclusion Guava leaf juice can increase the megakaryocyte count in the bone marrow of thrombocytopenic mice. This suggests a potential role of guava leaf juice in improving the platelet count in thrombocytopenic disorders.

Effect of Carica papaya Linn. Unripe and Ripe Fruit Pulp Juice in Augmenting Platelet Count in Rats: An Experimental Study

Effect of Carica papaya Linn. Unripe and Ripe Fruit Pulp Juice in Augmenting Platelet Count in Rats: An Experimental Study

Cenerimod, a novel selective S1P1 receptor modulator with unique signaling properties.

Sphingosine‐1‐phosphate receptor 1 (S1P1) modulators sequester circulating lymphocytes within lymph nodes, thereby preventing potentially pathogenic autoimmune cells from exiting into the blood stream and reaching inflamed tissues. S1P1 receptor modulation may thus offer potential to treat various autoimmune diseases. The first nonselective S1P1‐5 receptor modulator FTY720/fingolimod/Gilenya® has successfully demonstrated clinical efficacy in relapsing forms of multiple sclerosis. However, cardiovascular, hepatic, and respiratory side‐effects were reported and there is a need for novel S1P1 receptor modulators with better safety profiles. Here, we describe the discovery of cenerimod, a novel, potent and selective S1P1 receptor modulator with unique S1P1 receptor signaling properties and absence of broncho‐ and vasoconstrictor effects ex vivo and in vivo. Cenerimod dose‐dependently lowered circulating lymphocyte counts in rats and mice after oral administration and effectively attenuated disease parameters in a mouse experimental autoimmune encephalitis (EAE) model. Cenerimod has potential as novel therapy with improved safety profile for autoimmune diseases with high unmet medical need.

EDC IMPACT: Reduced sperm counts in rats exposed to human relevant mixtures of endocrine disrupters.

Human semen quality is declining in many parts of the world, but the causes are ill defined. In rodents, impaired sperm production can be seen with early life exposure to certain endocrine-disrupting chemicals, but the effects of combined exposures are not properly investigated. In this study, we examined the effects of early exposure to the painkiller paracetamol and mixtures of human relevant endocrine-disrupting chemicals in rats. One mixture contained four estrogenic compounds; another contained eight anti-androgenic environmental chemicals and a third mixture contained estrogens, anti-androgens and paracetamol. All exposures were administered by oral gavage to time-mated Wistar dams rats (n = 16–20) throughout gestation and lactation. In the postnatal period, testicular histology was affected by the total mixture, and at the end of weaning, male testis weights were significantly increased by paracetamol and the high doses of the total and the anti-androgenic mixture, compared to controls. In all dose groups, epididymal sperm counts were reduced several months after end of exposure, i.e. at 10 months of age. Interestingly, the same pattern of effects was seen for paracetamol as for mixtures with diverse modes of action. Reduced sperm count was seen at a dose level reflecting human therapeutic exposure to paracetamol. Environmental chemical mixtures affected sperm count at the lowest mixture dose indicating an insufficient margin of safety for the most exposed humans. This causes concern for exposure of pregnant women to paracetamol as well as environmental endocrine disrupters.

Sertoli cell specific knockdown of RAR-related orphan receptor (ROR) alpha at puberty reduces sperm count in rats

Globally, there is an alarming decline in sperm count. Very often hormonal supplementation fails to restore normal sperm count. Sertoli cells (Sc) present within seminiferous tubules provide appropriate niche and factors required for the differentiation of germ cells (Gc) into mature sperm (spermatogenesis). Functionally compromised Sc may be one of the reasons for failure of hormones to facilitate normal spermatogenesis. Although role of secretory proteins and signaling molecules of Sc has been studied well, role of transcription factors regulating sperm count has not been addressed appropriately. Retinoic acid receptor-related orphan receptor (ROR)-alpha is one of such transcription factors reported in testis but its role in testicular function is not yet known. In a separate study, we found abundant ROR-alpha binding sites on promoter regions of several genes upregulated in pubertal rat Sc as compared to infant Sc. Immunostaining studies also revealed presence of ROR alpha in nucleus of pubertal Sc. We generated a transgenic knockdown rat model expressing shRNA targeted to ROR-alpha under Sc specific promoter, which is transcriptionally active only at and after puberty. ROR-alpha knockdown animals were found to have abnormal association of Sc and Gc, including Gc sloughing and restricted release of sperm. The knockdown animals displayed compromised spermatogenesis leading to significant reduction in sperm count. This is the first report describing the Sc specific role of ROR-alpha in maintaining quantitatively normal sperm output. Identification of various such molecules can generate avenues to limit or reverse an alarmingly declining sperm count witnessed globally in men.

Intervention effect and mechanism of curcumin in chronic urinary tract infection in rats

To analyze the invention effect of curcumin on chronic urinary tract infection in rats and explore its possible mechanism of action. The experimental animals were randomly divided into three groups, normal, model and curcumin group. Chronic urinary tract infection models were built for model group and curcumin group by injecting coliform fluid into the cavity of bladder. From the first day of modeling, rats in the curcumin group were injected with 150mg/kg curcumin, while rats in normal group and model group were given no other treatment. The treatment lasted for 14d. The white blood cell counts in blood and urine, bacterial colony count in urine and renal tubular functional indexes of rats in all groups at day 1, 7, and 14 after treatment were detected. Urine β2-microglobulin (β2-MG), urinary N-acetyl-β-D glucosaminidase (NAG) levels were used to detected the inflammatory cytokines in serum after treatment including the contents of IL-6, IL-8, IL-10 and monocyte chemoattractant protein-1 (MCP-1), and real-time PCR was employed to determine the expression of mRNA of toll-like receptor 2 (TLR-2) and TLR-4 in renal tissues and bladder tissues of all groups after treatment. The white blood cell counts at day 1 and 7 after treatment in rats of model group and curcumin group were significantly higher than those of normal group at the same time points, while the white blood cell counts of the curcumin group were significantly lower than those of model group (P<0.05). The urine white blood cell counts in rats of model group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points; those in the curcumin group were significantly lower than those of the model group at day 1, 7 and 14 at the same time points (P<0.05). The bacterial colony counts of urine in rats of model group and curcumin group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points, while the counts of curcumin group were significantly lower than those of model group at the same time points (P<0.05). Levels of urine β2-MG, NAG, IL-6, IL-8, IL-10, MCP-1 and expression of TLR2 mRNA and TLR4 mRNA in renal and bladder tissues in rats of model group were significantly higher than those of the normal group, while these variables of the curcumin group were significantly higher than those of the normal group but lower than those of model group (P<0.05). Curcumin can significantly improve the symptoms of chronic urinary tract infections, protect renal tubular function, and also decline inflammatory responses by influencing the expressions of TLR2 mRNA and TLR4 mRNA so as to exert its curative effect on chronic urinary tract infections.

Population Pharmacokinetic–Pharmacodynamic Modeling of 5-Fluorouracil for Toxicities in Rats

Myelosuppression is a dose-limiting toxicity of 5-fluorouracil (5-FU). Predicting the inter- and intra-patient variability in pharmacokinetics and toxicities of 5-FU may contribute to the individualized medicine. This study aimed to establish a population pharmacokinetic-pharmacodynamic model that could evaluate the inter- and intra-individual variability in the plasma 5-FU concentration, 5-FU-induced body weight loss and myelosuppression in rats. Plasma 5-FU concentrations, body weight loss, and blood cell counts in rats following the intravenous administration of various doses of 5-FU for 4days were used to develop the population pharmacokinetic-pharmacodynamic model. The population pharmacokinetic model consisting of a two-compartment model with Michaelis-Menten elimination kinetics successfully characterized the individual and population predictions of the plasma concentration of 5-FU and provided credible parameter estimates. The estimates of inter-individual variability in maximal rate of saturable metabolism and residual variability were 8.1 and 22.0%, respectively. The population pharmacokinetic-pharmacodynamic model adequately described the individual complete time-course of alterations in body weight loss, erythrocyte, leukocyte, and lymphocyte counts in rats treated with various doses of 5-FU. The inter-individual variability of the drug effects in the pharmacodynamic model for body weight loss was 82.6%, which was relatively high. The results of the present study suggest that not only individual fluctuations in the 5-FU concentration but also the cell sensitivity would affect the onset and degree of 5-FU-induced toxicity. This population pharmacokinetic-pharmacodynamic model could evaluate the inter- and intra-individual variability in drug-induced toxicity and guide the assessments of novel anticancer agents in drug development.