Superoxide Dismutase in Extender Improves the in vitro Quality and in vivo Fertility of Cryopreserved Water Buffalo (Bubalus bubalis) Spermatozoa.

BACKGROUND: Superoxide dismutase (SOD) as an antioxidant in semen extender may be used for the cryopreservation of buffalo spermatozoa and in vivo fertility. OBJECTIVE: This study was aimed to evaluate the effects of SOD (SOD1, 100 IU/mL; SOD2, 200 IU/mL; SOD3, 300 IU/mL) and control (0.0) in Tris citric acid extender on in vitro quality and in vivo fertility of cryopreserved water buffalo bull spermatozoa. MATERIALS AND METHODS: Semen collection was carried out on a weekly basis (four bulls, three replicates, and n = 24 ejaculates). The conventional freezing of semen loaded straws (0.5 mL) was undertaken by placing them horizontally on a steel rack inside a Styrofoam box for 10 min containing liquid nitrogen (LN2) vapours, and plunging into a liquid nitrogen tank (-196 °C) for storage, followed by thawing at 37 °C for 30 s and analysis by computer-assisted sperm analyzer (CASA) and other assays. RESULTS: At post-dilution, the acrosome integrity (ACR-I, %) was significantly improved (P < 0.05) in extender supplemented with SOD3 as compared to other experimental groups. In addition, DNA integrity (DNA-I, %) was significantly higher (P < 0.05) in SOD1 and SOD3 compared to SOD2 and control. At post-thawing, the mean values of sperm progressive motility (PM, %), average path velocity (VAP, µm/s) and straight line velocity (VSL, µm/s) were significantly higher (P < 0.05) in extender supplemented with SOD3 compared to the control. At post-thawing, mean values of subjective motility (SM, %), plasma membrane integrity (PMI, %) and ACR-I were significantly higher (P < 0.05) in extender supplemented with SOD3 compared to the control. At post-thawing, sperm DNA-I was significantly higher (P < 0.05) in extender supplemented with all SOD doses compared to the control in a dose-dependent manner. The in vivo fertility rate (%) was significantly higher with SOD3 compared to the control (68.2 % vs. 49.5 %). CONCLUSION: The supplementation of SOD3 (300 IU/mL) in Tris citric acid extender improves both in vitro quality and in vivo fertility of buffalo bull spermatozoa.

Indicator microbes correlate with pathogenic bacteria, yeasts and helminthes in sand at a subtropical recreational beach site.

AIMS: Research into the relationship between pathogens, faecal indicator microbes and environmental factors in beach sand has been limited, yet vital to the understanding of the microbial relationship between sand and the water column and to the improvement of criteria for better human health protection at beaches. The objectives of this study were to evaluate the presence and distribution of pathogens in various zones of beach sand (subtidal, intertidal and supratidal) and to assess their relationship with environmental parameters and indicator microbes at a non-point source subtropical marine beach. METHODS AND RESULTS: In this exploratory study in subtropical Miami (Florida, USA), beach sand samples were collected and analysed over the course of 6 days for several pathogens, microbial source tracking markers and indicator microbes. An inverse correlation between moisture content and most indicator microbes was found. Significant associations were identified between some indicator microbes and pathogens (such as nematode larvae and yeasts in the genus Candida), which are from classes of microbes that are rarely evaluated in the context of recreational beach use. CONCLUSIONS: Results indicate that indicator microbes may predict the presence of some of the pathogens, in particular helminthes, yeasts and the bacterial pathogen Staphylococcus aureus including methicillin-resistant forms. Indicator microbes may thus be useful for monitoring beach sand and water quality at non-point source beaches. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of both indicator microbes and pathogens in beach sand provides one possible explanation for human health effects reported at non-point sources beaches.

Proteome analysis of soybean hypocotyl and root under salt stress.

To evaluate the response of soybean to salt stress, the related changes in protein expression were investigated using the proteomic approach. Soybean plants were exposed to 0, 50, 100, and 200 mM NaCl. Especially at 200 mM, the length and fresh weight of the hypocotyl and root reduced under salt stress, while the proline content increased. Proteins from the hypocotyl and root treated with 100 mM NaCl were extracted and separated by two-dimensional polyacrylamide gel electrophoresis; 321 protein spots were detected. In response to salt stress, seven proteins were reproducibly found to be up- or down-regulated by two to sevenfold: late embryogenesis-abundant protein, beta-conglycinin, elicitor peptide three precursor, and basic/helix-loop-helix protein were up-regulated, while protease inhibitor, lectin, and stem 31-kDa glycoprotein precursor were down-regulated. These results indicate that salinity can change the expression level of some special proteins in the hypocotyl and root of soybean that may in turn play a role in the adaptation to saline conditions.

Localization of a domain in the FimH adhesin of Escherichia coli type 1 fimbriae capable of receptor recognition and use of a domain-specific antibody to confer protection against experimental urinary tract infection.

The FimH subunit of type 1-fimbriated Escherichia coli has been implicated as an important determinant of bacterial adherence and colonization of the urinary tract. Here, we sought to localize the functionally important domain(s) within the FimH molecule and to determine if antibodies against this domain would block adherence of type 1-fimbriated E. coli to the bladder mucosa in situ and in vivo in an established mouse model of cystitis. We generated translational fusion proteins of disparate regions of the FimH molecule with an affinity tag MalE, and tested each of the fusion products in vitro for functional activity. The minimum region responsible for binding mouse bladder epithelial cells and a soluble mannoprotein, horseradish peroxidase, was contained within residues 1-100 of the FimH molecule. We validated and extended these findings by demonstrating that antibodies directed at the putative binding region of FimH or at synthetic peptides corresponding to epitopes within the binding domain could specifically block type 1 fimbriae-mediated bacterial adherence to bladder epithelial cells in situ and yeast cells in vitro. Next, we compared the ability of mice passively immunized intraperitoneally with antisera raised against residues 1-25 and 253-264 of FimH or 1-13 of FimA to resist bladder colonization in vivo after intravesicular challenge with type 1-fimbriated E. coli. Only the antibody directed at the putative binding region of FimH (anti- s-FimH1-25) significantly reduced E. coli bladder infections in the experimental mouse model of urinary tract infections. Similar results were obtained when the mice were actively immunized with synthetic peptides corresponding to residues 1-25 and 253-264 of FimH or 1-13 of FimA. The mechanism of protection was attributed, at least in part, to inhibition of bacterial adherence to the bladder surface by s-FimH1-25-specific antibody molecules that had filtered through the kidneys into the urine. The level of FimH antibodies entering the bladder from the circulatory system of the immunized mice was found to be markedly enhanced upon bacterial challenge. The potential broad spectrum activity of the protective FimH antibody was indicated from its serologic cross-reactivity with various urinary tract bacterial isolates bearing type 1 fimbriae. These findings could be relevant in the design of an efficacious and broadly reactive FimH vaccine against urinary tract infections.

The role of mental rotation and memory scanning on the performance of laparoscopic skills: a study on the effect of camera rotational angle.

OBJECTIVE: The rotational angle of the laparoscopic image relative to the true horizon has an unknown influence on performance in laparoscopic procedures. This study evaluates the effect of increasing rotational angle on surgical performance. METHODS: Surgical residents (group 1) (n = 6) and attending surgeons (group 2) (n = 4) were tested on two laparoscopic skills. The tasks consisted of passing a suture through an aperture, and laparoscopic knot tying. These tasks were assessed at 15 degrees intervals between 0 degrees and 90 degrees , on three consecutive repetitions. The participant's performance was evaluated based on the time required to complete the tasks and number of errors incurred. RESULTS: There was an increasing deterioration in suturing performance as the degree of image rotation was increased. Participants showed a statistically significant 20-120% progressive increase in time to completion of the tasks (p = 0.004), with error rates increasing from 10% to 30% (p = 0.04) as the angle increased from 0 degrees to 90 degrees. Knot-tying performance similarly showed a decrease in performance that was evident in the less experienced surgeons (p = 0.02) but with no obvious effect on the advanced laparoscopic surgeons. CONCLUSIONS: When evaluated independently and as a group, both novice and experienced laparoscopic surgeons showed significant prolongation to completion of suturing tasks with increased errors as the rotational angle increased. The knot-tying task shows that experienced surgeons may be able to overcome rotational effects to some extent. This is consistent with results from cognitive neuroscience research evaluating the processing of directional information in spatial motor tasks. It appears that these tasks utilize the time-consuming processes of mental rotation and memory scanning. Optimal performance during laparoscopic procedures requires that the rotation of the camera, and thus the image, be kept to a minimum to maintain a stable horizon. New technology that corrects the rotational angle may benefit the surgeon, decrease operating time, and help to prevent adverse outcomes.

Omeprazole as adjuvant therapy to endoscopic combination injection sclerotherapy for treating bleeding peptic ulcer.

BACKGROUND: Therapeutic endoscopy has provided a new means of treating bleeding peptic ulcers. Additional medical therapy may enhance the therapeutic benefit. Hemostasis is highly pH dependent and is severely impaired at low pH. Proton pump inhibitors, by achieving a significantly higher inhibition of gastric acidity, may improve the therapeutic outcomes after endoscopic treatment of ulcers. PATIENT AND METHODS: We enrolled 166 patients with hemorrhage from duodenal, gastric, or stomal ulcers and signs of recent hemorrhage, as confirmed by endoscopy. Twenty-six patients had ulcers with an arterial spurt, 41 patients had active ooze, 37 had a visible vessel, and 62 patients had an adherent clot. All patients received endoscopic injection sclerotherapy using 1:10,000 adrenaline and 1% polidocanol and were randomly assigned to receive omeprazole (40 mg orally) every 12 hours for 5 days or an identical-looking placebo. The outcome measures used were recurrent bleeding, surgery, blood transfusion, and hospital stay. RESULTS: Six (7%) of 82 patients in the omeprazole group had recurrent bleeding, as compared with 18 (21%) in the placebo group (P = 0.02). Two patients in the omeprazole group and 7 patients in the placebo group needed surgery to control their bleeding (P = 0.17). One patient in the omeprazole group and 2 patients in the placebo group died (P = 0.98). Twenty-nine patients (35%) in the omeprazole group and 61 patients (73%) in the placebo group received blood transfusions (P <0.001). The average hospital stay was 4.6 +/- 1.1 days in the omeprazole group and 6.0 +/- 0.7 days in the placebo group (P <0.001). CONCLUSION: The addition of oral omeprazole to combination injection sclerotherapy decreases the rate of recurrent bleeding, reduces the need for surgery and transfusion, and shortens the hospital stay for patients with stigmata of recent hemorrhage.

Evaluation of the genotoxic, cytotoxic, and antitumor properties of Commiphora molmol using normal and Ehrlich ascites carcinoma cell-bearing Swiss albino mice.

The genotoxic, cytotoxic and antitumor properties of Commiphora molmol (oleo gum resin) were studied in normal and Ehrlich ascites carcinoma cell-bearing mice. In normal mice, the genotoxic and cytotoxic activity was evaluated on the bases of the frequency of micronuclei and the ratio of polychromatic to normochromatic cells in bone marrow, which was substantiated by the biochemical changes in hepatic cells. The antitumor activity of C. molmol was evaluated from the total count and viability of Ehrlich ascites carcinoma cells and their nucleic acid, protein, malondialdehyde, and elemental concentrations in addition to observations on survival and the trend of changes in body weight. The tumors at the site of injection were evaluated for histopathological changes. Treatment with C. molmol (125-500 mg/kg) showed no clastogenicity but was found to be highly cytotoxic in normal mice. The results obtained in the Ehrlich ascites carcinoma cell-bearing mice revealed the cytotoxic and antitumor activity of C. molmol which was found to be equivalent to those of the standard cytotoxic drug cyclophosphamide. On the basis of the nonmutagenic, antioxidative, and cytotoxic potential of C. molmol as observed in the present study, its use in cancer therapy seems to be appropriate and further investigations are suggested.

Influence of anethole treatment on the tumour induced by Ehrlich ascites carcinoma cells in paw of Swiss albino mice.

The anticarcinogenic potential of anethole was studied in Ehrlich ascites tumour (EAT) in the paw of Swiss albino mice. The antitumour activity was evaluated from the cytotoxicity of EAT-cells in the paw and their biochemical changes were determined from nucleic acids, protein, malondialdehyde (MDA) and glutathione (NP-SH) concentrations. Furthermore, the observations on survival rate, tumour weight, its volume and body weight of EAT-bearing mice were made. The EAT-bearing paws were also evaluated for histopathological changes. Additional studies were undertaken on the cytological effects of anethole in order to establish its clastogenic and mitodepressive activity in normal mice. The results obtained in the present study revealed anethole to increase the survival time, reduce the tumour weight and volume and body weight of the EAT-bearing mice. It caused a significant cytotoxic effect in EAT cells in the paw, reduced the levels of nucleic acids and MDA, and increased NP-SH concentrations. The histopathological changes observed after treatment with anethole were comparable to the standard cytotoxic drug cyclophosphamide. The results on the frequency of micronuclei and the ratio of polychromatic erythrocytes to normochromatic erythrocytes showed anethole to be mitodepressive and non-clastogenic in the femoral cells of mice. Our results indicate the anticarcinogenic, cytotoxic and non=clastogenic nature of anethole. Further studies are warranted to explore the mode of action and safety of anethole for its possible use in cancer therapy.

Uric acid as an inhibitor of biochemical changes induced by cyclophosphamide in mice.

The biochemical changes induced by uric acid, cyclophosphamide and uric acid plus cyclophosphamide were evaluated in Swiss albino male mice. Uric acid dissolved in water was administered orally in different doses for 7 days. Some mice from each group were injected intraperitoneally with cyclophosphamide (25 mg/kg) and sacrificed after 30 h. The blood of all animals was analyzed for uric acid levels. Uric acid was found not to affect the biosynthesis of nucleic acids and proteins in liver, testes and brain at 10-100 mg/kg/d. Pretreatment with uric acid provided significant protection against cyclophosphamide-induced impairment of DNA, RNA and protein biosynthesis.

Uric acid as an inhibitor of cyclophosphamide-induced micronuclei in mice.

Swiss albino male mice, 6-8 weeks old, were treated orally with different doses of uric acid dissolved in water for 7 days. Some of the mice in each group were injected i.p. with cyclophosphamide (25 mg/kg) and killed after 30 h. The blood of all animals was analyzed for uric acid levels. The femoral cells of the mice in different groups were collected and studied. Uric acid was found to be devoid of mitodepressant or clastogenic activity at 10-100 mg/kg/day. Pretreatment with uric acid was found to provide significant protection against cyclophosphamide-induced bone marrow depression and micronucleated polychromatic erythrocytes.

A study of uric acid pretreatment for the protection of rat gastric mucosa against toxic damage.

Uric acid was evaluated for its potential to protect the gastric mucosa against the injuries caused by 80% ethanol, 0.6 m-HCl and 0.2 M-NaOH in rats. Uric acid at doses of 50, 100 or 300 mg/kg body weight provided dose-dependent protection against the ulcerogenic effects of all three agents. Other effects caused by ethanol only were studied. Serum uric acid concentrations were statistically significantly increased by both uric acid and ethanol treatments. Treatments of rats by gavage with 1 ml 80% ethanol was found to cause depletion of stomach-wall mucus, to lower the concentrations of protein, nucleic acids and non-protein sulphydryl groups in the stomach wall, and to cause histopathological lesions, including necrosis, erosions, congestion and haemorrhage, of the stomach wall. Treatment with uric acid, at doses of 50, 100 or 300 mg/kg body weight, by gavage, provided some measure of protection against all of these effects, and the protection was generally dose dependent. The protective effects of uric acid against damage to the gastric-wall mucosa may be mediated through its effects on mucus production and non-protein sulphydryl concentrations, and/or its free-radical scavenging properties.

Effect of Caralluma tuberculata on the cytological and biochemical changes induced by cyclophosphamide in mice.

Treatment with Caralluma tuberculata extract induced complex biochemical and cytological changes in mice. Its cytotoxicity in the bone marrow cells of mice was comparable with that of the standard drug cyclophosphamide (CP); however, unlike CP, C. tuberculata was not clastogenic (as shown by the micronucleus assay). A dose-dependent decrease in the RNA content of liver and testes was produced by C. tuberculata treatment whereas there was no effect on the content of nucleic acid and protein in the brain. In the extract-treated animals there was a significant and dose-dependent increase in the DNA content of the liver, with a negligible effect on the protein content. Combined treatment with C. tuberculata and CP showed that C. tuberculata diminished the effect of CP on DNA levels; however, RNA levels were further suppressed, resulting in increased cytotoxicity. Pretreatment with C. tuberculata extract significantly reduced the clastogenicity of CP. These results indicated the involvement of different phytoconstituents acting by different routes.

Toxicity studies in mice of ethanol extracts of Foeniculum vulgare fruit and Ruta chalepensis aerial parts.

Acute (24-h) and chronic (90-day) oral toxicity studies on the ethanolic extracts of Foeniculum vulgare fruit and Ruta chalepensis aerial parts were carried out in mice. Acute dosages were 0.5, 1.0 and 3 g/kg while the chronic dosage was 100 mg/kg per day of extract. All external morphological, haematological and spermatogenic changes, in addition to body and vital organ weights were recorded. The extracts caused no significant acute or chronic mortality as compared to controls during this investigation. The treated male mice gained significant weight during chronic treatment while a loss or no significant change in weight was noticed in the female mice treated with the same extracts. Haematological studies revealed a significant fall in RBC level of R. chalepensis-treated animals. Both the extracts failed to show spermatotoxic effects.

Effect of Allium sativum on epididymal spermatozoa, estradiol-treated mice and general toxicity.

A water extract of Allium sativum (garlic) was given to male and female mice in drinking water (100 mg/kg/day) for three months. There was a significant increase in the weight of seminal vesicles and epididymides of male animals as compared to controls and the sperm count was significantly elevated. There was no increase in the body weight of the test animals, and weights of heart, liver and spleen were reduced as compared to controls. Haematological studies revealed an increase in WBC and a decrease in RBC levels of the test animals. Three-day intraperitonial treatment (500 mg/kg) with the A. sativum extract failed to exhibit any estrogenic or antiestrogenic activity.

Toxicity studies in mice of Commiphora molmol oleo-gum-resin.

Acute (24 h) and chronic (90 days) oral toxicity studies on Commiphora molmol (oleo-gum-resin) were carried out in mice. Dosages in acute study were 0.5, 1.0 and 3 g/kg, while in chronic study dosage was 100 mg/kg per day. All external morphological, biochemical and haematological changes, in addition to body and vital organ weights were recorded. There was no significant difference in mortality in acute or chronic treatment as compared to controls. At the end of the treatment, weight gain in the treated as well as control group was significant. There was a significant increase in weight of testes, caudae epididymides and seminal vesicles in C. molmol treated group. Biochemical studies revealed no differences in C. molmol treated animals, however, haematological studies revealed a significant increase in RBC and haemoglobin levels as compared to the control group. C. molmol failed to show any spermatotoxic effects.

Mitodepressive, clastogenic and biochemical effects of (+)-usnic acid in mice.

Mice were treated orally with aqueous suspensions of (+)-usnic acid in a single dose of either 100 or 200 mg/kg. The effects on femur cells and proteins and on nucleic acids of liver cells were studied 24-72 h after treatment. (+)-Usnic acid was found to affect the proliferation of polychromatic erythrocytes possibly by interference with RNA biosynthesis. The slight increase in the micronucleated polychromatic erythrocytes without affecting DNA synthesis suggests an effect of usnic acid on spindle apparatus.

Gastric antiulcer and cytoprotective effect of Commiphora molmol in rats.

The aqueous suspension of Commiphora molmol (oleo-gum resin) has been screened for its potential to protect gastric mucosa against the ulcers caused by 80% ethanol, 25% NaCl, 0.2 M NaOH, indomethacin and combined indomethacin-ethanol treatment. C. molmol pretreatment at doses of 250, 500 and 1000 mg/kg provided dose-dependent protection against the ulcerogenic effects of different necrotizing agents used. The effects caused by ethanol were further investigated. Treatment of rats with 1 ml of 80% ethanol was found to cause depletion of stomach wall mucus, reduction in the concentration of protein, nucleic acids and NP-SH groups in the stomach wall. Ethanol treatment also caused histopathological lesions including necrosis, erosion, congestion and haemorrhage of the stomach wall. Pretreatment with C. molmol offered a dose-dependent protection against all these effects. In the same manner it affected the malondialdehyde concentration altered by ethanol treatment. C. molmol also offered protection against mucosal damage caused by indomethacin and its combination with ethanol. The protective effect of C. molmol observed in the present study is attributed to its effect on mucus production, increase in nucleic acid and non-protein sulfhydryl concentration, which appears to be mediated through its free radical-scavenging, thyroid-stimulating and prostaglandin-inducing properties.

Effect of Trigonella foenum-graecum and Ammi majus on calcium oxalate urolithiasis in rats.

The present study was undertaken to investigate the effect of Trigonella foenum-graecum seed and Ammi majus fruit on experimentally-induced kidney stones. Oxalate urolithiasis in male rats was produced by the addition of 3% glycolic acid to their diet. After 4 weeks, highly significant deposition in the kidneys was noticed and changes in water intake and body weight recorded. Daily oral treatment with T. foenum-graecum significantly decreased the quantity of calcium oxalate deposited in the kidneys thus supporting its use in Saudi folk medicine. The effects obtained by A. majus were, however, not significant.

Effect of camel urine on the cytological and biochemical changes induced by cyclophosphamide in mice.

Camel urine treatment was found to cause a significant cytotoxic effect in the bone marrow cells of mice. This cytotoxicity at higher doses was comparable with that of standard drug cyclophosphamide (CP). However, unlike CP, the camel urine treatment failed to induce any clastogenicity. The cytotoxicity induced by camel urine treatment was substantiated by the reduction of liver nucleic acids and glutathione levels and increased malondialdehyde (MDA) contents in the same animals. CP treatment was found to be highly clastogenic, cytotoxic and it reduced the levels of nucleic acids, proteins, glutathione and increased malondialdehyde concentration due to its prooxidant nature. The non-clastogenic nature of camel urine was attributed to the antioxidant and antimutagenic compounds present in camel urine. Pretreatment with camel urine increased the cytotoxicity of CP and intensified the CP induced reduction of liver nucleic acids, glutathione and increased the MDA concentration. The increase of CP induced cytotoxicity appears to be partly due to the additive effect of the two treatments on cellular lipid peroxidation.

Preliminary toxicity studies on ethanol extracts of the aerial parts of Artemisia abyssinica and A. Inculta in mice.

Ethanolic extracts of the aerial parts of Artemisia abyssinica and A. inculta were subjected to acute toxicity observations in mice for 24 h and chronic toxicity evaluation for 3 months. External morphological changes, visceral toxicity, haematological changes, spermatogenic dysfunction and effect on body weight and vital organ weight were recorded. In both the chronically treated groups, no significant acute mortality was observed up to 3 g/kg p.o. There was no weight gain in A. abyssinica chronically-treated mice while the weight gain of A. inculta-treated animals matched that of the control group. Significant sperm damage was observed in A. abyssinica-treated mice while A. inculta failed to produce any significant spermatotoxic effect.