Triclabendazole (Anthelmintic Drug) Effects on the Excretory- Secretory Proteome of Fasciola hepatica in Two Dimension Electrophoresis Gel

Background

The aim of this study was to evaluate the protein spots of excretory - secretory products of Fasciola hepatica using two dimension electrophoresis method in the presence and absence of triclabendazole drug which can be considered to detect the target protein of the drug.

Methods

F. hepatica parasites were collected from infected cattle livers, divided in two groups and cultivated in RPMI 1640 medium. First group was treated with triclabendazole (TCBZ) and second group considered as control. The excretory-secretory (ES) products of each group were separated and total protein determined by Bradford method. To provide proteome spots, the ES proteins were precipitated and two dimension electrophoresis (2-DE) gel prepared. Protein amounts of two groups were compared using the statistical t-test and protein spots from 2-DE in test and control groups were also statistically analyzed. The protein spots of gels were identified by using protein database.

Results

The t-test showed a significant increase of total proteins in treated group (P<0.5). The protein spots count in the control group was less than test group however statistically not significant (p>0.05). Cathepsin L- protein (MW 36.7 pH 5.34), 14-3-3 epsilon 2 isoform (MW 28.2 pH 5.36), Cathepsin L1D (MW 36.5 pH 5.8) and Cathepsin L1D (MW 36.6 pH 6.26) were identified in test group.

Conclusion

It seems that, these results can be considered to determine the proteins which the drug acts as a target on them.     

Frequency of Toxoplasma gondii in HIV Positive Patients from West of Iran by ELISA and PCR

Background

Toxoplasma gondii, the obligate intracellular parasite is life threatening in AIDS patients. Diagnosis of toxoplasmosis is based on serological methods especially increasing of IgM and IgG titers, but finding of parasite or its components (antigenemia) may be beneficial method in order to detection of acute toxoplasmosis in immunocompromised patients.

Methods

Ninety-four serum samples from HIV positive patients were collected from Sanandaj, Kordistan west of Iran. These patients were lived in Sanandaj of whom 26 were prisoners infected with HIV virus in prison. Toxoplasma gondii antibodies were determined by IgG ELISA. T. gondii antigen was identified by capture-ELISA. PCR was performed on samples with T. gondii antigenemia. CD4+ T cells counts had been determined by flowcytometry and were obtained from records of each patient.

Results

Among the examined HIV seropositive individuals, 19.1% (18/94) and 5.3% (5/94) were positive for Toxoplasma-IgG and antigenemia, respectively. Besides, one of the samples was positively detected by PCR method. Mean age of participants was 37.9 ± 9.5 year. Prevalence of IgG antibody and antgenemia was higher in age group of 40-50 years old. The Mean of CD4+ T cells counts of participants (total of HIV+ patients, IgG positive patients and patients with antigenemia) was 699.2 ± 345.2, 655.1 ± 237.9 and 620.2 ± 215.1 respectively.

Conclusion

Capture-ELISA and PCR could confirm the T. gondii acute infection in HIV positive patients. For precise diagnosis of acute toxoplasmosis in HIV positive patient, performance of more studies based on more sensitive types of PCR is suggested.     

P-glycoprotein A Gene Expression in Glucantime-Resistant and Sensitive Leishmania major (MRHO/IR/75/ER)

Background

Leishmaniasis is a parasitic disease caused by different species of Leishmania parasites with a wide range of clinical manifestations. Antimonial compounds such as meglumine antimoniate (glucantime) are the first line drugs for the treatment of leishmaniasis. However, according to reports of the drug resistance of parasites, the efficacy of antimonial compounds is low. The ATP-binding cassette (ABC) proteins are present in all organisms and mediate the transport of vital elements through biological membranes. One of the important mechanisms of resistance in Leishmania parasites is the overexpression of ABC efflux pumps. P-glycoprotein A (pgpA) is a related gene for ABC transporter in Leishmania species. The aim of this study was to compare the pgpA expression in laboratory-induced resistant L. major (MRHO/IR/75/ER) and sensitive parasites.

Methods

RNA extraction of promastigotes of sensitive and resistant clones was performed and total RNA was reverse transcribed. The real-time quantitative polymerase chain reaction (PCR) was used to assess RNA expression profiles and the expression levels were calculated using 2^-ΔCt method.

Results

The mean expression level of pgpA mRNA was 2.70 ± 0.51 in in sensitive Leishmania clone and 6.08 ± 1.50 in resistant Leishmania clone (P = 0.021).

Conclusion

The expression of pgpA gene in resistant strains of L. major was almost fivefold higher than those in susceptible strains. Therefore, this can be used in field isolates, i.e. overexpression of the gene can prove resistance in wild type field isolates.     

Helminth Infections in Rattus ratus and Rattus norvigicus in Tehran,Iran

Background

The aim of this study was to determine intestinal and liver helminth infections in Rattus rodents in Tehran Iran.

Methods

Overall, 306 traps were put in 39 different regions in Tehran from 2009 to 2010. Rodents, including R. rattus and R. norvegicus were caught by live-traps. They become unconscious and the spinal cords were cut, afterwards the body was dissected and the stomach, small intestine, large intestine, liver, and cecum were studied separately. The dominant type and the prevalence rate of parasites in the rodents were determined based on the infected parts of their body.

Results

After recognition of the helminthes’ types, among the 120 total number of rodents, 39 belonged to males, while among the infected rodents, 57(47.5%) were female and 18(15%) were male. The prevalence of infection in Tehran was 62.5%. Seventy cases (58.33%) of helminth infections were observed in R. rattus and 5 cases (4.16%) were observed in R. norvegicus. The maximum prevalence (15.5%) was seen in the center and east part of Tehran, while the minimum (9.16%) was in the north part of the city. The helminthes types and the corresponding percentages were Hymenolepis nana fraterna (35.8%), Heterakis spumosa (17.5%), Hymenolepis diminuta (7.5%) and Capillaria annulosa (1.6%). The dominant rodent was Rattus rattus and among the identified helminthes, Hymenolepis diminuta and Hymenolepis nana fraterna are zoonotic ones.

Conclusion

The information presented here improves our understanding of the major parasitic infections that rodents harbor and can transmit to human and animal populations in Iran. To prevent infectivity of human, the hazard of the identified zoonotic species needs to be contemplated.     

Novel in Vitro Efficiency of Chitosan Biomolecule against Trichomonas gallinae

Background

Development of new natural agents for parasitic diseases treatment has unexpectedly increased to overcome effectively against emergence and re-emergence of parasitic diseases, the appearance of drug resistant organisms and toxic side effects of current agents. The aim of the study was to evaluate antiprotozoal activities of chitosan biomolecule on trophozoites of Trichomonas gallinae.

Methods

The antitrichomonal activity of various low molecular weight chitosan concentrations including 125, 250, 500 and 1250 µg ml⁻¹ against T. gallinae trophozoites cultured in trypticase-yeast extract-maltose medium supplemented with heat-inactivated cold horse serum was evaluated in vitro. Samples containing medium without chitosan were also assayed as controls.

Results

The mortality rates at 0, 3 and 6 h post treatment with all concentrations were significantly different from control group (P<0.05). Treated trophozoites showed more susceptibility to the highest concentration reaching mortality rate of 100% at 3h post inoculation. However, at this time, results for 125, 250 and 500 µg ml⁻¹ were 93%, 95% and 96.7%, respectively.

Conclusion

The results demonstrate that the application of chitosan biomolecule is a promising option for treatment of trichomoniasis in pigeons.     

Pathogenic Assays of Acanthamoeba Belonging to the T4 Genotype

Background

Acanthamoeba genus is introduced as opportunistic and cosmopolitan parasite. Monkey and wistar rat are appropriate models for experimental study on Acanthamoeba infection. In this study Acanthamoeba spp. were isolated from hot spring (HS), windows dust (WD) and a corneal sample of keratitis patient (KP) and their pathogenicity surveyed by in vitro and in vivo tests.

Methods

Isolates of Acanthamoeba were cultivated axenically for 12 months in PYG medium. Overall, 30 wistar rats, in 6 equal groups were used for developing experimental Acanthamoeba keratitis (AK) and Granulomatous Amoebic Encephalitis (GAE). The Keratitis and Granulomatous Encephalitis experiments were performed by intrastromal and intranasal inoculation of Acanthamoeba cysts, respectively. Pathogenicity of the three isolates was also evaluated by in vitro test using osmotolerance and temperature tolerance assays. Identification of genotypes were performed by PCR technique and sequencing.

Result

None of the isolates could perform AK and GAE in wistar rats, although all isolates were described as T4 genotype. Isolates obtained from KP and WD could grow only in 30 °C, but not in 37 °C and 40 °C. On the other hand, HS isolate grew in 30 °C and 37 °C but not in 40 °C. Moreover, all of isolate grew in 0.5 M mannitol but not in 1 M and 1.5 M.

Conclusion

T4 isolates with a long-term axenic culture and different factors related to host and parasite may play role in pathogenicity of these free-living amoebae.     

Axenic Cultivation and Pathogenic Assays of Acanthamoeba Strains Using Physical Parameters

Background

The main goal of the present study was to set up an axenic cultivation of Acanthamoeba and assess the pathogenic ability of T4 genotypes from different clinical and environmental strains of Acanthamoeba using two physical assays.

Methods

Sixteen Acanthamoeba isolates including 10 environmental and 6 clinical strains were cultured axenically. Axenic cultivation was performed using Proteosepepton, yeast extract and glucose medium and TY-I-S33culture. Pathogenic survey was done using osmotolerance and thermotolerance assay. Briefly, differentosmolarity (0.5 M and 1 M) of non-nutrient agar plates were performed. One hundred fifty µl of axenic culture were collected and were inoculated in 1% agar medium. For thermotolerance assay 150 µl of amoebas from axenic culture were divided into fresh culture mediums. Cultures were incubated at 37°C and 42 °C. All plates were monitored for 24 h, 48 h and 72 h.

Results

Overall, 16 strains of Acanthamoeba isolates previously genotyped as T4 were cultivated axenically after several months. Thermotolerance and osmotolerance assay revealed that all of clinical strains, soil and animal feces strains were highly pathogenic isolates. Two dust and water strains did not grow at high temperature (42 °C) and osmolarity (1.5 M) and thus they were classified as weak pathogens.

Conclusion

Most of T4 genotypes are highly pathogenic organisms. This is an important finding since Acanthamoeba belonging to T4 type is the predominate genotype in environmental and clinical samples. The presence of highly pathogenic Acanthamoeba may pose a risk within susceptible people.     

Evaluation of Alum-Naltrexone Adjuvant Activity,on Efficacy of Anti-Leishmania Immunization with Autoclaved Leishmania major (MRHO/IR/75/ER) Antigens in BALB/C Mice

Background

Naltrexone, an opioid receptor antagonist shifts the immune response toward a Th1 profile. In the current study, we evaluated the efficacy of the mixture of NTX and alum, as a new adjuvant, to enhance immune response and induce protection against Leishmania major in a mouse model.

Methods

BALB/c mice were immunized three times either autoclaved L. major promastigotes’ antigens alone or in combination with the adjuvant alum, naltrexone or the alum–naltrexone mixture. Both humoral and cellular immune responses were assessed two weeks after the last immunization and compared with control mice.

Results

The administration of alum- NTX in combination with the parasite antigen, significantly increased production of IFN-γ IFN-γ /IL-5 ratio, lymphocyte proliferation and improved DTH response against L. major. There was no significant difference in survival following challenge among groups.

Conclusion

Immunization with the alum– naltrexone mixture as an adjuvant, in combination with the autoclaved L. major promastigotes antigens, can enhance cellular immunity and shift the immune responses to a Th1 pattern.     

Interleukin-10 and Transforming Growth Factor-β in Early and Late Lesions of Patients with Leishmania major induced Cutaneous Leishmaniasis

Background

Cutaneous leishmaniasis is a neglected parasitic disease, which imposes massive human distress and financial costs to the endemic countries. Better understanding of host immune response to the parasite leads to helpful strategies for disease control. Interleukin (IL)-10 and transforming growth factor (TGF)-β are important immune regulatory cytokines, which appear to develop non-healing forms of leishmaniasis. However, there is little information about the function of IL-10 and TGF-β in old world cutaneous leismaniasis. The aim of this study was to analyze the role of IL-10 and TGF-β in human cutaneous leishmaniasis due to Leishmania major infection.

Methods

Biopsies were obtained from lesions of twenty proven cases of L. major induced cutaneous leishmaniasis. IL-10 and TGF-β positive cells were detected by immunofluorescence staining of frozen sections and compared between two groups of patients with early and late lesions.

Results

The mean percentage of IL-10 positive cells were significantly (P= 0.035) higher in late lesions (0.51±0.24) than early ones (0.15±0.07). Similar results were obtained for TGF-β with mean percentages of 0.16±0.05 and 0.53±0.28 in early and late lesions respectively (P= 0.008).

Conclusion

IL-10 and TGF-β are present in lesions of L. major induced cutaneous leishmaniasis and contribute to the pathogenesis of long lasting disease forms.     

Haemolymph Components of Infected & None Infected Lymnaea snails with Xiphidiocercariae

Background

In this study the haemolymph components of infected and none infected Lymnaea gedrosiana with xiphidiocercaria larvae was compared.

Methods

Five hundred Fifty Lymnaea snails were collected from Ilam and Mazandaran provinces, Iran, during 2008–2009. The snails were transported to the lab at Tehran University of Medical Sciences and their cercarial sheddings were studied. Haemolmyphs of snails were extracted and cells were counted using haemocytometer and cell-surface carbohydrate were recognized by conjugated lectin (Lentil). Haemolymph protein concentrations were measured by Bradford protein assay method and soluble protein compositions were determined on sodium dodecyl sulphate polyacrilamide gel electrophoresis (SDS-PAGE).

Results

From the 550 examined Lymnaea snails for cercariae, 27 snails were infected with xiphidiocercariae. Mean of haemolymph cells (haemocyte) number were obtained 93480±2.43 (cells/ml) for none infected snails (25 snail) and 124560±2800 (cells/ml) for infected snails (25 snail). Mannose carbohydrate was recognized on haemocyte of none infected and infected snails. Mean of protein concentration of haemolymph plasma was obtained as 1354±160 µg/ml (1.4 mg/ml) for none infected snails (25 snails) and 1802±138 µg/ml (1.8 mg/ml) for infected snail (25 snails). Comparing to none infected snails, the SDS-PAGE results of haemolymph plasma of infected snails, showed an extra protein band (70 kDa). The results showed a significant difference between the amounts and the kinds of proteins in haemolymph of infected and none infected snails.

Conclusion

This information might be useful to understand of parasite detection, adhesion, engulfment and antigen agglutination by snail.     

Parasitic Infection of an Endemic Fish (Blicca bjoerkna) and an Exotic Fish (Hemiculter beucisculus) In Anzali Lagoon,Caspian Sea,Iran

Background

In Anzali Lagoon, there are some endemic and exotic fishes. The present study was conducted to compare the parasitic fauna of Blicca bjeorkna, as an endemic fish and Hemiculter leucisculus, as an introduced fish to the lagoon.

Methods

A parasitological investigation was done on 78 specimens of B. bjoerkna and 114 of H. leucisculus. The fishes were collected from August 2009 to April 2010 by the electro fishing from Anzali Lagoon.

Results

Eleven parasites species were found in 192 fish samples. The prevalence and mean intensity of parasites in each host were as follows: Parasites from B. bjorkna were Trichodina perforata (53.85%); Myxobolus musayevi (27.19%, 1±0.79); Dactylogyrus difformis (88.05%, 8±7.24) and D. sphyrna (5.18%, 0.95±0.51), Diplostomum spataceum (98.72%, 9.51±9.01), Posthodiplostomum cuticula (15.38%, 4.25±2.5), Ripidocotyle sp. (1.28%, 2±0.74); Contracaecum osculatum (17.95%, 1.64±0.79), Philometra rischta (12.8%, 1.4±0.54), and Raphidascaris acus (1.04%, 0.03±0.26). The H. leucisculus were infected with T. perforata (27.19%), D. spataceum (7.89%, 1.33±0.54), Ps. tomentosa (7.02%, 1.62±0.49) and R. acus (0.88%, 3±0.28). B. bjoerkna was presented as a new host for M. musayevi and C. osculatum, while H. leucisculus was introduced as a new host for T. perforata and Ps. tomentosa.

Conclusion

The prevalence of parasites was significantly more in native fish than that of exotic fish (P<0.05). This reduction in parasitic infection in H. leucisculus may be due to its immune system resistance, well adaptation to the new environment, host-specific limitation for endemic parasites and disability of introduced parasite to complete its life cycle in the new host as well.     

Effect of St. John's Wort (Hypericum perforatum) on obesity,lipid metabolism and uterine epithelial proliferation in ovariectomized rats

BACKGROUND/OBJECTIVES

This study was conducted to assess the potential of St. John''s Wort (Hypericum perforatum) to prevent obesity and abnormalities in lipid metabolism induced by ovariectomy in a rat model without stimulatory activity on uterus.

MATERIALS/METHODS

Ovariectomized (OVX) rats were treated for 6 weeks with 70% ethanol extracts of Hypericum perforatum [HPEs: whole plant (WHPE) and flower and leaves (FLHPE)], β-estradiol-3-benzoate at a dose of 50 µg/kg/day (E2) or vehicle (distilled water).

RESULTS

As expected, OVX increased body weight gain and adiposity and showed higher food efficacy ratio. OVX also increased the serum cholesterol as well as insulin resistance, while reducing uterus weight and uterine epithelial proliferation rate. HPEs (WHPE and FLHPE) showed estrogen-like effect on body weight gain, adipose tissue weight and food efficacy ratio in OVX rats. HPEs prevented hypercholesterolemia induced by OVX more effectively than E2. E2 increased uterus weight and epithelial proliferation rate in OVX rats, while HPEs maintained them at the level of the sham-operated animals.

CONCLUSIONS

Our finding demonstrates that HPEs can be considered as an effective agent to prevent OVX-induced obesity without stimulatory activity on uterus.     

Cadmium Alters the Biotransformation of Carcinogenic Aromatic Amines by Arylamine N-Acetyltransferase Xenobiotic-Metabolizing Enzymes: Molecular,Cellular, and in Vivo Studies

Background

Cadmium (Cd) is a carcinogenic heavy metal of environmental concern. Exposure to both Cd and carcinogenic organic compounds, such as polycyclic aromatic hydrocarbons or aromatic amines (AAs), is a common environmental problem. Human arylamine N-acetyltransferases (NATs) are xenobiotic-metabolizing enzymes that play a key role in the biotransformation of AA carcinogens. Changes in NAT activity have long been associated with variations in susceptibility to different cancers in relation with exposure to certain AAs.

Objective

We explored the possible interactions between Cd and the NAT-dependent biotransformation of carcinogenic AAs.

Methods

We exposed purified enzymes, lung epithelial cells, and mouse models to Cd and subsequently analyzed NAT-dependent metabolism of AAs.

Results

We found that Cd, at biologically relevant concentrations, impairs the NAT-dependent acetylation of carcinogenic AAs such as 2-aminofluorene (2-AF) in lung epithelial cells. NAT activity was strongly impaired in the tissues of mice exposed to Cd. Accordingly, mice exposed to Cd and 2-AF displayed altered in vivo toxicokinetics with a significant decrease (~ 50%) in acetylated 2-AF in plasma. We found that human NAT1 was rapidly and irreversibly inhibited by Cd [median inhibitory concentration (IC₅₀) ≈ 55 nM; rate inhibition constant (k_inact) = 5 × 10⁴ M⁻¹ · sec⁻¹], with results of acetyl coenzyme A (acetyl-CoA) protection assays indicating that Cd-mediated inhibition was due to the reaction of metal with the active-site cysteine residue of the enzyme. We found similar results for human NAT2, although this isoform was less sensitive to inactivation (IC₅₀ ≈ 1 μM; k_inact = 1 × 10⁴ M⁻¹ · sec⁻¹).

Conclusions

Our data suggest that Cd can alter the metabolism of carcinogenic AAs through the impairment of the NAT-dependent pathway, which may have important toxicological consequences.     

Interleukin-10 and Transforming Growth Factor-β in Early and Late Lesions of Patients with Leishmania major Induced Cutaneous Leishmaniasis

Background

Cutaneous leishmaniasis is a neglected parasitic disease, which imposes massive human distress and financial costs to the endemic countries. Better understanding of host immune response to the parasite leads to helpful strategies for disease control. Interleukin (IL)-10 and transforming growth factor (TGF)-β are important immune regulatory cytokines, which appear to develop non-healing forms of leishmaniasis. However, there is little information about the function of IL-10 and TGF-β in old world cutaneous leismaniasis. The aim of this study was to analyze the role of IL-10 and TGF-β in human cutaneous leishmaniasis due to Leishmania major infection.

Methods

Biopsies were obtained from lesions of twenty proven cases of L. major induced cutaneous leishmaniasis. IL-10 and TGF-β positive cells were detected by immunofluorescence staining of frozen sections and compared between two groups of patients with early and late lesions.

Results

The mean percentage of IL-10 positive cells were significantly (P= 0.035) higher in late lesions (0.51±0.24) than early ones (0.15±0.07). Similar results were obtained for TGF-β with mean percentages of 0.16±0.05 and 0.53±0.28 in early and late lesions respectively (P= 0.008).

Conclusion

IL-10 and TGF-β are present in lesions of L. major induced cutaneous leishmaniasis and contribute to the pathogenesis of long lasting disease forms.     

Expression of Recombinant pET22b-LysK-Cysteine/Histidine-Dependent Amidohydrolase/Peptidase Bacteriophage Therapeutic Protein in Escherichia coli BL21 (DE3)

Objectives

Bacteriophage-encoded endolysins are a group of enzymes that act by digesting the peptidoglycan of bacterial cell walls. LysK has been reported to lyse live staphylococcal cultures. LysK proteins containing only the cysteine/histidine-dependent amidohydrolase/peptidase (CHAP) domain has the capability to show lytic activity against live clinical staphylococcal isolates, including methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to clone and express LysK-CHAP domain in Escherichia coli BL21 (DE3) using pET22b as a secretion vector. The pET22b plasmid was used, which encoded a pelB secretion signal under the control of the strong bacteriophage T7 promoter.

Methods

The E. coli cloning strains DH5α and BL21 (DE3) were grown at 37°C with aeration in the Luria-Bertani medium. A plasmid encoding LysK-CHAP in a pET22b backbone was constructed. The pET22b vector containing LysK-CHAP sequences were digested with NcoI and HindIII restriction enzymes. Cloning accuracy was confirmed by electrophoresis. The pET22b-LysK plasmid was used to transform the E. coli strain BL21. Isopropyl β-d-1-thiogalactopyranoside (IPTG) was added to a final concentration of 1mM to induce T7 RNA polymerase-based expression. Finally, western blot confirmed the expression of target protein.

Results

In this study, after double digestion of pEX and pET22b vectors with HindIII and NcoI, LysK gene was cloned into two HindIII and NcoI sites in pET22b vector, and then transformed to E. coli DH5α. Cloning was confirmed with double digestion and analyzed with agarose gel. The recombinant pET22b-LysK plasmid was transformed to E. coli BL21 and the expression was induced by IPTG. The expression was confirmed by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting method. Observation of a 28.5 kDa band confirmed LysK protein expression.

Conclusion

In the present study, LysK-CHAP domain was successfully cloned and expressed at the pET22b vector and E. coli BL21 (DE3).     

Padina arborescens extract protects high glucose-induced apoptosis in pancreatic β cells by reducing oxidative stress

BACKGROUND/OBJECTIVES

This study investigated whether Padina arborescens extract (PAE) protects INS-1 pancreatic β cells against glucotoxicity-induced apoptosis.

MATERIALS/METHODS

Assays, including cell viability, lipid peroxidation, generation of intracellular ROS, NO production, antioxidant enzyme activity and insulin secretion, were conducted. The expressions of Bax, Bcl-2, and caspase-3 proteins in INS-1 cells were evaluated by western blot analysis, and apoptosis/necrosis induced by high glucose was determined by analysis of FITC-Annexin V/PI staining.

RESULTS

Treatment with high concentrations of glucose induced INS-1 cell death, but PAE at concentrations of 25, 50 or 100 µg/ml significantly increased cell viability. The treatment with PAE dose dependently reduced the lipid peroxidation and increased the activities of antioxidant enzymes reduced by 30 mM glucose, while intracellular ROS levels increased under conditions of 30 mM glucose. PAE treatment improved the secretory responsiveness following stimulation with glucose. The results also demonstrated that glucotoxicity-induced apoptosis is associated with modulation of the Bax/Bcl-2 ratio. When INS-1 cells were stained with Annexin V/PI, we found that PAE reduced apoptosis by glucotoxicity.

CONCLUSIONS

In conclusion, the present study indicates that PAE protects against high glucose-induced apoptosis in pancreatic β cells by reducing oxidative stress.     

In Vitro Study on Cytotoxic Effects of ZnO Nanoparticles on Promastigote and Amastigote Forms of Leishmania major (MRHO/IR/75/ER)

Background

Although pentavalent antimony compounds are used as antileishmanial drugs but they are associated with limitations and several adverse complications. Therefore, always effort to find a new and effective treatment is desired. In this study, the effect of ZnO nanoparticles with mean particle size of 20 nanometers (nm) on Leishmania major promastigotes and amastigotes was evaluated.

Methods

Viability percentage of promastigotes after adding different concentrations of ZnO nanoparticles (30, 60, 90 and 120 μg/ml) to the parasite culture was evaluated by MTT assay. In the flow cytometry study, Annexin V-FITC Apoptosis detection Kit was used to study the induced apoptosis and necrotic effects.

Result

IC50 after 24 hours of incubation was 37.8 μg/ml. ZnO nanoparticles exert cytotoxic effects on promastigotes of L. major through the induction of apoptosis. A concentration of 120 μg/ml of ZnO nanoparticles induced 93.76% apoptosis in L. major after 72 hours.

Conclusion

ZnO NPs can induce apoptosis in L. major by dose and time-depended manner in vitro condition.     

Prevalence of Antibiotic Resistance in Escherichia coli Fecal Isolates From Healthy Persons and Patients With Diarrhea

Objectives

This study aimed to investigate the prevalence of antibiotic resistance in fecal Escherichia coli isolates from healthy persons and patients with diarrhea.

Methods

E. coli isolates (n = 428) were obtained from fecal samples of apparently healthy volunteers and hospitalized patients with diarrhea. Susceptibility patterns of isolates to 16 antimicrobial agents were determined by agar disc diffusion.

Results

Most E. coli isolates exhibited less than 10% resistance against imipenem, cefotetan, aztreonam, cefepime, cefoxitin, amikacin and netilamicin, although greater than 65% were resistant to ampicillin and tetracycline. No significant difference in resistance rates for all tested antibiotics was found between isolates from the healthy-and diarrheal-patient groups, including for multi-drug resistance (p = 0.22). The highest number of resistant antibiotics was 12 antibiotics. No significant differences in antibiotic resistance were found among the sex and age strata for isolates from healthy individuals. However, antibiotic resistance rates to cefoxitin, cefotaxime, amikacin, and netilamicin were significantly higher in the isolates of men than those of women (p < 0.05) in isolates from patients with diarrhea. Furthermore, isolates from patients with diarrhea older than 40-years of age showed higher resistance to cefepime and aztreonam (p < 0.05).

Conclusion

High resistance to the antibiotics most frequently prescribed for diarrhea was found in isolates from patients with diarrhea and apparently healthy individuals without any significant difference.     

Comparison of Resistant and Susceptible Strains of Trichomons vaginalis to Metronidazole Using PCR Method

Background

Metronidazole is drug of choice recommended by WHO for treatment of trichomoniasis, however, some reports claims drug resistance in Trichomonas vaginalis isolates recently. The objective of this study was to determine the minimum lethal concentration (MLC) of metronidazole in resistant and sensitive strains, as well as genetic patterns of these stains by PCR method.

Methods

From February 2006 to March 2007, in a cross sectional study, clinical and wet mount examination of vaginal smear along with culture were performed on 683 women attending to public and private outpatient clinics in Hamadan. Trichomoniasis marked based on major clinical symptoms. Diagnosis confirmed using wet mount microscopically and culture in Diamond medium. A serial concentration of metronidazole was provided and all isolated Trichomonas strains (resistant and sensitive) tested by standard method. Finally, all sensitive and resistant strains examined by PCR technique.

Results

Only 15/683, (2.2%) of patients clinically diagnosed trichomonal vaginitis were positive for T. vaginalis by wet smear and culture. The minimum lethal concentration (MLC) for clinically sensitive isolates was 25 µg/ml; however, this concentration for resistant isolates was 200 µg/ml after 24 h and 100 µg/ml after 50 h. The results of PCR examination of DNA from sensitive and resistant isolates had same pattern. The lanes appeared by two primers were 98 bp and 261 bp for both clinically sensitive and resistant strains.

Conclusion

Resistance to metronidazole in T. vaginalis has not relation to genetic variations and might be related to some physiologic pathways of organism.     

In-Vitro Assessment of the Acaricidal Properties of Artemisia annua and Zataria multiflora Essential Oils to Control Cattle Ticks

Background

The aim of this study was to investigate the ‘acaricidal effect’ of Zataria multiflora and Artemisia annua essential oils on Rhipicephalus (Boophilus) annulatus.

Methods

This study was carried out in 2009 in the Laboratory of Parasitology of the Faculty of Veterinary Medicine of Shahrekord University, west central Iran. Six dilutions (5, 10, 20, 40, 60 and 80 µL/cm³) of both essential oils were used against engorged female R. (Boophilus) annulatus ticks using an in vitro immersion method. The mortality rates for each treatment were recorded 6, 15 and 24 hours post inoculation (hpi). Mortality rate was analyzed using Repeated Measures Analysis of Variance, and comparison of means was carried out using General Linear Models Procedure.

Results

The mortality rate caused by different dilutions of Z. multiflora essential oil ranged from 26.6% (using 10 µL/cm³) to 100% (using 40 µL/cm³) and for A. annua essential oil it was 33.2 to 100% (using 20 and 80 µL/cm³, respectively) by the end of the experiment (36 hpi). No mortality was recorded for the non-treated control group or for dilutions less than 5 and 10 µL/cm³ using Zataria and Artemisia essential oils, respectively. For Z. multiflora mortality peaked at 15 hpi for all concentrations other than 20 µL/cm³ and took 24 h to achieve its maximum effect while for A. annua the two highest concentrations needed 24 hpi to reach their full effect. In addition, essential oils applied at more than 20 and 60 µL/cm³ caused 100% egg-laying failure in engorged female ticks by Zataria and Artemisia, respectively while no failure was observed for the non-treated control group. The mortality rate in both botanical acaricides was dose-dependent.

Conclusion

Both these medicinal plants have high potential acaricidal effects on the engorged stage of R. (Boophilus) annulatus in vitro.