The Effect of Dual Bioactive Compounds Artemisinin and Metformin Co-loaded in PLGA-PEG Nano-particles on Breast Cancer Cell lines: Potential Apoptotic and Anti-proliferative Action.

The most prevalent malignancy among women is breast cancer. Phytochemicals and their derivatives are rapidly being recognized as possible cancer complementary therapies because they can modify signaling pathways that lead to cell cycle control or directly alter cell cycle regulatory molecules. The phytochemicals' poor bioavailability and short half-life make them unsuitable as anticancer drugs. Applying PLGA-PEG NPs improves their solubility and tolerance while also reducing drug adverse effects. According to the findings, combining anti-tumor phytochemicals can be more effective in regulating several signaling pathways linked to tumor cell development. The point of the study was to compare the anti-proliferative impacts of combined artemisinin and metformin on cell cycle arrest and expression of cyclin D1 and apoptotic genes (bcl-2, Bax, survivin, caspase-7, and caspase-3), and also hTERT genes in breast cancer cells. T-47D breast cancer cells were treated with different concentrations of metformin (MET) and artemisinin (ART) co-loaded in PLGA-PEG NPs and free form. The MTT test was applied to assess drug cytotoxicity in T47D cells. The cell cycle distribution was investigated using flow cytometry and the expression levels of cyclin D1, hTERT, Bax, bcl-2, caspase-3, and caspase-7, and survivin genes were then determined using real-time PCR. The findings of the MTT test and flow cytometry revealed that each state was cytotoxic to T47D cells in a time and dose-dependent pattern. Compared to various state of drugs (free and nano state, pure and combination state) Met-Art-PLGA/PEG NPs demonstrated the strongest anti-proliferative impact and considerably inhibited the development of T-47D cells; also, treatment with nano-formulated forms of Met-Art combination resulted in substantial downregulation of hTERT, Bcl-2, cyclin D1, survivin, and upregulation of caspase-3, caspase-7, and Bax, in the cells, as compared to the free forms, as indicated by real-time PCR findings. The findings suggested that combining an ART/MET-loaded PLGA-PEG NP-based therapy for breast cancer could significantly improve treatment effectiveness.

Bacteriological profiling of diphenylureas as a novel class of antibiotics against methicillin-resistant Staphylococcus aureus.

Bacterial resistance to antibiotics remains an imposing global public health challenge. Of the most serious pathogens, methicillin-resistant Staphylococcus aureus (MRSA) is problematic given strains have emerged that exhibit resistance to several antibiotic classes including ß-lactams and agents of last resort such as vancomycin. New antibacterial agents composed of unique chemical scaffolds are needed to counter this public health challenge. The present study examines two synthetic diphenylurea compounds 1 and 2 that inhibit growth of clinically-relevant isolates of MRSA at concentrations as low as 4 µg/mL and are non-toxic to human colorectal cells at concentrations up to 128 µg/mL. Both compounds exhibit rapid bactericidal activity, completely eliminating a high inoculum of MRSA within four hours. MRSA mutants exhibiting resistance to 1 and 2 could not be isolated, indicating a low likelihood of rapid resistance emerging to these compounds. Bacterial cytological profiling revealed the diphenylureas exert their antibacterial activity by targeting bacterial cell wall synthesis. Both compounds demonstrate the ability to resensitize vancomycin-resistant Staphylococcus aureus to the effect of vancomycin. The present study lays the foundation for further investigation and development of diphenylurea compounds as a new class of antibacterial agents.

Dual-target-directed 1,3-diphenylurea derivatives: BACE 1 inhibitor and metal chelator against Alzheimer's disease.

Dual-target-directed 1,3-diphenylurea derivatives were designed by hybridizing BACE 1 inhibitor 1 with metal chelator LR-90. A database consisted of 1,3-diphenylurea derivatives was built and screened by the pharmacophore model (Hypo 1) of BACE 1 inhibitor. Based on the predicted results, 11 compounds (6a-d, 9a-g) with favorable Fitvalues were selected, synthesized and evaluated for their BACE 1 inhibitory activities, which showed that the predicted results were in good agreement with the experimental values. Besides, the synthesized compounds also displayed the ability to chelate metal ions. The most effective BACE 1 inhibitor 9f (27.85+/-2.46 micromol/L) was selected for further receptor-binding studies, the result of which indicated that an essential hydrogen bonds was formed between the urea group of 9f and the catalytic aspartate Asp228.

In vivo assays for drug resistance in Babesia divergens using the Mongolian gerbil, Meriones unguiculatus.

Two in vivo drug resistance assays were developed using gerbils. Cross resistance, involving related babesicides as well as the chemically unrelated antibiotic, oxytetracycline, was demonstrated, but the suggestion that imidocarb may select for pathogenic strains of parasites was not supported. Limited tests of field strains did not detect resistance. It is suggested that an in vitro assay would be more appropriate for surveys through in vivo assays would be essential for confirmatory studies.

Vaccination against bovine babesiosis with drug-controlled live parasites.

Live Babesia divergens derived from gerbils were used to vaccinate cattle that had previously been treated with imidocarb dipropionate. Drug doses ranged from 1 to 2 mg/kg and animals were infected subcutaneously three to seven days later. After a further 35 days, vaccinated and control animals were given a heavy heterologous intravenous challenge. This regimen was effective for both avirulent and virulent strains in 12- to 18-month-old cattle. However, at low drug doses some animals reacted to the virulent vaccine strain and at high doses animals infected with the avirulent strain failed to seroconvert although they were still resistant to challenge. The variable infectivity of vaccine strains was a minor problem which can be overcome by strain selection and optimisation of infectivity using gerbils as experimental animals. Gerbils would also be useful as a source of parasites for the further development of in vitro cultures which could ultimately produce the vaccine.

[Importance of an antiseptic solution, Solubacter, in vulvo-vaginal infections]. / Intérêt d'une solution antiseptique, Solubacter, dans les infections vulvo-vaginales.

Solubacter was prescribed, in combination with etiological treatment, for local application in 275 women with vulvovaginitis by 19 gynecologists in general practice as part of an open, multicentre study. The condition most frequently being treated was candidosis. A rapid improvement in vulvar symptoms was observed, right from the first few days of treatment. The treatment was well accepted by the patients and showed good tolerance, both in the judgement of the patients as well as that of the doctors. Solubacter thus proved to be a suitable additional aid in the treatment of vulvovaginitis.

Cationic antitrypanosomal and other antimicrobial agents in the therapy of experimental Pneumocystis carinii pneumonia.

Cationic compounds used in the treatment of veterinary African trypanosomiasis have structural properties similar to those of pentamidine, which has been used in the therapy of human trypanosomiasis and infection with Pneumocystis carinii. We have compared the activities of these drugs and other antimicrobial agents in an immunosuppressed rat model of P. carinii pneumonia. Diminazene, imidocarb, amicarbalide, quinapyramine, and isometamidium showed efficacy greater than or equal to that of pentamidine in the therapy of P. carinii infection, whereas ethidium and methylglyoxal bis(guanylhydrazone) were only slightly active against the organism. Diminazene and pentamidine also exhibited comparable efficacy in P. carinii prophylaxis, alpha-Difluoromethylornithine (DFMO), a polyamine inhibitor, was ineffective therapy when used alone and did not improve the effectiveness of pentamidine or diminazene. Quinine, quinidine, quinacrine, chlorpromazine, spiramycin, Pentostam, Astiban, dehydroemetine, ampicillin, gentamicin, chloramphenicol, and spectinomycin also showed little or no activity against the organism. Thus, in this model anti-P. carinii activity appears to be a common property of veterinary cationic trypanocidal compounds. This should be important in studying structure-activity relationships and in developing new drugs for the treatment of P. carinii infection in humans.

Prevention of intraoperative wound contamination with chlorhexidine shower and scrub.

In a prospective, controlled, clinical trial, we found that preoperative showering and scrubbing with 4% chlorhexidine gluconate was more effective than povidone-iodine or triclocarban medicated soap in reducing skin colonization at the site of surgical incision. Mean log colony counts of the incision site were one half to one log lower for patients who showered with chlorhexidine compared to those who showered with the other regimens. No growth was observed on 43% of the post shower skin cultures from patients in the chlorhexidine group compared with 16% of the cultures from patients who had povidone-iodine showers and 5% of those from patients who used medicated soap and water. The frequency of positive intraoperative wound cultures was 4% with chlorhexidine, 9% with povidone-iodine and 14% with medicated soap and water. This study demonstrates that chlorhexidine gluconate is a more effective skin disinfectant than either povidone-iodine or triclocarban soap and water and that its use is associated with lower rates of intraoperative wound contamination.

Anticoccidial activity of combinations of narasin and nicarbazin.

The efficacy of mixtures of narasin and nicarbazin were evaluated by comparing broiler performance, susceptibility to heat stress, and the mode of action against Eimeria. In a floor pen trial, narasin (70 ppm) alone or in combination with nicarbazin at levels between 10/10 and 50/50 ppm gave significantly better performance than unmedicated birds or birds given nicarbazin at 125 ppm alone. Amelioration of nicarbazin-associated mortality with heat as a stressor was observed in birds given the 50/50 ppm mixture of narasin and nicarbazin: mortality in these birds was similar to that of unmedicated birds and was reduced by 15 to 20% of that occurring in birds in the nicarbazin (125 ppm) treatment. The narasin/nicarbazin mixture (50/50) appears primarily to prevent further development of sporozoites. However, in birds treated with 25/25 ppm of narasin and nicarbazin, both the deleterious action of nicarbazin on merogeny and the antisporozoite activity of narasin were observed.

[Coccidiosis in broiler chicks: the prevalence of oocysts in feces in relation to necropsy findings in (sub)clinical coccidiosis and the effect of nicarbazin on these findings]. / Coccidiose bij slachtkuikens: het vóórkomen van oöcysten in faeces in relatie tot sektiebevindingen bij (sub)klinische coccidiose en het effekt hierop van nicarbazin.

Faecal samples were also collected from broiler farms presenting birds for autopsy. 126 Samples were examined, using two methods. Oocysts were detectable 3.5 days previously on an average before the presence of coccidiosis was determined at autopsy. Of the faecal samples 65.9 per cent were positive, and 46.0 per cent of the findings at autopsy were positive for coccidiosis. Examination of the faeces showed that more cases of mixed infection were present than could be concluded from autopsy. The largest number of oocysts was observed in E. acervulina infection, though large numbers of oocysts were also detected in cases of mixed infection. Treatment with nicarbazin during te first three weeks of life was found to delay the appearance of coccidiosis. The frequent use of withdrawal periods in the administration of anticoccidial drugs in view of 'thinning' showed high excretion rates.

Imidocarb and parvaquone in the treatment of piroplasmosis (Babesia equi) in equids.

The therapeutic efficacies of imidocarb and parvaquone were tested against Babesia equi of European origin in carrier horses and for induced acute infections in splenectomized ponies. Imidocarb, at a dosage of 4 mg/kg of body weight, given IM at 72-hour intervals 4 times, was ineffective in eliminating B equi-carrier infection in 9 mature geldings. A single IM administration of 4 mg/kg was not therapeutic in acutely infected splenectomized ponies. When given at 3 different dosages and treatment schedules, parvaquone was ineffective in clearing carrier infection. Parvaquone given IM once at a dosage of 20 mg/kg was effective for acute B equi infections in splenectomized ponies; parasitemia began to decrease within 24 hours after treatment. Infections were not eliminated however, and within 4 weeks, secondary parasitemia and anemia developed. Of 4 ponies, 3 died of acute piroplasmosis.

Processing yields and meat flavor of broilers fed a mixture of narasin and nicarbazin as an anticoccidial agent.

Processed yields (percent hot carcass) and cooked meat flavor of broilers fed 100 ppm of an anticoccidial agent (a mixture of 50 ppm narasin and 50 ppm nicarbazin) were compared with yields of birds fed a ration without the anticoccidial agent. Broilers were processed at 7 wk of age (49 days) after a 4-day withdrawal from the anticoccidial agent for the treated birds. The flavor of meat was evaluated by a 12-member sensory panel. Meat was either deep fat-fried or oven roasted. Sensory evaluations were made on freshly cooked samples and on cooked meat refrigerated for 24 h and reheated. The anticoccidial agent did not produce a difference (P greater than .05) in the hot carcass yields of the broilers as compared with control birds fed the nonmedicated diet. Analyses of triangle test data for flavor evaluations by two statistical methods indicated that there were no detectable differences (P greater than .05) in flavor between broilers fed the anticoccidial agent in the diet and those fed the control diet.

Efficacy, toxicity and metabolism of imidocarb dipropionate in the treatment of Babesia ovis infection in sheep.

An intramuscular dose of 1.2 mg kg-1 of imidocarb dipropionate (IMDP) was effective in controlling fatal infections with Babesia ovis in sheep. The sheep were infected by the intravenous injection of sheep blood containing B ovis. A severe recrudescence of infection occurred in most sheep 10 to 14 days after therapy but this could be controlled by a second dose of 1.2 mg kg-1 IMDP. Studies on the toxicology, residues and metabolism of IMDP showed this to be a safe dosage regimen. Transient or mild signs of toxicity were seen at 2.4 and 4.8 mg kg-1 IMDP. Severe toxicity was observed in sheep treated with 9.6 mg kg-1.

Chemoprophylactic activity of imidocarb, diminazene and oxytetracycline against Babesia bovis and B. bigemina.

Splenectomized calves treated with imidocarb, diminazene, and oxytetracycline were exposed to Babesia bigemina and B. bovis stabilates at various time intervals following treatment to evaluate prophylactic efficacy. Diminazene showed no residual activity against a B. bigemina challenge given 54 days after treatment. Oxytetracycline appeared responsible for an increased incubation time when given 2 days prior to B. bigemina exposure. Imidocarb showed marked prophylactic efficacy against both B. bigemina and B. bovis. Treatment with 1 or 2 mg kg-1 imidocarb, followed by Babesia exposure on the day of treatment, 7 days after treatment, then every 14 days for 91 days, delayed patent B. bigemina infections for 49 days and patent B. bovis infections for 42 days. Imidocarb at 4 or 5 mg kg-1, followed by similar Babesia exposures, delayed patent B. bovis infections for 68 days, and delayed B. bigemina for 81-103 days, and in some instances prevented infections. The delayed onset of infection due to either B. bigemina or B. bovis, following imidocarb treatment was accompanied by a significantly milder clinical response. Calves not responding to the primary challenge were fully susceptible to stabilate challenge 196 days after treatment. Calves experiencing a mild clinical response to B. bovis following imidocarb treatment and exposure failed to show any signs of response to a 196-day challenge exposure. Calves experiencing a mild clinical response to B. bigemina following imidocarb treatment and exposure did, in some instances, show a second mild response when challenged 196 days after initial treatment.

Effect of imidocarb dipropionate prophylaxis on the infectivity and immunogenicity of a Babesia bovis vaccine in cattle.

Imidocarb dipropionate (IDP), a potent prophylactic drug against Babesia bovis infections in cattle, was tested for its effect on the infectivity and immunogenicity of a live B. bovis vaccine marketed in Australia. At the recommended prophylactic dose rate of 3 mg/kg, IDP suppressed infectivity of the vaccine for at least 6 weeks. The vaccine infected all cattle inoculated either 8 weeks after treatment with 3 mg/kg, or 4 weeks after treatment with the recommended therapeutic dose of 1.2 mg/kg. These infections were, however, partially suppressed and the level of immunity to a subsequent heterologous virulent challenge was reduced. Cattle that failed to become infected after vaccination were fully susceptible to the challenge. It is concluded that where B. bovis vaccination is contemplated, prophylactic use of IDP should be avoided.

Dynamics of Anaplasma marginale in splenectomised calves treated with either imidocarb or oxytetracycline.

Neither imidocarb dipropionate (at 5 mg/kg body weight intramuscularly once or twice at a seven-day interval) nor oxytetracycline hydrochloride (at 20 mg/kg body weight intramuscularly once or twice at a seven-day interval) given to animals in the prepatent period of anaplasmosis, prevented the eventual onset of infection. Both compounds delayed the onset of detectable parasitaemia but this action was more noticeable with oxytetracycline. Oxytetracycline given twice significantly reduced the severity of the primary infection but the secondary infection was more severe. Neither compound showed prophylactic properties even though both are effective in extending the prepatent period and, in the case of oxytetracycline, reducing the severity of infection.

Chemotherapeutic efficacy of imidocarb dipropionate on experimental Eperythrozoon ovis infection in sheep.

Non-splenectomised Cheviot and Soay sheep experimentally infected with Eperythrozoon ovis were treated subcutaneously with two doses of 4 mg/kg of imidocarb dipropionate 24 hours apart. A rapid reduction in E. ovis parasitaemias following the first injection of imidocarb was observed and E. ovis organisms were not demonstrable 48 hours after the first treatment. However, recrudescences of infection were observed 14 and 28 days after treatment in the Soay and Cheviot sheep respectively. Side-effects associated with treatment were registered in the Cheviot sheep but not in the Soay sheep.