Pharmacokinetics of Two Ingredients of the Compound Chinese Herbal Medicine Transdermal Preparation in Cows

The topical administration of the compound Chinese herbal medicine transdermal preparation has been widely used in treating the mastitis of cows. In order to understand the metabolic process, four cows suffering from clinical mastitis were selected for the pharmacokinetic study. The transdermal preparation was applied to the diseased part of breast. Then the plasma and milk samples were collected respectively at 0, 0.5, 1, 2, 4, 6, 8, 10, 12, 24, and 36 h, The concentrations of baicalin and phillyrin in plasma and milk were determined by HPLC and the data of time-concentrations were analyzed by the pharmacokinetic software. The results showed that two ingredients in the plasma were not detectable, but they in the milk had linear relationship with the time. The peak concentration of baicalin [（2.84 ±0.15）μg mL^-1] appeared at （4.93 ± 0.35） h, AUC0.1OQ was （27.32 ± 7.91） mg L^-1 h^-1, and the mean retention time was （28.31 ±0.49） h. The peak concentration of phillyrin [（0.49 ± 0.41） μg mL^-1] achieved at （3.68 ± 3.14） h, AUC0-LOQ was （4.10± 0.31） mg L^-1 h^-1, and the mean retention time was （14.52 ± 0.02） h. These suggested that two ingredients had slow absorbing speed and high absorbing degree. They could not be eliminated in a short time, thus exerted the topical curative effect.

AcrAB Efflux Pump in Fluoroquinolone Resistant Salmonella gallinarum Induced by Ciprofloxacin Selective Pressure

Salmonella gallinarum has shown multiple drug resistance(MDR),especially high level fluoroquinolone(FQ)resistance in recent years.To determine whether the active efflux system was responsible for high-level FQ resistance,this research studied AcrAB efflux pump in Salmonella gallinarum on molecular level.The resistant strains were induced by standard strain C79-13 with ciprofloxacin in vitro.With carbonylcyanide-p-chlorophenyl hydrazone(CCCP)as an energy inhibitor,efflux inhibition test initially showed the potential impact of efflux pump on drug resistance.Sequence analysis of acrA gene indicated that gene mutation of AcrAB efflux pump was not definitely associated with MDR and drug resistance level of Salmonella gallinarum.Detected by competitive RT-PCR,the mRNA expression of acrA and acrB genes in the resistant strains significantly increased(p<0.01)compared with that of the control strain C79-13.The mRNA expression level of acrB gene(increased from 1.6-to 2.9-folds)was consistent with that of acrA gene(increased from 1.6-to 2.8-folds),which increased with the drug resistance level.However,gene mutation of acrA gene showed no correlation with its mRNA expression level,indicating that gene mutation did not affect the expression of AcrAB pump itself.The results suggested that the overexpression rather than the gene mutation of AcrAB efflux pump was an important factor causing the high level drug resistance of Salmonella gallinarum.

Effects of p27^(Kip1)- and p53- shRNAs on kanamycin damaged mouse cochlea

AIM： To study the effects of adeno-associated virus （AAV） delivered short hairpin RNAs （shRNAs） on adult CD-1 mouse cochlea damaged by aminoglycoside anti-biotic kanamycin. METHODS： Three different shRNAs were designed （p27^Kip1, p53 and p27^Kip1＋p53） and tested in COS cells. A total of 20 adult CD-1 mice were used in the experiment. Mice were divided into fve different groups （four animals/group） depending on the AAV-shRNA construct they received and whether they received kanamycin or not. Saline and AAV-EGFP injected animals were used as controls. All constructs were injected through the round window membrane （RWM） into the cochlea. Cochleae were harvested after 1 mo. Apoptosis was detected with Tunel labeling from paraffin-embedded cochlear tissue sections.RESULTS： AAV2/2-p27^Kip1-shRNA and AAV2/2-p53-shRNA were tested in COS cells. Western blotting analysis confirmed that both constructs silenced their target genes effectively in the cell culture. AAV2/2-shRNA constructs were injected into the cochlea of CD-1 mice through the intact RWM. Cotransductionof Cotransduction of individual AAV2/2-shRNAs with AAV2/2-EGFP resulted in EGFP expression in the organ of Corti.Kanamycin Kanamycin treatment had no effect on the expression pattern of the EGFP. AAV2/2-shRNA treated mice （either with p53 or p27Kip1and p53 together） showed fewer apop-totic hair cells in the cochlea than the control group （P 〈 0.05; AAV2/2-p53-shRNA vs saline P = 0.00014; AAV2/2-p27＋p53-shRNA vs saline P = 0.0011）. AAV2/2-p27-shRNA injected cochleae showed no significant difference in the number of apoptotic cells when compared to the saline injected cochleae.CONCLUSION： Silencing of p53 protein in the kana-mycin treated ears may decrease cell death in the organ of Corti.

Biochemical Composition and Anticancer Effect of Different Seaweed Species(In-vitro and In-vivo Studies)

Seaweed is an enormous resource comprised with natural bioactive compounds with several therapeutic effects including anticancer activity.In this context,the biochemical composition of seaweed plays a major role.Many biochemical compounds isolated from seaweed,fractions of seaweed and crude extracts has revealed ability of seaweed to fight against several cancer types.In this contrast seaweed extracts inhibit cancer cell growth and proliferation by inducing apoptosis and by inhibiting metastasis activity.In this review,biochemical and anticancer properties of seaweeds are discussed and this will provide the basic information to develop a novel chemotherapeutic drug to challenge the cancers.

Diarrhoea-predominant irritable bowel syndrome distinguishable by 16S rRNA gene phylotype quantification

AIM:To study whether selected bacterial 16S ribosomal RNA(rRNA)gene phylotypes are capable of disting- uishing irritable bowel syndrome(IBS). METHODS:The faecal microbiota of twenty volunteers with IBS,subdivided into eight diarrhoea-predominant (IBS-D),eight constipation-predominant(IBS-C)and four mixed symptom-subtype(IBS-M)IBS patients,and fifteen control subjects,were analysed at three time-points with a set of fourteen quantitative real-timepolymerase chain reaction assays.All assays targeted 16S rRNA gene phylotypes putatively associated with IBS,based on 16S rRNA gene library sequence analysis. The target phylotypes were affiliated with Actinobac-teria,Bacteroidetes and Firmicutes.Eight of the target phylotypes had less than 95%similarity to cultured bacterial species according to their 16S rRNA gene sequence.The data analyses were made with repeated-measures ANCOVA-type modelling of the data and principle component analysis(PCA)with linear mixed-effects models applied to the principal component scores. RESULTS:Bacterial phylotypes Clostridium cocleatum 88%,Clostridium thermosuccinogenes 85%,Coprobacillus catenaformis 91%,Ruminococcus bromii-like, Ruminococcus torques 91%,and R.torques 93%were detected from all samples analysed.A multivariate analysis of the relative quantities of all 14 bacterial 16S rRNA gene phylotypes suggested that the intestinal microbiota of the IBS-D patients differed from other sample groups.The PCA on the first principal component(PC1),explaining 30.36%of the observed variation in the IBS-D patient group,was significantly altered from all other sample groups(IBS-D vs control, P=0.01;IBS-D vs IBS-M,P=0.00;IBS-D vs IBS-C, P=0.05).Significant differences were also observed in the levels of distinct phylotypes using relative values in proportion to the total amount of bacteria.A phy- lotype with 85%similarity to C.thermosuccinogenes was quantified in significantly different quantities among the IBS-D and control subjects(-4.08±0.90 vs -3.33±1.16,P=0.04)and IBS-D and IBS-M subjects (-4.08±0.90 vs-3.08±1.38,P=0.05).Furthermore,a phylotype with 94%similarity to R.torques was more prevalent in IBS-D patients'intestinal micro- biota than in that of control subjects(-2.43±1.49 vs -4.02±1.63,P=0.01).A phylotype with 93%simi- larity to R.torques was associated with control sam- ples when compared with IBS-M(-2.41±0.53 vs -2.92±0.56,P=0.00).Additionally,a R.bromii-like phylotype was associated with IBS-C patients in com- parison to control subjects(-1.61±1.83 vs-3.69± 2.42,P=0.01).All of the above mentioned phylotype specific alterations were independent of the effect of time. CONCLUSION:Significant phylotype level alterationsin the intestinal microbiotas of IBS patients were observed,further emphasizing the possible contribution of the gastrointestinal microbiota in IBS.

Molecular study on methicillin-resistant Staphylococcus aureus strains isolated from dogs and associated personnel in Jordan

Objective: To determine the prevalence, genetic relatedness, and pattern of antimicrobial susceptibility in methicillin-resistant Staphylococcus aureus(S. aureus)(MRSA) isolated from household dogs, farm dogs, and stray dogs, compared to isolates from their associated personnel.Methods: MRSA was isolated from 250 nasal swabs(150 swabs from dogs and 100 swabs from humans). PCR assays were used to detect the presence of both the nuc and mec A genes,which con firmed the identity of S. aureus isolates and the presence of methicillin resistance,respectively. Disk diffusion was used to determine the antibiotic susceptibility against 15 antimicrobial agents along with an E-test that determined the minimum inhibitory concentration for oxacillin. Pulsed field gel electrophoresis was conducted to determine the genetic relatedness of MRSA isolates from dogs to those from associated and unassociated personnel.Results: The prevalence of S. aureus in dogs and humans was 12.7% and 10.0%respectively, while the prevalence of MRSA isolates in dogs and humans was 5.3% and5.0%, respectively. The prevalence of MRSA isolates in household dogs, farm dogs, and stray dogs was 7.8%, 4.7%, and 0.0%, respectively. MRSA isolates demonstrated a significantly higher rate of multi-resistance against three or more antimicrobial agents than methicillin-susceptible S. aureus(MSSA). Trimethoprim-sulphamethoxazole and chloramphenicol were the most effective antibiotics against all MRSA isolates. Pulsed field gel electrophoresis revealed a strong association between dog MRSA isolates and MRSA isolates from strongly associated personnel.Conclusions: MRSA is prevalent in house dogs, as well as in dog rearing centers and among their strongly associated personnel. A strong association was found between the MRSA isolates from dogs and those from humans who are in close contact. In addition,MRSA isolates showed a high rate of multi-resistance compared to MSSA isolates.

Bovine trypanosomosis: A threat to cattle production in Chena district, southwest Ethiopia

A cross-sectional study to determine the prevalence of bovine trypanosomosis was carried out in Chena district, Kefa zone, southwest Ethiopia from September 2010 to January 2011. Blood samples were collected from 391 randomly selected local (zebu) breed cattle in three representative peasant associations (PAs). The buffy coat and Giemsa stained thin blood films examination techniques were used for parasite detection and identification. The packed cell volume (PCV) estimation was also conducted. From a total of 391 cattle, 6.9% (n = 27) of the animals were found positive for trypanosome infection. The trypanosome species observed across the study animals were T. congolense (4.89%), T. vivax (1.54%), and T. b. brucei (0.51%) as single infections. The infection rate of T. congolense and T. b. brucei varied significantly (P 0.05) in infection rate was found between male (7.79%) and female (5.62%) animals. The prevalence was 2.24% and 8.29% in young and adult cattle, respectively with significant difference (P < 0.05) between them. The prevalence rate in good, medium and poor body conditioned animals were 7.28%, 0.78% and 13.39%, respectively with a statistical significant difference (P < 0.05) among them. The mean PCV of the infected animals (17.56%) appeared significantly (P < 0.05) lower than the non-infected (25.4%). Even if, the present study indicated a low prevalence of bovine trypanosomosis in the study areas, its impact on production and potential transmission to others should not be neglected.

H5N1 influenza outbreak during March 2006 in Jordan

The highly pathogenic avian influenza H5N1 pandemic motivated countries around the World to be prepared for outbreaks within their borders. Well ahead of the outbreak, in Jordan contingency plans were prepared involving all relevant bodies in both the private and public sectors and a practice was simulated to test the effectiveness of the task force and the applicability of the plan. The highly pathogenic H5N1 influenza virus was detected and confirmed in one turkey in a backyard flock comprising 12 turkeys and 25 chickens in Kufranjah, 6 km from Ajloun city, on 23 March 2006. The samples were confirmed by the following day. Control measures were implemented immediately according to the national contingency plan and complete clean up achieved by 27 March 2006. A 3 km area around the index case was designated as a control zone, with a 10 km area around the holding further designated as a surveillance zone. The objective was to stamp out the infection and approximately 20,000 birds were destroyed including all commercial and backyard flocks in the control zone. All the human contacts were examined, treated and discharged from the hospital. The immediate response from all Jordanian sectors involved in the control strategy was positive as a result of increasing awareness and training programs that were implemented six months before the occurrence of the outbreak. A total estimated economic loss posed by this outbreak including compensation was almost $US169 million. Following the outbreak, passive surveil-lance for the virus was undertaken and training and capacity building continued. Efforts made in the preparation for an outbreak paid dividends and the need for a department responsible for monitoring migratory and wild birds in Jordan was recognized. Legislation regarding biosecurity of poultry farms and poultry transport vehicles should be revised.

Histopathological Characterization of Lungs from Slaughtered Swine in Selected Abattoirs in Laguna, Philippines and Its Relation to Serological Profiling for Porcine Pleuropneumonia (PPP), Porcine Reproductive and Respiratory Syndrome (PRRS) and Enzootic Pneumonia (EP)

Porcine Respiratory Disease Complex (PRDC) is one of the most important health concerns in swine production due to consumer demand for high quality pork from healthy animals. This study was conducted to characterize and compare the gross microscopic lesions and serological profiles of swine pneumonic lungs from selected slaughterhouses in Laguna, Philippines. Blood and lung samples were collected from the municipalities of Santa Cruz, San Pablo, Los Baños, and Cabuyao. Enzyme-Linked Immunosorbent Assay (ELISA) test was conducted for the blood samples and the histopathological changes in the lung tissues were observed microscopically. The results showed that the presence of Porcine Pleuropneumoniae (PPP), Enzootic Pneumoniae (EP) and Porcine Reproductive and Respiratory Syndrome (PRRS) virus were detected using ELISA. In Los Baños, Sta. Cruz and San Pablo the most prevalent disease was EP at 26.1%, 13.0% and 17.4% respectively. While in Cabuyao the diseases found were PRRS and EP, both at 8.7%. Based on the histopathological examination, it was found out that the most occurring types of lesions were hemorrhage, congestion and lymphoid hyperplasia of BALT (Bronchial Associated Lymphoid Tissue). However, the histopathological findings were found to be not significantly associated with the disease present in the sampled swine .