Carvacrol-Fabricated Chitosan Nanoparticle Synergistic Potential with Topoisomerase Inhibitors on Breast and Cervical Cancer Cells.

Breast and cervical cancers are the most common heterogeneous malignancies in women. Chemotherapy with conventional drug delivery systems having several limitations along with development of multidrug resistance compelled us to seek out targeted therapeutics. Nanoparticles are suitable substitutes to circumvent multidrug resistance for the targeted treatment of cancer. The current study was aimed to investigate the anticancer effect of carvacrol-loaded chitosan nanoparticles with topoisomerase inhibitors. The average size of carvacrol-loaded chitosan nanoparticles was found to be 80 nm with 24.7 mV ζ-potential, and maximum absorbance was observed at 275 nm. Among all drug combinations, the carvacrol nanoparticles with the doxorubicin combination group exerted greater dose-dependent growth inhibition of both MCF-7 and HeLa cells as compared to single carvacrol nanoparticles and doxorubicin. Combination index values of carvacrol nanoparticles and the doxorubicin combination group showed a strong synergistic effect as they were found to be between 0.2 and 0.4, 0.31 for MCF-7 and 0.34 for HeLa cells. The carvacrol nanoparticles in combination with doxorubicin on MCF-7 cells reduced the dose 16.32-fold for carvacrol nanoparticles and 4.09-fold for doxorubicin at 6.23 µg/mL IC50, while on HeLa cells, this combination reduced the dose 13.18-fold for carvacrol nanoparticles and 3.83-fold for doxorubicin at 9.33 µg/mL IC50. As the dose reduction values were greater than 1, they indicated favorable dose reduction. It was concluded that the combination of carvacrol-loaded chitosan nanoparticles with topoisomerase inhibitors may represent an innovative and promising strategy to improve the efficacy, resistance, and targeted delivery of chemotherapeutics in cancer.

Synergistic antibacterial activity of carvacrol loaded chitosan nanoparticles with Topoisomerase inhibitors and genotoxicity evaluation.

The increasing prevalence of antibiotic resistant bacteria is a significant healthcare crisis with substantial socioeconomic impact on global community. The development of new antibiotics is both costly and time-consuming prompting the exploration of alternative solutions such as nanotechnology which represents opportunities for targeted drug delivery and reduced MIC. However, concerns have arisen regarding genotoxic effects of nanoparticles on human health necessitating an evaluation of nanoparticle induced DNA damage. This study aimed to investigate the antibacterial potential of already prepared, characterized chitosan nanoparticles loaded with carvacrol and their potential synergism with Topoisomerase II inhibitors against S. aureus, E. coli and S. typhi using agar well diffusion, microdilution and checkerboard method. Genotoxicity was assessed through comet assay. Results showed that both alone and drug combinations of varying concentrations exhibited greater zones of inhibition at higher concentrations. Carvacrol nanoparticles combined with ciprofloxacin and doxorubicin significantly reduced MIC compared to the drugs used alone. The MIC50 values for ciprofloxacin were 35.8 µg/ml, 48.74 µg/ml, 35.57 µg/ml while doxorubicin showed MIC50 values of 20.79 µg/ml, 34.35 µg/ml, 25.32 µg/ml against S. aureus, E. coli and S. typhi respectively. The FICI of ciprofloxacin and doxorubicin with carvacrol nanoparticles found ≤ 0.5 Such as 0.44, 0.44,0.48 for ciprofloxacin and 0.45, 0.45, 0.46 for doxorubicin against S. aureus, E. coli and S. typhi respectively revealed the synergistic effect. The analysis of comet assay output images showed alteration of DNA at high concentrations. Our results suggested that carvacrol nanoparticles in combination with Topoisomerase inhibitors may prevent and control the emergence of resistant bacteria with reduced dose.

Lack of antiviral activity of ivermectin against foot-and-mouth disease virus serotype O in BALB/c mice.

Ivermectin is an FDA approved drug and showed in vitro antiviral activity against different serotypes of Foot-and-mouth disease virus (FMDV). We here assessed the effect of ivermectin in 12 day old female BALB/c mice infected with 50LD50 FMDV serotype O intraperitoneally. Initially FMDV was adopted on 3-day old BALB/c mice by blind passages. After successful adaptation of virus mice showed hind limb paralysis. Mice were divided in 6 different groups and each group has 6 mice. Ivermectin was given at clinically prescribed dose of 500 µg/kg subcutaneously at different time interval. Ivermectin was given at 0 h post infection (hpi) and 12 hpi. Moreover we compared commercially available ivermectin with purified ivermectin preparation in sterilized DMSO. Viral load was evaluated through RT-qPCR and ELISA in different groups. Results showed that positive control and negative control has CT-value 26.28 and 38 respectively. Treated groups at 0hpi, 12hpi, purified ivermectin and pre-post treatment group has CT values 24.89, 29.44, 27.26 and 26.69 respectively that showed there was no significant reduction in virus load in treated groups as compare to positive control. In histopathology of lung tissue perialveolar capillaries were congested and alveoli were altelactic. Some emphysema was seen in alveoli and mild thickening in the alveolar wall was observed. In the alveolar epithelium mononuclear cells infiltration was seen. There was discoloration haemorrhages and enlargement of heart. Degeneration, fragmentation and loss of sarcoplasm were seen in the cardiac muscle fibers. Above results showed that ivermectin did not lessen lung and heart viral load. This study contributes that ivermectin does not have a significant antiviral effect when used in mice against FMDV serotype O, according to a growing body of research.

Preparation of purified vaccine from local isolate of foot and mouth disease virus and its immune response in bovine calves.

Foot and Mouth Disease (FMD) is globally pandemic which badly affect the economics of livestock based countries like Pakistan. There are different types of Foot and Mouth Disease Virus (FMDV) among these types O is most prevalent in Pakistan. Recently Pakistan is producing approximately fifteen million doses of non-purified FMD vaccine against the demand of 160 million doses annually. More over the Pakistan is still striving for the development and optimization of concentration as well as purification of FMDV. The present project was designed to develop the technology for the purification of FMDV indigenously. The locally isolated and adapted FMDV type O virus was propagated on adherent culture of BHK-21cells to get final volume of virus one liter. This virus suspension was concentrated by peggylation as well as ultra-filtration method. The purification and quantification of concentrated virus was done by size exclusion chromatography. The results showed that peggylation is better method of concentration up to 603.75 µg/ml with 82.80 % recovery rate than ultra-filtration with 43.90 % followed by chromatography for purification. The PD50 was calculated in bovines at 24, 12, 6, 3 and 1.5 µg of FMDV Ag/dose and it revealed that antigen load of 1.98 µg is the dose, where the 50 % of inoculated animals showed the protective antibody level based upon percent inhibition through antibody detecting ELISA. According to the British pharmacopeia, the vaccine should contain 3PD50 which found equivalent to our findings about 6 µg/dose. The group of animal injected with 6/dose (3.23PD50) showed protective titer up to 20th week post priming.

Renessans Helps in Early Clearance of SARS-CoV-2: In-Vivo Activity of the Iodine Complex in Rhesus macaque.

Iodine complexes have known antimicrobial properties along with reported in-vitro antiviral activity for several viruses. Renessans is one such product with iodine complexes and ascorbic acid. The present study was designed to determine its efficacy for SARS-CoV-2 in Rhesus macaque. Rhesus macaque were assigned to: A) prophylactic group (n = 3), (B) treatment group (n = 3), (C) infection control group (n = 4), and (D) negative control group (n = 4). Groups A, B, and C were challenged with 2 × 106 TCID of SARS-CoV-2. The prophylactic group (A) was administered Renessans from 5 days before infection till 8 days postinfection (DPI). The treatment group (B) was administered Renessans from 3 till 8 DPI. Group C was administered water-insoluble fractions only. Nasal swabs from all monkeys of groups A, B, and C remained positive for SARS-CoV-2 till 2 and 7 DPI, while the swabs became negative for groups A and B at 14 DPI. Likewise, fecal matter of monkeys in group A returned negative results during the experiment, while that of group B had significantly decreased viral load (101.5 genome copies/mL) compared to group C (103 genome copies/mL). Hence, it is concluded that Renessans has in-vivo SARS-CoV-2 activity and may result in early clearance of SARS-CoV-2.

Optimizing factors for the efficient expression and purification of spao and lamb from Salmonella typhi

We optimized the expression and purification of outer membrane proteins SpaO and LamB from Salmonella typhi. We investigated various factors in the expression and purification processes, including the use of isopropyl ß-d-1 thiogalactopyranoside (IPTG), imidazole, and urea. First, PCR amplification was carried out on SpaO and LamB genes. The genes were then cloned in pTZ57R/T, and then expressed in pET28a vector and transformed into Escherichia coli BL21 (DE3). Gene insertion was confirmed by enzymatic digestion with NdeI and XhoI. Inclusion bodies expressing recombinant SpaO and LamB were induced with 200 and 400 µL 0.5 mM IPTG, respectively. The formed protein inclusion bodies were then isolated from the pellet and solubilized in IB buffer containing 8 M urea for SpaO and 6 M urea for LamB. Proteins were refolded by dialysis in 3M urea. Purified proteins with nickel-nitrilotriacetic acid affinity chromatography and eluted with buffer containing 250 mM imidazole for SpaO and 150 mM imidazole for LamB. The protein expression profiles were analyzed by SDS-PAGE, which identified the 33 and 49 kDa bands corresponding to rSpaO and rLamB. Western blotting Purification was carried out by nickel affinity resin with 250 mM and 150 mM imidazole for rSpaO and rLamB and refolded through stepwise dialysis with anti-His tag antibodies confirmed their expression. These optimized methods can be used to generate recombinant proteins for the development of future vaccines.

Effect of cell density on the biological titer and yield of 146S fraction of foot-and-mouth disease virus O in cell suspension.

Foot-and-mouth disease (FMD) is a highly infectious disease of cattle and other cloven-hoofed animals, causing huge economic losses annually worldwide. This disease is endemic in Pakistan where the serotypes of the foot-and-mouth disease virus (FMDV) are A, O and ASIA-1. At present, trivalent FMDV vaccines are being used to prevent FMD but the current production process is laborious and is unable to fulfill the needs of the meat and dairy industries. To meet the vaccine needs of Pakistan, the conventional method of using adherent cell lines to produce the vaccine could be replaced by suspension cell cultures which produce higher yields in less time and less volume. Therefore, the aim of this study was to investigate and optimize some of the factors that affect viable cell density and subsequent virus yield. The relationship between the yield of the 146S fraction and the TCID50 of the virus preparations obtained was also evaluated as a mean to control and check the quality of the vaccine product. The results provided optimized conditions for vaccine production using cell suspensions and showed that there was a linear relationship between TCID50 and 146S fraction yield. Either TCID50 or the 146S fraction yield, or both could be used as parameters for quality monitoring during vaccine production. Using TCID50 reduced the number of steps involved in virus production while measuring 146S fraction yield was useful for quality control. However, more studies are required to evaluate the relative effectiveness of vaccines produced by virus cultures using either TCID50 or 146S fraction as quality monitoring tools.

Anti-hyperlipidemic and anti-diabetic evaluation of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy

The use of plants in disease treatment is cost effective and relatively safe. This study was designed to investigate anti-hyperlipidemic and anti-diabetic activity of ethanolic leaf extract of Catharanthus roseus alone and in combination therapy in hyperlipidemic & diabetic mice. Eight groups comprising five mice each were used. Group A was hyperlipidemic control, group B, C, D received atorvastatin (20 mg/kg), leaf extract (200 mg/kg) and leaf extract in combination with atorvastatin (200 mg/kg and 20 mg/kg) orally for 15 days. Group E was diabetic control. Group F, G, H received sitagliptin (40 mg/kg), leaf extract (200 mg/kg) and extract in combination with sitagliptin (200 mg/kg and 40 mg/kg) orally for 7 days. Blood cholesterol levels were measured at 1st, 5th, 10th and 15th day and fasting blood sugar levels were measured at 2, 12, 24, 72 and 168 hours during treatment. One-way ANOVA with tukey- kramer multiple comparison test was used. The chemical characterization of ethanolic extract of Catharanthus roseus leaves showed presence of alkaloids, saponins, tannins and flavonoids. Ethanolic extract of Catharanthus roseus has significant anti-hyperlipidemic & anti-diabetic effects (p<0.05, p<0.01) when compared with control but had not cause significantly increase in anti-hyperlipidemic effects of atorvastatin. While significantly increased the antidiabetic effect of sitagliptin (p<0.05)

An in vitro antiviral activity of iodine complexes against SARS-CoV-2.

Since the emergence of COVID-19 pandemic in China in late 2019, scientists are striving hard to explore non-toxic, viable anti-SARS-CoV-2 compounds or medicines. We determined In vitro anti-SARS-CoV-2 activity of oral formulations (syrup and capsule)of an Iodine-complex (Renessans). First, cell cytotoxicity of Renessans on the Vero cells was determined using MTT assay. Afterwards, the antiviral activity of Renessans was determined using viral inhibition assays and TCID50. For this, nontoxic concentrations of the Renessans were used. The results showed that Renessans is nontoxic to the cells up to 50 µg/mL. At 1.5 µg/mL concentration, SARS-CoV-2 production was significantly reduced to 101.43 TCID50 and 101.58 TCID50 for the syrup and capsule, respectively, as compare to virus infected control cells 106.08 TCID50 and we found the dose dependent inhibition of virus replication in the presence of Renessans. Renessans inhibited SARS-CoV-2 with an EC50 value of 0.425 µg/mL and 0.505 µg/mL for syrup and capsule, respectively. Furthermore, there was no virus detected at concentration of 50 µg/mL of Renessans. This study indicates that Renessans, containing iodine, have potential activity against SARS-CoV-2 which needs to be further investigated in human clinical trials.

Antiviral potential of ivermectin against foot-and-mouth disease virus, serotype O, A and Asia-1.

Each year, foot-and-mouth disease leads to enormous economic losses to the livestock industry. Currently, the killed whole virus is widely using to control FMD. However, vaccination is constrained by lack of or incomplete protection. Therefore, along with vaccination, we need to find the antivirals against FMD. This study was conducted to investigate the antiviral potential of ivermectin against multiple serotypes of FMDV. Initially, an MTT assay was performed on the BHK-21 cell line to determine assay ivermectin cytotoxicity. Viral inhibition assays using the non-cytotoxic concentration of ivermectin were performed to check the antiviral potential of ivermectin on different stages of virus replication. At 2.5 µM and 5 µM concentrations of ivermectin, the virus titer was reduced significantly (p < 0.001) by two to three log in all three strains of viruses at both non-toxic concentrations (2.5 and 5 µM). The virus titer in strain O control was 106.0 TCID50/0.1 mL and was reduced to 104.1 TCID50/0.1 mL at a concentration of 2.5 µM and 103.10 TCID50/0.1 mL at 5 µM concentration. In the case of strain Asia-1, the virus titer was reduced to 103.8 TCID50/0.1 mL at 2.5 µM and 103.01TCID50/0.1 mL at 5 µM concentration. The titer of strain A was reduced from 105.8 TCID50/0.1 mL to 103.9 TCID50/0.1 mL at 2.5 µM concentration and 103.1 TCID50/0.1 mL at 5 µM concentration. Moreover, the virus titer was reduced more at the replication stage as compared to attachment and entry stages. This study showed the in vitro anti-FMDV potential of ivermectin for the first time and predicted its potential use against FMDV infections.

Role of surgical modality and timing of surgery as clinical outcome predictors following acute subdural hematoma evacuation.

BACKGROUND & OBJECTIVE: A Craniotomy (CO) or decompressive craniectomy (DC) are the two main surgical procedures employed for evacuation of acute traumatic subdural hematoma (ASDH). However, the optimal surgical procedure remains controversial. The beneficial effect of early surgical evacuation of acute subdural hematoma in improving outcome also remains unclear. Our objective was to study the role of these two parameters in determining the outcome in patients undergoing surgical evacuation of acute traumatic subdural hematoma. METHODS: A retrospective analysis of 58 patients presenting with acute traumatic subdural hematoma and with presenting Glasgow Coma Scale (GCS) ≤ 8 that had been operated in Lahore General Hospital between June 2014 and July 2015 was performed. The demographic data, preoperative GCS, type of surgical procedure performed and timing of surgery were analysed. RESULTS: Forty (69%) patients underwent CO, and eighteen (31%) patients underwent DC. The CO and DC groups showed no difference in the demographic data and preoperative GCS. Six patients survived in the craniotomy group, while none survived in the decompressive craniectomy group (p=0.083). The relationship of timing of surgery with survival in the craniotomy group was found not to be clinically significant (p=0.87). CONCLUSION: In this study craniotomy was associated with a better outcome as compared to decompressive craniectomy, however, the difference did not reach statistical significance. Early surgery was also found not to be associated with an improved outcome.

Anti-oxidative and anti-proliferative activities of extracted phytochemical compound thymoquinone.

OBJECTIVE: To evaluate the anti-oxidant and anti-proliferative potential of Thymoquinone extracted from the essential oil of indigenous herbs of Nigella sativa and Thymus vulgaris. METHODS: Extraction and quantification of Thymoquinone was carried out in July, 2017 in Department of Environmental Science, Lahore College for Women University (LCWU), Lahore. Thymoquinone was extracted from seeds of Nigella Sativa and aerial parts of Thymus vulgaris by employing soxhlet extraction with 1:4 ratios of nhexane and methanol. High Performance Liquid Chromatography was used to quantify Thymoquinone from the methanolic extracted oil of sample by applying calibration curve method. Extracted Thymoquinone was identified by sample peaks obtained at retention time were compared with peak of standard Thymoquinone at respective time. The Thymoquinone obtained from both samples was then subjected to Fourier-transform infrared spectroscopy for confirmation by identifying its functional groups. Anti-oxidant activities of both samples were measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) assay in Department of Environmental Science, LCWU. In-vitro anti-proliferative activities of extracted Thymoquinone were evaluated in HeLa cell cancer lines by cell proliferations Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay in Department of Microbiology, University of Veterinary and Animal Sciences (UVAS), Lahore. SPSS 18 and Graph pad prism 18 was used for data analysis. RESULTS: Soxhlet extraction with solvents ratios yielded 48.92% oil from Nigella sativa and 23.2 % from Thymus vulgaris. High Performance Liquid Chromatography peak of standard Thymoquinone was measured at retention time of 5.5 min which was then compared with the peak obtained from both samples at the similar retention time. The extracted Thymoquinone from both samples were quantified by calibration curve method showing 614.25 mg/L from Nigella sativa and 548.86 mg/L from Thymus vulgaris. The two anti-oxidant assays of both samples compared with standard Thymoquinone showed significant scavenging activities in dose amount manner. Cell proliferation of HeLa cancer significantly decreased with dose response manner (p<0.01), showing highest cell death in high concentration of Thymoquinone. Inhibitory concentration 50 (IC50) of cancer cell line treated with Nigella sativa oil was 0.5 µM and Thymus vulgaris was 18 µM compared to standard Thymoquinone, showing Inhibitory concentration50 (IC50) of 6 µM using Graph pad prism v.8.0. CONCLUSION: Both Nigella sativa and Thymus vulgaris were found to be the best source of Thymoquinone as chemotherapeutic drug expressed potent anti-oxidant and anti-proliferative activities.

Radiolological predictors of recurrence of chronic subdural hematoma.

OBJECTIVE: Chronic subdural hematoma is one of the most common clinical entities encountered in daily neurosurgical practice. Considerable recurrence rates have been reported for chronic subdural hematoma following surgical evacuation. Many studies have suggested various radiological factors that may be associated with the recurrence of CSDH. However, the results are inconsistent. This study focuses on determining the radiological factors predictive of chronic subdural hematoma recurrence. METHODS: A retrospective analysis of 113 patients diagnosed with chronic subdural hematoma who were surgically treated between August 2013 and December 2014 was performed. The radiological features were analyzed to clarify the correlation between these radiological factors and postoperative recurrence of chronic subdural hematoma. RESULTS: Twenty patients (17.7%) experienced recurrence. Chronic subdural hematoma recurrence was found to be significantly associated (p<0.05) with preoperative hematoma thickness ≥ 20 mm. Midline shift, hematoma density and bilaterality were not significantly associated with recurrence. Post operative drainage also significantly (p<0.05) reduced chronic subdural hematoma recurrence. CONCLUSION: Preoperative hematoma thickness ≥ 20 mm is an independent predictor of recurrence of chronic subdural hematoma. Postoperative drainage also significantly reduces chronic subdural hematoma recurrence.

Evaluation of cytotoxic and antiviral activities of aqueous leaves extracts of different plants against foot and mouth disease virus infection in farming animals.

Cytotoxic and antiviral activity of aqueous leaves extracts of three plants: Azadirachta indica, Moringa oleifera and Morus alba against Foot and Mouth disease virus (FMDV) were determined using MTT assay (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide). Eight different concentrations of each plant were evaluated. Cytotoxic and antiviral activity of each extract was evaluated as cell survival percentage and results were expressed as Means ± S.D. From the tested plant extracts, Azadirachta indica & Moringa oleifera exhibited cytotoxicity at 200 & 100 µ/ml respectively. In case of antiviral assay, Moringa oleifera showed potent antiviral activity (p<0.05) while Azadirachta indica showed significant antiviral activity in the range of 12.5-50 µ/ml & 50-100 µ/ml respectively. In contrast no anti-FMDV activity in the present study was observed with Morus alba, although all the tested concentrations were found to be safe.

Effect of calcium hypochlorite and chloramine on blood biochemistry and sodium pentobarbital induced sleeping time in mice.

Disinfectants are chemical agents used to eradicate, deactivate or kill microorganisms. Chemical disinfectants especially chlorine compound are extensively used for water sanitization. Among these calcium hypochlorite and chloramines are commonly used now a day. Large number of chemical compounds, drugs and endogenous substances are metabolized by hepatic enzymes known as cytochrome P450 enzyme system. Many chemicals are capable of enzyme induction. Enzyme induction may change the metabolism of other drugs and endogenous substances which may alter the plasma concentration of these chemicals. To evaluate the enzyme inducing ability of calcium hypochlorite and chloramine, sleeping time induced by sodium pentobarbital was noted in mice. Normal saline was taken as negative control. Rifampicin, chloramphenicol and grapefruit juice were taken as positive control group. On completion of dosing after 4 weeks, alteration in sleep induction and recovery times was noted and compared. Histological evaluation of liver was observed. A significant decrease in sleeping time was observed in calcium hypochlorite and chloramine treated groups. Both calcium hypochlorite and chloramine caused a significant change in liver enzymes and in the values of complete blood count. In histological evaluation both caused fat deposition in the hepatocytes. It was concluded from the study that both calcium hypochlorite and chloramine were hepatic microsomal enzyme inducer.

Microbiological Load of Ethylene Oxide Sterilized Medical Devices and its Elimination by Cobalt 60 Source.

OBJECTIVE: To determine the residing microbial flora of ethylene oxide (EtO) sterilized medical devices and optimization of safe dose of gamma radiation (Cobalt 60 source) for the complete elimination of microbial load. STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: Department of Biotechnology, Lahore College for Women University, Lahore, Pakistan from September 2014 to June 2015. METHODOLOGY: Thirty-six samples of EtO sterilized medical devices of same batch of three different companies were collected for this study. Isolation and enumeration of microbes were done by using different selective and differential media. Gram staining and biochemically characterization by API 20 (Bio Merieux, France) kit was done for identification of the microorganisms. The medical devices having high microbial load were sent to Pakistan Radiation Services (PARAS) for gamma irradiations at 3 different selected doses (20 KGy, 25 KGy, and 30 KGy). RESULTS: Different types of Gram positive bacteria (Staphylococcus epidermidis, Staphylococcus aureus andBacillus subtilis) were isolated from the EtO sterilized samples. Gram negative bacteria and fungi were not detected on these medical devices. Gamma irradiations results showed that 30 KGy was optimized dose for complete elimination of microbial flora on endotracheal, Nelaton, and tracheostomy tubes. CONCLUSION: Gamma radiations (Co 60 source) effectively decontaminate the microbial flora on the equipment previously sterilized by the ethylene oxide gas; and 30 KGy is the optimized dose for all these medical devices.

Toxicity Appraisal of Untreated Dyeing Industry Wastewater Based on Chemical Characterization and Short Term Bioassays.

Characterizing wastewaters only on a chemical basis may be insufficient owing to their complex nature. The purpose of this study was to assess toxicity of textile dyeing wastewater based on analytical techniques and short term toxicity based bioassays. In this study, screening of the fractionated wastewater through GC-MS showed the presence of phenols, phthalic acid derivatives and chlorpyrifos. Metal analysis revealed that chromium, arsenic and mercury were present in amounts higher than the wastewater discharge limits. Textile dyeing wastewater was found to be highly mutagenic in the Ames test. DNA damage in sheep lymphocytes decreased linearly with an increase in the dilution of wastewater. MTT assay showed that 8.3 percent v/v wastewater decreased cell survival percentage to 50 %. It can be concluded from this study that short term toxicity tests such as Ames test, in vitro comet assay, and cytotoxicity assays may serve as useful indicators of wastewater pollution along with their organic and inorganic chemical characterizations.

Management of Cerebrospinal Fluid Leak following Posterior Cranial Fossa Surgery.

OBJECTIVE: Cerebrospinal fluid leakage remains a significant cause of morbidity following posterior fossa surgery, and its treatment remains a difficult problem. The aim of the study was to propose a treatment algorithm for its management. METHODS: A retrospective, single-center study was conducted on 147 patients who underwent elective posterior fossa surgery for a variety of diseases . Patients with post operative CSF leakage had either been treated initially with conservative measures including re-suturing of the wound, with CSF lumbar drainage to be employed in case the CSF leakage didn't stop, or the initial intervention was the institution of CSF lumbar drainage simultaneously with conservative measures. VP (ventriculo-peritoneal) shunt was done in patients with gross hydrocephalus on postoperative CT brain. RESULTS: There were 25 (17%) cases of CSF leakage, including 24 incisional CSF leaks and one case of CSF otorrhea. In eight patients with incisional CSF leakage treated initially with conservative measures including re-suturing of the wound, CSF leakage stopped in only two cases. CSF lumbar drainage instituted later on in six cases with persistent leakage stopped the CSF leakage. In fourteen patients managed initially with re-suturing of the wound and concomitant CSF lumbar drainage, CSF leakage settled in all the cases. Two patients with gross hydrocephalus on post operative CT were managed successfully with VP shunt. Re-suturing of the wound with concomitant CSF lumbar drainage was found to be significantly associated (p=0.003) with the stoppage of CSF leakage, and the settlement of meningitis (p= 0.014). CONCLUSION: Incisional CSF leaks after posterior fossa surgery should be managed with re-suturing of the wound and concomitant CSF lumbar drainage, instead of an initial trial of conservative therapy alone.

Pharmaceutical wastewater being composite mixture of environmental pollutants may be associated with mutagenicity and genotoxicity.

Pharmaceutical industries are amongst the foremost contributor to industrial waste. Ecological well-being is endangered owing to its facile discharge. In the present study, heavy metals and organic contaminants in waste water were characterized using atomic absorption spectrophotometer and GC-MS, respectively. Mutagenicity and genotoxic potential of pharmaceutical waste water were investigated through bacterial reverse mutation assay and in vitro comet assay, respectively. Ames test and comet assay of first sample were carried out at concentrations of 100, 50, 25, 12.5, 6.25 % v/v effluent with distilled water. Chromium (Cr), lead (Pb), arsenic (As), and cadmium (Cd) were found in high concentrations as compared to WHO- and EPA-recommended maximum limits. Arsenic was found to be the most abundant metal and its maximum concentration was 0.8 mg.L(-1). GC-MS revealed the presence of lignocaine, digitoxin, trimethoprim, caffeine, and vitamin E in waste water. Dose-dependent decrease in mutagenic index was observed in both strains. Substantial increase in mutagenicity was observed for TA-100, when assay was done by incorporating an enzyme activation system, whereas a slight increase was detected for TA-102. In vitro comet assay of waste water exhibited decrease in damage index and percentage fragmentation with the increase in dilution of waste water. Tail length also decreased with an increase in the dilution factor of waste water. These findings suggest that pharmaceutical waste water being a mix of different heavy metals and organic contaminants may have a potent mutagenic and genotoxic effect on exposed living organisms.

Evaluation of antioxidant and antimicrobial potential of two endangered plant species Atropa belladonna and Matricaria chamomilla.

BACKGROUND: Plants are the natural source of antioxidants as well as antimicrobial compounds that has great potentials in pharmaceutical industry. In the present study, two medicinal plants Atropa belladonna and Matricaria chamomilla were collected from Northern areas of Pakistan. MATERIALS AND METHODS: The extracts of the collected plants were obtained by microwave assisted extraction (MAE) with changing parameters, power level and time; methanol and ethanol were solvents used during extraction. The extracts of plants were tested against different bacterial strains. RESULTS: It was observed that ethanolic extracts of Atropa belladonna has more significant antimicrobial activity against S.aureus than E.coli. In parallel, methanolic extract of Matricaria chamomilla showed greater significant antibacterial activity against S.aureus when compared with E.coli. In comparison, ethanolic extracts of Matricaria chamomilla has shown more significant results against S. aureus than E.coli (p ≤ 0.05). Both plants had no antibacterial activity against S.typhi. The free radical scavenging activity observed by DPPH assay, indicate that both plants have antioxidant activity at all levels of concentrations in solvent tested during the present work. However, methanolic extracts had greater antioxidant activity when compared with ethanolic extracts. CONCLUSION: Present study is thus helpful in highlighting present potentials for antioxidant and antimicrobial properties in the selected plants.