Anthrax: A disease of biowarfare and public health importance

Bioterrorism has received a lot of attention in the first decade of this century. Biological agents are considered attractive weapons for bioterrorism as these are easy to obtain, comparatively inexpensive to produce and exhibit widespread fear and panic than the actual potential of physical damage. Bacillus anthracis(B. anthracis), the etiologic agent of anthrax is a Gram positive, spore forming, non-motile bacterium. This is supposed to be one of the most potent BW agents because its spores are extremely resistant to natural conditions and can survive for several decades in the environment. B.anthracis spores enter the body through skin lesion(cutaneous anthrax), lungs(pulmonary anthrax), or gastrointestinal route(gastrointestinal anthrax) and germinate, giving rise to the vegetative form. Anthrax is a concern of public health also in many countries where agriculture is the main source of income including India. Anthrax has been associated with human history for a very long time and regained its popularity after Sept 2001 incidence in United States. The present review article describes the history, biology, life cycle, pathogenicity, virulence, epidemiology and potential of B. anthracis as biological weapon.

Effect of α-Ketoglutarate on Cyanide-induced Biochemical Alterations in Rat Brain and Liver

Objective To investigate the biochemical changes in rat brain and liver following acute exposure to a lethal dose of cyanide, and its response to treatment of α-ketoglutarate （α-KG） in the absence or presence of sodium thiosulfate （STS）. Methods Female rats were administered 2.0 LD50 potassium cyanide （KCN; oral） in the absence or presence of pre-treatment （-10 rain）, simultaneous treatment （0 rain） or post-treatment （＋2-3 min） of α-KG （2.0 g/kg, oral） and/or STS （1.0 g/kg, intraperitoneal, -15 min, 0 rain or ＋ 2-3 min）. At the time of onset of signs and symptoms of KCN toxicity （2-4 min） and at the time of death （5-15 min）, various parameters particularly akin to oxidative stress viz. cytochrome oxidase （CYTOX）, superoxide dismutase （SOD）, glutathione peroxidase （GPx）, reduced glutathione （GSH） and oxidized glutathione （GSSG） in brain, and CYTOX, sorbitol dehydrogenase （SDH）, alkaline phosphatase （ALP）, GSH and GSSG in liver homogenate were measured. Results At both time intervals brain CYTOX, SOD, GPx, and GSH significantly reduced （percent inhibition compared to control） to 24%, 56%, 77%, and 65%, and 44%, 46%, 78%, and 57%, respectively. At the corresponding time points liver CYTOX and GSH reduced to 74% and 63%, and 44% and 68%, respectively. The levels of GSSG in the brain and liver, and hepatic ALP and SDH were unchanged, Pre-treatment and simultaneous treatment of α-KG alone or with STS conferred significant protection on above variables. Post-treatment was effective in restoring the changes in liver but failed to normalize the changes in the brain. Conclusions Oral treatment with α-KG alone or in combination with STS has protective effects on cyanide-induced biochemical alterations in rat brain and liver.

Effect of Pre-treatment of α-Ketoglutarate on Cyanide-induced Toxicity and Alterations in Various Physiological Variables in Rodents

Objective To investigate the effects of pre-treatment of α-ketoglutarate （α-KG） on cyanide-induced lethality and changes in various physiological parameters in rodents. Methods The LD50 of potassium cyanide （KCN） given orally （po）, intraperitoneally （ip）, subcutaneously （sc） or intravenously （iv） was determined in male mice, in the presence or absence α-KG given po, ip or iv. α-KG was administered 10, 20 or 40 min prior to KCN at 0.50, 1.0 or 2.0 g/kg by po or ip route, and at 0.10, 0.20 or 0.40 g/kg by iv route. Protection index （PI） was calculated as the ratio of LD50 of KCN in the presence of α-KG （protected animals） and LD50 of KCN in the absence of α-KG （unprotected animals）. In a separate experiment, several physiological variables viz. mean arterial pressure （MAP）, heart rate （HR）, respiratory rate （RR）, neuromuscular transmission （NMT） and rectal temperature （RT） were measured in anesthetized female rats pre-treated （-10 rain） with po （2.0 g/kg） or iv （0.125 g/kg） α-KG and then administered sub-lethal （0.75 LD50） or lethal （2.0, 4.0 or 8.0 LD50） doses of KCN （po）. Results PI of 4.52, 6.40 and 7.60 at -10 min, 3.20, 5.40 and 6.40 at -20 min, and 1.40, 3.20 and 5.40 at -40 min of po administration with α-KG was observed for 0.50, 1.0 and 2.0 g/kg doses, respectively, against KCN given by po route. When KCN was given ip, a PI of 3.38, 4.79 and 5.70 was observed for 0.50, 1.0 and 2.0 g/kg α-KG given ip （-10 min）, respectively. A lower PI of 3.37, 2.83 and 2.38 was observed when KCN given sc was challenged by 2.0 g/kg α-KG given ip at -10, -20 or -40 min, respectively. Similarly, a PI of 3.37, 2.83 and 2.0 was noted when KCN given sc was antagonized by 2.0 g/kg α-KG given po at -10, -20 or -40 rain, respectively. No appreciable protection was observed when lower doses of α-KG （ip or po） challenged KCN given by sc route. Pre-treatment of iv or po administration of α-KG did not afford any protection against KCN given po or iv route. Oral treatment of 0.75 LD50 KCN caused significant decrease in MAP and HR after 15 min, RR after 30 min and NMT after 60 min. There was no effect on RT. No reduction in MAP, HR, RR and RT was observed when rats received 2.0 or 4.0 LD50 KCN after pre-treatment of α-KG （po; 2.0 g/kg）. However, no protection was observed on NMT. Protective efficacy of α-KG was not observed on MAP, HR, RR, and NMT decreased by 8.0 LD50 KCN. Decrease in MAP and NMT caused by 2.0 LD50 KCN （po） was resolved by iv administration of α-KG Conclusions Cyanide antagonism by α-KG is best exhibited when both α-KG and KCN are given by po route. The protective effect of α-KG on cyanide-induced changes in several physiological parameters also indicates a promising role of α-KG as an alternative cyanide antidote.

Cyanide Intoxication in Mice Through Different Routes and its Prophylaxis by α-Ketoglutarate

Antagonising effects of α-ketoglutarate (α-KG) could be attributed to complexing of the reactive nucleophile (CN-) to form cyanohydrin in cyanide intoxication. However, an enormous protection obtained could not be delineated on account of possible in situ binding of α-KG given intraperitoneally (i.p.) in mice to cyanide administered through the same route. The present study was designed to see the efficacy of a-KG alone or in combination with sodium nitrite (SN) and/or sodium thiosulfate (STS) in male mice exposed to cyanide administered through subcutaneous (s.c.) or inhalation route. A technique for generation of hydrogen cyanide (HCN) is also discussed. On the basis of protection index (PI), defined here as the LD50 of cyanide in protected mice/LD50 of cyanide in unprotected mice and survival time, STS + α-KG regimen was equipotent to the conventional SN + STS regimen. This is further substantiated by effect of α-KG in reducing plasma cyanide levels. The efficacy of α-KG remains undeterred irrespective of the route of cyanide intoxication, while the magnitude of protection varies.

Andrographolide inhibits chikungunya virus infection by up-regulating host innate immune pathways

Objective: To investigate the therapeutic efficacy of andrographolide, a plant derived compound, against chikungunya virus(CHIKV) infection. Methods: Using flow cytometry and immunoblotting assay, in vitro viral protein expression was studied in THP-1 cells line. In Balb/c mouse neonates, viral RNA copy number was determined by real time PCR. Results:The results showed reduced CHIKV protein expression on andrographolide treatment in CHIKV-infected human peripheral blood mononuclear cells, Vero cells and THP-I cell line.In vivo, andrographolide treatment to CHIKV-infected neonates reduced viral RNA copy number. Further. andrographolide also increased cytotoxic T lymphocytes both in vitro and in vivo. Andrographolide also activated host innate immune pathways, viz., protein kinase R.phosphorylated eukaryotic initiation factor 2 α, retinoic acid inducible gene-Ⅰ and interferon regulatory factor 3/7, thereby increasing IFN-a secretion. CHIKV-induced nuclear factor κlight chain enhancer of activated B cells and tumor necrosis factor-a was also reduced on andrographolide treatment. Conclusion: Andrographolide inhibits CHIKV by suppressing viral protein expression and up-regulating host innate immunity and hence could be an effective therapeutic agent against CHIKV infection.

Acute Toxicity and Cardio-Respiratory Effects of 2-Deoxy-D-Glucose: A Promising Radio Sensitiser

Objective To evaluate the acute toxicity of 2-deoxy-D-glucose （2DG） by oral （p.o.） and intravenous （i.v.） routes, and also the cardio-respiratory effects following high doses of 2DG in animal models. Methods The LD50 of 2DG （in water） was determined in rats and mice by p.o. route and in mice by i.v. route. The effect of 2-DG （250 mg/kg, 500 mg/kg, and 1000 mg/kg, i.v.） was studied on various cardio-respiratory parameters viz., mean arterial blood pressure, heart rate and respiratory rate in anaesthetised rats. The effect of 2DG （500 mg/kg, 1000 mg/kg, and 2000 mg/kg, p.o.） was also studied on various respiratory parameters viz., respiratory rate and tidal volume in conscious rats and mice using a computer program. Results The p.o. LD50 of 2DG was found to be 〉8000 mg/kg in mice and rats, and at this dose no death was observed. The LD50 in mice by i.v. route was found to be 8000 mg/kg. At this dose 2 out of 4 mice died and the death occurred within 6 h. A significant increase in the body weight was observed after p.o. administration of 2DG in rats at 500 mg/kg, 1000 mg/kg, and 2000 mg/kg doses. There was no significant change in the body weight at 4000 mg/kg and 8000 mg/kg by the p.o. route in rats and up to 8000 mg/kg by p.o. as well as i.v. routes in mice. Intravenous administration of 2DG （250 mg/kg, 500 mg/kg, and 1000 mg/kg） in anaesthetised rats showed a time-dependent decrease in the mean arterial blood pressure. There was no change in the heart rate in any of the treatment groups. The tidal volume was not changed significantly by p.o administration in conscious rats, but a significant decrease in the respiratory frequency at 500 mg/kg and 1000 mg/kg doses was observed. In the mice also there was no change in the tidal volume after p.o, administration, but the respiratory frequency decreased significantly at 2000 mg/kg dose. Conclusion 2DG is a safe compound but can cause a fall in the blood pressure and a decrease in respiratory frequency at high doses.

Arsenic Induced Inhibition of δ-aminolevulinate DehydrataseActivity in Rat Blood and its Response To Meso2,3-dimercaptosuccinic Acid andMonoisoamyl DMSA

Objective The objective of this study was to investigate arsenic induced changes in blood 8-aminolevulinic acid dehydratase (ALAD) after in vitro and in vivo exposure to this element and its response to co-administration of meso 2,3-dimercaptosuccinic acid (DMSA) and monoisoamyl DMSA (MiADMSA) either individually or in combination. Methods Rat whole blood was exposed to varying concentrations (0.1, 0.2 and 0.5 mmol/L) of arsenic (III) or arsenic (V), to assess their effects on blood ALAD activity. Varying concentrations of MiADMSA and DMSA (0.1,0.5 and 1.0 mmol/L) were also tried in combination to determine its ability to mask the effect of arsenic induced (0.5 mmol/L) inhibition of blood ALAD in vitro. In vitro and in vivo experiments were also conducted to determine the effects of DMSA and MiADMSA either individually or in combination with arsenic, on blood ALAD activity and blood arsenic concentration. Results In vitro experiments showed significant inhibition of the enzyme activity when 0.1-0.5 mmol/L of arsenic (III and V) was used. Treatment with MiADMSA increased ALAD activity when blood was incubated at the concentration of 0.1 mmol/L arsenic (III) and 0.1 mmol/L MiADMSA. No effect of 0.1 mmol/L MiADMSA on ALAD activity was noticed when the arsenic concentration was increased to 0.2 and 0.5 mmol/L. Similarly, MiADMSA at a lower concentration (0.1 mmol/L) was partially effective in the turnover of ALAD activity against 0.5 mmol/L arsenic (III), but at two higher concentrations (0.5 and 1.0 mmol/L) a complete restoration of ALAD activity was observed. DMSA at all the three concentrations (0.1,0.5 and 1.0 mmol/L) was effective in restoring ALAD activity to the normal value. Conclusions The results thus suggest that arsenic has a distinct effect on ALAD activity. Another important toxicological finding of the present study, based on in vivo experiments further suggests that combined administration of DMSA and MiADMSA could be more beneficial for reducing blood ALAD inhibition and blood arsenic concentration than the individual treatment.

Arsenic-induced abnormalities in glucose metabolism: Biochemical basis and potential therapeutic and nutritional interventions

Health hazards due to the consumption of heavy metals such as arsenic have become a worldwide problem. Metabolism of arsenic produces various intermediates which are more toxic and cause toxicity. Arsenic exposure results in impairment of glucose metabolism, insulin secretion in pancreatic β-cells, altered gene expressions and signal transduction, and affects insulinstimulated glucose uptake in adipocytes or skeletal muscle cells. Arsenic toxicity causes abnormalities in glucose metabolism through an increase in oxidative stress. Arsenic interferes with the sulfhydryl groups and phosphate groups present in various enzymes involved in glucose metabolism including pyruvate dehydrogenase and α-ketoglutarate dehydrogenase, and contributes to their impairment. Arsenic inhibits glucose transporters present in the cell membrane, alters expression of genes involved in glucose metabolism, transcription factors and inflammatory cytokines which stimulate oxidative stress. Some theories suggest that arsenic exposure under diabetic conditions inhibits hyperglycemia. However, the exact mechanism behindthe behavior of arsenic as an antagonist or synergist on glucose homeostasis and insulin secretion is not yet fully understood. The present review delineates the relationship between arsenic and the biochemical basis of its relationship to glucose metabolism. This review also addresses potential therapeutic and nutritional interventions for attenuating arsenic toxicity. Several other potential nutritional supplements are highlighted in the review that could be used to combat arsenic toxicity.

Topical Preparation of Newer and Safer Analogs of N,N-diethyl-2-phenylacetamide (DEPA) against <i>Aedes aegypti</i>Mosquitoes

Cosmetic acceptability and primary skin irritation are the two main parameters for assessing the suitability of any topical formulation meant for protection against the painful bites of mosquitoes. In the present study four newer analogs of N,N-diethyl-2-phenylacetamide (DEPA), were synthesized and formulated for topical application as insect repellent. They were assessed for their irritant behavior on rabbit’s skin for erythema and edema. The topical formulations of the analogs were also assessed for their protection time at varying concentrations against Aedes aegypti mosquitoes.

Prophylactic Effect of Gossypin Against Percutaneously Administered Sulfur Mustard

Objective To evaluate the protective efficacy of gossypin （3,3＇,4＇,5,7,8-hexahydroxyflavone 8-glucoside） by administering it intraperitoneally, for dose, time, and vehicle dependent effects against sulphur mustard （SM）, administered through percutaneous route in mice. Methods SM （diluted in PEG-300） was administered percutaneously. The protective efficacy of gossypin was evaluated by administering it intraperitoneally （50, 100, 200, and 400 mg/kg）, in various vehicles （water, PEG-300 and DMSO）, and time intervals （30 min prior, simultaneous and 2 h post）. The time dependent protection of gossypin （200 mg/kg in PEG-300; i.p.） was also evaluated using selected biochemical variables （GSH, GSSG, MDA, total antioxidant status, Hb, WBC count, RBC count, glutathione peroxidase, glutathione reductase, and superoxide dismutase） and liver histology. The protection of gossypin by oral route was also evaluated against percutaneously administered SM. Results The protection against systemic toxicity of SM （LD50 8.1 mg/kg） was better when gossypin was given with PEG-300 （8.0 folds） than DMSO （5.7 folds）. No protection was observed when gossypin was administered with water. Good protection （8.0 folds） was observed when gossypin was administered （200 mg/kg in PEG-300; i.p.） at 30 min prior or simultaneous to SM exposure, but no protection was observed when gossypin was administered 2 h post to SM exposure. A significant weight loss was observed 7 days after SM administration （2 LD50）, with a significant increase in RBC and Hb. A significant decrease in total antioxidant status of plasma, liver GSH and GSSG levels, and in the activities of glutathione peroxidase, glutathione reductase and superoxide dismutase was also observed 7 days after SM administration. SM treated mouse liver also showed necrosis. A significant protection was observed when gossypin （200 mg/kg in PEG-300; i.p.） was administered either as a pretreatment （30 min before） or simultaneous treatment, and not as a post treatment （2 h）. The protective efficacy of gossypin was better through oral route when administered with DMSO （4.8 folds） than with PEG-300 （2.4 folds）. No protection was observed when gossypin was administered orally with water. Conclusion Percutaneous administration of SM induces oxidative stress and gossypin can protect it as a prophylactic agent by intraperitoneal or oral routes.

Altered Host Resistance to Listeria monocytogenes In Mice Exposed to 1-Chloroacetophenone (CN) Vapours

Short term repeated exposure of 1-chloroacetophenone (CN) vapours at a concentration of 0.153 mg per litre for 15 minutes daily on 10 consecuitve days in Swiss albino male mice resulted in increased mortality to Listeria monocytogenes. Significantly elevated bacterial growth was observed in the spleen and liver of the CN exposed animals. The increased bacterial count in these organs was evident within 4-6 days post challenge as compared to vehicle exposed infected and unexposed infected animals. Increased susceptibility to infection has been considered to be the function of immune alteration due to cumulative short term effects ofCN vapour inhalation. This may be attributed to immunotoxic effects of CN on Tcells mediated macrophage functions.

Acknowledgments to reviewers of World Journal of Pharmacology

We acknowledge our sincere thanks to our reviewers.Many reviewers have contributed their expertise and time to the peer review,a critical process to ensure the quality of our World Series Journals.Both the editors of the journals and authors of the manuscripts submitted to the journals are grateful to the following reviewers for reviewing the

Acknowledgments to reviewers of World Journal of Pharmacology

Many reviewers have contributed their expertise and time to the peer review,a critical process to ensure the quality of World Journal of Pharmacology.The editors and authors of the articles submitted to the journal are grateful to the following reviewers for evaluating the articles(including those published in this issue and those rejected for this issue)during the last editing time period.Tzyh-Chyuan Hour,Ph D,Associate Professor,Institute of Biochemistry,Kaohsiung Medical University,100 Shih-Chuan 1st Road,Kaohsiung 807,Taiwan。

Acknowledgments to reviewers of World Journal of Pharmacology

We acknowledge our sincere thanks to our reviewers.Many reviewers have contributed their expertise and time to the peer review,a critical process to ensure the quality of our World Series Journals.Both the editors of the journals and authors of the manuscripts submitted to the journals are grateful to the following reviewers for reviewing the articles(either published or rejected)over the past period of time.Tzyh-Chyuan Hour,Ph D,Associate Professor,Institute of Biochemistry,Kaohsiung Medical University,100 Shih-Chuan 1st Road,

Are malaria elimination efforts on right track?An analysis of gains achieved and challenges ahead

Background:Malaria causes significant morbidity and mortality each year.In the past few years,the global malaria cases have been declining and many endem ic countries are heading towards malaria elimination.Nevertheless,reducing the number of cases seems to be easy than sustained elimination.Therefore to achieve the objective of com plete elimination and maintaining the elimination status,it is necessary to assess the gains made during the recent years.Main text:With inclining global support and World Health Organisation(WHO)efforts,the control programmes have been im plem ented effectively in many endemic countries.Given the aroused interest and investments into malaria elimination program m es at global level,the am bitious goal o f elimination appears feasible.Sustainable interventions have played a pivotal role in malaria contraction,however drug and insecticide resistance,social,dem ographic,cultural and behavioural beliefs and practices,and unreformed health infrastructure could drift back the progress attained so far.Ignoring such im peding factors coupled with certain region specific factors may jeopardise our ability to abide righteous track to achieve global elimination of malaria parasite.Although support beyond the territories is important,but well managed integrated vector managem ent approach at regional and country level using scrupulously selected area specific interventions targeting both vector and parasite along with the com m unity involvem ent is necessary.A brief incline in malaria during 2016 has raised fresh perturbation on whether elimination could be achieved on time or not.Conclusions:The intervention tools available currently can most likely reduce transmission but clearing of malaria epicentres from where the disease can flare up any time,is not possible without involving local population.Nevertheless maintaining zero malaria transmission and checks on malaria import in declared malaria free countries,and further speeding up of interventions to stop transmission in elimination countries is most desirable.Strong collaboration backed by adequate political and financial support am ong the countries with a common objective to eliminate malaria must be on top priority.The present review attempts to assess the progress gained in malaria elimination during the past few years and highlights some issues that could be important in successful malaria elimination.

Preclinical investigation of the pharmacokinetics, metabolism, and protein and red blood cell binding of DRDE-07: a prophylactic agent against sulphur mustard

DRDE-07,a newly synthesized amifostine analog currently under clinical investigation in a phase I trial,is a potent antidote against sulfur mustard toxicity.The purpose of this research was to evaluate the pharmacokinetic profile of DRDE-07 in female Swiss Albino mice after a single oral dose of 400 or 600 mg/kg.The physicochemical properties of DRDE-07,including solubility,pK a,Log P,plasma protein binding and plasma/blood partitioning,were determined to support the pharmacokinetic characterization.DRDE-07 concentration was determined by an HPLC-UV method.The profile of plasma concentration versus time was analyzed using a non-compartmental model.Plasma protein binding was assessed using ultrafiltration.DRDE-07 appeared rapidly in plasma after oral administration with peak plasma levels(C max)observed in less than 15 min.There was a rapid decline in the plasma levels followed by a smaller second peak about 90 min after dosing.The plasma protein binding of DRDE-07 was found to be less than 25%at all concentrations studied.Plasma clearance of DRDE-07 is expected to be?1.5 fold higher than the blood clearance of DRDE-07.The probable metabolite of DRDE-07 was identified as phenyl-S-ethyl amine.