Alveolar Soft Part Sarcoma Metastasizing to the Brain: A Rare Entity Revisited with Review of Recent Literature.

A 20-year-old male patient presented with a recent history of two episodes of seizures and Glasgow Coma Scale of E3V3M6. He was drowsy at presentation with bradycardia. On further examination, he was found to be having a 10 × 6cm well-circumscribed mass in the right medial thigh whose fine needle aspiration cytology was suggestive of soft tissue sarcoma. Contrast-enhanced magnetic resonance imaging brain showed enhancing lobulated lesions in bilateral hemispheres with significant edema. A decision to decompress the intracranial space occupying lesion was taken. Left-sided larger brain lesions underwent total gross excision followed by excision of the thigh mass. Histopathology of brain lesion was suggestive of metastatic deposits of alveolar soft part sarcoma, whereas the thigh mass was suggestive of alveolar soft part sarcoma grade 2 according to Fédération Nationale des Centres de Lutte Contre Le Cancer grading system. Postoperative course of the patient was unremarkable and his general condition improved remarkably (Functional Assessment of Cancer Therapy-Brain version 4 score improvement >50% at 1 month). He is in follow-up for the last 12 months and currently has completed chemoradiotherapy and is living a self-sufficient life. Though patients with aforementioned rare diagnosis have overall poor prognosis, decompressive craniotomy with metastasectomy may improve the overall quality of life of such young patients, and further chemoradiotherapy may help in prolonging life with acceptable level of comorbidity. This case is being discussed here in terms of the rarity of the final diagnosis, the management strategy used, and the quality of life improvement.

Randomised prospective study of using intraoperative, intraincisional and intraperitoneal ropivacaine for the early discharge of post-laparoscopic cholecystectomy patients as a day case in a cost-effective way in government setup of low-income and middle-income countries: Opening new horizons.

BACKGROUND: Pain is the main reason for staying overnight at hospital after an uncomplicated laparoscopic cholecystectomy. OBJECTIVES: A randomised prospective study was planned to compare the efficacy of intraincisional and intraperitoneal use of 0.2% ropivacaine so that patients undergoing an uncomplicated laparoscopic cholecystectomy can be discharged as a day case in a cost-effective way. METHODS: 191 patients were operated by elective four-port laparoscopic cholecystectomy. They were randomised into three groups after triple blinding according to location of 0.2% ropivacaine use. All patients were given ~23 mL of solution (drug or normal saline depending on the group), 20 mL of which was given at intraperitoneal location and ~1 mL/cm of incision intraincisionally. Pain scores (Visual Analogue Scale (VAS), Numeric Rating Scale (NRS) and Faces Pain Scale-Revised (FPS-R)) were evaluated at 4 and 8 hours postoperatively. Only those patients with a VAS ≤3, NRS ≤3 and FPS-R ≤2, no requirement of rescue analgesia, no shoulder pain, ambulated at least once, passed urine and taking oral sips were offered discharge as a day case. RESULTS: 31% of patients in intraperitoneal group (n=62) could be discharged as a day case as compared with 48% in intraincisional group (n=68) (p>0.05) and 89% in combined group (n=61) (p<0.05, with respect to both other groups). CONCLUSION: The combined use of intraincisional and intraperitoneal ropivacaine is a cost-effective way of discharging approximately 9 in 10 patients as a day case. This study is unique as this is the first study in which only a local anaesthetic has been used to predict discharges as a day case.

Traumatic Vertebral Body Second Lumbar over Third Lumbar Retrolisthesis in a Child: Reporting the First Case along with the Review of Relevant Literature.

Traumatic retrolisthesis of the lumbar vertebrae is a rare entity in children. Only four such cases, two cases each of first lumbar-second lumbar (L1-L2) and L5-S1 retrolisthesis in children, have been reported so far in the English scientific literature. Here, we report a traumatic retrolisthesis of the L2 vertebra in an 8-year-old male child. He was injured when he lost control while playing, skidded, and fell into a 1-m deep drainage system hole. He presented with backache and urinary retention. His plain radiographs and noncontrast computed tomography of the lumbosacral spine revealed Meyerding Grade II retrolisthesis of the L2 vertebra over the third. The magnetic resonance imaging of the affected area revealed no significant canal narrowing, and there was no spinal cord compression or contusion. A urodynamic study was done which revealed a normal bladder function. The patient was given a trial of spontaneous urination by removing the Foley's catheter after 5 days of injury, and he passed urine normally. The patient was managed conservatively. He was discharged on day 7 with the advice of complete bed rest of 6 weeks and thoracolumbosacral orthoses. The patient has been in follow-up for the past 15 months, and his listhesis has completely resolved. The patient is ambulatory with no neurodeficit. This case is being presented in view of rarity. This is the first case report of L2 over L3 retrolisthesis in a child.

A prospective study of evaluation of operative duration as a predictor of mortality in pediatric emergency surgery: Concept of 100 minutes laparotomy in resource-limited setting.

INTRODUCTION: Operative duration is an important but under-studied predictor of mortality in emergency laparotomies. AIMS AND OBJECTIVES: The primary objective of this study was to quantify the effect of duration of emergency laparotomy in children on mortality; and to identify a rough cut-off duration of laparotomy to serve as a guide so that a laparotomy can be planned to optimize pediatric surgical patient outcome. MATERIALS AND METHODS: This is a prospective study conducted in a government tertiary teaching institution over a period of 24 months. All children in the age group of 5-10 years presenting in the emergency department with Pediatric Risk of Mortality III score ≤8, undergoing emergency laparotomy in emergency operation theater, were included. OBSERVATIONS AND RESULTS: In all, 213 children were included in the study. The mean time from presentation to shifting to the operating room was 3.7 h. The mean operative duration was 108 min. The mean operative time in survived patients was 102 min as compared to 135 min in expired patients (P < 0.05). The 30-day in-hospital mortality rate was 17.4%. After application of binary logistic regression analysis, it was found that time to laparotomy and operative duration were significant risk factors (<0.05) predicting post-operative mortality. Kaplan-Meier survival curve showed a decrease at a mean weighted operative duration of approximately 100 min. Receiver operating characteristic curve analysis yielded operative duration of 123.5 min at which Youden's index maximized. CONCLUSION: This 100-min duration of laparotomy might appear a long duration but in casualty setup of a government hospital with limited resources, there are so many hurdles for optimal working that completion of an emergency laparotomy in children in 100 min can be considered a realistic target for improving post-operative outcome. At an operative duration of <123.50 min, mortality rates within acceptable limits can be achieved.

Abdominal herniation associated with bullhorn injury as a separate entity from traumatic abdominal wall hernias.

Bullhorn injury is a rare mechanism causing traumatic abdominal wall hernia (TAWH). Bullhorn injury needs to be discussed as a separate sub-entity among TAWHs as the mechanism of injury is such that the great force is generated at a relatively small area of impact for a short duration of time which may lead to muscle defect without compromising integrity of overlying skin (referred to as sheathed goring) leading to herniation of abdominal viscera. The purpose of this review was to discuss abdominal herniation's associated with bullhorn injury as a separate entity from TAWHs; recognize the common presentations, mechanism of injury, and modalities of treatment currently utilized for this rare condition. A comprehensive online English, Spanish, Portuguese, and French language medical literature search was done using various electronic search databases. Different search terms including MeSH related to bullhorn-injury associated injuries including abdominal wall hernias were used. An advanced search was further conducted by combining all the search fields in abstracts, keywords, and titles. We summarized the data from the searched articles and found 12 cases who underwent emergency or elective herniorrhaphy with or without the use of mesh. We have proposed a treatment algorithm for such cases in light of the present era of laparoscopy and propose the usage of the term "bullhorn-injury associated traumatic hernia" for such cases. We present here the first most comprehensive discussion of all such cases reported till date.

Primary cecal pathologies presenting as acute abdomen and critical appraisal of their current management strategies in emergency settings with review of literature.

BACKGROUND: The importance of cecal pathologies lie in the fact that being the first part of the large intestine, any disease involving the cecum affects the overall functioning of the large bowel. Primary cecal pathologies presenting as acute abdomen have not been described in any previous study in terms of presentation, management, and outcome. OBJECTIVES: The objective of this study was to identify the reported causes of primary cecal pathologies presenting as acute abdomen and the various causes presenting in our setting, the to discuss morbidity and mortality associated with cecal pathologies, and to critically analyze the various management modalities employed in an emergency setting. MATERIALS AND METHODS: This is a retrospective analysis of a prospectively collected data of all adult patients admitted to our unit in the last 10 years for acute abdomen, in whom cecal pathology was identified as the primary offending agent. RESULTS: Our analysis of 43 patients revealed cecal perforation secondary to amebic colitis along with the simultaneous presence of liver abscess to be the most common primary cecal entity met in practice as acute abdomen. Other pathologies that were encountered included volvulus, diverticulitis, and idiopathic typhlitis. Primary acute cecal pathologies are associated with high mortality rates (≈42%). Delay in diagnosis seems to be the most important factor contributing to high mortality as these conditions are commonly misdiagnosed as appendicular pathology. Most of our patients were managed by conservative colonic resection with proximal diversion. This seems to be a more plausible option in current practice in an emergency setting (mortality rate ≈ 36%) as hemicolectomy is associated with proportionately higher mortality rates (67%). CONCLUSION: A high index of suspicion, timely and adequate pre-operative workup, optimal resuscitation, and surgical conservatism with proximal diversion might help in early accurate identification of these conditions and possibly improved outcome.

Pneumohernios due to perforated cecum in left Amyand's hernia.

We report a case of incarcerated left indirect inguinal hernia in a male child which on exploration revealed the presence of free air and fecal matter containing fluid in the hernial sac. This is the second reported case of the presence of cecal perforation in left Amyand's hernia in pediatric age group and unique in the sense of the form of abnormal anatomy encountered per-operatively.

Randomized controlled study of intraincisional infiltration versus intraperitoneal instillation of standardized dose of ropivacaine 0.2% in post-laparoscopic cholecystectomy pain: Do we really need high doses of local anesthetics-time to rethink!

BACKGROUND: Earlier studies done to compare the efficacy of use of local anesthetics at intraperitoneal location versus intraincisional use had utilized equal amount of drugs at the two locations, usually 10-20 ml. Using this large amount of drug in the small space of intraincisional location as compared to similar amount of drug in large intraperitoneal space created an inadvertent bias in favor of patients receiving the drug intraincisionally so these patients naturally experienced less pain. AIMS AND OBJECTIVES: To conduct a randomized, triple-blind, placebo-controlled study by standardizing dose of local anesthetic, to compare the effectiveness of intraperitoneal against intraincisional use of ropivacaine 0.2% for post-laparoscopic cholecystectomy pain relief. MATERIALS AND METHODS: 294 patients underwent elective 4-port laparoscopic cholecystectomy. Patients were triple blindly randomized. All patients received ~ 23 ml of solution, of which 20 ml was given intraperitoneally (1 ml/cm; 16 ml along right hemi-dome and 4 ml in gall bladder fossa) and ~ 3 ml intraincisionally (1 ml/cm of length of incision). Solution was either normal saline or drug (0.2% ropivacaine) depending on the group [controls (n = 86), intraperitoneal group (n = 100), and intraincisional group (n = 108)]. 5 different pain scales were used for assessment of overall pain. Pain scores were assessed at 5 points of time. RESULTS: Patients in intraincisional group showed significantly less overall pain and rescue analgesia requirement (p < 0.05). Intraincisional group showed significantly less overall pain (p < 0.05) as compared to intraperitoneal group; however, use of rescue analgesia was comparable in the two groups (p > 0.05); and shoulder pain was significantly less in intraperitoneal group (p < 0.05). CONCLUSION: The intraincisional use of injection ropivacaine at its minimum concentration of 0.2% in minimal doses of 1 ml/cm at the end of procedure provides significantly more post-operative analgesia as compared to intraperitoneal group and controls. However, for controlling shoulder pain, the use of intraperitoneal ropivacaine is desirable.

Negotiating the Curve of Laparoscopic Hepatopancreaticobiliary Procedures (Basic to Advanced) at a Tertiary Rural Teaching Institute.

BACKGROUND: We present here our experience of laparoscopic hepatopancreaticobiliary (HPB) surgeries at our rural institute and the hurdles we faced overcoming the various challenges. AIMS AND OBJECTIVES: The objectives of this study were to assess the feasibility, successful completion, operative time, conversions/requirement of assistance, duration of hospital stay, and postoperative complications on the HPB procedures performed laparoscopically in our surgical unit; and to help young surgeons for smooth navigation through their laparoscopic career. MATERIALS AND METHODS: All the patients admitted under our unit over the past 9 years for elective HPB surgeries operated by a single surgeon were included in this study. RESULTS: Total 1304 basic laparoscopic biliary procedures were successfully completed laparoscopically. After getting well versed with the standard procedure, we switched over to difficult cases involving densely adhered gallbladder, frozen Calot's, Mirizzi's syndrome, use of intraoperative cholangiogram, and take down of cholecystoduodenal fistulas. Next step in evolution was doing laparoscopic common bile duct exploration and biliary procedures with decreased number of ports. Five hundred and sixty-eight procedures were advanced HPB surgeries. With time, we also started performing a variety of complex advanced laparoscopic procedures such as cystogastrostomy, hepaticojejunostomy, choledochoduodenostomy, and pancreaticojejunostomy. All these procedures have been discussed with respect to operative duration, conversion rates, blood loss, hospital stay, and complication rates in the initial and later parts of the learning curves and further compared with previous standard large case studies on specific surgeries. CONCLUSION: Several hurdles are met in a new institute, that too, a rural one. The present discussion will help the budding surgeons to identify their deficiencies and chart a way forward in a systematic scientific manner.

Evolution of Various Components of Pain After Laparoscopic Cholecystectomy: Importance of Its Prognostication for Effective Pain Control Using a Local Anesthetic and for Making a Valid Practical "Discharge Criteria" Model Predicting Early Discharge of Patients.

OBJECTIVES: Pain continues to be the most important limiting factor affecting the early discharge of patients after laparoscopic cholecystectomy (LC). Our aim was to conduct a randomized case controlled study to predict the evolution of various components of postoperative pain by using ropivacaine 0.2% at intraperitoneal and intraincisional locations; and to further use this information to make a model predicting early discharge of patients. METHODS: Two hundred forty-four patients underwent elective four-port LC. Patients were triple blindly randomized. All patients received ∼23 mL of solution, of which 20 mL was given intraperitoneally and ∼3 mL was given intraincisionally. Solution was either normal saline or drug (0.2% ropivacaine) depending on the group (controls [n = 77], intraperitoneal group [n = 80], and intraincisional group [n = 87]). Five different pain scales were used for assessment of overall pain. Only those patients with a Visual Analog Scale (VAS) ≤3, Numeric Rating Scale (NRS) ≤3, Visual Descriptor Scale (VDS) ≤ "Slight Pain," Faces Pain Scale-Revised (FPS-R) ≤2, and Activity Tolerance Scale (ATS) ≤ "Can Be Ignored" along with absence of use of rescue analgesia and shoulder pain were considered for "Discharge Criteria." RESULTS: Incisional component of pain was found to be the main component of pain that predominated in the immediate postoperative period. However, it declined rapidly over 12 hours and was then dominated by the visceral component. Shoulder component peaked around the eighth postoperative hour. Seven percent of patients in controls could be discharged at the 12th postoperative hour and 18% at the 24th hour. In the intraperitoneal group, 18% and 61% patients could be discharged at the 12th and 24th hour, respectively, as compared with 57% and 78% in the intraincisional group using the "Discharge Criteria." "Discharge Criteria" was 100% effective in predicting patients' acceptance to go home. CONCLUSION: The effect of local anesthetic at intraincisional and intraperitoneal sites is additive with drug catering to different components of pain. We recommend using the "Abbreviated Discharge Criteria" routinely in practice to check for patients' eligibility to be discharged.

A Humorously Serious Take on Plight of Postgraduate Medical Entrance Examinees in India.

Following is a satire on the plight of postgraduate medical entrance examinees in India. It highlights the questions they have to face during their entrance examination preparations and the annoying explanations they have to read in the entrance preparation books. Though the system of examination is improving in India, few loopholes do exist and need to be rectified. This article, thus, attracts the attention of readers towards one such backdrop regarding controversial/wrong/misprinted questions which has come a long way but still has scope of improvement.

Concomitant Presence of Carcinoma of Breast in a Patient with Early-Onset Cataract: First Case Report of a Potential Association.

A recent nation-wide population based retrospective cohort study from Taiwan in 2014 suggested a propensity of developing breast cancer in young females with early-onset cataract. We report here a case of a young non-obese female who presented to us with a large lump in the right breast with skin nodules and bilateral painless progressive diminution of vision. Patient was diagnosed to be suffering from locally advanced carcinoma of right breast with axillary metastasis (Stage III B) and bilateral early-onset nuclear cataract. Patient was started on taxane and anthracycline based neoadjuvant chemotherapy and after three cycles of chemotherapy patient showed partial response (≈50% reduction of tumour mass). Patient underwent small incision cataract surgery in left eye after the first cycle of chemotherapy. However, the patient was lost before any operative intervention could be done as she died in a road traffic accident. Possible mechanism of development of breast cancer in patients with early-onset cataract is discussed. This is the first reported case of a patient of early-onset cataract developing breast cancer as well as the first reported case of concomitant presence of early-onset bilateral cataract with breast cancer.

Characterization of immunoreactive proteins of Setaria cervi isolated by preparative polyacrylamide gel electrophoresis.

Filarial parasites are complex mixtures of antigenic proteins and characterization of these antigenic molecules is essential to identify the diagnostically important filaria-specific antigens. In the present study, we have fractionated the somatic extracts from adults of Setaria cervi (bovine filarial parasite) on preparative SDS-polyacrylamide gel electrophoresis and tested the immunoreactivity of the separated gel fractions with polyclonal antibodies against filarial excretory-secretory antigens as well as filarial patients sera. The SDS-PAGE analysis of gel eluted fractions revealed 1 protein band in F-1 fraction, 2 protein bands in F-2 fraction and 2-3 protein bands in all other fractions (F3- F11). Seven gel eluted fractions (F1, F2, F3, F4, F5, F6 and F11) showed high ELISA reactivity with the polyclonal antibody (against excretory-secretory antigen) and four of these fractions (F-1, F-2, F3 and F6) exhibited high ELISA reactivity with antibodies present in filarial patient sera. The reactivities of the gel fractions (F1 and F2), recognized by filarial patients sera, were also tested with the monoclonal antibody (detecting the filarial circulating antigen). The F1 and F2 gel eluted fractions were found to have the target antigen of monoclonal antibody as evident by high reactivity with the monoclonal antibody in ELISA and immunoblotting. The S. cervi gel eluted F1 fraction (containing single antigen) could detect antibodies in filarial patients sera and not in non-filarial sera thereby suggesting its usefulness for specific serodiagnosis of human filariasis.

Immunogenicity of a plasmid DNA vaccine encoding 42kDa fragment of Plasmodium vivax merozoite surface protein-1.

Plasmodium vivax is the second major human malaria parasite that inflicts debilitating morbidity and consequent economic impact in South-East Asian countries. The relapsing nature of P. vivax along with the emergence of drug-resistant P. vivax strains has emphasized the urgent need for a vaccine. However, the development of an effective vivax vaccine is seriously hampered due to the diversity and variation in parasite antigens and non-availability of suitable animal models. DNA based vaccines represent an alternative approach in inducing immunity to multiple targets from different stages of malaria parasite. DNA prime-boosting strategies induce both antibody mediated and cell-mediated immune responses that are the major mechanisms of protection against malaria parasites. We have earlier studied the immunogenicity and protective efficacy of the soluble and refolded forms of recombinant 42kDa fragment of Plasmodium vivax merozoite surface protein-1 (PvMSP-142) using P. cynomolgi rhesus monkey model. In the present study, we have constructed a recombinant DNA vaccine encoding 42kDa fragment of P. vivax MSP-1 and studied the immunogenicity of PvMSP-142 DNA vaccine construct in mice. The 42kDa gene fragment of PvMSP-1 was PCR amplified using gene specific primers and subcloned into pcDNA 3.1 (+) eukaryotic expression vector. In vitro expression of PvMSP-142 plasmid construct was checked by transfection in COS-1 cell line. Indirect immunofluorescence of transfected COS-1 cells probed with monoclonal antibodies against PvMSP-142 exhibited positive fluorescence. Immunization of BALB/c mice with PvMSP-142-pcDNA vaccine construct revealed the immunogenicity of recombinant vaccine plasmid that can be enhanced by prime boosting with recombinant protein corresponding to the DNA vaccine as evidenced by significant elevation of antibody and the cytokines responses.

Isolation and characterization of endochitinase and exochitinase of Setaria cervi.

Chitin metabolism has been shown to have a role in the development of parasitic nematodes including filarial parasites and the enzymes associated with chitin metabolism have been considered as potential vaccine and drug target. Chitinases are members of the enzyme superfamily of glycoside hydrolases, which are characterized by the ability to hydrolyze glycosidic bonds in chitin chain by either an endolytic or an exolytic mechanism. In the present study, we have demonstrated the chitinase (exochitinase and endochitinase) activity in different stages of Setaria cervi (bovine filarial parasite) and have also purified and characterized the endochitinase from microfilarial stage of the parasite. The chitinase activity has been detected in adult and microfilarial stages of S. cervi using the fluorescent substrates. The S. cervi adult stage was found to have high activity of exochitinase (28.72±0.25 nmol/min/mg) while microfilarial stage showed high activity of endochitinase (24.40±0.25 nmol/min/mg). Native polyacrylamide gel electrophoresis, followed by staining of enzyme activity with fluorescent substrates, revealed single isoenzymic form of exochitinase in adults and endochitinase in microfilariae of S. cervi. The endochitinase from S. cervi microfilariae was purified employing chitin affinity matrix and DEAE-Sephacel ion-exchange chromatography. The enzyme was purified about 55 fold with an enzyme recovery of 22.33%. The purified enzyme exhibited a doublet of protein bands on SDS-PAGE at 65-70 kDa. The closantel (chitinase inhibitor) strongly inhibited the enzyme activity of S. cervi microfilariae endochitinase with a Ki value of 4.3±0.18 µM.

Cloning, overexpression and characterization of soluble 42kDa fragment of merozoite surface protein-1 of Plasmodium vivax.

Plasmodium vivax represents the second most prevalent malaria species of major public health importance and the global eradication of malaria requires the development of vaccines to prevent infection. The lack of in vitro culture and a suitable animal model for P. vivax malaria are the major problems for the delay in developing a functional vivax vaccine. A number of antigens have been identified for P. vivax as potential malaria vaccine candidates and among these 42kDa fragment of merozoite surface protein-1 (MSP-142) is one of most promising antigen of asexual blood stage. In most of the earlier studies, the MSP-142 of malaria parasites was expressed as insoluble protein in inclusion bodies and it is difficult to get purified protein in conformation form. In the present study, we have cloned, overexpressed and characterized the 42kDa fragment of P. vivax MSP-1 as soluble protein in Escherichiacoli. The 42kDa gene fragment of P. vivax MSP-1 was PCR amplified using specific primers, sequenced and subcloned into pTriEx-4 expression vector. The optimum expression of recombinant P. vivax protein was obtained in SOC growth medium by inducing with 0.2mM IPTG at 37°C for 4h. The SDS-PAGE analysis showed a fusion protein of 55kDa and about 80% was present in soluble form. The purified P. vivax MSP-142 was characterized and found to be correctly folded and in conformation form as evident by CD spectroscopy, presence of 1 free -SH group and the reactivity with reduction sensitive conformational monoclonals against P. vivax MSP-142.

Production and characterization of monoclonal antibodies against substrate specific loop region of Plasmodium falciparum lactate dehydrogenase.

Plasmodial lactate dehydrogenase, terminal enzyme of the glycolytic pathway, has been shown to be biochemically, immunologically and structurally different from the mammalian enzyme. The substrate specific loop region of plasmodial lactate dehydrogenase (pLDH) has 5 amino acids insert (DKEWN) important for anti-malarial drug targeting. In the present study, we have produced six monoclonal antibodies, which are against three different epitopes of Plasmodium falciparum LDH (PfLDH). Two of these monoclonal antibodies (10C4D5 and 10D3G2) are against the substrate specific loop region of PfLDH (residues 98-109, AGFTKAPGKSDKEWNR). The 10C4D5 and 10D3G2 monoclonals bind to substrate specific loop region resulting in inhibition of PfLDH activity. A Microplate Sandwich ELISA was developed employing high affinity non-inhibitory (10A5H5, Kaff 1.272 ± 0.057 nM) and inhibitory (10C4D5, Kaff 0.306 ± 0.011 nM) monoclonal antibodies and evaluated using gossypol, a well known inhibitor of pLDH. The binding of gossypol to substrate specific loop region resulted in inhibition of binding of 10C4D5 monoclonal. This Microplate Sandwich ELISA can be utilized for identification of compounds inhibitory to PfLDH (binding to substrate specific loop region of parasite LDH) from combinatory chemical libraries or medicinal plants extracts. The Microplate Sandwich ELISA has also shown potential for specific diagnosis of malaria using finger prick blood samples.

Chronic inhibition of central Angiotensin-converting enzyme ameliorates colchicine-induced memory impairment in mice.

Preclinical and clinical studies indicated involvement of the central renin-angiotensin system (RAS) in memory functions. However, the role of central angiotensin-converting enzyme (ACE) in memory function is still unclear. The present study investigated the involvement of central ACE in colchicine-induced memory impairment in the context of cholinergic function and oxidative stress. Memory impairment was induced by intracerebral colchicine administration in mice. The ACE inhibitor, perindopril (0.05 and 0.1 mg/kg/day), was administered orally for 14 days. Memory function was evaluated by the Morris water maze (MWM) test from the 14(th) day on after colchicine injection. Donepezil was used as a standard. Parameters of oxidative stress and cholinergic function, ACE activity in serum and the brain were estimated after the completion of behavioral studies. Colchicine caused memory impairment as revealed by no significant change in latency to reach a hidden platform in the MWM test. Furthermore, there was a significant increase in MDA, ROS, and nitrite levels with a reduction in GSH level and acetylcholinesterase (AChE) activity in the brain of colchicine-treated mice. Colchicine significantly increased brain ACE activity without affecting serum ACE. Donepezil prevented colchicine-induced memory impairment in mice. The antidementic effect of perindopril may be attributed to reduced oxidative stress and improvement in cholinergic function. Moreover, the elevated brain ACE activity was also inhibited by perindopril. The study showed that central ACE plays an important role in colchicine-induced memory deficit, corroborating a number of studies that show that treatment with ACE inhibitors could be neuroprotective.

Cloning, overexpression, purification and characterization of Plasmodium knowlesi lactate dehydrogenase.

Plasmodial lactate dehydrogenase, key enzyme of anaerobic glycolysis, has been shown to be a potential immunodiagnostic marker as well as a novel target for chemotherapy. We have cloned, overexpressed and immunochemically characterized the recombinant lactate dehydrogenase of Plasmodium knowlesi, the fifth human malaria parasite. The P. knowlesi lactate dehydrogenase (PkLDH) gene was PCR amplified and 0.9 kb PCR product was cloned into pGEM-T Easy vector. Sequencing and BLAST analysis revealed open reading frame of 316 amino acids of PkLDH showing 96.8% homology with Plasmodium vivax LDH and around 90% with Plasmodium falciparum, Plasmodium malariae and Plasmodium ovale LDHs. The PkLDH gene was subcloned into pGEX-6P1 expression vector and the SDS-PAGE analysis revealed that about 70% of fusion protein was present in the soluble fraction. The fusion protein was cleaved with PreScission protease and recombinant PkLDH (34 kDa) was affinity purified to homogeneity. The purified PkLDH exhibited high reactivity with polyclonal and monoclonal antibodies against plasmodial LDH. The polyclonal antibody produced against purified recombinant PkLDH in rabbits showed high ELISA reactivity with both native and recombinant PkLDH and could detect parasite LDH in malaria infected blood samples by sandwich ELISA. The purified recombinant PkLDH can be used to produce P. knowlesi specific monoclonal antibodies for specific diagnosis of P. knowlesi infection in humans.

Isolation of an antigen fraction from Setaria cervi adults having potential for immunodiagnosis of human filariasis.

Crude antigenic preparations from heterologous filarial parasites gave false positive results because of complex nature of these antigens and their cross-reactivity with other helminth parasites. In the present study, efforts have been made to isolate and characterize the antigens from Setaria cervi important for diagnostic purposes. The fractionation of S. cervi somatic antigenic preparation on Sephacryl S-200 resulted in separation of three major antigenic peak fractions. Crossed immunoelectrophoretic analysis, using immune rabbit serum, revealed 13-14 antigens in SFP-I pool fraction, which showed high reactivity with filarial patients sera as compared to other two pool fractions. This SFP-I fraction was further purified by DEAE-Cellulose column chromatography. Out of the 4 antigen pool fractions, DFP-IV fraction showed high ELISA reactivity with filarial patient serum pool (Wuchereria bancrofti and Brugia malayi) as compared to other fractions. The SDS-PAGE analysis of DFP-IV fraction revealed 2 major and 1 minor protein bands (mol. wt. range 65-70 kDa). Crossed immunoelectrophoresis also showed the presence of 3 antigenic peaks in DFP-IV fraction. The purified DFP-IV fraction showed high reactivity with filarial patients sera but did not cross-react with sera from ascaris and hookworm infections thereby suggesting the filaria-specificity and potential for immunodiagnosis of human filariasis.