Microscopic and molecular investigation of vector borne haemoprotozoan diseases in dromedary camel of North Gujarat, India.

BACKGROUND OBJECTIVES: Vector-borne haemoprotozoan diseases comprise diverse group of single celled organism transmitted by haematophagus invertebrates. The current study was aimed at the identification of major haemoprotozoan (Babesia, Theileria and Trypanosoma) in dromedary camel of North Gujarat region in India using microscopy and Polymerase Chain Reaction (PCR). METHODS: A total of 234 blood samples were screened by the microscopic and molecular detection assays. Molecular prevalence studies of Theileria, Trypanosoma spp and Babesia was undertaken using 18s ribosomal DNA, RoTat 1.2 and SS rRNA gene respectively. The data relating to microscopic and molecular prevalence along with associated risk factors were analysed by statistical methods. RESULTS: The overall prevalence of hamoprotozoan disease based on microscopic and molecular investigation was 23.50%. The sensitivity and specificity (95% Confidence Interval) of PCR assay was 100% in comparison to microscopy (45.45 % sensitive and 100 % specific). The kappa coefficient between PCR and microscopy indicated good level of agreement with a value of 0.704 and SE of 0.159. INTERPRETATION CONCLUSION: Despite holding much significance to the animal sector, little work has been undertaken in regional parts of India regarding camel parasites. The present study offers first preliminary research data investigating haemoprotozoan disease using parasitological and molecular methods in camels in the region.

Microscopic and molecular investigation of vector borne haemoprotozoan diseases in dromedary camel of North Gujarat.

BACKGROUND OBJECTIVES: Vector borne haemoprotozoan diseases comprise diverse group of single celled organism transmitted by haematophagus invertebrates. The current study was aimed at identification of major haemoprotozoan (Babesia, Theileria and Trypanosoma) in dromedary camel of North Gujarat region using microscopy and Polymerase Chain Reaction (PCR). METHODS: A total of 234 blood samples were screened by the microscopic and molecular detection assays. Molecular prevalence studies of Theileria, Trypanosoma spp and Babesia was undertaken using 18s ribosomal DNA, RoTat 1.2 and SS rRNA gene respectively. The data relating to microscopic and molecular prevalence along with associated risk factors were analysed by statistical methods. RESULTS: The overall prevalence of hamoprotozoan disease based on microscopic and molecular investigation was 23.50%. The sensitivity and specificity (95% Confidence Interval) of PCR assay was 100% in comparison to microscopy (45.45% and 100%). The kappa coefficient between PCR and microscopy indicated good level of agreement with a value of 0.704 and SE of 0.159. INTERPRETATION CONCLUSION: Despite holding much significance to the animal sector, little work has been undertaken in regional part of India regarding camel parasites. The present paper offers the first preliminary research data investigating haemoprotozoan disease using parasitological and molecular methods in camels in the region.

Hemato-biochemical and ultrasonographic evaluation of hepatic lipidosis in dairy buffaloes.

Around 60% dairy animals developed moderate to severe hepatic lipidosis at the time of parturition or during early lactation stage. Most of clinician suspect the hepatic lipidosis during above time window only. However, negative energy balance or feeding of high concentrate diet can lead to hepatic lipidosis at any phase of life. The aim of the present study was to evaluate the potential for diagnosis of hepatic lipidosis by means of hemato-biochemical parameters and ultrasonography of the liver at any stage of life. Here, ultrasonographic back fat thickness measurement was correlated with ultrasonographic features of hepatic lipidosis. A total 60 buffaloes were included under the study and sampled for hematological and biochemical parameters. Hematological parameters did not exhibit any significant difference between healthy and hepatic lipidosis-affected buffaloes. Biochemical parameters like beta hydroxy butyric acid, non esterified fatty acid, aspartate amino transferase, gamma glutamyl transferase and alkaline phosphatase revealed a significant increase, while triglyceride, cholesterol, and glucose declined significantly in hepatic lipidosis-affected buffaloes. Total protein, albumin, and total bilirubin levels did not exhibit any significant difference. Based on ultrasonographic findings, the hepatic lipidosis-affected buffaloes were further sub divided into mild, moderate, and severe groups. Portal vein diameter and depth of portal vein were also estimated in current study. Ultrasonographic examination could diagnose 53.33% hepatic lipidosis cases in buffaloes. Among it, 37.50% buffalo had mild hepatic lipidosis, 33.33% had moderate hepatic lipidosis, and 29.16% had severe hepatic lipidosis. Depth of portal vein significantly increased in hepatic lipidosis cases. However, portal vein diameter exhibited a non-significant difference in mild, moderate, and severe groups of hepatic lipidosis. Back fat thickness also revealed a non-significant difference in mild, moderate, and severe hepatic lipidosis. Above study indicate that B mode ultrasonography of the liver can be employed to differentiate various grades of hepatic lipidosis in buffaloes. Biochemical parameters like NEFA, BHBA, AST, GGT, ALP, TG, cholesterol, and glucose can be helpful to screen the hepatic lipidosis at farm level.

Draft genome sequence of a thermostable, alkaliphilic α-amylase and protease producing Bacillus amyloliquefaciens strain KCP2.

Bacillus amyloliquefaciens strain KCP2 was isolated from municipal food waste samples collected in Vallabh Vidyanagar, Gujarat, India. Strain KCP2 is noteworthy due to its ability to produce a thermostable, alkaliphilic α-amylase and a protease. These enzymes have importance in several industrial processes including bread making, brewing, starch processing, pharmacy, and textile industries. Whole genome sequencing of strain KCP2 showed that the estimated genome size was 3.9 Mb, the G + C content was 46%, and it coded for 4113 genes.

Comparative efficacy of anthelmintics and their effects on hemato-biochemical changes in fasciolosis of goats of South Gujarat.

AIM: Fasciolosis is a parasitic disease caused by Fasciola spp. of the family Fasciolidae (trematodes) characterized by bottle jaw, anemia, progressive debility, and potbelly condition. There are many aspects of fasciolosis remaining unknown thus hemato-biochemical alterations in closantel, triclabendazole + ivermectin, and oxyclozanide + levamisole treated goats were studied. MATERIALS AND METHODS: A total of 40 naturally fasciolosis infected goats having egg per gram more than 100 were randomly divided into four groups. Goats of Group I-III were treated with three different anthelmintics, whereas, goats of Group-IV were kept as control or untreated. Whole blood, serum, and fecal samples were collected on 0, 7(th), and 30(th) day of treatment. RESULTS: During the study, values of hemoglobin, total erythrocyte count, pack cell volume, and total protein were significantly elevated to their normal levels in anthelmintics treated groups. Whereas, values of total leukocyte count, aspartate transaminase (AST), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) were significantly reduced to their normal level in anthelmintics treated groups. The efficacy of closantel (T1), triclabendazole + ivermectin (T2), and oxyclozanide + levamisole (T3) was 99.63%, 100%, and 94.74% and 100%, 100%, and 97.38% on 7(th) and 30(th) day of treatment, respectively. CONCLUSIONS: Fasciolosis in goats can be diagnosed on the basis of fecal sample examination, but alterations in important biomarkers such as AST, GGT, and LDH are also helpful for early diagnosis. The use of newer anthelmintic either alone or in combination showed a higher therapeutic response in fasciolosis of goats.

Cortical Activation Through Passive-Motion Functional MRI.

BACKGROUND AND PURPOSE: Functional brain mapping is an important technique for neurosurgical planning, particularly for patients with tumors or epilepsy; however, mapping has traditionally involved invasive techniques. Existing noninvasive techniques require patient compliance and may not be suitable for young children. We performed a retrospective review of our experience with passive-motion functional MR imaging in anesthetized patients to determine the diagnostic yield of this technique. MATERIALS AND METHODS: A retrospective review of patients undergoing passive-motion fMRI under general anesthesia at a single institution over a 2.5-year period was performed. Clinical records were evaluated to determine the indication for fMRI, the ability to detect cortical activation, and, if present, the location of cortical activation. RESULTS: We identified 62 studies in 56 patients in this time period. The most common indication for fMRI was epilepsy/seizures. Passive-motion fMRI identified upper-extremity cortical activation in 105 of 119 (88%) limbs evaluated, of which 90 (86%) activations were in an orthotopic location. Lower-extremity cortical activation was identified in 86 of 118 (73%) limbs evaluated, of which 73 (85%) activations were in an orthotopic location. CONCLUSIONS: Passive-motion fMRI was successful in identifying cortical activation in most of the patients. This tool can be implemented easily and can aid in surgical planning for children with tumors or candidates for epilepsy surgery, particularly those who may be too young to comply with existing noninvasive functional measures.

Vascular neoplasm or pseudovascular nevus? Potential pitfalls in diagnosis.

Melanocytic nevi, on histopathologic evaluation, occasionally contain slit-like clefts or spaces that may resemble vascular or lymphatic spaces. The spaces may contain blood or, perhaps more concerning, nests of melanocytes that could suggest lymphatic invasion of melanoma. When lined by melanocytes rather than true endothelium, these pseudovascular spaces within melanocytic nevi are generally attributable to tissue processing artifact. When the space in question is pronounced, a proper diagnostic work-up is prudent in order to exclude a true vascular neoplasm or melanoma. In this case series we present several melanocytic lesions with prominent vascular-appearing spaces that warranted further investigation.

Study of 35 Cases of Hansen's Disease, which Required Treatment beyond Fixed Duration--Multi Drug Therapy.

Multi Drug Therapy (MDT) is the main weapon against leprosy since its inception in 1981. India achieved the level of elimination (< 1 case/10,000) on 31st December 2005. It has been proved in few studies that despite 2 years of regular therapy 10% of the patients continue to harbour viable persisters. There are many problems related with FD-MDT. Many cases have residual disease activity after completion of treatment. Aims of the present study was to study the profile of RFT cases in leprosy treated with FD-MDT, who required extended MDT, duration between completion of FD -MDT and clinical presentation, Acid Fast Bacilli (AFB) status, histopathology and type of leprosy at the time of presentation. A prospective study of 35 RFT (Released FromTreatment) cases with signs of activity were recruited in period betveen May 2007 to November 2001. All cases were diagnosed clinically and investigations were done for AFB smear, histopathological examination and Fite Faraco staining. We found that all the 35 cases, which required extended MDT, age group ranged from 10 to 65 years. Majority (71.4%) had taken previous Multi-Bacillary (MB) treatment for 1 year duration. Eleven (31.42%) of cases came within one year, 17 (48.57%) between one to two years and 7 (20%) cases after two years of stopping FD-MDT. AFB smear was positive in 36.84% of cases in which done. Majority of previously diagnosed MB cases presented as BT/TT in histopathology. Thus there is need to search for reliable prognostic markers for therapeutic purposes.

A cross-sectional multicenter study of osteogenesis imperfecta in North America - results from the linked clinical research centers.

Osteogenesis imperfecta (OI) is the most common skeletal dysplasia that predisposes to recurrent fractures and bone deformities. In spite of significant advances in understanding the genetic basis of OI, there have been no large-scale natural history studies. To better understand the natural history and improve the care of patients, a network of Linked Clinical Research Centers (LCRC) was established. Subjects with OI were enrolled in a longitudinal study, and in this report, we present cross-sectional data on the largest cohort of OI subjects (n = 544). OI type III subjects had higher prevalence of dentinogenesis imperfecta, severe scoliosis, and long bone deformities as compared to those with OI types I and IV. Whereas the mean lumbar spine area bone mineral density (LS aBMD) was low across all OI subtypes, those with more severe forms had lower bone mass. Molecular testing may help predict the subtype in type I collagen-related OI. Analysis of such well-collected and unbiased data in OI can not only help answering questions that are relevant to patient care but also foster hypothesis-driven research, especially in the context of 'phenotypic expansion' driven by next-generation sequencing.

Early feeding factors associated with exclusive versus partial human milk feeding in neonates receiving intensive care.

OBJECTIVE: To evaluate early feeding factors associated with exclusive human milk (EHM) feeding at discharge in a cohort of human milk-fed infants admitted to the neonatal intensive care unit (NICU). STUDY DESIGN: Retrospective cohort of consecutively discharged infants from two NICUs over a 12-month period who received any human milk during the 24 h before hospital discharge. We used logistic regression to evaluate early feeding factors associated with EHM feeding at discharge. RESULT: We evaluated a total of 264 infants. EHM-fed infants were twice as likely to receive human milk at the first feeding compared with partial human milk-fed infants (65% vs 32%; P<0.01). In multivariable analysis, including adjustment for race and type of maternal insurance, infants receiving human milk as the initial feeding, compared with formula, had a greater odds of EHM feeding at hospital discharge (adjusted odds ratio (OR)=3.41; 95% confidence interval (CI)=1.82 to 6.39; P<0.001). CONCLUSION: Among infants admitted to the NICU whose mothers provide human milk, those receiving human milk as the first feeding were more likely to receive EHM feeding at discharge.

The short stem: promises and pitfalls.

Conventional uncemented femoral implants provide dependable long-term fixation in patients with a wide range of functional requirements. Yet challenges associated with proximal-distal femoral dimensional mismatch, preservation of bone stock, and minimally invasive approaches have led to exploration into alternative implant designs. Short stem designs focusing on a stable metaphyseal fit have emerged to address these issues in total hip replacement (THR). Uncemented metaphyseal-engaging short stem implants are stable and are associated with proximal bone remodeling closer to the metaphysis when compared with conventional stems and they also have comparable clinical performances. Short stem metaphyseal-engaging implants can meet the goals of a successful THR, including tolerating a high level of patient function, as well as durable fixation.

Pain, motor and gait assessment of murine osteoarthritis in a cruciate ligament transection model.

OBJECTIVE: The major complaint of Osteoarthritis (OA) patients is pain. However, due to the nature of clinical studies and the limitation of animal studies, few studies have linked function impairment and behavioral changes in OA animal models to cartilage loss and histopathology. Our objective was to study surrogate markers of functional impairment in relation to cartilage loss and pathological changes in a post-traumatic mouse model of OA. METHOD: We performed a battery of functional analyses in a mouse model of OA generated by cruciate ligament transection (CLT). The changes in functional analyses were linked to histological changes graded by OARSI standards, histological grading of synovitis, and volumetric changes of the articular cartilage and osteophytes quantified by phase contrast micro-computed tomography (µCT). RESULTS: OA generated by CLT led to decreased time on rotarod, delayed response on hotplate analysis, and altered gait starting from 4 weeks after surgery. Activity in open field analysis did not change at 4, 8, or 12 weeks after CLT. The magnitude of behavioral changes was directly correlated with higher OARSI histological scores of OA, synovitis in the knee joints, cartilage volume loss, and osteophyte formation. CONCLUSION: Our findings link functional analyses to histological grading, synovitis, comprehensive three-dimensional assessment of cartilage volume and osteophyte formation. This serves as a reference for a mouse model in predicting outcomes of OA treatment.

Early caffeine therapy and clinical outcomes in extremely preterm infants.

OBJECTIVE: To determine if early caffeine (EC) therapy is associated with decreased bronchopulmonary dysplasia (BPD) or death, decreased treatment of patent ductus arteriosus (PDA), or shortened duration of ventilation. STUDY DESIGN: In a retrospective cohort of 140 neonates ≤1250 g at birth, infants receiving EC (initiation <3 days of life) were compared with those receiving late caffeine (LC, initiation ≥3 days of life) using logistic regression. RESULT: Of infants receiving EC, 25% (21/83) died or developed BPD compared with 53% (30/57) of infants receiving LC (adjusted odds ratio (aOR) 0.26, 95% confidence interval (CI) 0.09 to 0.70; P<0.01). PDA required treatment in 10% of EC infants versus 36% of LC infants (aOR 0.28, 95%CI 0.10 to 0.73; P=0.01). Duration of mechanical ventilation was shorter in infants receiving EC (EC, 6 days; LC, 22 days; P<0.01). CONCLUSION: Infants receiving EC therapy had improved neonatal outcomes. Further studies are needed to determine if caffeine prophylaxis should be recommended for preterm infants.

Regeneration of stevia plant through callus culture.

Stevia rebaudiana Bertoni that conventionally propagated by seed or by cuttings or clump division which has a limitation of quality and quantity seed material. In present study, callus culture technique was tried to achieve rapid plant multiplication for quality seed material. Callus induction and multiplication medium was standardized from nodal as well as leaf sagments. It is possible to maintain callus on Murashige and Skoog medium supplemented with 6-benzyl amino purine and naphthalene acetic acid. Maximum callus induction was obtained on Murashige and Skoog medium incorporated with 6-benzyl amino purine (2.0-3.0 mg/l) and naphthalene acetic acid (2.0 mg/l) treatments. However, Murashige and Skoog medium containing 2.0 mg/l 6-benzyl amino purine+2.0 mg/l naphthalene acetic acid was found to be the best for callus induction. Higher regeneration frequency was noticed with Murashige and Skoog medium supplemented with 2.0 mg/l 6-benzyl amino purine+0.2 mg/l naphthalene acetic acid. Regenerated plants were rooted better on (1/4) Murashige and Skoog strength supplemented with 0.1 mg/l indole-3-butyric acid. The rooted plantlets were hardened successfully in tera care medium with 63 per cent survival rate. The developed protocol can be utilized for mass production of true to type planting material on large scale independent of season, i.e. external environmental conditions.

RNA-mediated gene silencing of superoxide dismutase (bcsod1) in Botrytis cinerea.

Gene silencing is a powerful tool utilized for identification of gene function and analysis in plants, animals, and fungi. Here, we report the silencing of superoxide dismutase (bcsod1) in Botrytis cinerea through sense and antisense-mediated silencing mechanisms. Because superoxide dismutase (SOD) is a virulence factor, transformants were tested for phenotypic silencing in vitro and reduction in pathogenicity in planta. Plate-based assays with and without paraquat were performed to screen initial silencing efficiency, and a subset of transformants was used for in planta studies of virulence. Transformants exhibiting strongly decreased transcripts levels were recovered with both constructs but none of those exhibited a reduction in virulence in planta. Our investigations may help optimize a high-throughput gene silencing system useful for identifying potential gene targets for future fungal control.

Biosorption of heavy metals by red algae (Palmaria palmata).

The biosorption of heavy metals from aqueous solutions was investigated, using a cheap and abundant dry biomass of red algae P. palmata. The Freundlich, Langmuir and Brunauer Emmer and Teller (BET) models were used to describe the uptake of lead (pb2+), copper (Cu2+), nickel (Ni2+), cadmium (Cd 2+) and zinc (Zn2+) on P. palmata. The good fits of the Langmuir and BET models to the experimental data reflected that the sorption on P. palmata was a multi-layer sorption, in which a Langmuir equation could be applied to each layer. The highest maximum sorption capacity q(max), derived from the Langmuir model was 15.17 mg g(-1) for lead and 6.65 mg g(-1) for copper (dry weight metal/dry weight biosorbent) at a pH of 5.5-6. The affinity of metals for P. palmata was found to decrease in the order: Pb2+ > Cd2+ > Cu2+ > Ni2+. The factors influencing copper and lead uptake were found to be contact time, pH, initial concentration and temperature. Biosorption of copper and lead was a rapid process, with 70% and 100% of the respective uptakes occurring within the first 10 minutes.

Surface-active properties of rhamnolipids from Pseudomonas aeruginosa GS3.

Pseudomonas aeruginosa GS3 produced rhamnolipid biosurfactants during growth on carbohydrates, higher chain length n-alkanes and l-alkenes, petroleum crude oil and vegetable oils. With glucose as the substrate, maximum surfactant production (0.44 g/l) was observed during the stationary phase of growth. Partially purified rhamnolipids showed excellent surface-active properties in terms of reduction in the interfacial tension between them and a variety of hydrocarbons, hydrocarbon mixtures and vegetable oils and formation of stable emulsion.

Regulatory aspects of modifications to innovator bronchodilator metered dose inhalers and development of generic substitutes.

Regulatory requirements for modifications to an approved innovator metered dose inhaler (pressurized MDI; USP nomenclature: inhalation aerosol) and for development of a new generic product are discussed. Although many of the requirements apply generally to MDI's, they are discussed with specific reference to albuterol. Changes to the container and closure system may impact on the dosimetry of the redesigned product, as well as upon toxicologic and chemistry, manufacturing and controls (CMC) concerns. Changes to the formulation, including the use of alternate propellants, may raise issues requiring both clinical and in vivo performance evaluation. In view of the level of interest of a number of firms in approval requirements for generic Albuterol Inhalation Aerosol products, the article discusses in considerable detail the CMC and bioequivalence requirements for a generic product. Similarities in the CMC requirements for innovator and generic products are evident. Three comparative in vivo bioequivalence tests, particle size distribution, spray pattern and plume geometry, and unit spray content, established by the Division of Bioequivalence are discussed. Similarities and differences in the in vivo requirements for innovator and generic products are evident. Differences are the result of U.S. statute, which requires safety and efficacy testing for a product approved under a new drug application (NDA), but documentation of bioequivalence for a product approved under an abbreviated new drug application (ANDA). The advantages and disadvantages of three pharmacodynamic study designs which have potential usefulness for documentation of in vivo bioequivalence are discussed.