Expression and activity of key lipases during the larval development of walking catfish (Clarias magur).

The study was conducted to investigate the expression and activity of key lipolytic enzymes during the ontogenetic development of Clarias magur. After partial characterization, the messenger RNA (mRNA) expression analysis of lipoprotein lipase (LPL), pancreatic triacylglycerol lipase (PL), and bile salt-activated lipase (BAL) genes along with the specific lipase activity were performed in larvae from Day 1 after hatching till 34-day posthatch (dph). Heterogeneous patterns of mRNA expression were shown by the important lipolytic enzymes and were detected before first exogenous feeding during the yolk-sac stage. LPL started increasing from 13 dph and peaked at 16 dph followed by a declining trend till 34 dph. However, the PL observed to be peaking at 9, 22, and 30 dph. Similarly, BAL showed an increasing trend from 11 to 22 dph with a significantly high level of mRNA expression at 16 dph. Later, the specific lipase activity was evaluated which appears at Day 1 after hatching with a progressive increase from 7 to 16 dph and a further declining trend afterwards with a peak at 22 dph. The results indicated the development of exocrine pancreas at 16 dph. Furthermore, the transcript levels and the activity of lipases were regulated with the age. Hence, the present study can be helpful in devising different strategies containing optimum lipid levels at a suitable stage of development for improving the survival during larval rearing. Furthermore, the study could be a baseline for elucidating the optimized dietary lipid levels of this catfish during its larval rearing.

Effects of dietary synbiotic on innate immunity, antioxidant activity and disease resistance of Cirrhinus mrigala juveniles.

The dietary supplementation of synbiotic in Cirrhinus mrigala juvenile (with initial body weight ranging from 2.87 ±â€¯0.01 g to 3.26 ±â€¯0.05 g) was evaluated in terms of changes in innate immunity, antioxidant activity and disease resistance against Aeromonas hydrophilla infection. One hundred eighty acclimatized juveniles of mrigal were randomly distributed in the three replicates of each of four experimental groups i.e. control (without Probiotic and Prebiotic), T1 (High Probiotic + Low Prebiotic), T2 (Low Probiotic + High Prebiotic) and T3 (High Probiotic + High Prebiotic), using completely randomized design (CRD). At the end of the feeding trial for 60 days, fish were challenged by Aeromonas hydrophila and survival rate was recorded for the next 15 days. Bacillus subtilis used as a probiotic source and MOS used as a prebiotic source in the experiment. Results showed that innate immunity was comparatively improved in T3 group. Lysozyme activity and respiratory burst activity (NBT) were significantly (P < 0.05) affected in T3 group. Highest activities of antioxidant enzymes (P < 0.05) were reported in T3 group. Cumulative mortality % was found to be lower in the fish fed dietary synbiotic on T3 group after challenging with Aeromonas hydrophilla infection. The results of this study showed that under the experimental conditions, dietary supplementation of synbiotic had a synergestic effect on enhancing innate immunity and disease resistance of Cirrhinus mrigala (P < 0.05).

Haemato-biochemical, non-specific immunity, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolate.

A 60-day feeding trial was conducted to evaluate the haemato-biochemical, innate immune response, antioxidant capacity and histopathological changes in Labeo rohita fingerlings fed rubber protein isolates (RPI). One hundred and eighty fingerlings (average weight 4.45 ± 0.01 g) were distributed into five experimental groups in triplicate and fed with isonitrogenous and isocaloric diets. Soybean protein isolate (SPI) served as the reference diet (Control), and the treatment diets were formulated as RPI25, RPI50, RPI75 and RPI100 replacing 25, 50, 75 and 100% of SPI protein, respectively. The growth performance indices like final body weight (9.54-10.27 g), net weight gain (5.09-5.84 g), metabolic growth rate (4.54-5.02) and feed efficiency ratio (0.60-0.65) among the various groups were not significantly different (P > 0.05). All the haematological parameters, except red blood cells, showed no significant differences compared with the control group (P > 0.05). The immuno-biochemical parameters like albumin, globulin, total immunoglobulin, respiratory burst and lysozyme activities among the various groups did not differ significantly (P > 0.05). The stress enzyme such as superoxide dismutase (SOD), catalase (CAT) and oxidative stress marker malondialdehyde (MDA) showed no significant difference (P > 0.05). Histopathological examination of the liver revealed no marked changes. In summary, the results showed that RPI was well utilised by the fish and its inclusion did not generate any oxidative-induced stress, thus, RPI may be suggested as a potential replacement for SPI in fish diets without any detrimental effects. Hence, protein isolation offers a unique opportunity for the utilisation of rubber seed meal.

Dietary protein enhances non-specific immunity, anti-oxidative capability and resistance to Aeromonas hydrophila in Labeo rohita fingerlings pre-exposed to short feed deprivation stress.

Present experiment was conducted to study the effect of dietary protein levels on growth, immunity and anti-oxidative status of Labeo rohita fingerlings during feed deprivation followed by refeeding. Fish (5.44 ± 0.10 g) were deprived of feed for 3 weeks and then re-fed to satiation for 5 weeks with one of the diets containing 25 (25P), 30 (30P), 35 (35P) or 40 (40P) percent crude protein (CP) level. In addition to these groups, a control group (C) was also maintained by feeding to satiation level twice daily with a diet containing 30% CP throughout the experimental period. At the end of 8-weeks' trial, fish were challenged with Aeromonas hydrophila and survival was recorded for the next 7 days. Complete recovery of growth in terms of weight gain percentage was achieved in the fish fed 35 and 40% protein during refeeding. The body indices (condition factor and hepatosomatic index), haematological parameters and serum protein contents at the end of the experimental trial were not significantly different (P > 0.05) among different groups suggesting that the overall health of the fish was not compromised. However, respiratory burst activity and serum lysozyme activity were indicative of a better immune function in the higher protein fed groups (35P and 40P) than the lower protein groups (25P and 30P). Following challenge with Aeromonas hydrophila, survival rate, blood monocyte%, respiratory burst activity, serum lysozyme activity, serum protein and globulin were significantly higher (P < 0.05) in the 35P and 40P groups compared to the other groups. Further, fish fed lower dietary protein were not able to restore the activities of anti-oxidative enzymes (superoxide dismutase and catalase) in the liver. Conclusively, an improved disease resistance capability and immune status was observed in the fish fed a higher dietary protein (35-40%), even out-performing the daily-fed fish.

Prospective glioma grading using single-dose dynamic contrast-enhanced perfusion MRI.

AIM: To evaluate the sensitivity and specificity of single-dose dynamic contrast-enhanced (DCE) perfusion magnetic resonance imaging (MRI) in prospective evaluation of glioma grading and to correlate the relative cerebral blood volume (rCBV) values with mitotic and ki-67 indexes obtained at histopathology. MATERIALS AND METHODS: A total of 53 histologically proven patients with glioma were included in this study. DCE-MRI perfusion with a single dose of contrast medium was included in brain tumour protocol and prospective grading of glioma into low and high grade was done based on a previously reported rCBV cut-off value of 3. Tumours with rCBV ≥ 3 were considered to be high grade and rCBV < 3 were considered to be low grade. The sensitivity and specificity of the cut-off value were estimated. Ki-67 and mitotic indexes were also obtained on histopathological analysis along with histological grading. RESULTS: Based on pre-defined rCBV cut-off values, prospective grading of low- and high-grade glioma was achieved with a sensitivity and specificity of 97.22% and 100%, respectively. Significant correlation was found between the mitotic/ki-67 indexes and rCBV values when data for high- and low-grade tumours was combined. CONCLUSION: DCE-MRI performed with a single dose of contrast medium is as effective as a protocol with a double-dose of contrast medium for glioma grading using 3 T MRI and could be added to the routine evaluation protocol of brain tumours.

Synthetic biology and personalized medicine.

Synthetic biology, application of synthetic chemistry to biology, is a broad term that covers the engineering of biological systems with structures and functions not found in nature to process information, manipulate chemicals, produce energy, maintain cell environment and enhance human health. Synthetic biology devices contribute not only to improve our understanding of disease mechanisms, but also provide novel diagnostic tools. Methods based on synthetic biology enable the design of novel strategies for the treatment of cancer, immune diseases metabolic disorders and infectious diseases as well as the production of cheap drugs. The potential of synthetic genome, using an expanded genetic code that is designed for specific drug synthesis as well as delivery and activation of the drug in vivo by a pathological signal, was already pointed out during a lecture delivered at Kuwait University in 2005. Of two approaches to synthetic biology, top-down and bottom-up, the latter is more relevant to the development of personalized medicines as it provides more flexibility in constructing a partially synthetic cell from basic building blocks for a desired task.

Advances in use of functionalized carbon nanotubes for drug design and discovery.

INTRODUCTION: As a part of increasing interest in nanobiotechnology, nanoparticle-based drug discovery as well as development and drug delivery constitute an important area in nanomedicine, and it is also driven by search for new drugs by the pharmaceutical industry. Nanomaterials for pharmaceutical application include carbon nanotubes (CNTs). AREAS COVERED: This article describes the properties of CNTs, both single-walled CNTs (SWCNTs) and multiwalled CNTs (MWCNTs) with relevance to drug discovery and development. Pharmacokinetics of CNTs as well as CNT-based drug delivery is discussed. The article also looks at how the scope for pharmaceutical applications of CNTs is broadened by conjugation with other molecules and presents the potential therapeutic applications. Finally, the toxicology of CNTs is considered with measures under investigation for reducing it. Literature on CNTs, from the past 5 years, was reviewed and selected publications relevant to drug discovery, development, and delivery were included in the bibliography. EXPERT OPINION: Carbon nanotubes combine more properties relevant to drug development and delivery than any other nanomaterial. Although a tremendous amount of basic research has been done on CNTs during the past decade, little of this is nearing translation into human applications. No CNT-based medicine has reached clinical trials. Nevertheless, CNT conjugation with other molecules has extended the horizons for their potential therapeutic applications. The most promising of these is PEGylation, which extends the survival of CNTs in circulation. Potential future applications of CNTs include combination of diagnostics and therapeutic drug delivery as well as a component of multimodal therapies for tissue regeneration.

Water soaking and exogenous enzyme treatment of plant-based diets: effect on growth performance, whole-body composition, and digestive enzyme activities of rohu, Labeo rohita (Hamilton), fingerlings.

A 2 × 2 × 2 factorial experiment was conducted to delineate the main effect of water soaking of plant ingredients, phytase, cellulase, and their interactions on the growth and digestive enzyme activities of Labeo rohita fingerlings. Two basal diets were prepared using water-soaked (S) or unsoaked (US) plant-based ingredients. Feed of US ingredients was supplemented with phytase (U kg(-1)) and cellulase (%) at the level of 0, 0 (C(us)); 500, 0 (T(1)); 0, 0.2 (T(2)); 500, 0.2 (T(3)), and feed of S ingredients at 0, 0 (C(s)); 500, 0 (T(4)); 0, 0.2 (T(5)), and 500, 0.2 (T(6)), respectively. Three hundred and sixty fingerlings were randomly distributed into eight treatments, each with three replicates. Soaking of the ingredients for 24 h significantly reduced the tannin content. However, feeding of S diets did not improve the fish growth. Highest performance was recorded in the T(3) group. A significant interaction between dietary phytase and cellulase was observed for apparent net protein utilization. Tissue crude protein, ether extract, and ash content of the fingerlings were observed highest in the T(3) group. Activities of amylase, protease, and lipase were recorded highest in the T(3) group. Results suggested that soaking of plant-based ingredients reduces tannin content; however, growth and digestive enzyme activities of group fed soaked diet were not improved, possibly due to leaching of soluble nutrients. Probably, a shorter duration soaking may be effective in reducing tannin content and avoiding nutrients leaching.

Role of biological therapies in the development of personalized medicine.

Development of personalized medicine involves integration of several biotechnologies. This editorial stresses the important role that biological therapies such as cell and gene therapies, recombinant proteins and vaccines play in personalization of treatment. Cell-based therapies, particularly vaccines made from the patient's own tumor cells, were the first therapeutic vaccines for cancer. Adoptive cell therapy is an immunotherapy based on ex vivo expansion of autologous T lymphocytes and subsequent administration to cancer patients. Stem cells as well as genetic modification of cells has been used for in vivo production of therapeutic substances best suited for individual patients. Besides cell therapy, RNAi has been used for personalized therapy of cancer. Monoclonal antibodies, designed to bind specifically to receptors in certain tumors, are also personalized medicines.

Effect of alternate day feeding strategy of sub-optimal protein level on haemato-biochemical responses in Labeo rohita (Hamilton) juveniles.

A feeding trial was conducted for 60 days to delineate the effect of alternate day feeding strategy of sub-optimal protein level on haematological parameters, serum parameters and phagocyte respiratory burst activity (NBT) in Labeo rohita juveniles. One hundred and thirty-five fingerlings (1.87 ± 0.01-2.26 ± 0.05 g) were distributed in triplicate groups of each treatment, and fish were fed at 5% body weight daily. Three experimental isocaloric (401.32-410.28 kcal/100 g) diets of 30%, 25%, and 20% crude protein designated as diet A, diet B, and diet C respectively, were prepared, using locally available feed ingredients. Three different feeding schedules of normal protein diet continuously (diet A-30%), alternate feeding of 1-day diet A followed by 1-day diet B (1A/1B) and alternate feeding of 1-day diet A followed by 1-day diet C (1A/1C) were tested. The total erythrocyte count and haemoglobin content was significantly (p < 0.05) enhanced in the group T1 fed (1A/1B), and the lowest count was recorded in the group T2 fed (1A/1C). Total leucocyte counts, total serum protein, and serum globulin were higher in the group T1 fed (1A/1B) and lower in the group T2 fed (1A/1C) as compare to control. The respiratory burst activity (NBT) of blood phagocytes and serum A-to-G ratio was recorded significantly difference in among the treatment groups. Based on the results of the present study, it is concluded that alternate feeding of 1-day diet A followed by 1-day diet B (1A/1B) is equally effective and promote the immunity in Labeo rohita juveniles.

Nanobiotechnology and personalized medicine.

This chapter will start with a definition and scope of personalized medicine and describe how various nanobiotechnologies will contribute to its development. Nanodiagnostics and its combination with therapeutics as well as nanoparticle-based drug delivery will play an important role. The most important applications of nanobiotechnology will be personalized management of cancer, neurological disorders, and cardiovascular diseases.

Advances in the field of nanooncology.

Nanooncology, the application of nanobiotechnology to the management of cancer, is currently the most important chapter of nanomedicine. Nanobiotechnology has refined and extended the limits of molecular diagnosis of cancer, for example, through the use of gold nanoparticles and quantum dots. Nanobiotechnology has also improved the discovery of cancer biomarkers, one such example being the sensitive detection of multiple protein biomarkers by nanobiosensors. Magnetic nanoparticles can capture circulating tumor cells in the bloodstream followed by rapid photoacoustic detection. Nanoparticles enable targeted drug delivery in cancer that increases efficacy and decreases adverse effects through reducing the dosage of anticancer drugs administered. Nanoparticulate anticancer drugs can cross some of the biological barriers and achieve therapeutic concentrations in tumor and spare the surrounding normal tissues from toxic effects. Nanoparticle constructs facilitate the delivery of various forms of energy for noninvasive thermal destruction of surgically inaccessible malignant tumors. Nanoparticle-based optical imaging of tumors as well as contrast agents to enhance detection of tumors by magnetic resonance imaging can be combined with delivery of therapeutic agents for cancer. Monoclonal antibody nanoparticle complexes are under investigation for diagnosis as well as targeted delivery of cancer therapy. Nanoparticle-based chemotherapeutic agents are already on the market, and several are in clinical trials. Personalization of cancer therapies is based on a better understanding of the disease at the molecular level, which is facilitated by nanobiotechnology. Nanobiotechnology will facilitate the combination of diagnostics with therapeutics, which is an important feature of a personalized medicine approach to cancer.

Cell therapy for CNS trauma.

Cell therapy plays an important role in multidisciplinary management of the two major forms of central nervous system (CNS) injury, traumatic brain injury and spinal cord injury, which are caused by external physical trauma. Cell therapy for CNS disorders involves the use of cells of neural or non-neural origin to replace, repair, or enhance the function of the damaged nervous system and is usually achieved by transplantation of the cells, which are isolated and may be modified, e.g., by genetic engineering, when it may be referred to as gene therapy. Because the adult brain cells have a limited capacity to migrate to and regenerate at sites of injury, the use of embryonic stem cells that can be differentiated into various cell types as well as the use of neural stem cells has been explored. Preclinical studies and clinical trials are reviewed. Advantages as well as limitations are discussed. Cell therapy is promising for the treatment of CNS injury because it targets multiple mechanisms in a sustained manner. It can provide repair and regeneration of damaged tissues as well as prolonged release of neuroprotective and other therapeutic substances.

Characterization of tumefactive demyelinating lesions using MR imaging and in-vivo proton MR spectroscopy.

BACKGROUND AND OBJECTIVES: Diagnosis of tumefactive demyelinating lesions (TDLs) is challenging to both clinicians and radiologists. Our objective in this study was to analyze and characterize these lesions clinically, biochemically, electrophysiologically, and on imaging. METHODS: A retrospective analysis with prospective follow-up of 18 cases of TDLs was performed. Imaging included T2-, T1-weighted, fluid-attenuated inversion recovery (FLAIR), post-contrast T1-weighted, diffusion weighted imaging (DWI), and proton magnetic resonance spectroscopy (PMRS). RESULTS: All the lesions appeared hyperintense on T2 and FLAIR images. Increased Apparent diffusion coefficient (ADC) (0.93-2.21 x 10(-3) mm(2)/s) in centre of the lesion was seen in 14/18 cases; however, peripheral restriction (ADC values 0.55-0.64 x 10(-3) mm(2)/s) was noted in 11/18 cases. In all, 13/18 cases showed contrast enhancement with open ring (n = 5), complete ring (n = 1), minimal (n = 4), and infiltrative (n = 3) pattern of enhancement. Nine of these 13 cases also showed venular enhancement. On PMRS, nine showed glutamate/glutamine (Glx) at 2.4 ppm. CONCLUSION: Clinical features along with several MRI characteristics such as open ring enhancement, peripheral restriction on DWI, venular enhancement, and presence of Glx on spectroscopy may be rewarding in differentiating TDLs from neoplastic lesions.

Cell therapy for pain.

BACKGROUND: Management of chronic pain remains a challenge in spite of numerous drugs that are either approved or still in development. Apart from inadequate relief, there are concerns about adverse effects and addiction. Cell therapy is being explored for relief of pain. OBJECTIVE: To address the rationale for cell therapy for treatment of pain and its advantages over conventional pharmaceuticals. The prospects of translation of these techniques from experimental animals to clinical use are discussed. METHODS: This review is based on the literature on cell therapy in relation to pain and is confined to experimental work as there are no approved therapies in this category. RESULTS/CONCLUSIONS: A number of promising cell therapy technologies have been identified. These provide targeted approaches to delivery of antinociceptive molecules, avoiding subjecting the patient to systemic toxicity of drugs. There has been considerable progress in treating degenerative joint diseases causing pain. Management of neuropathic pain is a challenge and a number of ongoing studies are addressing it. Overall the future of cell therapy for pain is promising.

Gene therapy for pain.

BACKGROUND: Management of chronic pain remains a challenge in spite of the numerous drugs either approved or still in development. Apart from inadequacy of relief, there are concerns about adverse effects and addiction in the case of drugs such as opioids. Gene therapy is being investigated for improving management of pain. OBJECTIVE: To addresses the rationale of gene therapy for treatment of pain and its advantages over drugs. The prospects of translation of these techniques from experimental animals to clinical use are discussed. METHODS: The review is based on the available literature and is confined to experimental work, as there are no approved therapies in this category. RESULTS/CONCLUSION: A number of promising gene therapies as well as antisense- and RNA interference-based approaches have been identified. These provide targeted approaches to delivery of antinociceptive molecules or interruption of pain pathways without subjecting the patient to systemic toxicity of drugs. Some of these approaches are aimed at correcting the underlying pathology of the diseases (e.g., treating degenerative joint diseases causing pain). Management of neuropathic pain is a challenge and a number of studies are addressing it. Overall the future of gene therapy for pain is promising.

Neuroprotection in traumatic brain injury.

The management of traumatic brain injury (TBI) is challenging and there is a need for neuroprotective therapies. A better understanding of the pathomechanism of TBI, particularly of the evolution of secondary damage, is providing targets for new approaches and selected ones in clinical development are described. Clinical trials that have been discontinued in the past for lack of efficacy or other reasons are also listed. One of the problems has been the translation of promising animal experimental results into clinically successful therapies. The complexity of sequelae of TBI requires a multifaceted approach. In addition to the investigation of drugs for neuroprotective effect in TBI, new technologies based on cell/gene therapies, biomarkers and nanobiotechnology are being employed for the integration of neuroprotection with neuroregeneration and are promising.