Transient receptor potential (TRP) channels: a metabolic TR(i)P to obesity prevention and therapy.

Cellular transport of ions, especially by ion channels, regulates physiological function. The transient receptor potential (TRP) channels, with 30 identified so far, are cation channels with high calcium permeability. These ion channels are present in metabolically active tissues including adipose tissue, liver, gastrointestinal tract, brain (hypothalamus), pancreas and skeletal muscle, which suggests a potential role in metabolic disorders including obesity. TRP channels have potentially important roles in adipogenesis, obesity development and its prevention and therapy because of their physiological properties including calcium permeability, thermosensation and taste perception, involvement in cell metabolic signalling and hormone release. This wide range of actions means that organ-specific actions are unlikely, thus increasing the possibility of adverse effects. Delineation of responses to TRP channels has been limited by the poor selectivity of available agonists and antagonists. Food constituents that can modulate TRP channels are of interest in controlling metabolic status. TRP vanilloid 1 channels modulated by capsaicin have been the most studied, suggesting that this may be the first target for effective pharmacological modulation in obesity. This review shows that most of the TRP channels are potential targets to reduce metabolic disorders through a range of mechanisms.

Immunoprotective responses of T helper type 1 stimulatory protein-S-adenosyl-L-homocysteine hydrolase against experimental visceral leishmaniasis.

It is well known that a patient in clinical remission of visceral leishmaniasis (VL) remains immune to reinfection, which provides a rationale for the feasibility of a vaccine against this deadly disease. In earlier studies, observation of significant cellular responses in treated Leishmania patients as well as in hamsters against leishmanial antigens from different fractions led to its further proteomic characterization, wherein S-adenosyl-L-homocysteine hydrolase (AdoHcy) was identified as a helper type 1 (Th1) stimulatory protein. The present study includes immunological characterization of this protein, its cellular responses [lymphoproliferation, nitric oxide (NO) production and cytokine responses] in treated Leishmania-infected hamsters and patients as well as prophylactic efficacy against Leishmania challenge in hamsters and the immune responses generated thereof. Significantly higher cellular responses were noticed against recombinant L. donovani S-adenosyl-L-homocysteine hydrolase (rLdAdoHcy) compared to soluble L. donovani antigen in treated samples. Moreover, stimulation of peripheral blood mononuclear cells with rLdAdoHcy up-regulated the levels of interferon (IFN)-γ, interleukin (IL)-12 and down-regulated IL-10. Furthermore, vaccination with rLdAdoHcy generated perceptible delayed-type hypersensitivity response and exerted considerably good prophylactic efficacy (∼70% inhibition) against L. donovani challenge. The efficacy was confirmed by the increased expression levels of inducible NO synthase and Th1-type cytokines, IFN-γ and IL-12 and down-regulation of IL-4, IL-10 and transforming growth factor (TGF)-ß. The results indicate the potentiality of rLdAdoHcy protein as a suitable vaccine candidate against VL.

A novel cobiotic-based preventive approach against high-fat diet-induced adiposity, nonalcoholic fatty liver and gut derangement in mice.

BACKGROUND: High-fat diets (HFDs) induce systemic inflammation, gut microbial derangements and disturb metabolic homeostasis, resulting in weight gain, insulin resistance and nonalcoholic fatty liver (NAFL). Numerous antioxidants and prebiotic/probiotics per se may prevent HFD-associated comorbidities, but there are no reports related to their combination. OBJECTIVE: In the present study, we aim to evaluate a cobiotic combination of lycopene (antioxidant) and isomalto-oligosaccharides (IMOs, a prebiotic) for prevention of HFD-induced alterations. DESIGN: Male Swiss albino mice were fed either normal pellet diet (NPD) or HFD and lycopene (5 and 10 mg kg(-1)), IMOs (0.5 and 1 g kg(-1)) or their combination for 12 weeks. Systemic adiposity, glucose tolerance, insulin sensitivity, feeding regulators in hypothalamus, hepatosteatosis and liver inflammation, cecal short chain fatty acids (SCFAs), serum inflammatory cytokines, gut morphology and alterations in selected gut microbes were studied. RESULTS: Lycopene, IMOs and their combination prevented weight gain, adiposity, improved adipose tissue fat mobilization and reduced insulin resistance. Hypothalamic orexigenic and anorectic genes have also been modulated by these treatments. Dietary interventions prevented NAFL-like symptoms and improved glucose homeostasis. Improvement in selected gut microbial abundance and SCFA concentration along with reduced systemic inflammation, metabolic endotoxemia and improved ileal and colonic health were observed in mice supplemented with lycopene, IMOs and their combination. Interestingly, cobiotic combination synergistically improved many of the HFD-induced alterations. CONCLUSION: The present work provide evidence that new approach based on cobiotic combination (antioxidant plus prebiotic) can be employed to develop novel class of functional foods for their application against HFD-associated pathological complications.

Immunostimulatory potential and proteome profiling of Leishmania donovani soluble exogenous antigens.

Isolation of the soluble exogenous antigens (SEAgs), its immune response study and proteome profiling is an essential prerequisite for understanding the molecular pathogenesis of Leishmania donovani. The immunostimulatory potential of L. donovani SEAgs, purified from culture of L. donovani clinical isolate, was evaluated for their ability to induce cellular responses in treated/cured hamsters. SEAgs induced significant proliferative responses in lymphocytes (SI 5.6 ± 2.3; P < 0.01) isolated from cured hamster. In addition, significant NO production in response to SEAgs was also noticed in macrophages of hamsters, mouse and human cell lines (J774A-1 and THP1). Western blot analyses with antibodies against proteophosphoglycan (PPG; surface-expressed and secreted molecule) of L. donovani revealed that PPG molecules are also present in L. donovani SEAgs. Mass spectrometry (MS)-based proteome analysis of 12 protein bands of SEAgs through MALDI-TOF/TOF endorsed the identification of some Th1-stimulatory immunogenic proteins. These immunogenic proteins may offer increased hope for the discovery of new promising vaccine candidates against visceral leishmaniasis (VL). The overall results suggest that immunostimulatory molecules are present in the SEAgs, which may be further exploited, for developing a subunit vaccine against VL a fatal human disease.

Simulation of vertical concentration gradient of influenza viruses in dust resuspended by walking.

UNLABELLED: Particles are resuspended from the floor by walking and are subject to turbulent transport in the human aerodynamic wake. These processes may generate a vertical concentration gradient of particles. To estimate the magnitude of turbulence generated by walking, we measured the velocity field in the wake from floor to ceiling at 10-cm intervals with a sonic anemometer. The resulting eddy diffusion coefficients varied between 0.06 and 0.20 m(2) /s and were maximal at ~0.75-1 m above the floor, approximately the height of the swinging hand. We applied the eddy diffusion coefficients in an atmospheric transport model to predict concentrations of resuspended influenza virus as a function of the carrier particle size, height in the room, and relative humidity, which affects the resuspension rate coefficient and virus viability. Results indicated that the concentration of resuspended viruses at 1 m above the floor was up to 40% higher than at 2 m, depending on particle size. For exposure to total resuspended viruses, the difference at 1 vs. 2 m was 11-14%. It is possible that shorter people are exposed to higher concentrations of resuspended dust, including pathogens, although experimental evidence is needed to verify this proposition. PRACTICAL IMPLICATIONS: Forces generated by walking can resuspend particles from the floor and create higher concentrations close to the floor and lower concentrations above it. These particles may include pathogens, such as the influenza virus, that were previously emitted into the air by an infected individual and that settled to the ground. Due to particle resuspension and turbulent transport, it is possible that shorter people are exposed to higher concentrations of particles, including certain pathogens, than are taller people. This work could be used in support of epidemiological investigations into the incidence of influenza as a function of a person's height and to guide the design of more effective control strategies to reduce transmission of influenza.

Neutrophil:lymphocyte ratios and serum cytokine changes after hepatic artery chimeric antigen receptor-modified T-cell infusions for liver metastases.

Our phase I Hepatic Immunotherapy for Metastases (HITM) trial tested the safety of chimeric antigen receptor-modified T-cell (CAR-T) hepatic artery infusions (HAI) for unresectable carcinoembryonic antigen (CEA)+ liver metastases (LM). High neutrophil:lymphocyte ratios (NLR) predict poor outcome in cancer patients and we hypothesized that NLR changes would correlate with early responses to CAR-T HAI. Six patients completed the protocol. Three patients received CAR-T HAI in dose escalation (1 × 10(8), 1 × 10(9) and 1 × 10(10) cells) and the remainder received three doses (1 × 10(10) cells) with interleukin (IL)2 support. Serum cytokines and NLR were measured at multiple time points. The mean NLR for all patients was 13.9 (range 4.8-38.1). NLR increased in four patients following treatment with a mean fold change of 1.9. Serum IL6 levels and NLR fold changes demonstrated a trend towards a positive correlation (r=0.77, P=0.10). Patients with poor CEA responses were significantly more likely to have higher NLR level increases (P=0.048). Increased NLR levels were associated with poor responses following CAR-T HAI. NLR variations and associated cytokine changes may be useful surrogates of response to CAR-T HAI.

A report on pharmacognostical evaluation of four Adiantum species, Pteridophyta, for their authentication and quality control

Genus Adiantum L., Pteridaceae, is an important fern used in traditional systems of medicine. Different species of Adiantum are known as avenca in Brazil; hansraj/hanmspadi in India; maiden hair fern in English. The present study aims to develop the quality control parameters of four similar looking Adiantum species viz. A. capillus-veneris L., A. lunulatum Burm. f., A. peruvianum Klotzsch, and A. venustum D. Don. Standard methods for macro-microscopic evaluation, physico-chemical parameters and HPTLC were used for authentication and identification. The salient distinctive characters under the microscope are the presence of slightly wavy elongated epidermal cells in A. capillus-veneris; epidermal cells strongly wavy in A. lunulatum; star shaped epidermal cells and mixed spores of regular and irregular shaped in A. peruvianum; stomata on both the surfaces of pinnule, absence of spinulus spores only in A. venustum. In addition, rachis anatomy showed different cellular and stellar characteristics as identifying characters of aforesaid four Adiantum species. Physico-chemical parameters and HPTLC finger print profiles along with stigmasterol and lupeol play significant role for the quality evaluation of raw drugs. The above finding will serve the purpose of quality control and assurance for the future studies.

Comparison of two ferns (Adiantum capillus-veneris Linn. and Microsorium punctatum (Linn.) Copel) for their Cr accumulation potential and antioxidant responses.

Study was undertaken to compare Cr accumulation in two ferns (Adiantum capillus-veneris Linn. and Microsorium punctatum (Linn.) Copel) and the role of antioxidants were also investigated towards metal tolerance in order to assess the use of ferns in phytomediation/ phytostabilization. Different concentrations (0, 50, 100, 150 microg g(-1) dw) of Cr were added to fern planted in pot containing 1 kg soil. In both the ferns, Cr accumulation increased with increase in metal concentration and maximum accumulation of 800.5 microg g(-1) (fronds) and 1457.4 microg g(-1) (roots) in M. punctatum and 660.8 microg g(-1) (fronds) and 1259.6 microg g(-1) (roots) in A. capillus-veneris was recorded. The increase in the levels of malondialdehyde, antioxidants and antioxidant enzymes (superoxide dismutase, glutathione peroxidase) in A. capillus-veneris was less pronounced than M. punctatum under Cr exposure as compared to their respective controls. In view of less decrease in chlorophyll content and antioxidants along with higher accumulation of Cr in the fronds M. punctatum, is indicative of its higher tolerance towards Cr. However, bioaccumulation factor (concentration of Cr in fronds/concentration of Cr in the soil) of both the ferns was recorded > 1 which qualifies the plants as potential Cr hyperaccumulator and suitable for phytoremediaton.

In-vitro shoot regeneration via caulogenesis in fern, Pteris vittata L.

An attempt was made to standardize the protocol for the shoot regeneration via caulogenesis in Pteris vittata L. employing leaf primordium explants. Calli were induced on Murashige and Skoog's (MS) and Parkers and Thompson's (P and T) media supplemented with different combinations of 2, 4-dichlorophenoxyaceticacid (2, 4-D), 6-benzylaminopurine (BAP), alpha-naphthalene acetic acid (NAA) and Indole-3-acetic acid (IAA). A combination of full strength MS medium with 2, 4-D (2.26 microM) and BAP (2.22 microM) was found to be ideal for profuse callusing (80%) against other combinations. More shoot differentiation (2.8 +/- 0.06) was achieved from the calli on one-fourth strength of P and T media fortified with BAP (4.44 microM) and NAA (2.68 microM) when compared to other combinations but statistically not significant.

Validity of heart rate based nomogram fors estimation of maximum oxygen uptake in Indian population.

Maximal oxygen uptake (VO2max) during a graded maximal exercise test is the objective method to assess cardiorespiratory fitness. Maximal oxygen uptake testing is limited to only a few laboratories as it requires trained personnel and strenuous effort by the subject. At the population level, submaximal tests have been developed to derive VO2max indirectly based on heart rate based nomograms or it can be calculated using anthropometric measures. These heart rate based predicted standards have been developed for western population and are used routinely to predict VO2max in Indian population. In the present study VO2max was directly measured by maximal exercise test using a bicycle ergometer and was compared with VO2max derived by recovery heart rate in Queen's College step test (QCST) (PVO2max I) and with VO2max derived from Wasserman equation based on anthropometric parameters and age (PVO2max II) in a well defined age group of healthy male adults from New Delhi. The values of directly measured VO2max showed no significant correlation either with the estimated VO2max with QCST or with VO2max predicted by Wasserman equation. Bland and Altman method of approach for limit of agreement between VO2max and PVO2max I or PVO2max II revealed that the limits of agreement between directly measured VO2max and PVO2max I or PVO2max II was large indicating inapplicability of prediction equations of western population in the population under study. Thus it is evident that there is an urgent need to develop nomogram for Indian population, may be even for different ethnic sub-population in the country.

Metabolic adaptation of Pteris vittata L. gametophyte to arsenic induced oxidative stress.

The sporophyte and gametophyte of Pteris vittata are arsenic hyperaccumulators, however, little is known about the mechanism by which the gametophyte deals with this toxic element. An in vitro system (spores grown in arsenic amended nutrient media) was used to investigate the impact of arsenic on growth of the gametophyte and the role of antioxidative systems in combating As-stress. When mature spores of P. vittata were grown in medium amended with 0-50 mg kg(-1) of arsenic (as arsenate), the arsenic concentration in the gametophyte increased, with increasing arsenate in the media, but did not inhibit the spore germination and biomass development. Increases in the level of antioxidant enzymes, superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and glutathione-Stransferase) and of ascorbic acid and glutathione probably enabled the gametophyte to withstand the oxidative stress caused by arsenate.

Development and performance evaluation of alginate-capped amphotericin B lipid nanoconstructs against visceral leishmaniasis.

The present study was aimed to assess the improvement of existing treatment regimens of Amphotericin B nanoconstrcuts which synergises with alginate for immunostimulation against visceral leishmaniasis. The particle size of Lip-nano (Plain AmB nanoconstructs) and Alg-Lip-nano (alginate capped Lip-nano) was 108.3 +/- 4.3 and 134.2 +/- 5.1 while zeta potential was (+) 28.4 +/- 3.3 and (-) 19.8 +/- 2.1 respectively. Percentage of parasite inhibition (intramacrophagic amastigotes) at 0.1 microg/ml conc. of AmB in case of Alg-Lip-nano (58%) was significantly higher (P = 0.05) compared to Lip-nano (48%). This supports that alginate coating over particles can activate macrophages to synergistically act with AmB in effective killing of parasite. This observation generates interest that immunotherapy with chemotherapeutic activity of AmB can effectively increase cure rate in visceral leishmaniasis.

Arsenic accumulation pattern in 12 Indian ferns and assessing the potential of Adiantum capillus-veneris, in comparison to Pteris vittata, as arsenic hyperaccumulator.

The present study was undertaken to evaluate the ability of some Indian ferns to accumulate and tolerate arsenic. Twelve species of Indian ferns were exposed to 10 mg L(-1) arsenic as sodium arsenate for 15 days in hydroponic system. Depending on the arsenic uptake in the plant parts--Pteris vittata, Pteris cretica, Adiantum capillus-veneris and Nephrolepis exaltata may be categorised as arsenic accumulator. Further, A. capillus-veneris plants were grown in arsenic contaminated soil (200-600 mg kg(-1)) under green-house condition, to assess its arsenic accumulation and tolerance mechanism, in comparison to known As-hyperaccumulator--P. vittata Linn., growing in the same conditions. The experiment identified A. capillus-veneris having a potential to tolerate arsenic up to 500 mg kg(-1). The plants were analysed for the extent of oxidative stress, as a result of arsenic accumulation. A. capillus-veneris was able to detoxify the arsenic stress through induction of anti-oxidant defence system.

Potential approaches for drug delivery to the brain: past, present, and future.

The objective of this article is to provide the reader with an update on some research highlights from the past to the present, as well as future possibilities to achieve improved delivery of drugs across the blood-brain barrier (BBB). In the past, dye studies confirmed the presence of the BBB and blood-cerebrospinal fluid barriers, which seem to play a major role in transporting drug molecules for the treatment of life-threatening diseases such as brain cancer and Alzheimer's. Presently, transportation mechanisms such as simple diffusion, carrier-mediated, absorptive-mediated, and receptor-mediated transcytosis are extensively used for BBB uptake of drug molecules. The spectrum of future neuropharmaceuticals falling into these categories ranges from peptides to nucleotide-based drugs as well as gene and stem cell delivery agents, and is increasing at a rapid rate with promising results. There has also been considerable progress in the development of quantitative methods to examine BBB permeability in humans and animals. Currently, intravenous administration and in situ brain perfusion techniques are the most versatile and sensitive methods to measure transport into the brain. This article also reviews the various methodologies available for assessing the transfer of drug molecules undergoing significant uptake through the BBB in vivo.

In Vitro Antibacterial and Antifungal Properties of Aqueous and Non-Aqueous Frond Extracts of Psilotum nudum, Nephrolepis biserrata and Nephrolepis cordifolia.

Plants are an important source of neutraceuticals that have proved to be effective against important microbial infections of humans. Lower plants are gaining importance in this regard. The present study is aimed at investigating the antimicrobial properties of three selected ferns, Psilotum nudum, Nephrolepis biserrata and Nephrolepis cordifolia. The aerial parts of the selected ferns, P. nudum, N. biserrata and N. cordifolia, were fractionated in different solvents. These fractions were concentrated to obtain a powder and were tested against nine bacterial and three fungal strains according to disc diffusion method. The water and ethanol fractions were active against most of the tested bacterial and fungal strains, some of these were more effective than the controls tested. Present study suggests that the pteridophytes, P. nudum, N. biserrata and N. cordifolia could be good source of antimicrobials. These natural compounds might be more effective as the microbes may have lesser chance of developing resistant mutants.

Tumoral calcium pyrophosphate dihydrate crystal deposition disease: a rare diagnosis by fine-needle aspiration.

Calcium pyrophosphate dihydrate crystal deposition disease (CPPD) is a well-recognized inflammatory joint disorder characterized by presence of calcium pyrophosphate dihydrate crystals in intraarticular and periarticular tissue. We report here a case of a 48-year-old male who presented with painless right hand swelling. Clinical suspicion was that of malignant soft tissue tumor. Fine-needle aspiration (FNA) yielded chalky white gritty material. Microscopic examination showed large areas of basophilic calcified material, histiocytes, giant cells and characteristic rhomboid shaped crystals. At places, chondroid material was also identified, hence, diagnosis of CPPD was made. This was confirmed on histopathological examination. Tophaceous/ tumoral pseudogout is a rare form of CPPD and it is important to recognize that this form can be diagnosed in FNA cytology (FNAC) and misdiagnosis of benign or malignant cartilaginous lesions can be avoided.

Antimicrobial activity of some important Adiantum species used traditionally in indigenous systems of medicine.

Adiantum Linn. of Adiantaceae family is one of the most common and widely distributed species. Ethnomedicinally, the genus is important and popularly known as "Hansraj" in Ayurvedic System of Medicine. It has been used in cold, tumors of spleen, liver and other viscera, skin diseases, bronchitis and inflammatory diseases. It is also considered as tonic and diuretic. In the present study its four important species, i.e. Adiantum capillus-veneris, Adiantum peruvianum, Adiantum venustum and Adiantum caudatum were collected and extracted with methanol. These extracts were tested for their antimicrobial agents against five gram positive, six gram negative (including multiresistant bacteria Staphylococcus aureus) and eight fungal strains using standard microdilution assay. The maximum activity was exhibited by the methanolic extract of Adiantum venustum followed by Adiantum capillus-veneris, Adiantum peruvianum and Adiantum caudatum. The methanolic extract of Adiantum capillus-veneris had very low MIC value (0.48 microg/ml) against Escherichia coli whereas, Adiantum venustum extract against Aspergillus terreus with MIC of 0.97 microg/ml. Total phenolic constituents of Adiantum species viz. Adiantum venustum, Adiantum capillus-veneris, Adiantum peruvianum and Adiantum caudatum were 0.81% (w/w), 0.83% (w/w), 0.71% (w/w) and 0.52% (w/w), respectively (as gallic acid equivalent); implying that the observed activity could be related to the amount of phenolics.

Natural twilight phase-response curves for the cave-dwelling bat, Hipposideros speoris.

Phase-response curves (PRCs) for the circadian rhythm of flight activity of the microchiropteran bat (Hipposideros speoris) were determined in a cave, employing discrete natural dawn and dusk twilight pulses. These PRCs are reported for the first time for any circadian system and they are unlike other PRCs constructed for nocturnal mammals. Dawn and dusk twilight pulses evoked advance and delay phase shifts, respectively. Advance phase shifts were followed by 3 to 4 advancing transients and a subsequent shortening of free-running period (tau); whereas, the delay phase shifts were instantaneous without any transients but with a subsequent lengthening of tau.

Altitudinal variation in the circadian rhythm of oviposition in Drosophila ananassae.

The effect of altitude on four basic properties of the pacemaker controlling the circadian rhythm of oviposition in two strains of Drosophila ananassae was determined. The high altitude (HA) strain from Badrinath (5123 m above sea level) had a low amplitude peak in the forenoon while the low altitude (LA) strain from Firozpur (179 m a.s.l.) had a high amplitude peak after the lights-off of LD 12:12 cycles. Free running periods in continuous darkness were about 22.6 and 27.4 h in the HA and LA strains, respectively. The light pulse phase response curve (PRC) for the HA strain showed a low amplitude and a dead zone of 8h; the ratio for the advance to delay region (A/D) was less than 1, while the PRC for the LA strain had a high amplitude, which was devoid of a dead zone and showed a ratio of A/D > 1. The magnitude of the delay phase shifts at CT 18 evoked by light pulses of 1 h duration, but varying light intensity was significantly different in the HA and LA strain, which suggests that the photic sensitivity of the clock photoreceptors mediating the phase shifts had been affected by the altitude.

Effects of temperature, photoperiod, and light intensity on the eclosion rhythm of the high-altitude Himalayan strain of Drosophila ananassae.

Eclosion rhythm of the high-altitude Himalayan strain of Drosophila ananassae from Badrinath (altitude 5123 m) was temperature-dependent and at 21 degrees C, it was entrained by cycles of 12h light: 12h darkness (LD 12:12) and free-ran in constant darkness, however, it was arrhythmic at 13 degrees C or 17 degrees C under identical experimental conditions (Khare, P. V., Barnabas, R. J., Kanojiya, M., Kulkarni, A. D., Joshi, D. S. (2002). Temperature dependent eclosion rhythmicity in the high altitude Himalayan strains of Drosophila ananassae. Chronobiol. Int. 19:1041-1052). The present studies were designed to see whether or not these strains could be entrained at 13 degrees C, 17 degrees C, and 21 degrees C by two types of LD cycles in which the photoperiod at 100 lux intensity varied from 6h to 18h, and the light intensity of LD 14:10 cycles varied from 0.001 lux to 1000 lux. All LD cycles entrained this strain at 21 degrees C but not at 13 degrees C or 17 degrees C. These results demonstrate that the entrainment of eclosion rhythm depends on the ambient temperature and not on the photoperiod or light intensity of LD cycles. Thus the temperature has taken precedence over the light in the entrainment process of eclosion rhythm of the high altitude Himalayan strain of D. ananassae. This may be the result of natural selection in response to the environmental temperature at Badrinath that resembles that of the sub-Arctic region but the photoperiod or light intensity are of the subtropical region.