Leptospiral Infection, Pathogenesis and Its Diagnosis-A Review.

Leptospirosis is a perplexing conundrum for many. In the existing literature, the pathophysiological mechanisms pertaining to leptospirosis is still not understood in full. Considered as a neglected tropical zoonotic disease, leptospirosis is culminating as a serious problem worldwide, seemingly existing as co-infections with various other unrelated diseases, including dengue and malaria. Misdiagnosis is also common as non-specific symptoms are documented extensively in the literature. This can easily lead to death, as the severe form of leptospirosis (Weil's disease) manifests as a complex of systemic complications, especially renal failure. The virulence of Leptospira sp. is usually attributed to the outer membrane proteins, including LipL32. With an armament of virulence factors at their disposal, their ability to easily adhere, invade and replicate within cells calls for a swift refinement in research progress to establish their exact pathophysiological framework. As an effort to reconstitute the current knowledge on leptospirosis, the basis of leptospiral infection, including its risk factors, classification, morphology, transmission, pathogenesis, co-infections and clinical manifestations are highlighted in this review. The various diagnostic techniques are also outlined with emphasis on their respective pros and cons.

Elucidating raw material variability--importance of surface properties and functionality in pharmaceutical powders.

The purpose of this study is to illustrate, with a controlled example, the influence of raw material variability on the excipient's functionality during processing. Soluble starch was used as model raw material to investigate the effect of variability on its compaction properties. Soluble starch used in pharmaceutical applications has undergone a purification procedure including washing steps. In this study, a lot of commercially available starch was divided into two parts. One was left intact and the other was subjected to an extra washing step. The two resulting lots were subjected to a series of physical characterization tests typical of those used to qualify raw materials. The two resulting lots gave virtually identical results from the tests. From the physical testing point of view, the two lots can be considered as two equivalent lots of the same excipient. However, when tested for their functionality when subjected to a compaction process, the two lots were found to be completely different. The compaction properties of the two lots were distinctly different under all environmental and processing conditions tested. From the functionality point of view, the two lots are two very different materials. The similar physical testing results but different functionality can be reconciled by considering the surface properties of the powders. It was found that the washing step significantly altered the surface energetic properties of the excipient. The washed lot consistently produced stronger compacts. These results are attributable to the measurably higher surface energy of induced by the additional washing step.

Surface-Engineered Super-Paramagnetic Iron Oxide Nanoparticles For Chromium Removal.

BACKGROUND: Super-paramagnetic iron oxide nanoparticles (SPIONs) are widely used metal nanoparticles for various applications for its magnetic property and biocompatibility. In recent years, pollution of our environment especially with heavy metals in waterbodies has become a major threat and has left us very minimal sources of freshwater to drink. SPIONs or surface modified SPIONs can be used to remove these heavy metals. METHODS: SPIONs were synthesized by co-precipitation method and further coated with a biopolymer, chitosan. Chromium solution was treated with the synthesized SPIONs to study the efficiency of chromium removal by surface adsorption. Later, the adsorption was analysed by direct and indirect analysis methods using UV-VIS spectrophotometry and isotherm studies. RESULTS: Stable chitosan-coated SPIONs were synthesized and they adsorbed chromium better than the uncoated SPIONs, where it was adsorbing up to 100 ppm. Adsorption was found to be increasing with decrease in pH. CONCLUSION: The surface-modified SPIONs expressed cumulative adsorption action. Even after the adsorption studies, chitosan-coated SPIONs were possessing magnetic property. Thus, the surface-modified SPIONs can become an ideal nanotechnology tool to remove the chromium from groundwater.

Preparation, characterization and utilization of coreshell super paramagnetic iron oxide nanoparticles for curcumin delivery.

In this study, super paramagnetic iron oxide nanoparticles (SPIONs) were produced by chemical co-precipitation method, then it was constructed to be a core shell nanoparticle by functionalizing with SDS, loading with curcumin and coating with a biopolymer i.e. chitosan. Each step was analyzed microscopically and spectroscopically. The produced coreshell particles were between 40 and 45nm and these coreshell particles were utilized for drug delivery studies against cervical cancer cell line-HeLa cells. The coreshell SPIONs were found to be releasing curcumin in between 6 and 12 h, which was evidenced by increased apoptotic cells and increased caspase 3 expression in HeLa cells.

A novel method for strict intranasal delivery of non-replicating RSV vaccines in cotton rats and non-human primates.

Respiratory syncytial virus (RSV) is the most common viral cause of bronchiolitis and pneumonia in children twelve months of age or younger and a significant cause of lower respiratory disease in older adults. As various clinical and preclinical candidates advance, cotton rats (Sigmodon hispidus) and non-human primates (NHP) continue to play a valuable role in RSV vaccine development, since both animals are semi-permissive to human RSV (HRSV). However, appropriate utilization of the models is critical to avoid mis-interpretation of the preclinical findings. Using a multimodality imaging approach; a fluorescence based optical imaging technique for the cotton rat and a nuclear medicine based positron emission tomography (PET) imaging technique for monkeys, we demonstrate that many common practices for intranasal immunization in both species result in inoculum delivery to the lower respiratory tract, which can result in poor translation of outcomes from the preclinical to the clinical setting. Using these technologies we define a method to limit the distribution of intranasally administered vaccines solely to the upper airway of each species, which includes volume restrictions in combination with injectable anesthesia. We show using our newly defined methods for strict intranasal immunization that these methods impact the immune responses and efficacy observed when compared to vaccination methods resulting in distribution to both the upper and lower respiratory tracts. These data emphasize the importance of well-characterized immunization methods in the preclinical assessment of intranasally delivered vaccine candidates.

Calorimetric study and modeling of molecular mobility in amorphous organic pharmaceutical compounds using a modified Adam-Gibbs approach.

The purpose of this study is to provide a quantitative characterization of the thermal behavior of amorphous organic pharmaceutical compounds across their glass transition temperature, and to assess their molecular mobility as a function of temperature and time by combining theoretical simulations with experimental measurements using differential scanning calorimetry. A computational approach built on the Boltzmann superposition principle of nonexponential decay and the Adam-Gibbs theory of entropic-dependent structural relaxation is presented. The heat capacities of the crystalline and amorphous forms are incorporated into the simulation in order to accurately assess the entropic fictive temperature as functions of temperature and time under any arbitrary set of experimental conditions. Using this method, we evaluated properties of the glass former, D and T0, and the nonexponentiality index beta, for amorphous salicin, felodipine, and nifedipine, by fitting the simulated glass transition profile with the experimentally determined heat capacity across the glass transition region. From this fit, the evolution of the relaxation time of the model compounds following any thermal cycle, including heating, cooling, and isothermal holds can then be estimated a priori. This study reveals the profound and inextricable effect of thermal history on the molecular mobility of the amorphous materials, and the ability of the glass to undergo fast changes in its molecular motions over an aging process even at low temperatures.

A cationic peptide consists of ornithine and histidine repeats augments gene transfer in dendritic cells.

Condensing the plasmid with high molecular weight cationic polymers such as poly-L-lysine (PLL) and poly-L-ornithine (PLO) can enhance antigen-specific immunity generated from genetic vaccination with naked DNA encoding antigens. While these high molecular weight polymers are clearly effective in transfection experiments, clinical applications are limited by their physical heterogeneity and toxicity. Three chemically defined low molecular weight cationic peptides, K(16), K(10)H(6), and O(10)H(6), were examined in the context of DNA binding, toxicity, and efficiency of gene transfer in dendritic cells (DC). The results showed that while all three peptides can bind to a plasmid encoding a reporter gene with similar efficiency, in vitro transfection with DNA complexed with O(10)H(6) complexed resulted in the highest level of gene expression. Moreover, free O(10)H(6) was not toxic to DC, while the lysine-based peptides caused significant cell death in DC cultures. We also showed that DC transfected ex vivo with DNA complexed with O(10)H(6) was capable of eliciting antigen-specific INFgamma production in vivo. Taken together, these results indicate ornithine and histidine repeats are suitable building blocks of non-viral gene transfer vector for DC.

Gene delivery to dendritic cells facilitated by a tumor necrosis factor alpha-competing peptide.

Efficient gene delivery systems tailor-designed for dendritic cells (DCs) would allow the possibility of therapeutic manipulation of a wide spectrum of immune functions. Toward achieving this goal, we have identified a novel heptameric peptide (YTYQGKL) that functions as a localization moiety to mediate gene transfer in murine DCs. The sequence was identified by screening a phage display library against a DC cell line (JAWSII) using mouse TNFalpha as the eluting ligand. Alignment analysis reveals YTYQGKL resembles a solvent accessible region in mouse and human TNFalpha structures. A cyclized synthetic peptide bearing the sequence CYTYQGKLC binds to DCs in a concentration-dependent manner. Appending the cyclic peptide to a DNA binding domain (16 consecutive lysine residues) enhances transfection of reporter gene-encoding plasmids in JAWSII cells and in bone marrow derived primary DCs (BMDC). Further enhancement of gene transfer was observed when the peptide-DNA construct was anchored onto polymeric microspheres, with up to 25% of BMDC expressing the transgene. Exposing cells to the free peptide prior to transfection significantly diminished transgene expression. These results demonstrate that YTYQGKL can be used to facilitate gene transfer in DCs.

Theoretical and experimental considerations on the enthalpic relaxation of organic glasses using differential scanning calorimetry.

The enthalpy relaxation of amorphous salicin, used as model organic glass of pharmaceutical relevance, was investigated using a combination of DSC measurements and theoretical simulations. The combined approach makes it possible to discern between the effect of the glass forming properties of the material and the effects of the thermal history and experimental conditions. The approach also facilitates an unambiguous definition of the time scale of the experiment, such that objective comparison among relaxation time and glass transition temperature values can be made. The simulation provides accurate predictions of the DSC profiles obtained under a wide variety of experimental conditions. The effects of annealing time and the heating/cooling rate on the enthalpy recovery were explained by tracking the evolution of relaxation times as a function of temperature and time. The combined experimental and simulation approach also makes it possible to systematically explore the effect of specific glass forming properties, such as fragility and nonexponentiality, on the relaxation and associated thermal behavior of molecular organic glasses of pharmaceutical interest. To fully characterize these materials, it is necessary to go beyond the onset T(g) and include the early stages of the glass transition.

The effect of dehydration conditions on the functionality of anhydrous amorphous raffinose.

The purpose of this investigation is to study the effect of dehydration conditions of raffinose pentahydrate (RF.5H2O) on the physical properties and functionality of the resulting material. Crystalline RF.5H2O was dehydrated at two temperatures, 80 degrees C and 110 degrees C, producing the amorphous anhydrous form (RF.am). The dehydration temperature had no effect on a number of physical properties of the obtained RF.am, including X-ray powder diffraction, surface energy and water uptake. However, despite resulting on the same dynamics and extent of water sorption, different dehydration temperatures produced amorphous samples with drastically different recrystallization tendencies. Thermodynamic parameters show that despite the similarities on certain physical attributes, different dehydration temperature results in samples with significantly different free energy, hence stability. The difference in free energy produced by the dehydration temperature is attributed to differences in supramolecular structure that persist even in the liquid domain (above T(g)) of the amorphous samples. Evidence of such effects is observed as fluctuations in heat capacity present in RF.am but absent in the freshly prepared glass and also supported by the presence of molecular mobility modes observed using thermal polarization measurements.

Time-dependence of molecular mobility during structural relaxation and its impact on organic amorphous solids: an investigation based on a calorimetric approach.

PURPOSE: To develop a calorimetry-based model for estimating the time-dependence of molecular mobility during the isothermal relaxation of amorphous organic compounds below their glass transition temperature (Tg). METHODS: The time-dependent enthalpy relaxation times of amorphous sorbitol, indomethacin, trehalose and sucrose were estimated based on the nonlinear Adam-Gibbs equation. Fragility was determined from the scanning rate dependence of Tg. Time evolution of the fictive temperature was determined from Tg, the heat capacity of the amorphous and crystalline forms, and from the enthalpy relaxation data. RESULTS: Relaxation time changes significantly upon annealing for all compounds studied. The magnitude of the increase in relaxation time does not depend on any one parameter but on four parameters: Tg, fragility, and the crystal-liquid and glass-liquid heat capacity differences. The obtained mobility data for indomethacin and sucrose, both stored at Tg-16 K, correlated much better with their different crystallization tendencies than did the Kohlrausch-Williams-Watts (KWW) equation. CONCLUSIONS: The observed changes in relaxation time help explain and address the limitations of the KWW approach. Due consideration of the time-dependence of molecular mobility upon storage is a key element for improving the understanding necessary for stabilizing amorphous formulations.

A calorimetric method to estimate molecular mobility of amorphous solids at relatively low temperatures.

PURPOSE: To present a calorimetry-based approach for estimating the initial (at the onset of annealing) relaxation time (tau (0)) of organic amorphous solids at relatively low temperatures, and to assess the temperature where molecular mobility of the amorphous drug is reduced to a level comparable with the desired shelf-life of the product. MATERIALS AND METHODS: Values of tau (0) for six amorphous pharmaceutical compounds were estimated based on the nonlinear Adam-Gibbs equation. Fragility was determined from the scanning rate-dependence of the glass transition temperature (T (g)). The initial enthalpic and entropic fictive temperatures were obtained from the T (g) and the heat capacities (C (p)) of the amorphous and crystalline forms. RESULTS: At a relatively low temperature ( approximately 40 degrees C or more below T (g)), tau (0) for the different compounds varies by over an order of magnitude. For some materials, the practical storage temperature at T (g) - 50 K was found to be still too high to ensure long-term stability. The estimated tau (0) is highly sensitive to the fragility of the material and the C (p) of the crystalline and amorphous forms. Materials with high fragility or greater C (p) differences between crystalline and amorphous forms tend to have longer tau (0). CONCLUSIONS: The proposed method can be used to estimate molecular mobility at relatively low temperatures without having to conduct enthalpy recovery experiments. An accurate tau (0) determination from this method relies on faithful fragility measurements.