Diagnosis of rabies using reverse-transcription polymerase chain reaction on post-mortem skin tissue specimens of the nasolabial plate in a rabies suspected cow: a case study.

In India, rabies in cattle is under-reported. Religious sentiments hamper its diagnosis, discouraging post-mortem examination, particularly opening the cranium. Specimens of peripheral tissue innervated by the cranial nerves could potentially be used as alternative diagnostic specimens to the brain. Herein, we present a case study of a novel approach for diagnosing rabies in a cow suspected of having rabies, using skin tissue specimens of the nasolabial plate obtained post-mortem. Brain and nasolabial tissue specimens tested positive for rabies using conventional reverse-transcription polymerase chain reaction. This approach has been previously shown to have a high diagnostic sensitivity in animals. We encourage further studies with more nasolabial plate skin specimens for both post- and antemortem diagnosis of rabies in cattle.

Toxoplasma gondii in Chickens (Gallus domesticus) from North India.

PURPOSE: Toxoplasma gondii, an important food-borne parasite has been reported in a wide range of food animals globally. Poultry meat production is on rise in several developing nations including India. Several studies on status of T. gondii conducted globally had shown substantial risk of this parasite through poultry meat. Such information is lacking in the north India, where poultry meat is a preferred non-vegetarian food over other meats; therefore, this study determined the seroprevalence (based on detection of antibodies) and tissue prevalence (based on detection of DNA) of T. gondii in poultry meat intended for human consumption in north India. METHODS: Samples (skeletal muscle and/heart as well as blood) were collected from slaughter poultry intended for human consumption reared in Punjab state (n = 366), Himachal Pradesh (n = 44) state, and Chandigarh union territory (n = 112) in North India. Serum samples were tested using commercially available Chicken T. gondii IgG ELISA kit to determine the serostatus. The DNA of T. gondii was detected by nested PCR. To determine the agreement between ELISA and PCR, Kappa value was estimated. RESULTS: Overall, study reports 2.3% (95% CI 1.3%-4.1%) tissue prevalence and 6.5% (95% CI 4.6%-9.1%) seroprevalence from chickens reared in north India. Backyard poultry had significantly higher proportion of T. gondii (24.4%, 10/41; p < 0.001) than caged (0%, 0/284) and deep litter poultry (16.2%, 32/197). The sequence alignment score of three sequences obtained in this study showed 97-100% nucleotide identity with Type I RH strain and VEG type III strain. The agreement between ELISA and PCR was poor (Kappa = 0.145; 95% CI 0.07-0.22). Thus, a combination of serological and molecular tests can improve the diagnosis of T. gondii. CONCLUSION: A low prevalence indicates that T. gondii in poultry presents a lower food safety risk for public health in north India; however, this study recommend cooking of poultry meat before consumption. Further studies are required to check the viability of T. gondii in poultry meat intended for human consumption, and to determine the risk factors associated and the genotypes circulating in poultry in north India.

Prevalence, molecular detection and risk factors investigation for the occurrence of Toxoplasma gondii in slaughter pigs in North India.

BACKGROUND: Toxoplasma gondii, an important food borne zoonotic parasite, infects almost all warm-blooded animals including pigs. People primarily become infected with T. gondii via consuming meat of infected animals. Status of T. gondii is largely unknown in pigs in India including northern regions. We, therefore, determined the prevalence of T. gondii infection in pigs from North India. RESULTS: DNA of T. gondii was detected in 6.7% (54/810) of the tested slaughter pigs. Highest prevalence was observed in pigs from Punjab (8.2%) followed by Chandigarh (5.3%) and Uttarakhand (4.8%). Phylogenetic analysis revealed that the isolates from pigs had 96-100% nucleotide identity with Type I RH strain (AF179871), 96-99.7% with VEG type III strain (LN714499) and 67-72% with type II ME 49 strain (XM002370240). However, low level of polymorphism in the targeted B1 gene did not allow the determination of the clonal lineages of the isolates. Antibodies against T. gondii was reported in 48.3% (73/151) of the sera obtained from pigs slaughtered at Chandigarh abattoir, and scavenging by pigs was a significant risk factor. CONCLUSION: Prevalence of T. gondii DNA was low in pigs in North India, however, presence of the parasite warrants food safety concerns. Further studies are required to identify the clonal lineage of T. gondii circulating in pigs reared in North India. Pig farmers should be educated about the hygienic management practices.

The Clean India Mission: Public and animal health benefits.

The Clean India Mission is a national campaign that aims for complete elimination of open defecation from the country. In India, 564 million people do not have access to toilets and defecate in the open environment. The 'Millennium development goals' have given increased weightage to elimination of open defecation for improving health, nutrition and productivity of developing country populations. The Indian economy bears an estimated annual total loss (in terms of health, education, access time and tourism) of US$ 54 billion due to lack of toilets, poor hygiene and over US$ 38.5 billion in treatment costs for diseases occurring due to poor hygiene. Out of 1415 human pathogens, at least more than 10% of pathogens are transmitted through the faecal-oral route. The practice of open defecation helps pathogens persist in the environment and cause diseases. This review focuses on the current status and harms of open defecation, as well as the public and animal health benefits of implementing 'The Clean India Mission' in India.

Social feeding in Caenorhabditis elegans is modulated by antipsychotic drugs and calmodulin and may serve as a protophenotype for asociality.

Here, we define a protophenotype as an endophenotype that has been conserved during evolution. Social feeding in Caenorhabditis elegans may be an example of a protophenotype related to asociality in schizophrenia. It is regulated by the highly conserved neuropeptide Y receptor, NPR-1, and we speculated that social feeding should be affected by antipsychotic drugs. The social feeding strain, npr-1(g320), was exposed to antipsychotic drugs, dopamine or calmodulin antagonists on plates with bacterial lawns, and the number of aggregates on the plates was counted as a measure of social feeding. First-generation antipsychotics, chlorpromazine, trifluoperazine, fluphenazine, and haloperidol, and the second-generation drug, olanzapine, inhibited social feeding. Dopamine accelerated aggregation, whereas selective D2 dopamine receptor antagonists, sulpiride and raclopride, were inhibitory. Calmodulin antagonists effectively inhibited social feeding, as did RNAi knockdown of calmodulin (cmd-1) expression. In addition, gap junction inhibitors prevented aggregation, which is consistent with the hub-and-spoke arrangement of neurons that regulate social feeding via functional gap junctions. The studies described here revealed novel connections between dopaminergic signaling, the NPY receptor, calmodulin, and gap junctions in the regulation of social behavior in C. elegans. These pathways are evolutionarily-conserved, and have also been implicated in the pathogenesis of schizophrenia.

Impact of polymers on dissolution performance of an amorphous gelleable drug from surface-coated beads.

A non-ionic amorphous active API ((RR)-3((1R)-3-oxocyclopentyl)-2-[3-chloro-4-methyl sulfonyl]phenyl-N-pyrozin-2ylpropanamide) with a glass transition temperature of 60 degrees C and aqueous solubility of 0.8 mg/mL was layered on the cellulose beads by the help of an anionic (Eudragit L100) and a non-ionic (polyvinylpyrrolidone) PVP K30 polymer respectively. An "immediate" and complete release of API from the anionic (Eudragit L100), and "sustained" but incomplete release from the hygroscopic non-ionic polymer coatings were observed. The effect of the PVP K30, and delivery patterns were investigated. Water uptake of the polymers and flow properties of API upon exposure to humidity as well as moisture sorption of beadlets were determined. Drug-polymer interactions and coating morphologies that were examined via near infrared imaging (NIR), microscopy and FTIR, enlightened any possible drug-polymer interaction. From the anionic polymer coating 93.5% API was dissolved in 50 min whereas the non-ionic polymer coating released 60% drug within 5 h. There were no API-polymer interactions as demonstrated by FTIR, implying that, this factor did not play any role in the differences observed in the release profiles. However, gelling, clumping and agglomeration was observed on the surface of the particles coated with PVP which resulted in slow and incomplete release of the drug. The anionic polymer protected API, by preventing its gelling and clumping in situ while the non-ionic polymer promoted gelling. Because API gels at a critical moisture level and at an associated critical time interval, any delivery system that can protect the drug from reaching to the critical moisture level can control API release. The drug was released via surface erosion from the Eudragit L100 coating, whereas PVP K30, the non-ionic polymer, released API via diffusion process. The results indicate that polymer properties can play a critical role in the release mechanism and kinetics of gelleable drugs. The anionic polymers may protect drugs of similar nature from gelling when exposed to the dissolution media. An understanding of mechanisms involved in drug-polymer interactions will be useful to screen the polymers that are useful in engineering suitable delivery systems for such drugs.

Transdermal and buccal delivery of methylxanthines through human tissue in vitro.

We examined the in vitro permeation of central nervous stimulants - caffeine, theophylline, and theobromine across human skin with the aid of six chemical enhancers. It was found that oleic acid was the most potent enhancer for all three methylxanthines. Further optimization studies with different solvents showed that caffeine transport could be enhanced to give flux values up to 585 microg/cm2.hr-1. Theobromine and theophylline delivery rates proved insufficient. An additional study involving a buccal tissue equivalent showed that this membrane was more permeable than skin for all model actives tested and would offer an alternate way of delivery.

Transdermal iontophoresis.

Iontophoresis is a technique used to enhance the transdermal delivery of compounds through the skin via the application of a small electric current. By the process of electromigration and electro-osmosis, iontophoresis increases the permeation of charged and neutral compounds, and offers the option for programmed drug delivery. Interest in this field of research has led to the successful delivery of both low (lidocaine) and high molecular drugs, such as peptides (e.g., luteinising hormone releasing hormone, nafarelin and insulin). Combinations of iontophoresis with chemical enhancers, electroporation and sonophoresis have been tested in order to further increase transdermal drug permeation and decrease possible side effects. In addition, rapid progress in the fields of microelectronics, nanotechnology and miniaturisation of devices is leading the way to more sophisticated iontophoretic devices, allowing improved designs with better control of drug delivery. Recent successful designing of the fentanyl E-TRANS iontophoretic system have provided encouraging results. This review will discuss basic concepts, principles and applications of this delivery technique.

Transdermal iontophoresis: combination strategies to improve transdermal iontophoretic drug delivery.

For several decades, there has been interest in using the skin as a port of entry into the body for the systemic delivery of therapeutic agents. However, the upper layer of the skin, the stratum corneum, poses a barrier to the entry of many therapeutic entities. Given a compound, passive delivery rate is often dependent on two major physicochemical properties: the partition coefficient and solubility. The use of chemical enhancers and modifications of the thermodynamic activity of the applied drug are two frequently employed strategies to improve transdermal permeation. Chemical enhancers are known to enhance drug permeation by several mechanisms which include disrupting the organized intercellular lipid structure of the stratum corneum , 'fluidizing' the stratum corneum lipids , altering cellular proteins, and in some cases, extracting intercellular lipids . However, the resulting increase in drug permeation using these techniques is rather modest especially for hydrophilic drugs. A number of other physical approaches such as iontophoresis, sonophoresis, ultrasound and the use of microneedles are now being studied to improve permeation of hydrophilic as well as lipophilic drugs. This article presents an overview of the use of iontophoresis alone and in conjunction with other approaches such as chemical enhancement, electroporation, sonophoresis, and use of microneedles and ion-exchange materials.

Mucosal drug delivery: membranes, methodologies, and applications.

In recent years, extensive research into novel forms of drug delivery has suggested that mucosal approaches offer a promising therapeutic alternative, especially for systemically acting drugs. Transmucosal drug delivery offers many benefits, including noninvasive administration, convenience, rapid onset, as well as elimination of hepatic first-pass metabolism. The investigated absorptive surfaces consist of the nasal, buccal, ocular, vaginal, and rectal mucosae. Among these, the nasal and buccal routes have proved the most promising to date. The bioavailability achieved mainly depends upon the pathophysiological state of the mucosa and the properties of both the drug and delivery systems. Various agents can increase the efficacy of transmucosal drug delivery. These include cyclodextrins, bile salts, surfactants, fusidic acid derivatives, microspheres, liposomes, and bioadhesive agents. The mechanisms of action, effectiveness, and toxicity profiles of these enhancers have been investigated extensively in both animal and human models.