Recent Studies on Serotonin 5-HT2A Receptor Antagonists in Medicinal Chemistry: A Last Decades Survey.

In the last decades, much attention has been paid to the functioning of receptors to understand better how they work with various chemical motifs. Among different families, G-proteincoupled receptor (GPCR) families have drawn much attention in the twenty-first century. They are the most prominent signal transducer across the cell membrane, comprising thousand-odd proteins. One of the members of GPCRs is the serotonin 2A (5-HT2A) receptor, which has been associated with complex etiological mental illnesses. In this survey, we collected data on 5-HT2A, i.e., the role of 5- HT2A receptors in human and animal analogy, various binding site functionalities, advanced effects, and synthetic aspects.

Health risk assessment of pesticide residues in fruits and vegetables from farms and markets of Western Indian Himalayan region.

The extensive use of pesticides in agriculture has become a very common practice in developing countries like India. Consequently, the increased concentration of residues of these hazardous pesticides in fruits and vegetables is manifested. The study aimed to assess the health hazards associated with the presence of pesticide residues in fruits and vegetables sampled from farms and markets of Kinnaur district of Himachal Pradesh (India). Residues of predominant pesticides used in the region, belonging to the group of organophosphates, pyrethroid and phthalimide, were analysed using gas chromatograph quadrupole mass spectrometer (GC-MS/MS). The pesticide extraction from the matrix was done following the modified QuEChERS method. Results indicated varying concentrations of pesticide residue in market and farm samples with farm samples more contaminated than market samples. Chronic health hazards prediction indicated that organophosphorus groups (methyl parathion and triazophos) posed health risk to children in the study area.

Predictors and Prevalence of Pain and Its Management in Four Regional Cancer Hospitals in India.

PURPOSE: More than 1 million new occurrences of cancer are diagnosed in India annually. Among patients with cancer, pain is a common and persistent symptom of the disease and its treatment. However, few studies to date have evaluated the prevalence of pain and the adequacy of pain management in Indian hospitals. This cross-sectional study aimed to assess the prevalence and sociodemographic patterns of cancer pain and pain management among a sample of inpatients and newly registered outpatients at four large regional cancer centers in India. METHODS: A sample of 1,600 patients with cancer who were current inpatients or newly registered outpatients were recruited and administered a questionnaire that was based on the Brief Pain Inventory. The survey tool included questions on demographics, medical history, and extent of clinical pain experienced. In addition, a pain management index score was created to link the severity of cancer pain with medication prescribed to treat it. RESULTS: A total of 88% of patients reported pain in the past 7 days, and approximately 60% reported that their worst pain was severe. Several demographic and medical characteristics of the study population predicted severe pain, including the following: lower educational level, outpatient status, and debt incurred as a result of illness. A total of 67% of patients were inadequately treated with analgesics. Inadequate pain management was associated with both treatment hospital and patient type, and patients who reported debt as a result of their illness were more likely to have inadequate pain management. CONCLUSION: A majority of Indian patients with cancer experience significant pain and receive inadequate pain management. Improvement of pain management for Indian patients with cancer is needed urgently.

Antiplasmodial efficacy of Calotropis gigantea (L.) against Plasmodium falciparum (3D7 strain) and Plasmodium berghei (ANKA).

BACKGROUND & OBJECTIVES: Malaria is a deadly parasitic disease, having a high rate of incidence and mortality across the world. The spread and development of resistance against chemical insecticides is one of the major problems associated with malaria treatment and control. Hence, plant based formulations may serve as an alternative source towards development of new drugs for treatment of malaria. The present study was aimed to evaluate the in vitro antiplasmodial activities of leaf, stem and flower of Calotropis gigantea against chloroquine-sensitive Plasmodium falciparum (3D7 strain) and its cytotoxicity against THP-1 cell lines. The plant extract which showed highest potency, in the in vitro antimalarial activity was further tested in vivo against P. berghei (ANKA strain) for validating its efficacy. METHODS: The crude extracts of methanol, ethyl acetate and chloroform from leaves, stem and flowers of C. gigantea were prepared using Soxhlet apparatus. These extracts were screened for in vitro antimalarial activity against P. falciparum 3D7 strain. The cytotoxicity studies of crude extracts were conducted against THP-1 cell line. Phytochemical analysis of these extracts was carried out by following the standard methods. The damage to erythrocytes due to the plant extracts was tested. The in vivo study was conducted in P. berghei (ANKA) infected BALB/c albino mice by following the 4-day suppressive test. RESULTS: The phytochemical screening of the crude extracts showed the presence of alkaloids, flavonoids, triterpenes, tannins, carbohydrates, phenols, coumarins, saponins, phlobatannins and steroids. Out of all the extracts, the methanolic extract of leaves showed highest antimalarial activity with IC50 value of 12.17 µg/ml. In cytotoxicity evaluation, none of the crude extracts, showed cytotoxicity on THP-1 cell line. Since, methanolic leaf extract of C. gigantea showed good antimalarial activity in vitro, it was tested in vivo. In the in vivo results, the methanolic leaf extract of C. gigantea exhibited an excellent activity against P. berghei malaria parasite, wherein the decrement of parasite counts was moderately low and dose-dependent (p < 0.05) in comparison to the P. berghei infected control group, which showed a daily increase of parasitaemia unlike the chloroquine-treated group. INTERPRETATION & CONCLUSION: The methanolic leaf extract of C. gigantea may act as potent alternative source for development of new medicines or drugs for the treatment of drug-resistant malaria. Thus, further research is needed to characterize the bioactive molecules of the extracts of C. gigantea that are responsible for inhibition of malaria parasite.

Nutritional composition and antioxidant activities of 18 different wild Cantharellus mushrooms of northwestern Himalayas.

A total of 18 wild edible mushrooms of Cantharellus species were collected from northwestern Himalayan region of India. The basic composition (moisture, total carbohydrates, dietary fiber, crude fat, ash, nitrogen and protein) and amino acid contents (by high-performance liquid chromatography) of these wild edible mushrooms were determined. The macronutrient profile in general revealed that the wild mushrooms were rich sources of protein and carbohydrates, and had low amounts of fat. Total phenolics and antioxidant activity from water and methanolic extracts of these mushrooms were also determined. These wild mushrooms also had significant amount of phenol content and antioxidant capacity. Studies also provide the precise antioxidant status of 18 indigenous species of mushrooms, which can serve as a useful database for the selection of mushrooms for the function of preparation of mushroom-based nutraceutics.

Sensitization to blackgram in patients with bronchial asthma and rhinitis: clinical evaluation and characterization of allergens.

BACKGROUND: Legumes are important causative agents of type I hypersensitivity in south Asia and Europe but such studies are lacking in Indian population. The present study investigates blackgram sensitization in asthma and rhinitis patients and identifies immunoglobulin E (IgE)-binding proteins. METHODS: Respiratory allergy patients were evaluated using standard questionnaire and skin prick tests (SPT) with common foods and aeroallergens. Blackgram-specific IgE level was estimated by enzyme-linked immunosorbent assay (ELISA) and sensitization was established by a double-blind, placebo-controlled food challenge (DBPCFC). The cross-reactivity of blackgram with other legumes was studied by immunobiochemical methods. RESULT: Of 816 patients, 35 gave history of blackgram hypersensitivity. From these, 16 patients were SPT positive and 14 showed elevated specific IgE (three times of negative control) to blackgram. DBPCFC established blackgram sensitivity in four of 14 patients. Immunoblotting with individual patient's sera recognized eight most prevalent allergens of 78, 56, 47, 43, 40, 30, 28 and 16 kDa. Roasted blackgram showed six major allergens whereas 47, 43 and 28 kDa proteins retained IgE reactivity upon boiling. Blackgram extract required 14 ng of self protein for 50% ELISA inhibition whereas roasted and boiled blackgram required 16 and 120 ng protein. ELISA and immunoblot inhibition show partial inhibition to blackgram proteins by lentil, limabean and pea. CONCLUSION: Blackgram induces IgE-mediated reactions in 1.7% of asthma and rhinitis patients and contains eight major IgE-binding components, of which six retained IgE reactivity after roasting. Blackgram shares allergenicity with lentil and limabean.

Transcription defects induced by repeat expansion: fragile X syndrome, FRAXE mental retardation, progressive myoclonus epilepsy type 1, and Friedreich ataxia.

Fragile X mental retardation syndrome, FRAXE mental retardation, Progressive myoclonus epilepsy Type I, and Friedreich ataxia are members of a larger group of genetic disorders known as the Repeat Expansion Diseases. Unlike other members of this group, these four disorders all result from a primary defect in the initiation or elongation of transcription. In this review, we discuss current models for the relationship between the expanded repeat and the disease symptoms.

Instability of the fragile X syndrome repeat in mice: the effect of age, diet and mutations in genes that affect DNA replication, recombination and repair proficiency.

Repeat expansion diseases such as fragile X syndrome (FXS) result from increases in the size of a specific tandem repeat array. In addition to large expansions, small changes in repeat number and deletions are frequently seen in FXS pedigrees. No mouse model accurately recapitulates all aspects of this instability, particularly the occurrence of large expansions. This may be due to differences between mice and humans in CIS and/or TRANS-acting factors that affect repeat stability. The identification of such factors may help reveal the expansion mechanism and allow the development of suitable animal models for these disorders. We have examined the effect of age, dietary folate, and mutations in the Werner's syndrome helicase (WRN) and TRP53 genes on FXS repeat instability in mice. WRN facilitates replication of the FXS repeat and enhances Okazaki fragment processing, thereby reducing the incidence of processes that have been suggested to lead to expansion. p53 is a protein involved in DNA damage surveillance and repair. We find two types of repeat instability in these mice, small changes in repeat number that are seen at frequencies approaching 100%, and large deletions which occur at a frequency of about 10%. The frequency of these events was independent of WRN, p53, parental age, or folate levels. The large deletions occur at the same frequency in mice homozygous and heterozygous for the repeat suggesting that they are not the result of an interallelic recombination event. In addition, no evidence of large expansions was seen. Our data thus show that the absence of repeat expansions in mice is not due to a more efficient WRN protein or p53-mediated error correction mechanism, and suggest that these proteins, or the pathways in which they are active, may not be involved in expansion in humans either. Moreover, the fact that contractions occur in the absence of expansions suggests that these processes occur by different mechanisms.

Deletion analysis of the dystrophin gene in Duchenne and Becker muscular dystrophy patients: use in carrier diagnosis.

The dystrophin gene was analyzed in 8 Duchenne muscular dystrophy (DMD) and 10 Becker muscular dystrophy (BMD) unrelated families (22 subjects: 18 index cases and 4 sibs) for the presence of deletions by multiplex polymerase chain reaction (mPCR; 27 exons) and Southern hybridization using 8 cDMD probes. Deletions were identified in 5 DMD and 7 BMD patients (6 index cases and 1 sib). The concordance between the clinical phenotype and "reading frame hypothesis" was observed in 11/12 patients (92%). The female relatives of DMD/BMD patients with identifiable deletions were examined by quantitative mPCR. Carriers were identified in 7 families. We also describe a variation in the HindIII pattern with cDNA probe 8 and 11-14. Molecular characterization of the dystrophin gene in this study has been helpful in advising the patients concerning the inheritance of the condition, and carrier diagnosis of female relatives, and should also prove useful for prenatal diagnosis.

Fragile X syndrome and Friedreich's ataxia: two different paradigms for repeat induced transcript insufficiency.

DNA repeat expansion is the genetic basis for a growing number of neurological disorders. While the largest subset of these diseases results in an increase in the length of a polyglutamine tract in the protein encoded by the affected gene, the most common form of inherited mental retardation, fragile X syndrome, and the most common inherited ataxia, Friedreich's ataxia, are both caused by expansions that are transcribed but not translated. These expansions both decrease expression of the gene in which the expanded repeat is located, but they do so by quite different mechanisms. In fragile X syndrome, CGG. CCG expansion in the 5' untranslated region of the FMR1 gene leads to hypermethylation of the repeats and the adjacent CpG-rich promoter. Methylation prevents the binding of the transcription factor alpha-Pal/NRF-1, and may indirectly affect the binding of other factors via the formation of transcriptionally silent chromatin. In Friedreich's ataxia, GAA. TTC expansion in an intron of the FRDA gene reduces expression by interfering with transcription elongation. The model that best describes the available data is transcription-driven formation of a transient purine. purine. pyrimidine DNA triplex behind an advancing RNA polymerase. This structure lassoes the RNA polymerase that caused it, trapping the enzyme on the template.

Interaction of the transcription factors USF1, USF2, and alpha -Pal/Nrf-1 with the FMR1 promoter. Implications for Fragile X mental retardation syndrome.

Hypermethylation of the FMR1 promoter reduces its transcriptional activity, resulting in the mental retardation and macroorchidism characteristic of Fragile X syndrome. How exactly methylation causes transcriptional silencing is not known but is relevant if current attempts to reactivate the gene are to be successful. Understanding the effect of methylation requires a better understanding of the factors responsible for FMR1 gene expression. To this end we have identified five evolutionarily conserved transcription factor binding sites in this promoter and shown that four of them are important for transcriptional activity in neuronally derived cells. We have also shown that USF1, USF2, and alpha-Pal/Nrf-1 are the major transcription factors that bind the promoter in brain and testis extracts and suggest that elevated levels of these factors account in part for elevated FMR1 expression in these organs. We also show that methylation abolishes alpha-Pal/Nrf-1 binding to the promoter and affects binding of USF1 and USF2 to a lesser degree. Methylation may therefore inhibit FMR1 transcription not only by recruiting histone deacetylases but also by blocking transcription factor binding. This suggests that for efficient reactivation of the FMR1 promoter, significant demethylation must occur and that current approaches to gene reactivation using histone deacetylase inhibitors alone may therefore have limited effect.

Detection of deletion in the dystrophin gene of a patient with quadriceps myopathy.

A 43 year old male presented with slowly progressive weakness of limbs and hypertrophy of triceps, brachioradialis and calf muscles for four years. There was thinning of quadriceps muscles in both thighs. Histological study was compatible with Becker muscular dystrophy (BMD). Genomic DNA analysis showed a deletion of the Hind III fragments, spanning exons 45-47. A junction fragment of 11.0 kb was observed along with a deletion of a 3.4 kb PstI fragment containing exon 51 in the patient, and in one of his two sisters. The clinical and laboratory characteristics in this patient are in keeping with what has been described 'quadriceps myopathy' and fall within the phenotypic variants of BMD as has been shown by others.

Ventilation grilles as a potential source of methicillin-resistant Staphylococcus aureus causing an outbreak in an orthopaedic ward at a district general hospital.

The spread of methicillin-resistant Staphylococcus aureus (MRSA) in a hospital is thought to be mainly by direct contact. Environmental sources such as exhaust ducting systems have been increasingly recognized as a source for MRSA outbreaks in intensive therapy units. We describe an outbreak of MRSA related to ventilation grilles in an orthopaedic ward. Six patients and one nurse were involved in an outbreak with EMRSA-15 during March 1996. The index case was transferred from a large university hospital in Leeds. One of the patients had shared the same bay with the index case. The rest of the patients were in another bay of the same ward and had no direct contact with the index patient. An environmental source was suspected and the ventilation grilles in boys 1 and 2 were found to be harbouring EMRSA-15. The ventilation system at that time was working on an intermittent cycle from 4 p.m.-8 a.m. Daily shut-down of the system temporarily created a negative pressure, sucking air in from the ward environment into the ventilation system and probably contaminating the outlet grilles. It is likely that contaminated air was blown back into the ward when the ventilation system was started. The system was thoroughly cleaned, appropriate infection control measures were instituted and the ventilation system was put back on a continuous running cycle and the outbreak terminated. Six months after the outbreak no isolates of EMRSA-15 had been made on the ward.

Molecular characterisation of Duchenne muscular dystrophy and phenotypic correlation.

Dystrophin gene was analysed in 32 unrelated DMD families (46 subjects: 32 index cases and 14 sibs) for the presence of deletions by mPCR for 27 exons and cDNA probes for the entire gene. Deletions were identified in 32 patients (25 index cases and seven sibs) from 25 families. The concordance between the clinical phenotype and 'reading frame' hypothesis was observed in 24 (75%) cases. Of these, nine patients were wheelchair bound between 8-12 years of age, nine (age range 5-10 years) showed progressive difficulty in walking and six (age range 1.6-4 years) had onset of muscle weakness. One patient (CH), who was wheelchair bound at 12 years, the effect of mutation on the ORF could not be ascertained due to the presence of a junction fragment. Seven patients had inframe deletions of which four were wheelchair bound by the age of 13 years, and three (age range 5-7 years) although, ambulatory had difficulty in walking. There were eight patients who showed no deletion, of which four became wheelchair bound by the age of 12 years, four, though still ambulatory, were unable to run and tired easily. Correlation between phenotype and genotype of these DMD patients demonstrates that genetic studies of lymphocyte DNA may not always reflect the situation in the tissue involved in dystrophin, i.e. muscle. We describe a common dystrophin gene polymorphism in the Indian population with cDNA 11-14 that alters the Hind III restriction sites. Novel RFLPs were observed in 26 patients and their family members. Whether this is a polymorphism or, related to the diseased phenotype needs confirmation.

Comparison and application of ribosome spacer DNA amplicon polymorphisms and pulsed-field gel electrophoresis for differentiation of methicillin-resistant Staphylococcus aureus strains.

Analysis of sequences in the fragments of the 16S-23S rRNA intergenic spacer region by the ribosome spacer PCR (RS-PCR) can differentiate strains of methicillin-resistant Staphylococcus aureus (MRSA). We compared this technique with pulsed-field gel electrophoresis (PFGE) for typing MRSA strains and its application during an investigation of an outbreak. A total of 180 isolates of MRSA collected from various hospital laboratories within the United Kingdom and elsewhere were typed by PFGE and RS-PCR. PFGE identified 17 different types among the 180 strains examined, and RS-PCR generated 13 different types. PFGE could detect minor genetic variations among the isolates and could identify the variants which were not discriminated by RS-PCR. Four unique strain types detected by PFGE were not detected by RS-PCR. When applied to typing the outbreak-related strains from the vascular surgery unit at the General Infirmary at Leeds, the results of RS-PCR were identical to those of PFGE. Our results have shown that RS-PCR is a rapid, inexpensive technique that is highly reproducible and almost as discriminatory as PFGE for typing MRSA isolates and should be useful in the local investigation of MRSA outbreaks.

Effect of retinol on ochratoxin-produced genotoxicity in mice.

Ochratoxin (1 microgram/kg body weight/day), when administered orally daily to albino Swiss mice for 14 continuous days, increased the incidence of abnormalities in mitotic and meiotic metaphase chromosomes, and the gross morphology of the sperm head; the sperm count per unit volume of caput epididymal suspension also decreased. These genotoxic effects were substantially reduced by concurrent oral administration of retinol at double the clinically therapeutic dose. It is possible that the electrophilic metabolites of ochratoxin form adducts with DNA or produce replacement-type mutations. Retinol may achieve its antigenotoxic effect by means of blocks and shunts in the ochratoxin metabolic pathway. Vitamin A-mediated cellular repair and scavenging of the mutagenic radicals can also take place. The vitamin itself has some genotoxic potential.

Red cell membrane alterations in human chronic fluoride toxicity.

Red cells from humans exposed chronically to toxic levels of fluoride through drinking water showed significant increase in lipid peroxidation and membranous cholesterol and phospholipids. Additionally, electrophoretic patterns of ghost membrane proteins revealed the presence of a new band in the range of congruent to 66 Kd and increase in the high molecular weight protein and predominance of bands with a molecular weight of congruent to 93 Kd and congruent to 20 Kd. The activities of total, Na(+)-K(+)-, Mg(2+)- and Ca(2+)-ATPases were significantly decreased in the red cell ghosts of fluorotic patients.