Carotenoid Extraction from Plant Tissues.

Carotenoids are the natural pigments available in nature and exhibit different colors such as yellow, red, and orange. These are a class of phytonutrients that have anti-cancer, anti-inflammatory, anti-oxidant, immune-modulatory, and anti-aging properties. These were used in food, pharmaceutical, nutraceutical, and cosmetic industries. They are divided into two classes: carotenes and xanthophylls. The carotenes are non-oxygenated derivatives and xanthophylls are oxygenated derivatives. The major source of carotenoids are vegetables, fruits, and tissues. Carotenoids also perform the roles of photoprotection and photosynthesis. In addition to the roles mentioned above, they are also involved and act as precursor molecules for the biosynthesis of phytohormones such as strigolactone and abscisic acid. This chapter briefly introduces carotenoids and their extraction method from plant tissue. Proposed protocol describes the extraction of carotenoid using solvents chloroform and dichloromethane. Reverse-phase HPLC can be performed with C30 columns using gradient elution. The column C30 is preferred to the C18 column because the C30 column has salient features, which include selective nature in the separation of structural isomers and hydrophobic, long-chain compounds, and shows the best compatibility with highly aqueous mobile phases. A complete pipeline for the extraction of carotenoids from plant tissue is given in the present protocol.

Sample Preparation from Plant Tissue for Gas Chromatography-Mass Spectrometry (GC-MS)we.

Metabolites are intermediate products formed during metabolism. Metabolites play different roles, including providing energy, supporting structure, transmitting signals, catalyzing reactions, enhancing defense, and interacting with other species. Plant metabolomics research aims to detect precisely all metabolites found within tissues of plants through GC-MS. This chapter primarily focuses on extracting metabolites using chemicals such as methanol, chloroform, ribitol, MSTFA, and TMCS. The metabolic analysis method is frequently used according to the specific kind of sample or matrix being investigated and the analysis objective. Chromatography (LC, GC, and CE) with mass spectrometry and NMR spectroscopy is used in modern metabolomics to analyze metabolites from plant samples. The most frequently used method for metabolites analysis is the GC-MS. It is a powerful technique that combines gas chromatography's separation capabilities with mass spectrometry, offering detailed information, including structural identification of each metabolite. This chapter contains an easy-to-follow guide to extract plant-based metabolites. The current protocol provides all the information needed for extracting metabolites from a plant, precautions, and troubleshooting.

Sustaining yield and nutritional quality of peanuts in harsh environments: Physiological and molecular basis of drought and heat stress tolerance.

Climate change is significantly impacting agricultural production worldwide. Peanuts provide food and nutritional security to millions of people across the globe because of its high nutritive values. Drought and heat stress alone or in combination cause substantial yield losses to peanut production. The stress, in addition, adversely impact nutritional quality. Peanuts exposed to drought stress at reproductive stage are prone to aflatoxin contamination, which imposes a restriction on use of peanuts as health food and also adversely impact peanut trade. A comprehensive understanding of the impact of drought and heat stress at physiological and molecular levels may accelerate the development of stress tolerant productive peanut cultivars adapted to a given production system. Significant progress has been achieved towards the characterization of germplasm for drought and heat stress tolerance, unlocking the physiological and molecular basis of stress tolerance, identifying significant marker-trait associations as well major QTLs and candidate genes associated with drought tolerance, which after validation may be deployed to initiate marker-assisted breeding for abiotic stress adaptation in peanut. The proof of concept about the use of transgenic technology to add value to peanuts has been demonstrated. Advances in phenomics and artificial intelligence to accelerate the timely and cost-effective collection of phenotyping data in large germplasm/breeding populations have also been discussed. Greater focus is needed to accelerate research on heat stress tolerance in peanut. A suits of technological innovations are now available in the breeders toolbox to enhance productivity and nutritional quality of peanuts in harsh environments. A holistic breeding approach that considers drought and heat-tolerant traits to simultaneously address both stresses could be a successful strategy to produce climate-resilient peanut genotypes with improved nutritional quality.

Anti-microbial Peptides against Methicillin-resistant Staphylococcus aureus: Promising Therapeutics.

BACKGROUND: Multidrug-resistant (MDR) methicillin-resistant Staphylococcus aureus (MRSA) has become a prime health concern globally. These bacteria are found in hospital areas where they are regularly dealing with antibiotics. This brings many possibilities for its mutation, so drug resistance occurs. INTRODUCTION: Nowadays, these nosocomial MRSA strains spread into the community and live stocks. Resistance in Staphylococcus aureus is due to mutations in their genetic elements. METHODS: As the bacteria become resistant to antibiotics, new approaches like antimicrobial peptides (AMPs) play a vital role and are more efficacious, economical, time, and energy saviours. RESULTS: Machine learning approaches of Artificial Intelligence are the in silico technique which has their importance in better prediction, analysis, and fetching of important details regarding AMPs. CONCLUSION: Anti-microbial peptides could be the next-generation solution to combat drug resistance among Superbugs. For better prediction and analysis, implementing the in silico technique is beneficial for fast and more accurate results.

Characterization and Anti-Cancerous Effect of Putranjiva roxburghii Seed Extract Mediated Silver Nanoparticles on Human Colon (HCT-116), Pancreatic (PANC-1) and Breast (MDA-MB 231) Cancer Cell Lines: A Comparative Study.

INTRODUCTION: A comparative study of Putranjiva roxburghii Wall. seed extract and developed silver nanoparticles (PJSNPs) for improving bioavailability that enhance their anti-cancer activity against HCT-116 (colon carcinoma), PANC-1 (pancreatic carcinoma), MDA-MB 231 (breast carcinoma) cell lines was performed. MATERIALS AND METHODS: The green synthesis of PJSNPs (Putranjiva silver nanoparticles) was performed using PJ (Putranjiva) extract, and characterization of synthesized nanoparticles was accomplished through UV-Vis spectrum, X-ray diffraction (XRD), transmission electron microscopy, energy-dispersive X-ray spectroscopy (TEM-EDAX), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), atomic force microscopy (AFM), and Raman spectroscopy. RESULTS: The results revealed that PJSNPs are homogeneous, spherical in shape, ~8±2 nm in size, and negatively charged with a zeta potential of about -26.71 mV. The cytotoxicity pattern observed was AgNO3 > PJSNPs > PJ extract. The morphological changes of the cells were observed by flow cytometry and also by the DNA ladder pattern on gel electrophoresis, which indicated that the process of cell death occurred via the apoptosis mechanism and PJSNPs were exerting late-stage apoptosis in all the tested cell lines. The small size and negative value of zeta potential could be the factors responsible for greater bioavailability and thus increased uptake by the tumor cells. CONCLUSION: The MTT assay and morphological changes observed by various methods indicate that the novel PJSNPs are a better anticancer agent than PJ extract. All the above properties make biologically synthesized PJSNPs an important target in the field of anti-cancer drug discovery.

Analysis of the effects of mEHT on the treatment-related toxicity and quality of life of HIV-positive cervical cancer patients.

Introduction: HIV infection is associated with increased treatment-related toxicity and worse outcomes in locally advanced cervical cancer patients (LACC), especially in resource-constrained settings. Local control (LC) in a phase III randomized, controlled trial investigating modulated electro-hyperthermia (mEHT) on LACC patients in South Africa (ethics registration: M120477/M190295), was significantly higher in participants randomized to receive chemoradiotherapy (CRT) with mEHT compared to CRT alone (stratum: HIV status, accounting for age and stage). This analysis investigates whether mEHT adds to the toxicity profile of CRT in HIV-positive LACC participants.Methods: Inclusion criteria: signed informed consent; International Federation of Gynecology and Obstetrics stages IIB to IIIB squamous cell carcinoma of the cervix; HIV-positive patients: CD4 count >200 cell/µL/on antiretroviral treatment for >6 months; eligible for CRT with radical intent. Recruitment: January 2014 to November 2017 (ClinicalTrials.gov: NCT03332069). Acute toxicity (evaluated using CTCAE v4 criteria) and quality of life (according to EORTC forms) in 206 participants randomized for treatment were evaluated alongside the LC results to determine safety and efficacy in HIV-positive participants.Results: Compliance to mEHT treatment was high (97% completed ≥8 treatments) with no significant differences in CRT-related toxicity between treatment groups or between HIV-positive and -negative participants. Adverse events attributed to mEHT were minor, even in obese patients, and did not affect CRT compliance. Participants treated with mEHT reported improved fatigue, pain, emotional and cognitive functioning.Conclusion: mEHT did not cause unexpected CRT-related toxicities and is a safe treatment modality for HIV-positive patients, with minor limitations regarding body weight, even in a low-resource setting.

The effect of modulated electro-hyperthermia on local disease control in HIV-positive and -negative cervical cancer women in South Africa: Early results from a phase III randomised controlled trial.

BACKGROUND: The global burden of cervical cancer remains high with the highest morbidity and mortality rates reported in developing countries. Hyperthermia as a chemo- and radiosensitiser has shown to improve treatment outcomes. This is an analysis of the local control results at six months post-treatment of patients enrolled in an ongoing study investigating the effects of the addition of modulated electro-hyperthermia (mEHT) to chemoradiotherapy for the treatment of HIV-positive and -negative cervical cancer patients in a low-resource setting. METHODS: This ongoing Phase III randomised controlled trial, conducted at a state hospital in Johannesburg, South Africa, was registered with the appropriate ethics committee. After signing an informed consent, participants with FIGO stages IIB to IIIB squamous cell carcinoma of the cervix were randomised to receive chemoradiotherapy with/without mEHT using a secure online random-sampling tool (stratum: HIV status) accounting for age and stage. Reporting physicians were blind to treatment allocation. HIV-positive participants on antiretroviral treatment, or with a CD4 count >200cell/µL were included. mEHT was administered 2/weekly immediately before external beam radiation. The primary end point is local disease control (LDC) and secondary endpoints are toxicity; quality of life analysis; and two year survival. We report on six month LDC, including nodes visualised in the radiation field on 18F-FDG PET/CT (censored for six month survival), and six month local disease free survival (LDFS) (based on intention to treat). Trial status: Recruitment closed (ClinicalTrials.gov: NCT03332069). RESULTS: 271 participants were recruited between January 2014 and November 2017, of which 210 were randomised for trial and 202 were available for analysis at six months post-treatment (mEHT: n = 101; Control: n = 101). Six month LDFS was higher in the mEHT Group (n = 39[38.6%]), than in the Control Group (n = 20[19.8%]); p = 0.003). LDC was also higher in the mEHT Group (n = 40[45.5%]) than the Control Group (n = 20[24.1%]); (p = 0.003). CONCLUSION: Our results show that mEHT is effective as a chemo-radiosensitiser for cervical cancer, even in high risk a patients and resource-constrained settings.

Evaluation of polyherbal ayurvedic formulation 'Peedantak Vati' for anti-inflammatory and analgesic properties.

ETHNOPHARMACOLOGICAL RELEVANCE: Peedantak Vati (PV) is a polyherbal ayurvedic formulation, which is regularly prescribed by the ayurvedic practitioner for the inflammatory disorders and joints pain in India. It is composed of 23 different herbs and minerals, described in ayurvedic text for their anti-inflammatory and analgesic properties. AIM OF THE STUDY: To investigate anti-inflammatory and anti-nociceptive potential of 'Peedantak Vati' using in vitro and in vivo methods. MATERIALS AND METHODS: In-vitro anti-inflammatory activity of PV was studied by estimating nitric oxide (NO) and LPS-induced pro-inflammatory cytokines IL-6 and TNF-α, using murine macrophage RAW264.7 and human monocyte THP-1 cell lines. PV's anti-inflammatory potential was studied in vivo using carrageenan-induced rat paw edema model. Similarly, anti-nociceptive property of PV was evaluated using hot plate, tail flick, formalin and writhing tests on CD-1 mice. Phytochemical profiling of hydro-alcoholic extract of PV was done using HPLC and HPTLC techniques to identify different marker compounds. These identified marker compounds were confirmed using LC-MS/MS analysis. RESULTS: In vitro results strongly suggest that, PV significantly (p < 0.001) inhibited NO release and LPS-stimulated pro-inflammatory cytokines IL-6 and TNF-α, in murine RAW264.7 and human THP-1 cells. Further, PV demonstrated significant (p < 0.05) anti-inflammatory activity at different time points after carrageenan injection with maximum effect at 2 h (40.4 ±â€¯5.2% at 400 mg/kg). Similarly, PV significantly (p < 0.05) decreased nociceptive pain, studied using hot plate, tail flick, formalin and writhing tests. Moreover, HPLC and HPTLC methods were developed for the standardization of PV. Five marker phytocompounds viz. rutin, caffeic acid, colchicine, withaferin A and curcumin were identified and quantified by HPLC and HPTLC methods. The presence of these phytoconstituents was confirmed by LC-MS/MS analysis. CONCLUSION: The findings of the study strongly suggest that, the polyherbal ayurvedic formulation 'Peedantak Vati' possesses remarkable anti-inflammatory and analgesic property, providing potent alternative for currently available allopathic medicines such as non steroidal anti-inflammatory drugs (NSAIDs).

Therapeutic Potential of the Medicinal Plant Aegle Marmelos (Linn.) Correa: Insight.

Various plant species are used globally for therapeutic purposes and have been authenticated by the World Health Organization. Aegle marmelos (L.) Corr., one of only three species in the genus Aegle, is a subtropical, fruit-bearing, deciduous tree that grows throughout the hills and plains of sub-Himalayan countries. Plants with medicinal importance have been used in almost every culture since ancient times. Various studies are underway to understand more about the qualities and components of medicinal plants, including drug preparation, phytochemical analysis, cultivation, toxicology, and pharmacology. According to the Indian conventional system of medicine, A. marmelos can successfully treat many diseases and conditions; for example, its extracts have been found to reduce the intensities of hepatic lipid peroxidation and augment the levels of hepatic antioxidants such as catalase, superoxide dismutase, and glutathione. The essential oils present in the foliage (leaves), fruits, and bark of A. marmelos provide strong antifungal action. A. marmelos is thus cytoprotective, works against ulcers and diarhea, promotes skin and bone healing, and acts as a hypoglycemic, antidiabetic, hepatoprotective, antimicrobial, analgesic, antiinflammatory, antipyretic, and anticancer agent. Such qualities have been well documented with scientific evidence.

Influence of cancer treatment on the Candida albicans isolated from the oral cavities of cancer patients.

PURPOSE: Cancer treatment causes mucositis and the manifestation of oral candidiasis. This study investigated the virulence properties and antifungal susceptibilities of Candida albicans isolated from cancer patients undergoing therapy. METHODS: C. albicans were isolated from 49 patients on cancer treatment and 21 healthy individuals and their virulence attributes measured. A correlation was determined between the length of treatment and the fungal counts and their virulence factors. RESULTS: Although Candida carriage was similar in all the study groups, high quantities of C. albicans and variety of Candida were found in cancer patients. Germ tubes were produced by all the strains. Significantly high number of yeast isolated from radiotherapy and chemotherapy produced large quantities of phospholipase compared to healthy individuals (p < 0.01). The length of chemotherapy was associated with an increase in the phospholipase production (p = 0.03) by the C. albicans. Proteinase production was seen in a significant number of isolates from the radiotherapy group (p < 0.01). Type of cancer treatment had no effect. Resistance to antifungal agents was low. CONCLUSIONS: High quantities of phospholipase were produced by C. albicans in cancer patients on therapy which also increased with the length of chemotherapy suggesting enhanced risk of oral and systemic infection. Therefore, during treatment, prophylactic topical antifungal therapy may be considered.

Different Advanced Therapeutic Approaches to Treat Vitiligo.

Vitiligo is a hypopigmentation disorder that is caused by the loss of melanocyte activity for melanin pigment generation. Vitiligo is distinguished by the existence of white macules. Vitiligo affects 0.1%-2% of individuals of different populations, irrespective of skin color, ethnic origin, race, or age. Although the actual mechanism behind this disease is not yet known, it is thought to be caused by a cumulative effect of various mechanisms (e.g., neurohormonal, genetic, cytotoxic, oxidative stress, autoimmune, and biochemical). This article reviews the published literature on various treatment modalities that might be effective in successfully treating patients with vitiligo, including phototherapies or some photochemotherapies, vitamin D analogs, topical and systemic corticosteroids, zinc treatment, anti-tumor necrosis factor agents, calcineurin inhibitors (tacrolimus, pimecrolimus), and surgical methods. This critical review also discusses a few herbal medications that may be worthy of future investigation because they have no significant side effects.

Quality control evaluation of Keshamasi, Keshanjana and Keshamasi eye ointment.

BACKGROUND: Keshanjana (collyrium) is a well known Ayurvedic preparation prepared out of Keshamasi (ash prepared by scalp hairs) mixed with Goghrita (cow's ghee). This medicine is indicated for the treatment of Shushkakshipaka (dry eye syndrome) in the classical literature of Ayurveda; hence, it was under taken for standardization and clinical evaluation in an extra-mural research project from Central Council for Research in Ayurvedic Sciences, Department of AYUSH, New Delhi. AIM: To develop standard quality parameters for the Keshamasi, Keshanjana and Keshamasi ointment. MATERIALS AND METHODS: Scalp hairs of male and females collected from saloons were converted to classical Masi Kalpana and mixed with cow ghee and petrolatum in the ratio of 1:5 to prepare the Keshanjana and Keshamasi ointment respectively. Standard Operation Procedure (SOP) were adopted and recorded accordingly. The raw material, furnished products and plain Goghrita were subjected for quality control parameters i.e., physico-chemical evaluation, anti-microbial study, particle size analysis, heavy metal analysis through inductive couple plasma spectroscopy with high performance thin layer liquid chromatography fingerprints. RESULTS: Rancidity was negative in all the samples, indicating that the physico-chemical parameters are in acceptable range. Lead and zinc were present in most of the samples; while all samples are were free from microbial contamination. CONCLUSION: As no standards are available to compare the results of the current study, the observations cannot be compared. Thus the profile generated in the current study can be considered as standard to refer in future studies.

Proteomic identification of Syzygium cumini seed extracts by MALDI-TOF/MS.

Syzygium cumini is traditionally used medicinal plant. The different part of the plant such as bark, leaves, seed and fruits are widely used as an alternative medicine in various diseases. Although the scientific community has a strong interest on S. cumini seed biochemistry focusing on metabolite composition, proteins have not yet been investigated. In the present study, we have applied a proteomic approach to study the proteome of the S. cumini seed using phenol extraction method for protein isolation, which were never analysed before. Fifteen brightly silver stained protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after resolving on two-dimensional gel electrophoresis. These proteins have been found to involve in various functions such as antifungal, sulphur metabolism, carbohydrate metabolism, fruit ripening and softening, dormancy breaking and seed germination, hormone signalling, secondary metabolite transport, defence and stress response, nitrogen metabolism, synthesis and stabilization. Amongst the identified protein, lactoferrin was a mammalian origin protein with high nutritious and pharmaceutical value, which was purified by different types of chromatographic techniques and confirmed by western blotting. The antibacterial activity of lactoferrin was assessed by disc diffusion assay. We suggest that the protein constituents of S. cumini may have role in various functions required for plant physiology and its dietary values.

Augmented primary humoral immune response and decreased cell-mediated immunity by Murraya koenigii in rats.

BACKGROUND: Murraya koenigii (Rutaceae) (curry patta: Hindi) of the family Rutaceae is used in the traditional Indian system of medicine for its immunomodulatory properties. The essential oil of the leaves of M. koenigii possesses antimicrobial, antifungal, and pesticidal activities and is used for the treatment of amebiasis, diabetes, and hepatitis. The present study was performed to evaluate the effect of M. koenigii on humoral and cell-mediated immune responses in rats. METHODS: Aqueous extract of M. koenigii leaves was administered orally in a dose of 350 mg/kg. Cell-mediated immunity was assessed by measuring foot pad thickness following sensitization by injection of keyhole limpet hemocyanin and subsequent challenge by the same. Humoral immunity was assessed by measurement of hemagglutination titer to sheep red blood cells (SRBCs). RESULTS: In the humoral immune response, the administration of M. koenigii [350 mg/kg per os (p.o.)] from day 1 to day 7 after sensitization with SRBC on day 0 caused a significant increase in the primary anti-SRBC titer. However, the secondary immune response was decreased significantly (p<0.05) as shown by a decrease in secondary anti-SRBC titer measured on day 11 following a booster dose of antigen on day 8. In the delayed-type hypersensitivity test, M. koenigii (350 mg/kg, p.o.), when administered for 14 days, produced a significant (p<0.05) decrease in foot pad thickness when compared with the control group. CONCLUSIONS: Thus, these results suggest that oral administration of M. koenigii augments primary humoral immune response and decreases cell-mediated immunity.

Hemostatic, antibacterial biopolymers from Acacia arabica (Lam.) Willd. and Moringa oleifera (Lam.) as potential wound dressing materials.

Acacia arabica and Moringa oleifera are credited with a number of medicinal properties. Traditionally gum of Acacia plant is used in the treatment of skin disorders to soothe skin rashes, soreness, inflammation and burns while Moringa seed extracts are known to have antibacterial activity. In the present study the potential of the polymeric component of aqueous extracts of gum acacia (GA) and the seeds of M. oleifera (MSP) in wound management was evaluated. The results revealed that both biopolymers were hemostatic and hasten blood coagulation. They showed shortening of activated partial thromboplastin time and prothrombin time and were non-cytotoxic in nature. Both showed antibacterial activity against organisms known to be involved in wound infections with MIC ranging from 500-600 microg mL(-1) for GA and 300-700 microg mL(-1) for MSP. They were biodegradable and exhibited water absorption capacity in the range of 415 to 935%. The hemostatic character coupled to these properties envisions their potential in preparation of dressings for bleeding and profusely exuding wounds. The biopolymers have been further analysed for their composition by Gas chromatography.

Jaggery protects hepatorenal injury induced by acute exposure to carbon tetrachloride in Wistar rats.

In this study, we evaluated the protective activity of aqueous extract of jaggery against CCl4-induced hepatic-renal damage in rats. Jaggery was administered in one group at doses of 250, 500 and 750 mg/ kg body weight (bwt) (p.o., once only), and CCL4 was administered in another group at a dose of 1.5 ml/kg bwt (i.p., once only) to evaluate the protective effect of jaggery on induced oxidative damage in rats. Various blood and tissue biochemical studies were performed. The administration of toxicant significantly altered blood biochemical variables. Hepatic and renal lipid peroxidation (LPO) levels increased significantly, whereas considerable depletion was observed in reduced glutathione (GSH) level after intoxication. A remarkable decrease was observed in the activities of adenosine triphosphatase (ATPase) and glucose-6-phosphatase (G-6-Pase) after induction of toxicity. Treatment with extract at three different altered all measured biochemical variables, but greater hepatic-renal protection was observed at higher doses (750 mg/kg bwt) than at lower does (250 and 500 mg/kg bwt). Jaggery also reversed histopathological alterations. Thus, it may be concluded that jaggery can be used to reduce hepatic and renal damage and may serve as an alternative medicine in hepatic and renal etiology.

Nephroprotective effect of jaggery against acute and subchronic toxicity of acetaminophen in Wistar rats.

The present investigation was planned to evaluate the nephroprotective activity of jaggery against acetaminophen (APAP)-induced renal damage in rats. The protective activity of jaggery at different doses (250, 500, and 750 mg/kg, orally) was evaluated against oxidative damage induced by APAP administration (2 g/kg, once orally in acute exposure; 20 mg/kg, orally for 21 days in subchronic exposure) in rats. APAP administration significantly increased the levels of serum urea, creatinine, and renal lipid peroxidation (LPO), whereas substantial decreases were observed in levels of glutathione (GSH), adenosine triphosphatase (ATPase), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPx) enzymatic activities after APAP administration. Administration of jaggery significantly moved the studied parameters toward normal levels and also reversed the histopathologic alterations. Thus, jaggery can be used to reduce renal damage and may serve as an alternative medicine in the treatment of renal etiologies.

Identification of indoline-2-thione analogs as novel potent inhibitors of α-melanocyte stimulating hormone induced melanogenesis.

Based on the hits, 3,4-dihydroquinazoline-2-thione (1) and benzimidazole-2-thione (2), a series of indole-2-thione (3) and indole-2-thiol inhibitors (4) of melanogenesis were designed, synthesized and evaluated in melanoma B16 cells under the stimulant of α-melanocyte stimulating hormone (α-MSH). The indole-2-thione compounds (3a-g) exhibited an effective inhibitory activity on melanin synthesis. The Structure-Activity Relationship (SAR) studies of 2 have revealed that in potent inhibitor 3a (>100% inhibition at 30 µM, IC50=1.40 µM) the role of nitrogen (3-N) at 3-position is insignificance. In addition, the hydrophobic substituents of 3 were better than the hydrophilic one. However, conversion of thione (-C=S, 3) to thiol (-C-SH, 4) led to decrease in the potency.

Genetic differentiation and diversity analysis of medicinal tree Syzygium cumini (Myrtaceae) from ecologically different regions of India.

This study represents the agro-ecological zone wise surveys of molecular variation of important medicinal tree Syzygium cumini Linn. (Jamun) which is native to India. It is used world wide in treatment of diabetes. Despite of its diverse medicinal properties no molecular data is available about the pattern of variation in its natural range. Populations of S. cumini in India are located in different habitats which differ from each other with regard to ecological factors. In this study, random amplified polymorphic DNA (RAPD) markers were used to detect inter and intra levels of genetic variations of sixteen S. cumini genotypes collected from three major agro-ecological zones of India. A total of 220 amplification products were scored of which 87.50 % were polymorphic. The level of polymorphism ranged from 47.69 % to 74.87 % polymorphic bands per population and was correlated with population size. Different measures of diversity: Shannon's index of phenotypic diversity (I) = 0.451 ± 0.230; Nei's genetic diversity (h) = 0.300 ± 0.172; effective number of alleles per locus (Ne) = 1.51 ± 0.347; total species diversity (Hsp) = 0.315 ± 0.031 and within population diversity (Hpop) = 0.158 ± 0.104 showed high genetic diversity at species level. Coefficient of genetic differentiation (Gst =0.498; Nm = 0.503) revealed significant genetic differentiation among the populations. Most of the genetic variations are contained among the populations. The results of cluster analysis and principal component analysis (PCA) give only little evidence for an ecotypic differentiation of S. cumini populations. Present genetic structure of population suggests ex situ conservation in seed banks in which seeds from at least five populations need to collected and conserved. Secondly, our study provides practical information to herbal drugs manufactures who use Jamun as a raw material.

Neoadjuvant chemotherapy followed by concurrent hyperfractionated radiation therapy and sensitizing chemotherapy for locally advanced (T3-T4) oropharyngeal squamous cell carcinoma.

BACKGROUND: Radiation therapy (RT) is commonly used in the management of patients with advanced (T3-T4) oropharyngeal squamous cell carcinomas. In recent years, based upon the meta-analyses of randomized trials, chemotherapy administered concurrently with RT (chemoradiotherapy) has become the standard of care. Twice-a-day hyperfractionated or accelerated-fractionated RT regimens have been shown in a number of randomized trials to significantly improve the rate of local control compared with conventional once-a-day fractionation. Concurrent chemotherapy administered along with hyperfractionated or accelerated RT has been shown to add significant additional benefit over hyperfractionated or accelerated RT alone. Neoadjuvant chemotherapy (usually consisting of cisplatin and fluorouracil) also produces favorable responses in most patients (approximately 75% partial or complete response rates) with advanced head and neck cancer, but its role remains controversial. METHODS: The results of treatment of 23 patients with T3 or T4 oropharyngeal squamous cell carcinomas who received neoadjuvant chemotherapy, followed by hyperfractionated RT (120 cGy twice-a-day to 74.4-76.8 Gy) were retrospectively reviewed. The 14 patients who were most recently treated also received concurrent sensitizing doses of single agent chemotherapy, usually cisplatin. No patient was seen with distant metastasis, and all were treated with curative intent. Ten patients had T3 and 13 patients had T4 primary tumors. Three patients (13%) had stage III disease and 20 patients (87%) had stage IV disease. Ten patients had base of tongue primaries, 12 had tonsillar primaries, and 1 had an oropharyngeal wall primary. Eighteen patients (78%) had clinically involved neck nodes. RESULTS: Seventy-four percent of patients had partial (>50%) or complete response at the primary site following neoadjuvant chemotherapy. One patient died of cardiorespiratory arrest after the first cycle. Thirteen percent of patients had unplanned interruptions of their RT courses secondary to severe mucositis. Local control at the primary site (minimum 2 years follow-up) was achieved in 17 of 19 (89%) patients. Two- and 5-year absolute survival rates were 71% and 55%, respectively. No patient was gastrostomy dependent beyond 18 months. Four patients developed neck failure after RT alone and none was successfully salvaged. CONCLUSION: This study is noteworthy in that it uses both neoadjuvant and concurrent sensitizing chemotherapy along with hyperfractionated RT. There is little information in the literature on this approach. Although the regimen is somewhat toxic, it is less so than many other regimens, which combined full-dose multiagent chemotherapy during the course of RT. The latter regimens also have a significant incidence of permanent dysphagia, gastrostomy dependence, and/or aspiration, complications that were not encountered in this group. The local control rate was high (89%). We continue to recommend this regimen for patients with locally advanced head and neck squamous cell carcinomas.