Influence of patch geometry, post-bark-extraction-treatment on bark recovery and standardizing number of sprouts for bark harvest from coppices in Cinnamomum zeylanicum blume: implications for sustainable harvesting.

Though Cinnamomum zeylanicum is a very important commercial aromatic bark yielding and oldest known tree spice of India, there are hardly any studies to understand the nature of wound healing and bark recovery. Further, optimal number of sprouts to be retained under coppice system to maximize the bark yield per tree in C. zeylanicum is not yet standardized. The present investigation was carried out to understand the influence of patch geometry and application of post-bark-extraction-protection treatments on bark regeneration in mature trees as well as to standardize an optimal number of sprouts to be retained under coppice system to maximize the bark yield per tree. In general, wound healing in C. zeylanicum occurred from the edge of the blaze and was quicker in the narrower patch than the broader patch; application of Bordeaux paste or neem seed kernel extract on the blazed area immediately after the bark extraction, resulted in significantly higher mean percent bark recovery and higher bark oil content than control. Allowing seven coppice sprouts per stem resulted in higher dry mass of bark per plant and higher bark oil than other treatments under coppice system. The results of the study would enable formulation of management strategies specifically for sustainable bark harvesting in Cinnamomum species.

Isolation and characterization of microsatellite markers in Garcinia gummi-gutta by next-generation sequencing and cross-species amplification.

Garcinia gummi-gutta (L.) Roxb. (Clusiaceae) is an endemic, semidomesticated, fruit-yielding tree species distributed in the Western Ghats of India and Sri Lanka. Various bioactive phytochemicals, such as garcinol, benzophenones and xanthones are isolated from G. gummi-gutta and have shown antibacterial, antiviral and antioxidant activities. We sequenced the total genomic DNA using Illumina Hiseq 2000 platform and examined 241,141,804 bp high quality data, assembled into 773,889 contigs. In these contigs, 27,313 simple-sequence repeats (SSRs) were identified, among which mononucleotide repeats were predominant (44.98%) followed by dinucleotide and trinucleotide repeats. Primers were designed for 9964 microsatellites among which 32 randomly selected SSR primer pairs were standardized for amplification. Polymerase chain reaction (PCR) amplification of genomic DNA in 30 G. gummi-gutta genotypes revealed polymorphic information content (PIC) across all 32 loci ranging from 0.867 to 0.951, with a mean value of 0.917. The observed and expected heterozygosity ranged from 0.00 to 0.63 and 0.896 to 0.974, respectively. Alleles per locus ranged from 12 to 27. This is the first report on the development of genomic SSR markers in G. gummi-gutta using next-generation sequencing technology. The genomic SSR markers developed in this study will be useful in identification, mapping, diversity and breeding studies.

Developmental temperature affects the expression of ejaculatory traits and the outcome of sperm competition in Callosobruchus maculatus.

The outcome of post-copulatory sexual selection is determined by a complex set of interactions between the primary reproductive traits of two or more males and their interactions with the reproductive traits of the female. Recently, a number of studies have shown the primary reproductive traits of both males and females express phenotypic plasticity in response to the thermal environment experienced during ontogeny. However, how plasticity in these traits affects the dynamics of sperm competition remains largely unknown. Here, we demonstrate plasticity in testes size, sperm size and sperm number in response to developmental temperature in the bruchid beetle Callosobruchus maculatus. Males reared at the highest temperature eclosed at the smallest body size and had the smallest absolute and relative testes size. Males reared at both the high- and low-temperature extremes produced both fewer and smaller sperm than males reared at intermediate temperatures. In the absence of sperm competition, developmental temperature had no effect on male fertility. However, under conditions of sperm competition, males reared at either temperature extreme were less competitive in terms of sperm offence (P(2)), whereas those reared at the lowest temperature were less competitive in terms of sperm defence (P(1)). This suggests the developmental pathways that regulate the phenotypic expression of these ejaculatory traits are subject to both natural and sexual selection: natural selection in the pre-ejaculatory environment and sexual selection in the post-ejaculatory environment. In nature, thermal heterogeneity during development is commonplace. Therefore, we suggest the interplay between ecology and development represents an important, yet hitherto underestimated component of male fitness via post-copulatory sexual selection.

Rohitukine, a chromone alkaloid and a precursor of flavopiridol, is produced by endophytic fungi isolated from Dysoxylum binectariferum Hook.f and Amoora rohituka (Roxb).Wight & Arn.

Rohitukine, a chromone alkaloid, has gained considerable international attention in recent years because of its novel semi-synthetic derivative, flavopiridol and P-276-00. Both these molecules are in advanced stages of clinical development and trial for cancer treatment. Recently, flavopiridol was approved as an orphan drug for treatment of chronic lymphocytic leukemia cancer. The natural occurrence of rohitukine is restricted to only four plant species, Amoora rohituka and Dysoxylum binectariferum (both from the Meliaceae family) and from Schumanniophyton magnificum and Schumanniophyton problematicum (both from the Rubiaceae family). Recently, an endophytic fungi isolated from D. binectariferum was reported to produce rohitukine in culture. In this study, we report the production of rohitukine and its subsequent attenuation by endophytic fungi, Fusarium oxysporum (MTCC-11383), Fusarium oxysporum (MTCC-11384) and Fusarium solani (MTCC-11385), all isolated from D. binectariferum and Gibberella fujikuroi (MTCC-11382) isolated from Amoora rohituka. The fungal rohitukine which was analyzed by HPLC, LC-MS and LC-MS/MS was identical to reference rohitukine and that produced by the plant. The rohitukine content in the mycelial samples ranged from 192.78µg to 359.55µg100g(-1) of dry weight of and in broth it ranged from 14.10 to 71.90µg100ml(-1). In all the fungal cultures, the production declined from first to fourth sub-culture. Studies are underway to unravel the mechanism by which the fungi produce the host metabolite in culture.

Cutaneous benign epithelioid peripheral nerve sheath tumour: a rare entity.

Benign epithelioid peripheral nerve sheath tumor, a rare entity is an umbrella term describing benign, neural origin tumors with epithelioid morphology. Clinically indistinguishable from other benign cutaneous lesions, histopathology offers the only source of accurate diagnosis. Morphologic mimics include many benign and malignant soft tissue lesions. Besides a predominant epithelioid component, the lesion can also show a fair share of spindle cells. A circumscribed nodular tumour of low mitotic activity, it often exhibits areas resembling schwannoma or neurofibroma. An awareness of this entity and its varied morphological aspects helps to arrive at the correct diagnosis and hence avoid unnecessary extensive surgical procedures. This case presents features of this benign tumor which occurred in a 47 years old man.

New plant sources of the anti-cancer alkaloid, camptothecine from the Icacinaceae taxa, India.

In this study, the production of camptothecine and its derivatives, in thirteen species of the family Icacinaceae, namely, Apodytes dimidiata, Codiocarpus andamanicus, Gomphandra comosa, Gomphandra coriacea, Gomphandra polymorpha, Gomphandra tetrandra, Iodes cirrhosa, Iodes hookeriana, Miquelia dentata, Miquelia kleinii, Natsiatum herpeticum, Pyrenacantha volubilis and Sarcostigma kleinii is reported. Seeds of M. dentata were found to produce the highest content of camptothecine (1.0-1.4% by dry weight of seeds). Full scan LC-MS and ESI-MS/MS analysis of M. dentata revealed, besides camptothecine, a number of other derivatives, namely, 10-hydroxycamptothecine, 9-methoxycamptothecine, 20-deoxycamptothecine. Crude extract preparations of the seeds of M. dentata were effective against a breast cancer cell line (IC50=3.82 µg/ml for MDA MB273 cell lines) and two ovarian cancer cell lines (IC50=2.8 µg/ml for NCI/ADR-RES and 4.5 µg/ml for SKOV). These results are the first reports of camptothecine and its derivatives in these species and offer rich alternative plant sources for the anticancer compound, camptothecine.

Endophytic fungal strains of Fusarium solani, from Apodytes dimidiata E. Mey. ex Arn (Icacinaceae) produce camptothecin, 10-hydroxycamptothecin and 9-methoxycamptothecin.

Camptothecin and 10-hydroxycamptothecin are two important precursors for the synthesis of the clinically useful anticancer drugs, topotecan and irinotecan. In recent years, efforts have been made to identify novel plant and endophytic fungal sources of camptothecin and 10-hydroxycamptothecin. In this study we have isolated endophytic fungi strains from Apodytes dimidiata (Icacinaceae), a medium sized tree from the Western Ghats, India. The fungi were identified as Fusarium solani using both ITS rDNA sequencing and spore morphology. Two strains, MTCC 9667 and MTCC 9668 were isolated, both of which produced camptothecin and 9-methoxycamptothecin in their mycelia; one of the strains, MTCC 9668 also produced 10-hydroxycamptothecin, though in small amounts. The yields of camptothecin in MTCC 9667 and MTCC 9668 were 37 and 53 microg/100g, respectively, after 4 days of incubation in broth culture. The yields of 10-hydroxycamptothecin and 9-methoxycamptothecin in MTCC 9668 were 8.2 and 44.9 microg/100g, respectively. Further research in optimizing the culture conditions of these fungal strains might permit their application for the production of camptothecin and 10-hydroxycamptothecin.

Development of polymorphic microsatellite loci in Nothapodytes nimmoniana, a medicinally important tree from the Western Ghats, India.

Nothapodytes nimmoniana is a medicinally important tree species that occur in the Western Ghats, a megadiversity hotspot in southern India. Inner stem bark of the tree contains an important anti-cancer alkaloid, camptothecin for which the natural population of the tree is heavily harvested. In this paper, we report the isolation and characterization of eight polymorphic microsatellite loci using enrichment hybridization protocol. Analysis of 36 individuals representing two populations revealed three to 12 alleles per locus. Observed heterozygosity ranged from 0.21 to 0.94 for the two populations. None of the loci tested showed linkage disequilibrium. These markers are invaluable for evaluating the genetic structure and assessing the genetic impacts of harvesting of N. nimmoniana in the Western Ghats to formulate strategies for conservation of the species.

Dispersion of circumferential waves in cylindrically anisotropic layered pipes in plane strain.

Dispersion spectra of circumferential waves along the periphery of circular pipes made of layered anisotropic materials do not seem to be available in literature. This note attempts to partially fill this gap by providing the dispersion spectra in two and three layered cylindrically anisotropic pipes in plane strain motion. The spectra for pipes executing time harmonic vibrations in plane strain condition are obtained as roots of a numerical characteristic equation derived extending a weighted residual method of solution of the governing equations for a single layer pipe [Towfighi et al., J. Appl. Mech. 69, 283-291 (2002)] to a general N layered pipe. The anisotropic elastic coefficients are considered to be independent of position coordinates and the bond condition at interfaces of the layers is assumed to be perfect. Numerical illustrations are presented for two and three layered pipes with anisotropy directions differing in adjacent layers. Increase in curvature of the pipe and inclination of the fiber orientation in the outermost layers to propagation direction are factors that seem to influence the mode number and pattern within the limited examples worked out.

Prospecting for Camptothecines from Nothapodytes nimmoniana in the Western Ghats, South India: identification of high-yielding sources of camptothecin and new families of camptothecines.

Camptothecin (CPT), a monoterpene alkaloid, is an important anti-cancer compound obtained from several plant sources including Camptotheca acuminta (from China) and Nothapodytes nimmoniana (from India). Currently, by far the highest levels of CPT (approximately 0.3% w/w) are reported from Nothapodytes nimmoniana, a small tree distributed in the Western Ghats, India. In recent years because of the heavy demand, there has been a serious threat of extinction of the populations of the tree in the Western Ghats forest of south India. Several studies have chemically profiled populations of the species in the Western Ghats to identify sources of high yield and therefore to enable the sustainable production and harvesting of CPT. In this study, using both high-performance liquid chromatography and liquid chromatography-mass spectrometry, we report for the first time the identification of trees that produce at least 5- to 8-fold more CPT than hitherto reported. Furthermore, we show for the first time the production of a few minor camptothecines, including 10-hydroxy camptothecin, in the stem and root bark extracts of the tree. These results have important implications for not only harnessing the high-yielding individuals for clonal multiplication but also for exploiting some of the minor camptothecines, which also have been shown to have important anti-cancer and anti-viral activity.

The effect of stimulant laxatives and polyethylene glycol-electrolyte lavage solution for colonoscopy preparation on serum electrolytes and hemodynamics.

We have previously shown that magnesium citrate or bisacodyl before polyethylene glycol electrolyte lavage solution (PEG-ELS) improves colonoscopy preparation. Patients prefer this to preparation with full-volume PEG-ELS alone. However, such combinations might cause undesirable changes in hemodynamics or serum electrolytes. This study examined the effects of these combinations on heart rate, blood pressure, and serum electrolytes. We randomized 68 consecutive patients undergoing colonoscopy to one of three preparations. Group 1 received 4 L of PEG-ELS, group 2 received 2 L of PEG-ELS with 20 mg bisacodyl, and group 3 received 2 L of PEG-ELS preceded by 296 mL of magnesium citrate. Heart rate, blood pressure, and serum electrolytes were measured before starting the preparation and immediately before colonoscopy. Mean serum magnesium fell by 0.01 mg/dL and 0.11 mg/dL in groups 1 and 2, respectively, and rose by 0.06 mg/dL in group 3 (p = 0.044). There were no other significant changes. Statistically significant changes in serum magnesium after preparation with PEG-ELS and a stimulant laxative are minor and probably of no clinical relevance. Two liters of PEG-ELS with a stimulant laxative is safe and effective for colonoscopy.

Colorectal cancer screening and surveillance practices by primary care physicians: results of a national survey.

OBJECTIVE: Screening for colorectal cancer reduces its morbidity and mortality and is cost-effective. Screening is usually the responsibility of primary care physicians who may be unsure about its implementation. We aimed to assess primary care physicians' knowledge and practice regarding colorectal cancer screening, and to compare their responses with those of three national experts and with published guidelines. METHODS: We mailed a postal questionnaire to 2,310 primary care physicians regarding demographics, nature of practice, use of screening tests, and six hypothetical patients who may have been candidates for screening or surveillance. We used published national guidelines and the collective opinions of the three external experts as the so-called "gold standard." RESULTS: Of all respondents, 85.1% offered colorectal cancer screening. Most used suitable tests, starting at an appropriate age; 49.8% continued screening indefinitely irrespective of patients' age and 43.6% performed fecal occult blood testing without appropriate dietary advice to patients. Also, respondents frequently performed this test for inappropriate indications. Only 51.8% would follow a positive fecal occult blood test with colonoscopy. CONCLUSIONS: Colorectal cancer screening practices by primary care physicians vary considerably from those recommended. Many offer screening to individuals in whom it is not appropriate, and continue it into advanced age. Frequent, inappropriate use of fecal occult blood tests will produce many false positives. Primary care physicians often do not appropriately follow a positive test. Further educational efforts are needed in an attempt to improve practice and further reduce the morbidity and mortality from colorectal cancer.

Ultrasound-assisted direct percutaneous endoscopic jejunostomy (DPEJ) tube placement.

Direct percutaneous endoscopic jejunostomy (DPEJ) is an effective method of enteral feeding. However, failure rates with this procedure remain high due to various technical problems. We describe a case where modifications in the technique and ultrasound guidance assisted in a difficult DPEJ placement. This technique has the potential to improve the success rate of this procedure in selected cases.

Changes in colorectal cancer over a 15-year period in a single United States city.

OBJECTIVE: The aim of this study was to identify colorectal cancer trends in different patient groups in one region. METHODS: We reviewed the records of all patients with colorectal cancer who were seen at two hospitals in Columbia, SC, between 1981 and 1995. Patients were divided into three 5-yr cohorts (cohort 1, 1981-1985; cohort 2, 1986-1990; and cohort 3, 1991-1995). We recorded demographic data, histological type, tumor stage, grade, and survival. RESULTS: A total of 1395 patients were diagnosed, of whom 1252 had adenocarcinoma. Mean age of diagnosis was 64.5 yr in black and 66 in white patients (p = 0.036). Adenocarcinoma proximal to the splenic flexure was found in 62% of women and 49% of men (p < 0.001), and in 51% of black and 44% of white patients (p = 0.08). Proximal cancers accounted for 61%, 48%, and 32% of cohorts 1, 2, and 3, respectively. The proportions of patients with advanced stage (III and IV) adenocarcinoma in the three cohorts were 54%, 47%, and 40%, respectively (p = 0.001). Median survival for adenocarcinoma was 23.7, 23.5, and 23.1 months in cohorts 1, 2, and 3 (p = 0.9). CONCLUSIONS: The prevalence and demographics of colorectal cancer have been fairly stable at this center. There has been a significant decrease in proximal cancers. Although adenocarcinoma is being diagnosed at an earlier stage, this has not affected survival. Colonoscopy may be the most appropriate screening method in women and in black patients, in whom proximal cancers are more often found.

A survey of gastroenterologists' perceptions and practices related to Helicobacter pylori infection.

OBJECTIVE: The aim of this study was to assess the current practice of gastroenterologists in the United States concerning Helicobacter pylori (H. pylori) infection. METHODS: We mailed a structured questionnaire to 1000 gastroenterologists chosen at random from a national database. We asked about personal and practice demographics and practices relating to testing for, and treating, H. pylori infection. RESULTS: A total of 922 questionnaires were delivered, from which we received 286 responses (31%). Respondents used many different tests for H. pylori infection, but only 10% each had used either the 13C- or 14C-urea breath test. Testing for H. pylori infection was usually for appropriate reasons, although 21% indicated that they might not treat a patient with a positive test result. Different multiple treatment regimens were used; the most frequent were combinations of a proton pump inhibitor, clarithromycin, and either amoxicillin or metronidazole. Estimates of the prevalence of antibiotic resistance were highly variable and often inaccurate. Most respondents would not check asymptomatic individuals for the infection; however, in the absence of symptoms, 38% would personally undergo testing and treatment if positive. CONCLUSIONS: Gastroenterologists usually test for H. pylori infection in appropriate conditions, but may not always treat the infection based on a positive test result. Most use efficacious regimens to treat the infection although many have inaccurate information on resistance rates, which may adversely influence prescribing. Many would have testing and, if positive, treatment in the absence of symptoms or a specific diagnosis, but do not recommend this for their patients.

The effects on intragastric acidity of per-gastrostomy administration of an alkaline suspension of omeprazole.

BACKGROUND: It may be difficult to administer proton pump inhibitors via gastrostomy. Previous studies have examined the effect of intact proton pump inhibitor granules in orange juice. This study examined the effect of an alkaline suspension of omeprazole (simplified omeprazole suspension (SOS)) on 24-h intragastric acidity. METHODS: Six men with an established gastrostomy had a baseline 24-h intragastric pH study using methodology we have previously described. They then received 20 mg SOS o.d. for 7 days and had a repeat pH study at the end of this period. Four of the patients then received 20 mg SOS with 30 cc of liquid antacid (Mylanta) per gastrostomy o.d. for a further 7 days and then underwent a third pH study. RESULTS: SOS raised mean pH from 2.2 to 4.1. Intragastric pH was above 3, 4 and 5 for 35, 28 and 17% of the 24-h period at baseline, respectively; corresponding values after SOS were 63, 51 and 39%, respectively. Addition of liquid antacid to SOS did not further increase its pH-controlling effect. CONCLUSIONS: We found a statistically significant effect of o.d. SOS on intragastric pH when administered via gastrostomy. We found no additional benefit of administering SOS with liquid antacid.

Simplified lansoprazole suspension--a liquid formulation of lansoprazole--effectively suppresses intragastric acidity when administered through a gastrostomy.

OBJECTIVE: Lansoprazole suppresses intragastric acidity when given as nonencapsulated intact granules. Because the administration of granules via small-bore tubes may still be problematic, we studied the effect of a liquid formulation of lansoprazole obtained by suspending the contents of a standard 30-mg capsule in 10 cc of 8.4% NaHCO3. METHODS: Six men with an established gastrostomy had a baseline 24-h intragastric pH study. Through the gastrostomy, they then received 7 days of once-daily dosing with 30 mg lansoprazole as intact granules in orange juice. After a 7-day washout period, they then received 7 days of once-daily dosing with the liquid formulation. Intragastric pH monitoring was repeated after each dosing period. RESULTS: Baseline mean intragastric pH was 1.8 +/- 0.5 (SD). This increased to 4.5 +/- 0.5 with lansoprazole granules in orange juice, and to 5.1 +/- 1.1 after the liquid formulation. At baseline, intragastric pH was >3, 4, and 5 for 19.5%, 12.7%, and 8.1%, respectively, of the 24-h recording period. Corresponding values after lansoprazole granules in orange juice were 77.5%, 67%, and 49.6% (p < 0.01 for each comparison with baseline). After the liquid formulation, the corresponding values were 84%, 77.9%, and 65.9% respectively (p < 0.01 for each comparison with baseline; p > 0.05 for each comparison with lansoprazole granules in orange juice). CONCLUSIONS: A liquid formulation of lansoprazole suppressed intragastric acidity when given through a gastrostomy. The degree of suppression was comparable to that obtained with intact nonencapsulated granules in orange juice.