Delineating the inherent functional descriptors and biofunctionalities of pectic polysaccharides.

Pectin is a plant-based heteropolysaccharide macromolecule predominantly found in the cell wall of plants. Pectin is commercially extracted from apple pomace, citrus peels and sugar beet pulp and is widely used in the food industry as a stabilizer, emulsifier, encapsulant, and gelling agent. This review highlights various parameters considered important for describing the inherent properties and biofunctionalities of pectins in food systems. These inherent descriptors include monosaccharide composition, galacturonic acid content, degree of esterification, molecular weight, structural morphology, functional group analysis, and functional properties, such as water and oil holding capacity, emulsification, foaming capacity, foam stability, and viscosity. In this study, we also delineate their potential as a nutraceutical, prebiotic, and carrier for bioactive compounds. The biofunctionalities of pectin as an anticancer, antioxidant, lipid-lowering, and antidiabetic agent are also conceptually elaborated in the current review. The multidimensional characteristics of pectin make it a potential candidate for use in food and biomedical science.

Non-invasive subcutaneous fat reduction: a review.

The risks, financial costs and lengthy downtime associated with surgical procedures for fat reduction have led to the development of a number of non-invasive techniques. Non-invasive body contouring now represents the fastest growing area of aesthetic medicine. There are currently four leading non-invasive techniques for reducing localized subcutaneous adipose tissue: low-level laser therapy (LLLT), cryolipolysis, radio frequency (RF) and high-intensity focused ultrasound (HIFU). To review and compare leading techniques and clinical outcomes of non-invasive subcutaneous fat reduction. The terms 'non-invasive', 'low-level laser', 'cryolipolysis', 'ultrasound' and 'radio frequency' were combined with 'lipolysis', 'fat reduction' or 'body contour' during separate searches in the PubMed database. We identified 31 studies (27 prospective clinical studies and four retrospective chart reviews) with a total of 2937 patients that had been treated with LLLT (n = 1114), cryolipolysis (n = 706), HIFU (n = 843) or RF (n = 116) or other techniques (n = 158) for fat reduction or body contouring. A majority of these patients experienced significant and satisfying results without any serious adverse effects. The studies investigating these devices have all varied in treatment regimen, body locations, follow-up times or outcome operationalization. Each technique differs in offered advantages and severity of adverse effects. However, multiple non-invasive devices are safe and effective for circumferential reduction in local fat tissue by 2 cm or more across the abdomen, hips and thighs. Results are consistent and reproducible for each device and none are associated with any serious or permanent adverse effects.

Discovery of a potent inhibitor of replication protein a protein-protein interactions using a fragment-linking approach.

Replication protein A (RPA), the major eukaryotic single-stranded DNA (ssDNA)-binding protein, is involved in nearly all cellular DNA transactions. The RPA N-terminal domain (RPA70N) is a recruitment site for proteins involved in DNA-damage response and repair. Selective inhibition of these protein-protein interactions has the potential to inhibit the DNA-damage response and to sensitize cancer cells to DNA-damaging agents without affecting other functions of RPA. To discover a potent, selective inhibitor of the RPA70N protein-protein interactions to test this hypothesis, we used NMR spectroscopy to identify fragment hits that bind to two adjacent sites in the basic cleft of RPA70N. High-resolution X-ray crystal structures of RPA70N-ligand complexes revealed how these fragments bind to RPA and guided the design of linked compounds that simultaneously occupy both sites. We have synthesized linked molecules that bind to RPA70N with submicromolar affinity and minimal disruption of RPA's interaction with ssDNA.

Inhibition of osteoclasts prevents cartilage loss and pain in a rat model of degenerative joint disease.

OBJECTIVE: To investigate the relationship between efficacy of a bisphosphonate, pain and extent of joint damage in the monosodium iodoacetate (MIA) model of painful degenerative joint disease. METHODS: Zoledronate treatment was initiated prior to and at various times following model induction, including late time points representing advanced disease. Radiographic and histological structural parameters were correlated with pain as measured by weight bearing. RESULTS: Intraarticular (IA) MIA resulted in a progressive loss of bone mineral density (BMD) and chondrocytes, thinning of cartilage, loss of proteoglycan, resorption of calcified cartilage and subchondral bone, as well as pain. This was completely prevented by pre-emptive chronic zoledronate treatment with joint sections being histologically indistinguishable from saline-injected controls. When initiation of treatment was delayed efficacy was reduced. In animals with advanced joint degeneration, treatment partially restored BMD and had a significant, but limited, effect on pain. We confirmed these radiographic and behavioral findings in the medial meniscal tear model. To understand the mechanism-of-action of zoledronate we investigated an early time point 4 days post-model induction when chondrocytes were histologically viable, with minor loss of proteoglycan and generalized synovitis. Osteoclast-mediated resorption of the calcified cartilage was observed and was prevented by two doses of zoledronate. CONCLUSION: Subchondral bone remodeling plays an important role in nociception and the pathobiology of the MIA model with osteoclasts being implicated in both bone and cartilage resorption. Inhibition of osteoclastic activity when initiated early leads to improved efficacy.

Total synthesis and biological evaluation of the marine bromopyrrole alkaloid dispyrin: elucidation of discrete molecular targets with therapeutic potential.

The first total synthesis of dispyrin, a recently reported bromopyrrole alkaloid from Agelas dispar with an unprecedented bromopyrrole tyramine motif, was achieved in three steps on a gram scale (68.4% overall). No biological activity was reported for dispyrin, so we evaluated synthetic dispyrin against>200 discrete molecular targets in radioligand binding and functional assays. Unlike most marine natural products, dispyrin (1) possesses no antibacterial or anticancer activity, but was found to be a potent ligand and antagonist of several therapeutically relevant GPCRs, the alpha1D and alpha2A adrenergic receptors and the H2 and H3 histamine receptors.

Dissipation kinetics of spiromesifen on tea (Camellia sinensis) under tropical conditions.

Spiromesifen (Oberon) is a new insecticide and miticide of chemical class ketoenol active against white flies (Bemisia spp., Trialeuroides spp.) and spider mites (Tetranychus and Panonychus spp.). Due to its potential significance in insect resistance management, it is important to establish its behaviour on crop and environment. In the present study, the degradation/dissipation of spiromesifen on tea crop under tropical environmental conditions was studied and its DT(50) (t(1/2)), and DT(90) (time to reduce to 90% of the initial value) were estimated. Spiromesifen was sprayed on tea crop after first rain flush at four different locations @ 96 and 192ga.i.ha(-1). Samples of tea leaves were drawn at 0, 1, 3, 5, 7, 10, 15, 21 and 30 days after treatment and that of soil at 10 days after treatment and at harvest from 0 to 15 and 15 to 30cm layers. After crude extraction of tea leaves for spiromesifen residues with acetone:water, the contents were partitioned with cyclohexane:ethyl acetate and cleaned up on Florosil column. Soil residues were also extracted similarly. Quantification of residues was done on GC-MS in Selected Ion Monitoring (SIM) mode in mass range 271-274m/z. The LOQ of this method was found to be 0.05microgg(-1) while LOD being 0.015microgg(-1). The DT(50) of spiromesifen when applied at recommended doses in tea leaves was found to be 5.0-8.5 days. Ninety-nine percent degradation was found to occur within 33-57 days after application. In soil, no residues of spiromesifen were detectable 10 days after treatment.

Applicability of pectate-entrapped Lactobacillus casei cells for L(+) lactic acid production from whey.

Lactic acid is a versatile organic acid, which finds major application in the food, pharmaceuticals, and chemical industries. Microbial fermentation has the advantage that by choosing a strain of lactic acid bacteria producing only one of the isomers, an optically pure product can be obtained. The production of L: (+) lactic acid is of significant importance from nutritional viewpoint and finds greater use in food industry. In view of economic significance of immobilization technology over the free-cell system, immobilized preparation of Lactobacillus casei was employed in the present investigation to produce L: (+) lactic acid from whey medium. The process conditions for the immobilization of this bacterium using calcium pectate gel were optimized, and the developed cell system was found stable during whey fermentation to lactic acid. A high lactose conversion (94.37%) to lactic acid (32.95 g/l) was achieved with the developed immobilized system. The long-term viability of the pectate-entrapped bacterial cells was tested by reusing the immobilized bacterial biomass, and the entrapped bacterial cells showed no decrease in lactose conversion to lactic acid up to 16 batches, which proved its high stability and potential for commercial application.

Encapsulated lactic acid bacteria for control of malolactic fermentation in wine.

The kinetics of both malolactic fermentation in Chardonnay wine by encapsulating Lactobacillus casei cells in pectate gel and lyophilized Oenococcus oeni culture has been carried out. The influence of acidity, sulfur dioxide content, and organic acid content on the malolactic activity of the bacteria has been controlled. Encapsulated bacteria degraded 30%, of malic acid in white wine, deacidifying it from pH 3.15 to 3.40, whereas the lyophilized culture degraded 48% of malic acid, deacidifying from pH 3.15 to 3.60. The degree of conversion of malic acid in wine by the encapsulated cells was twice as high as that obtained by the free Lactobacillus casei cells. The operational stability of calcium pectate gel capsules was 6 months. It has been proved that the encapsulated biocatalyst increases the rate of fermentation, and induces the fermentation to take place at high ethanol concentrations. The proposed encapsulated biocatalyst is an attractive material for industrial applications in continuous winemaking processes.

The distinctive isotopic signature of plant-derived chloromethane: possible application in constraining the atmospheric chloromethane budget.

Chloromethane (CH(3)Cl) is the most abundant halocarbon in the atmosphere. Although largely of natural origin it is responsible for around 17% of chlorine-catalysed ozone destruction. Sources identified to date include biomass burning, oceanic emissions, wood-rotting fungi, higher plants and most recently tropical ferns. Current estimates reveal a shortfall of around 2 million ty(-1) in sources versus sinks for the halocarbon. It is possible that emissions from green plants have been substantially underestimated. A potentially valuable tool for validating emission flux estimates is comparison of the delta13C value of atmospheric CH(3)Cl with those of CH(3)Cl from the various sources. Here we report delta13C values for CH(3)Cl released by two species of tropical ferns and show that the isotopic signature of CH(3)Cl from pteridophytes like that of CH(3)Cl from higher plants is quite different from that of CH(3)Cl produced by biomass burning, fungi and industry. delta13C values for CH(3)Cl produced by Cyathea smithii and Angiopteris evecta were respectively -72.7 per thousand and -69.3 per thousand representing depletions relative to plant biomass of 42.3 per thousand and 43.4 per thousand. The characteristic isotopic signature of CH(3)Cl released by green plants should help constrain their contribution to the atmospheric burden when reliable delta13C values for all other major sources of CH(3)Cl are obtained and a globally averaged delta13C value for atmospheric CH(3)Cl is available.

Effect of genetic merit and concentrate supplementation on grass intake and milk production with Holstein Friesian dairy cows.

A total of 48 high genetic merit (HM) and 48 medium merit (MM) cows, each given a low (LC), medium (MC), or high (HC) level of concentrate supplementation, were used in a split-plot design experiment, which was run in three consecutive years, to evaluate animal production responses. Individual cow intakes were estimated twice each year while at pasture; measurement period 1 (MP1) was in May/June, and measurement period 2 (MP2) was in early September, corresponding on average to d 110 and 200 of lactation, respectively. In MP1, cows were offered 0 (LC), 3 (MC), and 6 kg (HC), whereas in MP2 the levels were 0 (LC), 0 (MC), and 4 kg (HC) of concentrate daily. Genotype had a significant effect on all milk production parameters in MP1 and MP2. The HM cows had the highest yield of milk, fat, protein, and lactose, whereas the MM cows had the highest milk fat, protein, and lactose concentrations. The HM cows had significantly higher grass dry matter intake (GDMI) estimates. In MP1, the average responses, per kg concentrate dry matter, was +1.10 kg of milk, +0.038 kg of protein, +0.032 kg of fat. The corresponding values in MP2 were +0.94 kg of milk, +0.037 kg of protein, and +0.025 kg of fat. The response to concentrate was linear and independent of preexperimental milk yield. In MP1, the partial regression coefficients relating daily GDMI to an increase in 1 kg of preexperimental milk yield (PMY), preexperimental BW (PBW), and concentrate intake (CI) were 0.123, 0.006, and -0.54, respectively, whereas the corresponding values in MP2 were 0.190,0.007, and-0.444, respectively. This study indicates that with high yielding dairy cows, on gras only GDMI of 17 kg of supporting milk yield of 30-kg/d is achievable. In this scenario, concentrate supplementation will result in lower substitution rates, and higher milk yield response than previously published with lower yielding cows.

Comparative efficacy and pharmacokinetics of racemic bupivacaine and S-bupivacaine in third molar surgery.

PURPOSE: To compare the efficacy and pharmacokinetics of racemic bupivacaine (rac-bupivacaine) with S-bupivacaine as primary local anesthetic agent in bilateral impacted third molar extractions. METHOD: A randomised, double blind, two period cross-over design was employed. Six subjects (2 males, 4 females; age 19-25 years; weight 69.2+/-9.4 kg) received bupivacaine hydrochloride injection (6.6 ml) as rac-bupivacaine (0.5% as salt) or S-bupivacaine (0.5% as base) prior to extraction of impacted third molars on one side and three weeks later on the other side. Anesthesia, blood loss associated with surgery and post-operative pain experience were evaluated. Plasma samples were analysed for bupivacaine enantiomers by chiral HPLC. RESULTS: In 7/12 operations, anesthesia adequate for surgery was delayed (>10 min) or unsatisfactory requiring lidocaine rescue medication. Despite this, there were no significant differences in onset and duration of anesthesia, blood loss or post-operative pain experience between the two arms of the study. Pharmacokinetic parameters were not significantly different and there was no evidence of chiral inversion after dosing with S-bupivacaine. CONCLUSIONS: Both study drugs were inadequate as single anesthetic agent for third molar surgery. Any decision to use S-bupivacaine for oral surgery must rest on evidence that it is less toxic than the racemic drug.

A current review of olanzapine's safety in the geriatric patient: from pre-clinical pharmacology to clinical data.

OBJECTIVE: Olanzapine (OLZ) is unique among currently available antipsychotic medications in its antagonism of a range of receptor systems including dopamine, norepinephrine, serotonin, acetylcholine, and histamine. Olanzapine's mechanistic complexity provides a broad efficacy profile in patients with schizophrenia and acute, pure or mixed mania. Patients experience symptomatic relief of mania, anxiety, hallucinations, delusions, and agitation/aggression and reduced depressive, negative, and some cognitive symptoms. This paper will review the safety profile of OLZ, focusing on the elderly, where data are available. METHOD: Preclinical and clinical studies of OLZ are reviewed, with emphasis on its possible effects on the cholinergic system and the histamine H(1) receptor. Weight change and related metabolic considerations, cardiac and cardiovascular safety, and motor function during treatment with OLZ are also reviewed. RESULTS AND CONCLUSION: In vitro receptor characterization methods, when done using physiologically relevant conditions allow accurate prediction of the relatively low rate of anticholinergic-like adverse events, extrapyramidal symptoms, and cardiovascular adverse events during treatment with OLZ. Currently available clinical data suggest olanzapine is predictably safe in treating adult patients of any age with schizophrenia and acute bipolar mania, as well as in treatment of patients with some types of neurodegenerative disorders.

Genetic dissection of atypical antipsychotic-induced weight gain: novel preliminary data on the pharmacogenetic puzzle.

Atypical antipsychotics such as clozapine represent a significant improvement over typical antipsychotics in the treatment of schizophrenia, particularly regarding extrapyramidal symptoms. Despite their benefits, use is limited by the occurrence of adverse reactions such as sedation and weight gain. This article provides a comprehensive review and discussion of obesity-related pathways and integrates these with the known mechanisms of atypical antipsychotic action to identify candidate molecules that may be disrupted during antipsychotic treatment. Novel preliminary data are presented to genetically dissect these obesity pathways and elucidate the genetic contribution of these candidate molecules to clozapine-induced weight gain. There is considerable variability among individuals with respect to the ability of clozapine to induce weight gain. Genetic predisposition to clozapine-induced weight gain has been suggested. Therefore, genetic variation in these candidate molecules may predict patient susceptibility to clozapine-induced weight gain. This hypothesis was tested for 10 genetic polymorphisms across 9 candidate genes, including the serotonin 2C, 2A, and 1A receptor genes (HTR2C/2A/1A); the histamine H1 and H2 receptor genes (H1R/H2R); the cytochrome P450 1A2 gene (CYPIA2); the beta3 and alpha,alpha-adrenergic receptor genes (ADRB3/ADRAIA); and tumor necrosis factor alpha (TNF-alpha). Prospective weight gain data were obtained for 80 patients with schizophrenia who completed a structured clozapine trial. Trends were observed for ADRB3, ADRA1A, TNF-alpha, and HTR2C; however, replication in larger, independent samples is required. Although in its infancy, psychiatric pharmacogenetics will in the future aid clinical practice in the prediction of response and side effects, such as antipsychotic-induced weight gain, and minimize the current "trial and error" approach to prescribing.

Dietary supplement use in U.S. Army Special Operations candidates.

We administered a dietary supplement survey to 2,215 males (mean age, 25 years; range, 18-47 years) entering U.S. Army Special Forces and Ranger training schools. The survey contained questions on demographics and health as well as the use of vitamin, mineral, protein, pro-performance, and other cofactor supplements; answers were made on a five-point frequency scale. Eighty-five percent of the men reported past or present use of a supplement, 64% reported current use, and 35% reported daily use. Individuals using a supplement at least occasionally were significantly less likely to smoke (p < 0.05) and more likely to exercise on a daily basis. Recent U.S. Department of Agriculture survey data (1994) reported that 39% of males 20 to 39 years old used a dietary supplement at least occasionally. Our data indicate that the rate of supplement use in this study group was much higher than in the general population of young men. This observation supports the need to study more extensively the use and benefits or potential harm of dietary supplementation by otherwise healthy individuals.

Comparison of the Efficacies of Chloromethane, Methionine, and S-Adenosylmethionine as Methyl Precursors in the Biosynthesis of Veratryl Alcohol and Related Compounds in Phanerochaete chrysosporium.

The effect on veratryl alcohol production of supplementing cultures of the lignin-degrading fungus Phanerochaete chrysosporium with different methyl-(sup2)H(inf3)-labelled methyl precursors has been investigated. Both chloromethane (CH(inf3)Cl) and l-methionine caused earlier initiation of veratryl alcohol biosynthesis, but S-adenosyl-l-methionine (SAM) retarded the formation of the compound. A high level of C(sup2)H(inf3) incorporation into both the 3- and 4-O-methyl groups of veratryl alcohol occurred when either l-[methyl-(sup2)H(inf3)]methionine or C(sup2)H(inf3)Cl was present, but no significant labelling was detected when S-adenosyl-l-[methyl-(sup2)H(inf3)]methionine was added. Incorporation of C(sup2)H(inf3) from C(sup2)H(inf3)Cl was strongly antagonized by the presence of unlabelled l-methionine; conversely, incorporation of C(sup2)H(inf3) from l-[methyl-(sup2)H(inf3)]methionine was reduced by CH(inf3)Cl. These results suggest that l-methionine is converted either directly or via an intermediate to CH(inf3)Cl, which is utilized as a methyl donor in veratryl alcohol biosynthesis. SAM is not an intermediate in the conversion of l-methionine to CH(inf3)Cl. In an attempt to identify the substrates for O methylation in the metabolic transformation of benzoic acid to veratryl alcohol, the relative activities of the SAM- and CH(inf3)Cl-dependent methylating systems on several possible intermediates were compared in whole mycelia by using isotopic techniques. 4-Hydroxybenzoic acid was a much better substrate for the CH(inf3)Cl-dependent methylation system than for the SAM-dependent system. The CH(inf3)Cl-dependent system also had significantly increased activities toward both isovanillic acid and vanillyl alcohol compared with the SAM-dependent system. On the basis of these results, it is proposed that the conversion of benzoic acid to veratryl alcohol involves para hydroxylation, methylation of 4-hydroxybenzoic acid, meta hydroxylation of 4-methoxybenzoic acid to form isovanillic acid, and methylation of isovanillic acid to yield veratric acid.

Voluntary, named testing for HIV in a community based antenatal clinic: a pilot study.

Despite the increasing advantages of identifying HIV infection in pregnant women, only some 12% of HIV positive women attending antenatal clinics in London have been identified by named testing. As virtually all antenatal care will be community based within the next two to three years, we assessed the problems of introducing named HIV testing during pregnancy into the primary care setting. Planning the service took a considerable time and required the production of educational material for both staff and pregnant women and some reorganisation of procedures. Over a one year period an uptake of 44% was noted. Several problems were encountered including an average of 21 minutes needed to give information on AIDS and HIV, an adverse effect on the midwife-mother relationship, and anxiety (affecting both women and midwives). Possible solutions to this difficult problem are discussed.

Effect of interleukin-1 beta, tumour necrosis factor-alpha and interleukin-6 on the control of thyrotropin secretion.

Since serum concentrations of tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), and interleukin-6 (IL-6) are elevated in infectious and inflammatory illnesses, we examined their potential role in contributing to the low TSH concentrations associated with such conditions, both at the level of the pituitary and the hypothalamus. 20 hours exposure to recombinant murine TNF-alpha (10(-11) to 10(-10) mol/l) enhanced the basal and the TRH-stimulated release of TSH by cultured rat anterior pituitary cells, but 4 hours exposure increased only basal TSH secretion. Recombinant human (rh) IL-1 beta, at a dose of 10(-11) mol/l only, produced a very small increase in basal TSH secretion after 4h, but not 20h, exposure. TRH-stimulated TSH secretion was not affected by IL-1 beta in concentrations up to 10(-10) mol/l, at either exposure time. Rh IL-6 (10(-12) to 10(-9) mol/l), had no effect on basal or TRH-stimulated TSH secretion at either exposure time. TNF-alpha, IL-1 beta, and IL-6 all failed to modify the inhibitory response to triiodothyronine (T3) and thyroxine (T4) on TSH secretion, under basal or TRH-stimulated conditions. Indirect effects of the cytokines on the stimulation or inhibition of TSH secretion, via TRH or SRIF respectively, were tested in isolated rat hypothalamic slices. 30 min exposure to TNF-alpha, IL-1 beta, or IL-6 had no effect on the basal release of SRIF. However, IL-1 beta, from 2.5 x 10(-12) to 10(-10) mol/l, produced a dose-dependent enhancement of the SRIF released by 5 x 10(-2) mol/l extracellular K+. The effect appeared to be mediated via IL-1 receptors, and to involve prostanoid formation, since it was inhibited by IL-1 receptor antagonist protein, 10(-7) mol/l, and indomethacin, 2.8 x 10(-5) mol/l, respectively. Neither basal nor K(+)-stimulated TRH release was influenced by TNF-alpha, IL-1 beta, or IL-6. The results indicate that direct effects of these cytokines on the pituitary do not contribute to reduced circulating TSH concentrations during inflammation and infection, but that enhanced hypothalamic release of SRIF, in response to elevated IL-1 beta, could contribute to such a decrease in TSH. None of the cytokines tested decreased hypothalamic TRH release in vitro. However, further in vivo experiments would be required to determine whether a longer exposure to these agents could reduce TRH release either directly, or indirectly via inputs from outside the hypothalamus.