A simple, precise, rapid and accurate, binary-phase high performance liquid chromatographic method has been developed for the determination of oleanolic acid and ursolic acid contents in the Ziziphora clinopodioides Lam. with short run time. Chromatographic separation is achieved by using HPLC system consisting of a Shimadzu LC-6AD and Kromasil C(18) column (150 x 4.6 mm, 10 mum, with pre-column), the mobile phase consists of methanol and 0.03 M phosphate buffer (pH = 3, 90:10). Detection wavelength is 214 nm. The speed of flow is 0.5 ml/min. The specimen handing quantity is 10 mul. The oleanolic acid's linearity range is 0.4 ~ 1.2 mg/ml (r = 0.9996). The ursolic acid's linearity range is 0.6 ~ 1.8 mg/ml (r = 0.9996), and the linear relationship is accurate. The average recovery (n = 6) of oleanolic acid is 99.5% (RSD = 1.19%) and ursolic acid is 102.3% (RSD = 1.25%). The content of oleanolic acid and ursolic acid in Ziziphora clinopodioides are 0.76 mg/g and 1.176 mg/g, respectively. The developed HPLC method can therefore be applied to both in vitro studies of oleanolic acid and ursolic acid formulations as well as drug estimation in biological samples.

Hydrocarbon phytoremediation by Cyperus laxus Lam. growing on perlite and inoculated with hydrocarbon-degrading microorganisms was evaluated. Total petroleum hydrocarbons (TPH) were extracted from weathered soil (60.7 g of TPH kg(-1) of dry soil) and spiked on perlite at initial concentration of 5 g of TPH kg(-1) of dry perlite. Phenological characteristics, total microbial viable counts, hydrocarbon degraders and residual hydrocarbons were determined through 180 days of culture. Phenological characteristics of inoculated plants were improved as compared with non-inoculated plants: root biomass was 1.6 times greater, flowering time was reduced (13%), and the number of inflorescences was 1.5 times higher. The rhizospheric bacterial and fungi counts were higher for planted treatments (inoculated and not inoculated) than for unplanted pots. The maximum phytoremediation rate (0.51 mg of TPH g(-1) of dry plant d(-1)) for inoculated plants was reached at 60 days of culture, and was two times higher than for non-inoculated plants (55% TPH removal). Similar hydrocarbon phytoremediation extent values for inoculated (90%) and non-inoculated (85%) plants were obtained at 180 days of culture. The present study demonstrated that mutual benefits between C. laxus and inoculated hydrocarbon-degrading microorganisms are improved during phytoremediation. It is pertinent to note that this is the first report of hydrocarbon phytoremediation by Cyperus laxus Lam., a native plant growing in highly contaminated swamps.

This work aimed to study the process of stress adaptation in root and leaves of different developmental stages (apex, middle and basal regions) of Pluchea sagittalis (Lam.) Cabrera plants grown under exposure to five Pb levels (0, 200, 400, 600 and 1000 μM) for 30 days. Pb concentration and content in roots, stems, and leaves of different developmental stages increased with external Pb level. Consumption of nutrient solution, transpiration ratio, leaf fresh weight, leaf area, and shoot length decreased upon addition of Pb treatments. However, dry weight of shoot parts and roots did not decrease upon addition of Pb treatments. Based on index of tolerance, the roots were much more tolerant to Pb than shoots. δ-aminolevulinic acid dehydratase activity was decreased by Pb treatments, whereas carotenoid and chlorophyll concentrations were not affected. Lipid peroxidation and hydrogen peroxide concentration both in roots and leaves increased with increasing Pb levels. Pb treatments increased ascorbate peroxidase activity in all plant parts, while superoxide dismutase activity increased in leaves and did not change in roots. Catalase activity in leaves from the apex shoot was not affected by Pb, but in other plant parts it was increased. Pb toxicity caused increase in non-protein thiol groups concentration in shoot parts, whereas no significant difference was observed in roots. Both root and shoot ascorbic acid concentration increased with increasing Pb level. Therefore, it seems that Pb stress triggered an efficient defense mechanism against oxidative stress in P. sagittalis but its magnitude was depending on the plant organ and of their physiological status. In addition, these results suggest that P. sagittalis is Pb-tolerant. In conclusion, P. sagittalis is able to accumulate on average 6730 and 550 μg Pb g(-1) dry weight, respectively, in the roots and shoot, a physiological trait which may be exploited for the phytoremediation of contaminated soils and waters.

We report the unique association of a clear cell "sugar" tumor of the lung (CCTL) in a 32-year-old woman with tuberous sclerosis (TSC), lymphangioleiomyomatosis (LAM), and multifocal micronodular pneumocyte hyperplasia (MMPH). Chest radiographs demonstrated a peripheral solitary 1.0-cm lingular nodule, diffuse emphysematous changes, and bilateral pneumothorax. Microscopic examination of the coin lesion showed an unencapsulated tumor composed of round to oval to focally spindled cells with distinct cellular borders, abundant clear to eosinophilic granular cytoplasm, prominent thin-walled blood channels, and focal hyaline stroma. Rare multinucleated cells were identified, and neither necrosis nor mitotic figures were seen. Tumor cells contained abundant diastase-sensitive intracytoplasmic glycogen, as demonstrated with periodic acid-Schiff stains. Tumor cell immunoreactivity for HMB-45 and nonreactivity for cytokeratin supported the diagnosis. The lung tissue also contained MMPH and smooth muscle proliferations diagnostic of LAM. The histogenesis of CCTL remains controversial, and similarities between this lesion and both LAM and angiomyolipoma (AML) raise the possibility that these lesions are related not only to each other, but that CCTL should be added to the spectrum of pulmonary manifestations of TSC.

BACKGROUND

Tuberculosis is a major health problem in São Paulo, Brazil, which is the most populous and one of the most cosmopolitan cities in South America. To characterize the genetic diversity of Mycobacterium tuberculosis in the population of this city, the genotyping techniques of spoligotyping and MIRU were applied to 93 isolates collected in two consecutive years from 93 different tuberculosis patients residing in São Paulo city and attending the Clemente Ferreira Institute (the reference clinic for the treatment of tuberculosis).

RESULTS

Spoligotyping generated 53 different spoligotype patterns. Fifty-one isolates (54.8%) were grouped into 13 spoligotyping clusters. Seventy- two strains (77.4%) showed spoligotypes described in the international databases (SpolDB4, SITVIT), and 21 (22.6%) showed unidentified patterns. The most frequent spoligotype families were Latin American Mediterranean (LAM) (26 isolates), followed by the T family (24 isolates) and Haarlem (H) (11 isolates), which together accounted for 65.4% of all the isolates. These three families represent the major genotypes found in Africa, Central America, South America and Europe. Six Spoligo-International-types (designated SITs by the database) comprised 51.8% (37/72) of all the identified spoligotypes (SIT53, SIT50, SIT42, SIT60, SIT17 and SIT1). Other SITs found in this study indicated the great genetic diversity of M. tuberculosis, reflecting the remarkable ethnic diversity of São Paulo city inhabitants. The MIRU technique was more discriminatory and did not identify any genetic clusters with 100% similarity among the 93 isolates. The allelic analysis showed that MIRU loci 26, 40, 23 and 10 were the most discriminatory. When MIRU and spoligotyping techniques were combined, all isolates grouped in the 13 spoligotyping clusters were separated.

CONCLUSIONS

Our data indicated the genomic stability of over 50% of spoligotypes identified in São Paulo and the great genetic diversity of M. tuberculosis isolates in the remaining SITs, reflecting the large ethnic mix of the São Paulo city inhabitants. The results also indicated that in this city, M. tuberculosis isolates acquired drug resistance independently of genotype and that resistance was more dependent on the selective pressure of treatment failure and the environmental circumstances of patients.

SKG mice are a recently established experimental model for rheumatoid arthritis (RA). Although they spontaneously develop chronic autoimmune arthritis under conventional conditions, SKG mice failed to develop chronic arthritis in a strictly controlled specific pathogen-free (SPF) environment. Beta-glucan (BG) from Laminaria digitata, laminarin (LAM), induced arthritis under SPF conditions, thus BG would be a pathogenic factor for arthritis in SKG mice. Therefore, we prepared BG from Candida albicans, a pathogenic fungus and investigated whether BG from C. albicans induced arthritis in SKG mice under SPF conditions. SKG mice were injected intraperitoneally with particulate BG (oxidative-Candida albicans (OX-CA)), soluble BG (Candida soluble beta-glucan (CSBG)) from C. albicans and LAM as a positive control. In addition, schizophyllan (SPG) from Schizophyllum commune or Mycobacterium whole cells were injected into SKG mice to induce arthritis. Mice injected with OX-CA, CSBG and SPG had more severe arthritis than with LAM, and whole Mycobacterium cells. IL-6 concentration in sera from SKG mice injected with OX-CA or CSBG was high, whereas not detected in sera from mice treated with LAM. In histological analysis, infiltration of inflammatory cells was observed in SKG mice injected with BG. These results suggest that fungal infection may be a factor to induce and exacerbate autoimmune diseases such as RA.

OBJECTIVE

Hepatitis B recurrence after liver transplantation can be reduced to less than 10% by combination therapy with lamivudine (LAM) and hepatitis B immunoglobulin (HBIG). The aim of this study was to evaluate the efficacy and safety of prophylaxis with entecavir (ETV), which has higher efficacy and lower resistance rates than LAM, combined with HBIG in preventing hepatitis B recurrence after living-donor liver transplantation (LDLT).

METHODS

Twenty-six patients who received ETV plus HBIG (ETV group) after LDLT for hepatitis B virus (HBV)-related end-stage liver disease were analyzed by comparing with 63 control patients who had received LAM plus HBIG (LAM group).

RESULTS

The survival rates of the patients treated with ETV plus HBIG was 73% after both 1 and 3 years, and there was no statistical difference between the patients in the ETV group and LAM group. No HBV recurrence was detected during the median follow-up period of 25.1 months in the ETV group, whereas the HBV recurrence rate was 4% at 3 years and 6% at 5 years in the LAM group. No patients had adverse effects related to ETV administration.

CONCLUSIONS

ETV combined with HBIG provides effective and safe prophylaxis in preventing hepatitis B recurrence after LDLT.

Human alveolar macrophages obtained by bronchoalveolar lavage are usually poor accessory cells in in vitro lymphoproliferation assays. However, we recently described a subpopulation of pulmonary mononuclear cells, obtained from minced and enzyme-digested lung, which were potent stimulators of allogeneic T-lymphocyte proliferation. These cells were enriched in loosely adherent mononuclear cell (LAM) fractions, but further study of these accessory cells was hampered by the heterogeneous nature of LAM. It was observed that in the majority of lung tissue sections, most alveolar macrophages were autofluorescent, whereas most interstitial HLA-DR positive cells were not. Therefore autofluorescence was utilized to fractionate LAM in an attempt to remove alveolar macrophages and selectively purify interstitial accessory cells. LAM were separated by flow cytometry using forward and side scatter to exclude lymphocytes, and red autofluorescence to obtain brightly autofluorescent (A pos) and relatively nonautofluorescent (A neg) mononuclear cells. Although both populations contained over 80% HLA-DR positive cells, A pos cells were poor accessory cells, whereas A neg cells were extremely potent stimulators of a mixed leukocyte reaction at all stimulator ratios tested. When A pos cells were added to A neg cells, T-cell proliferation was markedly suppressed in the majority of experiments. Morphologically, A pos cells appeared similar to classical alveolar macrophages with 95% of the cells being large and intensely nonspecific esterase positive. In contrast, the majority of A neg were smaller, B-cell antigen-negative, nonspecific esterase negative, and had a distinctive morphology on Wright-stained smears. We conclude that fractionation of LAM based on autofluorescence is a powerful tool to isolate and characterize lung mononuclear cells that act either as stimulators or as suppressors of immune responses in the lung.

The mTORC1 pathway is a central regulator of cell growth, and defective mTORC1 regulation plays a causative role in a variety of human diseases, including cancer, tumor syndromes such as the tuberous sclerosis complex (TSC) and lymphangioleiomyomatosis (LAM), and metabolic diseases such as diabetes and obesity. Given the importance of mTORC1 signaling in these diseases, there has been significant interest in developing screening methods suitable for identifying inhibitors of mTORC1 activation. To this end, we have developed a high-throughput, cell-based assay for the detection of rpS6-phosphorylation as a measure of mTORC1 signaling. This assay takes advantage of the "In Cell Western" (ICW) technique using the Aerius infrared imaging system (LI-COR Biosciences). The ICW procedure involves fixation and immunostaining of cells in a manner similar to standard immunofluorescence methods but takes advantage of secondary antibodies conjugated to infrared-excitable fluorophores for quantitative detection by the Aerius scanner. In addition, the cells are stained with an infrared-excitable succinimidyl ester dye, which covalently modifies free amine groups in fixed cells and provides a quantitative measure of cell number. We present validation data and pilot screens in a 384-well format demonstrating that this assay provides a statistically robust method for both small molecule and siRNA screening approaches designed to identify inhibitors of mTORC1 signaling.

Lysine 5,6-aminomutase (5,6-LAM) catalyzes the interconversion of D-lysine with 2,5-diaminohexanoate and of L-beta-lysine with 3,5-diaminohexanoate. The coenzymes for 5,6-LAM are adenosylcobalamin (AdoCbl) and pyridoxal 5'-phosphate (PLP). In the proposed chemical mechanism, AdoCbl initiates the formation of substrate radicals, and PLP facilitates the radical rearrangement by forming an external aldimine linkage with the epsilon-amino group of a substrate, either D-lysine or L-beta-lysine. In the resting enzyme, an internal aldimine between PLP and an essential lysine in the active site facilitates productive PLP binding and catalysis. We present here biochemical, biophysical, and site-directed mutagenesis experiments, which document the existence of an essential lysine residue in the active site of 5,6-LAM from Porphyromonas gingivalis. Reduction of 5,6-LAM with NaBH(4) rapidly inactivates the enzyme and shifts the electronic absorption band from 420 to 325 nm. This is characteristic of the reduction of an aldimine linkage between the carbonyl group of PLP and the epsilon-amino group of a lysine residue. The reduced peptide was identified by Q-TOF/MS and further confirmed by Q-TOF/MS/MS sequencing. We show that lysine 144 in the small subunit of 5,6-LAM is the essential lysine residue. Lysine 144(beta) is separated by only 11 amino acids from histidine 133(beta), which forms a part of the "base-off"-AdoCbl binding motif. The sequence of the novel PLP-binding motif is conserved in 5,6-LAM from Clostridium sticklandii and P. gingivalis, and it is distinct from all known PLP-binding motifs. Mutation of lysine 144(beta) to glutamine led to K144Q(beta)-5,6-LAM, which displayed no enzymatic activity and no absorption band corresponding to an internal PLP-aldamine. In summary, we introduce a novel PLP-binding motif, the first to be discovered in an AdoCbl-dependent enzyme.

Lamivudine (LAM) is a nucleoside analogue widely used for the treatment of chronic hepatitis B virus (HBV) infection. Emergence of resistant strains with amino acid substitutions in the tyrosine-methionine-aspartate-aspartate (YMDD) motif of reverse transcriptase is a serious problem in patients on LAM therapy. The amount of covalently closed circular DNA in the serum is reported to be higher in patients who develop YMDD mutants than in those without mutants. However, there is no useful serum marker that can predict early emergence of mutants during LAM therapy. Analysis of patients who were treated with entecavir (n=7) and LAM (n=36) showed some patients had high serum levels of HBV RNA. Median serum levels of HBV RNA were significantly higher in patients in whom the YMDD mutant had emerged within 1 year (n=6, 1.688 log copies/ml) than in those in whom the YMDD mutant emerged more than 1 year after treatment (n=12, 0.456 log copies/ml, P=0.0125) or in whom the YMDD mutant never emerged (n=18, 0.688 log copies/ml, P=0.039). Our results suggest that HBV RNA is a valuable predictor of early occurrence of viral mutation during LAM therapy.

We evaluated the diagnostic performance of two tests based on the release of lipoarabinomannan (LAM) into the urine, the MTB-LAM-ELISA assay and the Determine TB-LAM-strip assay, in children with suspected tuberculosis (TB) in a high TB/HIV-prevalence setting.In a prospective study, 132 children with suspected active TB were assigned to diagnostic subgroups. Urine samples were subjected to testing by both assays to ascertain sensitivity and specificity. Host factors associated with positive LAM results were investigated and LAM excretion monitored after antituberculous treatment initiation.18 (13.6%) children had culture-confirmed pulmonary TB. The assays' sensitivity was higher in HIV-positive versus HIV-negative children: 70% (95% confidence interval 35-93%) versus 13% (0-53%) for MTB-LAM-ELISA and 50% (19-81%) versus 0% (0-37%) for Determine TB-LAM. In 35 (27%) children with excluded active TB, both assays showed a specificity of 97.1% (85-100%). Proteinuria and low body mass index were independently associated with LAM positivity. In most patients, LAM excretion declined to zero during or at conclusion of antituberculous treatment.HIV/TB co-infected children might benefit from LAM-based tests to aid early TB diagnosis and subsequent positive impact on morbidity and mortality. Using LAM as a rule-in and treatment-monitoring tool may also show further potential.

Interactions between plants and soil microbes are important determinants of both above- and belowground community composition, and ultimately ecosystem function. As exotic plants continue to invade and modify native plant communities, there has been increasing interest in determining the influence of exotic invasives on native soil microbial communities. Here, using highly sensitive molecular techniques, we examine fungal abundance and diversity in the soil surrounding a particularly aggressive invasive plant species in North America, Centaurea maculosa Lam. In mixed stands, we show that this invasive weed can alter the native fungal community composition within its own rhizosphere and that of neighboring native plants. At higher densities, the effect of C. maculosa on native soil fungal communities was even greater. Our results demonstrate that this invasive weed can have significant effects not only on visible aboveground biodiversity but also on the native soil microbial community that extends beyond its rhizosphere.

Estrogens affect the functioning of several non-reproductive tissues, the immune system in particular. In mammalian immunocytes, 17beta-estradiol (E2) has both dose- and cell-type specific effects and the responses to E2 seem to be mediated by rapid, non-genomic mechanisms; these may be initiated at either membrane or cytosolic locations, and can result in both direct local effects, such as modification of ion fluxes, and regulation of gene transcription secondary to activation of different kinase cascades, including mitogen activated protein kinases (MAPKs). In this work, the short-term effects of E(2) and the possible mechanisms of estrogen-mediated cell signaling were investigated in the hemocytes, the immune cells of the bivalve mollusc, the mussel Mytilus galloprovincialis Lam. The results show that E2 (25nM) caused a rapid and significant increase in hemocyte cytosolic [Ca2+]; lower concentrations (5 nM) showed a smaller, not significant effect. Both E2 concentrations affected the phosphorylation state of the components of tyrosine kinase-mediated signal transduction MAPK- and STAT- (signal transducers and activators of transcription) like proteins within 5-15 min from E2 addition. A greater effect and clearer time course were observed with 25 nM E2: in particular, E2 induced a transient increase in p-ERK2 MAPK and a persistent increase in p-p38 MAPK. Moreover, both STAT3 and STAT5 were tyrosine phosphorylated in response to E2. E2 (5 nM) induced both morphological (as evaluated by SEM) and functional changes (such as extracellular release of hydrolytic enzymes, lysosomal membrane destabilisation, and stimulation of the bactericidal activity) within 10-30 min from addition. Lysosomal membrane destabilisation induced by both E2 concentrations was abolished by hemocyte preincubation with the p38 MAPK inhibitor SB203580, and significantly reduced by PD98059 and Wortmannin (inhibitors of ERK MAPK and PI3-K, respectively), this suggesting that rapid activation of kinase cascades is involved in mediating the effects of E2 in mussel hemocytes. The antiestrogen Tamoxifen prevented or strongly reduced most, but not all, the effects of E2. Western blotting with heterologous anti-ERalpha-anti-ERbeta-antibodies revealed the presence of immunoreactive ERalpha- and ERbeta-like proteins in hemocyte protein extracts. Overall, our data support the hypothesis that the rapid effects and mechanisms of action of 17beta-estradiol are extremely conserved and that they may play a crucial role in endocrine-immune interactions in invertebrates.

The Equiguard is a dietary supplement comprised of standardized extracts from nine herbs, respectively, Herba epimedium brevicornum Maxim (stem and leaves), Radix morindae officinalis (root), Fructus rosa laevigatae michx (fruit), Rubus chingii Hu (fruit), Schisandra chinensis (Turz.) Baill (fruit), Ligustrum lucidum Ait (fruit), Cuscuta chinensis <em>Lam</em> (seed), Psoralea corylifolia L. (fruit), and Astragalus membranaceus (Fisch.) Bge (root). This proprietary product, formulated according to Chinese traditional medicinal concepts, is aimed at restoring harmony in the <primordial (original) ying-yang> of the kidney, an organ which Chinese medicinal principles consider to be vital for invigorating as well as maintaining balance of the entire urological system. As the prostate is an integral component of the urological system, we performed in vitro studies to test the effects of ethanol extracts of Equiguard to modulate prostate growth and gene expression. These studies used prostate cancer cells mimicking the androgen-dependent (AD) and androgen-independent (AI) states of prostate carcinogenesis. Results show that Equiguard significantly reduced cancer cell growth, induced apoptosis, suppressed expression of the androgen receptor (AR) and lowered intracellular and secreted prostate specific antigen (PSA), and almost completely abolished colony forming abilities of prostate cancer cells. These data support the interpretation that this herbal formulation contains ingredients that collectively may be efficacious in preventing or treating AD and AI prostate carcinoma. The anti-prostatic activities of Equiguard may stem from its complex composition capable of targeting multiple signal transduction/metabolic pathways, to effectively correct, counteract or circumvent the impaired or dysfunctional mechanisms accompanying different stages of prostate carcinogenesis.

Syzygium cumini (L.) SKEELS (syn. S. jambolanum DC, Eugenia jambolana LAM.) belongs to the medicinal plants most often recommended as an adjuvant therapy in type 2 diabetes. The plant was extensively studied during the last 125 years, approximately 100 case reports were reported already before the discovery of insulin. After the Second World War, research was concentrated on animal studies. Not all, but many of them reported some success in reducing type 2 diabetes symptoms. However, a state-of-the-art clinical study is still missing. In this review, historical literature dating back to the pre-insulin era was evaluated as were more recent in vitro-, animal-, and in vivo studies. Results were screened for information still useful today and compared to study results achieved in more recent decades. In view of the knowledge summarized here, a successful clinical study should use S. cumini seeds, seed kernels or fruit from India in fairly high doses. Reductions on blood sugar levels by about 30% seem reasonably to be expected. Adverse effects to be expected comprise gastrointestinal disturbances.

The parasitic protozoan Leishmania donovani is the causative organism for visceral leishmaniasis (VL) which persists in the host macrophages by deactivating its signaling machinery resulting in a critical shift from proinflammatory (Th1) to an anti-inflammatory (Th2) response. The severity of this disease is mainly determined by the production of IL-12 and IL-10 which could be reversed by use of effective immunoprophylactics. In this study we have evaluated the potential of Arabinosylated Lipoarabinomannan (Ara-LAM), a cell wall glycolipid isolated from non pathogenic Mycobacterium smegmatis, in regulating the host effector response via effective regulation of mitogen-activated protein kinases (MAPK) signaling cascades in Leishmania donovani infected macrophages isolated from BALB/C mice. Ara-LAM, a Toll-like receptor 2 (TLR2) specific ligand, was found to activate p38 MAPK signaling along with subsequent abrogation of extracellular signal-regulated kinase (ERKs) signaling. The use of pharmacological inhibitors of p38MAPK and ERK signaling showed the importance of these signaling pathways in the regulation of IL-10 and IL-12 in Ara-LAM pretreated parasitized macrophages. Molecular characterization of this regulation of IL-10 and IL-12 was revealed by chromatin immunoprecipitation assay (CHIP) which showed that in Ara-LAM pretreated parasitized murine macrophages there was a significant induction of IL-12 by selective phosphorylation and acetylation of histone H3 residues at its promoter region. While, IL-10 production was attenuated by Ara-LAM pretreatment via abrogation of histone H3 phosphorylation and acetylation at its promoter region. This Ara-LAM mediated antagonistic regulations in the induction of IL-10 and IL-12 genes were further correlated to changes in the transcriptional regulators Signal transducer and activator of transcription 3 (STAT3) and Suppressor of cytokine signaling 3 (SOCS3). These results demonstrate the crucial role played by Ara-LAM in regulating the MAPK signaling pathway along with subsequent changes in host effector response during VL which might provide crucial clues in understanding the Ara-LAM mediated protection during Leishmania induced pathogenesis.

We tested the hypothesis that laminarin (LAM), a beta (1-3) polysaccharide extracted from brown algae, can modulate the response to a systemic inflammation. Male Wistar rats (n=7 per group) were fed a standard diet (control) or a diet supplemented with LAM for 25 days (5% during 4 days followed by 10% during 21 days). Thereafter, Escherichia coli lipopolysaccharides (LPS; 10 mg/kg i.p.) were injected and the animals were sacrificed 24 h after LPS challenge. The hypothermia, hyperglycemia and hypertriglyceridemia occurring early after LPS administration were less pronounced in LAM-treated rats than in controls. The increase in serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) activities - reflecting hepatic alterations - was lessened after LPS injection in LAM-treated rats compared to control rats. LAM treatment decreased serum monocytes number, nitrite (NO2) and tumor necrosis factor-alpha (TNF-alpha). LAM also modulated intra-hepatic immune cells: it lowered the occurrence of peroxidase-positive cells (corresponding to monocytes/neutrophils) and, in contrast, it increased the number of ED2-positive cells, corresponding to resident hepatic macrophages, i.e. Kupffer cells. In conclusion, the hepatoprotective effect of marine beta (1-3) glucan during endotoxic shock may be linked to its immunomodulatory properties. We propose that both lower recruitment of inflammatory cells inside the liver tissue and lower secretion of inflammatory mediators play a role in the tissue protective effect of LAM. These effects could be due to a direct effect of beta-glucan on immune cells, or to an indirect effect through their dietary fibre properties (fermentation in the gut).

Amyloid consists of β-sheet polymers and is associated with disease and with functional assemblies. Amyloid-forming proteins differ widely in native structures and sequences. We describe here how conformational preferences of non-polar amino acid residues can affect amyloid formation. The most non-polar residues promote either β-strands (Val, Ile, Phe, and Cys, VIFC) or α-helices (Leu, Ala, and Met, LAM), while the most polar residues promote only α-helices. For 12 proteins associated with disease, the localizations of the amyloid core regions are known. Eleven of these contain segments that are biased for VIFC, but essentially lack segments that are biased for LAM. For the amyloid β-peptide associated with Alzheimer's disease and an amyloidogenic fragment of the prion protein, observed effects of mutations support that VIFC bias favors formation of β-sheet aggregates and amyloid, while LAM bias prevents it. VIFC and LAM profiles combine information on secondary structure propensities and polarity, and add a simple criterion to the prediction of amyloidogenic regions.

Different clinical outcomes of tuberculosis can be related to the balance between cell-mediated and humoral immunity. In this prospective study we examined the humoral immune responses to recombinant and native mycobacterial antigens in relation to clinical presentations of pulmonary TB. Two hundred and fifteen serum samples were examined including: non-cavitary (n=120), cavitary (n=65), caseous pneumonia (n=12), and disseminated TB (n=18). ELISA tests detecting IgG, IgA, and IgM against antigens: 38 kDa and 16 kDa, 38 kDa and lipoarabinomannan (LAM) were used. Univariate and multivariate logistic regression analyses were carried out to find the association between the antibody level and demographic or clinical characteristics. The relationships among specific antibody profiles and the phase of the disease in relation to demographic (age and sex) and clinico-radiological factors were investigated by measuring serum antibody levels (IgG, IgA, and IgM) to 38 kDa and 16 kDa recombinant M. tuberculosis antigens and to LAM - native mycobacterial antigen. The results show that the radiological extent of the disease is the strongest factor associated with IgG antibody production. Patients with more extensive pulmonary TB showed higher titers of IgG antibody to M. tuberculosis antigens (P<0.0001). The highest IgG and IgA level were observed in fibro-cavernous TB. The presence of cavity was associated only with IgG anti 38+16 kDa (P<0.001). IgA level was the highest in caseous pneumonia. IgM antibody production was not associated with any clinical and radiological factor, but only with the male gender. Age was independently and inversely associated with IgG anti 38 kDa+LAM level and IgM anti 38 kDa+LAM. We conclude that the humoral immune response to mycobacterial antigens is highly heterogeneous and varies with the stage of TB. IgG antibody level is higher in most advanced and extensive forms of the disease.

The aim of the present study was to evaluate the antioxidant, free radical scavenging and liver protective effects of friedelin isolated from Azima tetracantha Lam. leaves. In in vitro antioxidant study, the free radical scavenging effect of friedelin on 2,2-diphenyl-picrylhydrazyl (DPPH), hydroxyl, nitric oxide and superoxide radicals were evaluated. Friedelin showed very good scavenging effect on DPPH (IC50 21.1 mM), hydroxyl (IC50 19.8 mM), nitric oxide (IC50 22.1 mM) and superoxide (IC50 21.9 mM) radicals. Friedelin also showed strong suppressive effect on lipid peroxidation. In in vivo antioxidant study, CCl4 induced oxidative stress on rats produced significant increase in serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) and lactate dehydrogenase (LDH) levels along with reduction in liver superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH) and glutathione peroxidase (GPx) levels. Pre-treatment of rats with friedelin at 40 mg/kg for 7 days restored these levels to normality and showed liver protection, comparable to the standard, silymarin (25 mg/kg). These results clearly demonstrated that friedelin possessed marked antioxidant and liver protective effects.

The burden of neoplastic diseases is a significant global health challenge accounting for thousands of deaths. In Uganda, about 32,617 cancer cases were reported in 2018, accompanied by 21,829 deaths. In a view to identify some potential anticancer plant candidates for possible drug development, the current study was designed to compile the inventory of plants with reported anticancer activity used in rural Uganda and the evidences supporting their use in cancer therapy. An electronic survey in multidisciplinary databases revealed that 29 plant species belonging to 28 genera distributed among 24 families have been reported to be used in the management of cancer in Uganda. Anticancer plants were majorly from the families Bignoniaceae (7%), Caricaceae (7%), Fabaceae (7%), Moraceae (7%), and Rutaceae (7%). Most species occur in the wild (52%), though some are cultivated (48%). The growth habit of the plants is as trees (55%) or herbs (45%). Anticancer extracts are usually prepared from leaves (29%), bark (24%), roots (21%), and fruits (13%) through decoctions (53%), as food spices (23%) or pounded to produce ointments that are applied topically (10%). Prunus africana (Hook.f.) Kalkman, Opuntia species, Albizia coriaria (Welw. ex Oliver), Daucus carota L., Cyperus alatus (Nees) F. Muell., Markhamia lutea (Benth.) K. Schum., and Oxalis corniculata L. were the most frequently encountered species. As per global reports, Allium sativum L., Annona muricata L., Carica papaya L., Moringa oleifera Lam., Opuntia species, Prunus africana (Hook.f.) Kalkman, and Catharanthus roseus (L.) G. Don. are the most studied species, with the latter having vincristine and vinblastine anticancer drugs developed from it. Prostate, cervical, breast, and skin cancers are the top traditionally treated malignancies. There is a need to isolate and evaluate the anticancer potential of the bioactive compounds in the unstudied claimed plants, such as Cyperus alatus (Nees) F. Muell., Ficus dawei Hutch., Ficus natalensis Hochst., and Lovoa trichilioides Harms, and elucidate their mechanism of anticancer activity.

Sweetpotato [Ipomoea batatas (L.) Lam] is an important root crop that produces low molecular weight antioxidants such as carotenoids and anthocyanin. The sweetpotato orange (IbOr) protein is involved in the accumulation of carotenoids. To increase the levels of carotenoids in the storage roots of sweetpotato, we generated transgenic sweetpotato plants overexpressing IbOr-Ins under the control of the cauliflower mosaic virus (CaMV) 35S promoter in an anthocyanin-rich purple-fleshed cultivar (referred to as IbOr plants). IbOr plants exhibited increased carotenoid levels (up to 7-fold) in their storage roots compared to wild type (WT) plants, as revealed by HPLC analysis. The carotenoid contents of IbOr plants were positively correlated with IbOr transcript levels. The levels of zeaxanthin were ∼ 12 times elevated in IbOr plants, whereas β-carotene increased ∼ 1.75 times higher than those of WT. Quantitative RT-PCR analysis revealed that most carotenoid biosynthetic pathway genes were up-regulated in the IbOr plants, including PDS, ZDS, LCY-β, CHY-β, ZEP and Pftf, whereas LCY-ɛ was down-regulated. Interestingly, CCD1, CCD4 and NCED, which are related to the degradation of carotenoids, were also up-regulated in the IbOr plants. Anthocyanin contents and transcription levels of associated biosynthetic genes seemed to be altered in the IbOr plants. The yields of storage roots and aerial parts of IbOr plants and WT plants were not significantly different under field cultivation. Taken together, these results indicate that overexpression of IbOr-Ins can increase the carotenoid contents of sweetpotato storage roots.

The anti-inflammatory activity of Salacia oblonga rootbark powder and Azima tetracantha leaf powder was assayed in male albino rats using carrageenan-induced rat paw oedema (acute inflammation) and cotton pellet granuloma (chronic inflammation) methods. Both the crude drugs were maximally active at a dose of 1000 mg/kg. In the cotton pellet granuloma assay, these drugs were able to suppress the transudative, exudative and proliferative components of chronic inflammation. Furthermore, these drugs were able to lower the lipid peroxide content of exudate and liver, gamma-glutamyl transpeptidase activity in the exudate of cotton pellet granuloma. The increased acid and alkaline phosphatase activity and decreased serum albumin in cotton pellet granulomatous rats were normalised after treatment with these drugs. It is likely that these drugs may exert their activity by antiproliferative, antioxidative and lysosomal membrane stabilization.