Drug Standardization through Pharmacognostic Approaches and Estimation of Anticancer Potential of Chamomile (Matricaria chamomilla L.) using Prostate-Cancer cell lines: An In-vitro Study.

Cancer is the major challenge across world and the adenocarcinoma of prostate malignancy is the second most prevalent male cancer. Various medicinal plants are used for the treatment and management of various cancers. Matricaria chamomilla L., is one of the extensively used Unani medicament for the treatment of various type of diseases. In the current study we evaluated most of the parameters prescribed for drug standardization using pharmacognostic approaches. The 2,2 Diphenyl-1-picryl hydrazyl (DPPH) method was utilized for the analysis of antioxidant activity in the flower extracts of M. chamomilla. Moreover, we analyzed the antioxidant and cytotoxic activity of M. chamomilla (Gul-e Babuna) through in-vitro method. DPPH (2,2-diphenyl-1-picryl-hydrazl-hydrate) method was utilized for the analysis of antioxidant activity in the flower extracts of M. chamomilla. CFU and wound healing assay were performed to determine the anti-cancer activity. The results demonstrated that various extracts of M. chamomilla fulfilled most of the parameters of drug standardization and contained good antioxidant and anticancer activities. The ethyl acetate showed higher anticancer activity followed by aqueous, hydroalcoholic, petroleum benzene and methanol by CFU method. Also, the wound healing assay demonstrated that ethyl acetate extract has more significant effect followed by methanol and petroleum benzene extract on prostate cancer cell line (C4-2). The current study concluded that the extract of M. chamomilla flowers could act as good source of natural anti-cancer compounds.

The effect of not-anaerobicization and discolored bacteria on uranium reduction by Shewanella sp. RCRI7.

Shewanella sp. RCRI7 is a native strain capable of reducing uranium in anaerobic conditions. In order to employ this bacterium for the bioremediation, the mutual effects of uranium and the bacteria are studied in two different approaches. The optimal settings for the bacterial proliferation capacity and uranium reduction without anaerobicization of the environment, as well as the related effects of bioremediation and bacterial color under uranium-reducing conditions, have been investigated in this study. Uranium reduction procedure was analyzed using XRD, spectrophotometry and ICP-AES. In addition, the uranium's effect on the population of the first-generation of the bacteria as well as the color and growth of the second-generation were investigated using neobar lam and CFU (Colony Forming Unit), respectively. Uranium toxicity reduced the population of non-anaerobicized bacteria more than the anaerobicized bacteria after one day of incubation, while the amount of uranium extracted by the bacteria was almost the same. In both situations, the bacteria were able to reduce uranium after two weeks of incubation. In addition to the cell counts, uranium toxicity disrupts the growth and development of healthy second-generation anaerobicized bacteria, as created creamy-colored colonies grow slower than red-colored colonies. Furthermore, due to malfunctioning cytochromes, unlike red bacteria, creamy-colored bacteria were unable to extract the optimum amount of uranium. This study reveals that reduced uranium can be produced in a deprived environment without anaerobicization. Creamy-colored Shewanella can remove soluble uranium, however the most effective bacteria have red cytochromes. These findings represent a big step forward in the industrialization of uranium bioremediation.

Synergetic antibacterial activity of Vernonia auriculifera Hiern and Buddleja polystachya Fresen on selected human pathogenic bacteria.

Background: Medicinal plants have been used as traditional treatments for various human diseases for many years and they are still widely practiced throughout the world. Due to the long history of the practice, medicinal plants have become an integral part of the Ethiopian culture. This study aimed to evaluate the antibacterial activities of Vernonia auriculifera Hiern and Buddleja polystachya Fresen leaf extracts and their synergistic effect against some selected human pathogenic bacteria. Methods: Ethanol, methanol, and n-hexane crude extracts of Vernonia auriculifera, Buddleja polystachya, and a mixture of the two-plant respective of each solvent were evaluated against tested pathogenic bacteria using the agar well diffusion method; the inhibition zones were recorded in millimeters. Gentamycin was used as a positive control, while dimethyl sulfoxide served as a negative control. The minimum inhibitory concentration of the plant extracts against test bacteria was evaluated using two-fold broth dilution methods and then Minimum bactericidal concentration was determined by sub-culturing the test dilutions from minimum inhibitory concentration tubes onto fresh Muller Hinton Agar plates incubated at 37 °C for 24 h. Results: Maximum antibacterial inhibition zone was observed on methanol extracts of synergism against S. Typhimurium (ATCC 1333) (31.00 ± 1.73 mm) while, a minimum inhibition zone was observed on methanol extract of Buddleja polystachya, against E. coli (ATCC 35218) (5.67 ± 0.57). Minimum inhibitory concentration and minimum bactericidal concentration values of the crude extracts of Vernonia auriculifera, Buddleja polystachya, and their mixture lies between (3.125%-12.5%) and (6.25%-25%) respectively. The data were analyzed using the SPSS software package version 20 for windows. Conclusion: The present study revealed that ethanol and methanol extracts of Vernonia auriculifera and Buddleja polystachya possess significant inhibitory effects against tested pathogens and the antibacterial activity of both plants leaf extracts was greater than the activity of currently used antibiotics (Gentamycin) against some selected organisms.

Antimicrobial Effects of Inula viscosa Extract on the In Situ Initial Oral Biofilm.

Given the undesirable side effects of commercially used mouth rinses that include chemically synthesized antimicrobial compounds such as chlorhexidine, it is essential to discover novel antimicrobial substances based on plant extracts. The aim of this study was to examine the antimicrobial effect of Inula viscosa extract on the initial microbial adhesion in the oral cavity. Individual test splints were manufactured for the participants, on which disinfected bovine enamel samples were attached. After the initial microbial adhesion, the biofilm-covered oral samples were removed and treated with different concentrations (10, 20, and 30 mg/mL) of an I. viscosa extract for 10 min. Positive and negative controls were also sampled. Regarding the microbiological parameters, the colony-forming units (CFU) and vitality testing (live/dead staining) were examined in combination with fluorescence microscopy. An I. viscosa extract with a concentration of 30 mg/mL killed the bacteria of the initial adhesion at a rate of 99.99% (log10 CFU value of 1.837 ± 1.54). Compared to the negative control, no killing effects were determined after treatment with I. viscosa extract at concentrations of 10 mg/mL (log10 CFU value 3.776 ± 0.831; median 3.776) and 20 mg/mL (log10 CFU value 3.725 ± 0.300; median 3.711). The live/dead staining revealed a significant reduction (p < 0.0001) of vital adherent bacteria after treatment with 10 mg/mL of I. viscosa extract. After treatment with an I. viscosa extract with a concentration of 30 mg/mL, no vital bacteria could be detected. For the first time, significant antimicrobial effects on the initial microbial adhesion in in situ oral biofilms were reported for an I. viscosa extract.

Evaluation of a mouthrinse containing guava leaf extract as part of comprehensive oral care regimen- a randomized placebo-controlled clinical trial.

BACKGROUND: The control of biofilm adherence on tooth surface has always been the keystone of periodontal therapeutic systems. However, prevalence of gingivitis suggest inadequacy of self-performed oral hygiene measures and need for adjunctive aid for mechanical plaque control. Oral rinses containing chlorhexidine, has been widely used however, with certain limitations. Herbal products have been used widely reflecting its action as alternative and complementary remedy. Hence, the purpose of the present study was to evaluate the antimicrobial and antioxidant efficacy of a Guava leaf extract based mouthrinse in patients with chronic generalized gingivitis as an adjunct to oral prophylaxis. METHODS: Sixty subjects (n = 20) in compliance with the inclusion criteria were randomly assigned to one of the 3 study groups i.e. Group A- 0.15%Guava mouth rinse, Group B- 0.2% Chlorhexidine (CHX) mouth rinse, Group C- Distilled water (placebo). All the participants received professional oral prophylaxis and were dispensed with experimental mouth rinses and instructed to use for period of 30 days. Clinical parameters such as gingival index, plaque index along with microbial colony forming units using plaque samples and antioxidant levels in saliva were estimated at baseline, 30 and 90 days' time intervals. RESULTS: All 3 groups showed gradual reduction in GI, PI and microbial counts. Considering the mean scores of recorded parameters at the scheduled time intervals, notable changes were observed between chlorhexidine and guava mouth rinse compared to placebo group. Although there was improvement in the antioxidant status in all study participants, yet there was no statistically significant difference observed. CONCLUSION: Guava mouth rinse can be used as an empirical adjunct to professional oral prophylaxis owing to its multifactorial properties and favourable acceptance. However, long term studies need to be conducted to validate its use for an extended period of time. TRIAL REGISTRATION: The clinical trial has been prospectively registered on 17th February 2017 by the Clinical Trials Registry-India (CTRI/2017/02/007898).

Assessing the ability of silage lactic acid bacteria to incorporate and transform inorganic selenium within laboratory scale silos.

Selenium (Se) is a non-metallic trace element essential for normal cellular function, which has been linked with reduced risk of cancer, cardiovascular disease, cognitive decline and thyroid disease in humans. Se deficiency in livestock is associated with white muscle disease, retained placenta, ill-thrift and mastitis. Where Se status or bioavailability from the soil for plants is poor, livestock rely on supplemental Se in their diets predominantly as either sodium selenite (inorganic form) or selenised-yeast (organic form). As lactic acid bacteria (LAB) have been shown to incorporate Se as either organic or elemental (Nano-Se) there may be potential to use silage inoculant bacteria to improve the Se status of feed to provide the Se requirements of livestock. We screened twenty-seven LAB in MRS broth in the presence of sodium selenite for growth and uptake of Se as organic (selenocysteine and selenomethionine), inorganic (selenite and selenate) or/and Nano-Se, with the aim to identify potential candidates for a mini-silo study. Sodium selenite addition into the growth medium of LAB reduced growth rates but also resulted in the conversion of the inorganic sodium selenite into predominately Nano-Se and small quantities of organic-Se. Based on a rank analysis of growth and ability to take up (total Se content) and convert inorganic Se (Nano and organic Se content), three LAB were selected for further investigation as silage inoculants: L. brevis DSMZ (A), L. plantarum LF1 (B), and L. plantarum SSL MC15 (C). Each LAB was used as an inoculant within a grass mini-silo trial, either cultured in the presence of sodium selenite before inoculation or sodium selenite added to the inoculum at inoculation versus controls with no Se. The addition of sodium selenite either into the growth media of LAB or applied at inoculation of grass silage did not interfere with the ability of the LAB to act as a silage inoculant with no difference in silage fermentation characteristic between LAB with no Se added. The addition of sodium selenite either to the LAB growth medium or at inoculation resulted in the conversion of sodium selenite into Nano-Se and organic-Se (Nano-Se, ca. 103 higher than organic), as previously shown in the screening trial. There was no difference between the three LAB for incorporation of Se or in silage quality, indicating the potential to develop silage inoculants to increase the bioavailable form of Se (elemental and organic) to livestock through conversion of inorganic forms during ensiling.

Artemisinin and its derivatives prevent Helicobacter pylori-induced gastric carcinogenesis via inhibition of NF-κB signaling.

BACKGROUND: Gastric cancer has a high morbidity and is a leading cause of cancer-related mortality worldwide. Helicobacter pylori (H. pylori) infection is commonly found in the early stage of gastric cancer pathogenesis, which induces chronic gastritis. Artemisinin (ART) and its derivatives (ARTS, artesunate and DHA, dihydroartemisinin), a new class of potent antimalarials, have been reported to exert both preventive and anti-gastric cancer effects. However, the underlying mechanisms of the chemopreventive effects of ART and its derivatives in H. pylori infection induced-gastric cancer are not fully elucidated. PURPOSE: We investigated the effects of H. pylori infection in gastric cancer; and the preventive mechanisms of ART, ARTS and DHA. METHODS: The H. pylori growth was determined by the broth macro-dilution method, and its adhesion to gastric cancer cells was evaluated by using the urease assay. The protein and mRNA levels, reactive oxygen species (ROS) production, as well as the production of inflammatory cytokines were evaluated by Western blot, real-time PCR, flow cytometry and ELISA, respectively. Moreover, an in vivo MNU (N-methyl-N-nitroso-urea) and H. pylori-induced gastric adenocarcinoma mouse model was established for the investigation of the cancer preventive effects of ART and its derivaties, and the underlying mechanisms of action. RESULTS: ART, DHA and ARTS inhibited the growth of H. pylori and gastric cancer cells,suppressed H. pylori adhesion to the gastric cancer cells, and reduced the H. pylori-enhanced ROS production. Moreover, ART, DHA and ARTS significantly reduced tumor incidence, number of tumor nodules and tumor size in the mouse model. Among these three compounds, DHA exerted the most potent chemopreventive effect. Mechanistic studies showed that ART and its derivatives potently inhibited the NF-κB activation. CONCLUSION: ART, DHA and ARTS have potent preventive effects in H. pylori-induced gastric carcinogenesis. These effects are, at least in part, attributed to the inhibition of NF-κB signaling pathway. Our findings provide a molecular justification of using ART and its derivatives for the prevention and treatment of gastric cancer.

A New Method for Determination of Minimum Biofilm Eradication Concentration for Accurate Antimicrobial Therapy.

Antimicrobial susceptibility testing (AST) is an important technique to find the susceptibility pattern of clinical isolates in order to administer the appropriate drug. One such technique is minimum inhibitory concentration (MIC), which not only identifies the right drug but also suggests the appropriate concentration necessary to neutralize the organisms in planktonic form. MIC can vary in case of adherent organisms since they form biofilms and activate survival mechanisms like quorum sensing. Here we have strategized a new method which used an inoculator plate, a resazurin dye, and a standard plate to identify minimum biofilm eradication concentration (MBEC) of adherent organisms.

In vitro efficacy of eugenol in inhibiting single and mixed-biofilms of drug-resistant strains of Candida albicans and Streptococcus mutans.

BACKGROUND: Candida albicans is frequently associated with mixed infections of Streptococcus mutans in plaque biofilms. These pathogens under chemical interactions resulting in mixed biofilm development have turned it into a prevalent and costly oral disease, which is not successfully being treated by existing chemotherapeutics. HYPOTHESIS: Considering the need for newer drugs to overcome this challenge, the present study was aimed to investigate the efficacy of eugenol in inhibiting single and mixed biofilms of C. albicans and S. mutans. METHODS: The broth dilution assay was used to determine drug resistance in the test strains. Biofilm formation on polystyrene microtiter plate was studied by XTT reduction assay whereas biofilm development on glass coverslips was assessed using 0.1% crystal violet and visualised under light microscope. Single and mixed biofilms formed on glass coverslips in the presence and absence of eugenol was analysed by scanning electron microscopy. RESULTS: In our study, all the thirteen strains of C. albicans were resistant to fluconazole, itraconazole, ketoconazole, amphotericin B except C. albicans (CAJ-01) and C. albicans MTCC3017 which were sensitive to fluconazole. S. mutans MTCC497 was resistant to ampicillin, azithromycin, ceftriaxone and vancomycin. Among all the strains of C. albicans, CAJ-01, C. albicans ATCC90028 and C. albicans MTCC3017 formed strong biofilms and rest of the strains considered as moderate to weak biofilm formers. S. mutans MTCC497 was also formed strong biofilms. Eugenol showed concentration dependent anti-biofilm activity against single and mixed biofilms of C. albicans (CAJ-01) and S. mutans MTCC497. At sub-MIC of eugenol (100 µg/ml), the biofilm formation was 36.37% and 29.72% in CAJ-01 and S. mutans MTCC497, respectively, whereas 52.65% in mixed biofilms. The cell viability assay showed significant reduction (p < 0.05) in the log10 CFU/ml from 6.3 to 4.8 at 200 µg/ml of eugenol for CAJ-01, whereas, from 6.4 to 3.8 and 5.3 for S. mutans MTCC497 strains in single and mixed biofilms, respectively. Scanning electron microscopy showed the disruption of cell membrane and matrix structure in both single and mixed biofilms. CONCLUSIONS: Eugenol at sub-MICs effectively inhibited single and mixed biofilms formed by the drug resistant strains of two oral pathogens, C. albicans and S. mutans through multiple mode of action.

Lysozyme-rich milk mitigates effects of malnutrition in a pig model of malnutrition and infection.

Malnutrition remains a leading contributor to the morbidity and mortality of children under the age of 5 years and can weaken the immune system and increase the severity of concurrent infections. Livestock milk with the protective properties of human milk is a potential therapeutic to modulate intestinal microbiota and improve outcomes. The aim of this study was to develop an infection model of childhood malnutrition in the pig to investigate the clinical, intestinal and microbiota changes associated with malnutrition and enterotoxigenic Escherichia coli (ETEC) infection and to test the ability of goat milk and milk from genetically engineered goats expressing the antimicrobial human lysozyme (hLZ) milk to mitigate these effects. Pigs were weaned onto a protein-energy-restricted diet and after 3 weeks were supplemented daily with goat, hLZ or no milk for a further 2 weeks and then challenged with ETEC. The restricted diet enriched faecal microbiota in Proteobacteria as seen in stunted children. Before infection, hLZ milk supplementation improved barrier function and villous height to a greater extent than goat milk. Both goat and hLZ milk enriched for taxa (Ruminococcaceae) associated with weight gain. Post-ETEC infection, pigs supplemented with hLZ milk weighed more, had improved Z-scores, longer villi and showed more stable bacterial populations during ETEC challenge than both the goat and no milk groups. This model of childhood disease was developed to test the confounding effects of malnutrition and infection and demonstrated the potential use of hLZ goat milk to mitigate the impacts of malnutrition and infection.

Nutritional strategies to prevent gastrointestinal toxicity during pelvic radiotherapy.

Radiotherapy-induced damage to non-cancerous gastrointestinal mucosa has effects on secretory and absorptive functions and can interfere with normal gastrointestinal physiology. Nutrient absorption and digestion may be compromised. Dietary manipulation is an attractive option with sound rationale for intervention. The aim of this review was to synthesise published evidence for the use of elemental formulae, low or modified fat diets, fibre, lactose restriction and probiotics, prebiotics and synbiotics to protect the bowel from gastrointestinal side effects during long-course, radical pelvic radiotherapy. Thirty original studies (recruiting n 3197 patients) were identified comprising twenty-four randomised controlled trials, four cohort studies and two comparator trials. Endpoints varied and included symptom scales (Inflammatory Bowel Disease Questionnaire, Common Technology Criteria for Adverse Events, Radiation Therapy Oncology Group) and Bristol Stool Scale. Dietary and supplement interventions were employed with many studies using a combination of interventions. Evidence from RCT was weak for elemental, low or modified fat and low-lactose interventions and modestly positive for the manipulation of fibre during radiotherapy. Evidence for probiotics as prophylactic interventional agents was more promising with a number of trials reporting positive results but strength and strains of interventions vary, as do methodologies and endpoints making it difficult to arrive at firm conclusions with several studies lacking statistical power. This consolidated review concludes that there is insufficient high-grade evidence to recommend nutritional intervention during pelvic radiotherapy. Total replacement of diet with elemental formula could be effective in severe toxicity but this is unproven. Probiotics offer promise but cannot be introduced into clinical practice without rigorous safety analysis, not least in immunocompromised patients.

Effects of a new combination of plant extracts plus d-mannose for the management of uncomplicated recurrent urinary tract infections.

Urinary tract infections (UTIs) are an economic burden for public health. The increasing prevalence of resistant bacteria which cause UTIs may be related to the inappropriate prescription of antibiotics. The aim of this preliminary study was to evaluate whether three different combinations of plant extracts plus d-mannose are effective in preventing the recurrence of UTIs. Three groups of patients received three combinations of plant extracts in conjunction with d-mannose. These were: berberine, arbutin and birch (group A); berberine, arbutin, birch and forskolin (group B); and proanthocyanidins (group C). The clinical recurrence of cystitis at the end of treatment and during follow-up was determined by comparison with baseline measurements using the microbiological assessment of urine samples, vaginal swabs and vaginal smear slides. Patients in groups A and B had a lower incidence of episodes of recurrent cystitis during treatment and follow-up, samples with a significantly lower median bacterial load and a reduction of the grade of lactobacillary flora compared to patients in group C.

Supplementation with Lactobacillus paracasei or Pediococcus pentosaceus does not prevent diarrhoea in neonatal pigs infected with Escherichia coli F18.

Infectious diarrhoea is a worldwide problem in newborns. Optimal bacterial colonisation may enhance gut maturation and protect against pathogenic bacteria after birth. We hypothesised that lactic acid bacteria (LAB) administration prevents pathogen-induced diarrhoea in formula-fed newborns. Newborn caesarean-delivered, colostrum-deprived term piglets on parenteral nutrition for the first 15 h, were used as models for sensitive newborn infants. A commercially available probiotic strain, Lactobacillus paracasei F19 (LAP, 2·6×108 colony-forming units (CFU)/kg per d) and a novel LAB isolate, Pediococcus pentosaceus (PEP, 1·3×1010 CFU/kg per d), were administered for 5 d with or without inoculation of the porcine pathogen, Escherichia coli F18 (F18, 1010 CFU/d). This resulted in six treatment groups: Controls (n 9), LAP (n 10), PEP (n 10), F18 (n 10), F18-LAP (n 10) and F18-PEP (n 10). The pathogen challenge increased diarrhoea and density of F18 in the intestinal mucosa (P<0·05). LAB supplementation further increased the diarrhoea score, relative to F18 alone (P<0·01). Intestinal structure and permeability were similar among groups, whereas brush border enzymes were affected in variable intestinal regions with decreased activities in most cases after F18 and LAB inoculation. Bacterial density in colon mucosa increased after F18 inoculation (P<0·05) but was unaffected by LAB supplementation. In colon contents, acetic and butyric acids were increased by PEP (P<0·05). The LAB used in this study failed to reduce E. coli-induced diarrhoea in sensitive newborn pigs. In vulnerable newborns there may be a delicate balance among bacterial composition and load, diet and the host. Caution may be required when administering LAB to compromised newborns suffering from enteric infections.

Effect of rosuvastatin or its combination with omega-3 fatty acids on circulating CD34(+) progenitor cells and on endothelial colony formation in patients with mixed dyslipidaemia.

BACKGROUND AND AIMS: Hypercholesterolaemia is associated with a reduced number of circulating progenitor cells, a defect that is restored by statin therapy. We studied the effect of rosuvastatin monotherapy or its combination with omega-3 polyunsaturated fatty acids (ω-3 PUFAs) on progenitor cell number and function in patients with mixed dyslipidaemia. METHODS: Fifty patients with mixed dyslipidaemia and fifty-five normolipidaemic, apparently healthy, age- and sex-matched volunteers participated in the study. Patients were randomized to receive a daily dose of either 40 mg rosuvastatin (R group, n = 26) or 10 mg rosuvastatin plus 2 g of ω-3 PUFAs (R + O group, n = 24). The number of circulating CD34(+) and CD34(+)/KDR(+) progenitor cells as well as the number of colony-forming units-endothelial cells (CFU-ECs) at 10 days of culture were assessed at baseline and 3 months post-treatment. RESULTS: The number of CD34(+) and CD34(+)/KDR(+) cells per 10,000 leukocytes as well as the number of CFU-ECs were significantly lower in both patient groups compared with healthy volunteers (all p = 0.03). A 3-month treatment with either R or R + O significantly increased circulating CD34(+) and CD34(+)/KDR(+) cells as well as the number of CFU-ECs compared with baseline (all p = 0.03). Importantly, the increase in the above parameters was inversely correlated with therapy-induced reduction in lipid parameters in both patient groups. CONCLUSIONS: Patients with mixed dyslipidaemia exhibit low numbers of circulating CD34(+) and CD34(+)/KDR(+) cells as well as CFU-ECs in culture, a defect restored by 3-month treatment with either high-rosuvastatin dose or a combination of low-rosuvastatin dose with ω-3 PUFAs. The pathophysiological meaning of our results regarding the increased cardiovascular risk in these patients remains to be established.

Effectiveness of Lactobacillus helveticus and Lactobacillus rhamnosus for the management of antibiotic-associated diarrhoea in healthy adults: a randomised, double-blind, placebo-controlled trial.

Broad-spectrum antibiotic use can disrupt the gastrointestinal microbiota resulting in diarrhoea. Probiotics may be beneficial in managing this type of diarrhoea. The aim of this 10-week randomised, double-blind, placebo-controlled, parallel study was to investigate the effect of Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011 supplementation on antibiotic-associated diarrhoea in healthy adults. Subjects were randomised to receive 1 week of amoxicillin-clavulanic acid (875 mg/125 mg) once per day, plus a daily dose of 8×109 colony-forming units of a multi-strain probiotic (n 80) or placebo (n 80). The probiotic or placebo intervention was maintained for 1 week after completion of the antibiotic. Primary study outcomes of consistency and frequency of bowel movements were not significantly different between the probiotic and placebo groups. The secondary outcomes of diarrhoea-like defecations, Gastrointestinal Symptoms Rating Scale scores, safety parameters and adverse events were not significantly different between the probiotic intervention and the placebo. A post hoc analysis on the duration of diarrhoea-like defecations showed that probiotic intervention reduced the length of these events by 1 full day (probiotic, 2·70 (sem 0·36) d; placebo, 3·71 (sem 0·36) d; P=0·037; effect size=0·52). In conclusion, this study provides novel evidence that L. helveticus R0052 and L. rhamnosus R0011 supplementation significantly reduced the duration of diarrhoea-like defecations in healthy adults receiving antibiotics.

Local transplantation of bone marrow concentrated granulocytes precursors can cure without antibiotics infected nonunion of polytraumatic patients in absence of bone defect.

PURPOSE: Infected, long bone non-unions present a significant clinical challenge. New and alternative therapies are needed to address this problem. The purposes of this study were to compare the number of circulating granulocyte-macrophage colony-forming units (CFU-GM) in the peripheral blood of polytraumatic patients with infected tibial non-unions and in the peripheral blood of control patients with the hypothesis that their number was decreased in polytraumatic patients; and to treat their infection without antibiotics and with local transplantation of bone marrow concentrated granulocytes precursors. METHODS: Thirty (18 atrophic and 12 hyperthrophic ) infected tibial non-unions (without bone defect) that occurred after open fractures in polytraumatic patients were treated without antibiotics and with percutaneous injection of autologous bone marrow concentrate (BMC) containing granulocytes precursors (CFU-GM). CFU-GM progenitors were assessed in the bone marrow aspirate, peripheral blood, and fracture site of these patients. The number of these progenitors was compared with the CFU-GM progenitors of control patient samples (healthy donors matched for age and gender). Outcome measures were: timing of union, callus formation (radiographs and CT scan), and recurrence of clinical infection. RESULTS: As compared to control patients, the number of CFU GM derived colonies was lower at peripheral blood in patients with infected nonunions. The bone marrow graft injected in nonunions contained after concentration 42 621 ± 20 350 CFU-GM-derived colonies/cc. Healing and cure of infection was observed at six months for 25 patients and at one year follow up for 30 patients. At the median ten year follow-up (range: 5 to 15), only one patient had clinical recurrent infection after healing (between 6 months and last follow-up). CONCLUSION: The peripheral blood of these polytraumatic patients with infected nonunions had a remarkable decrease in CFU-GM-derived colonies as compared with normal controls. Local transplantation of concentrated CFU-GM-derived colonies aspirated from bone marrow allowed cure of infection and healing without antibiotics.

Probiotic strain Lactobacillus plantarum 299v increases iron absorption from an iron-supplemented fruit drink: a double-isotope cross-over single-blind study in women of reproductive age.

Iron deficiency is common, especially among young women. Adding probiotics to foods could be one way to increase iron absorption. The aim of this study was to test the hypothesis that non-haem iron absorption from a fruit drink is improved by adding Lactobacillus plantarum 299v (Lp299v). Iron absorption was studied in healthy women of reproductive age using a single-blind cross-over design in two trials applying the double-isotope (55Fe and 59Fe) technique. In Trial 1, iron absorption from a fruit drink containing 109 colony-forming units (CFU) Lp299v was compared with that from a control drink without Lp299v. Trial 2 had the same design but 1010 CFU were used. The test and control drinks contained approximately 5 mg of iron as ferrous lactate and were labelled with 59Fe (B) and 55Fe (A), respectively, and consumed on 4 consecutive days in the order AABB. Retention of the isotopes was measured with whole-body counting and in blood. Mean iron absorption from the drink containing 109 CFU Lp299v (28·6(sd 12·5) %) was significantly higher than from the control drink (18·5(sd 5·8) %), n 10, P<0·028). The fruit drink with 1010 CFU Lp299v gave a mean iron absorption of 29·1(sd 17·0) %, whereas the control drink gave an absorption of (20·1(sd 6·4) %) (n 11, P<0·080). The difference in iron absorption between the 109 CFU Lp299v and the 1010 CFU Lp299v drinks was not significant (P=0·941). In conclusion, intake of probiotics can increase iron absorption by approximately 50 % from a fruit drink having an already relatively high iron bioavailability.

Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella, Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic E. coli (EHEC) and Campylobacter jejuni.

In Part II we discuss the following bacterial pathogens: Shigella, Salmonella (non-typhoidal), diarrheogenic E. coli (enterotoxigenic and enterohemorragic) and Campylobacter jejuni. In contrast to the enteric viruses and Vibrio cholerae discussed in Part I of this series, for the bacterial pathogens described here there is only one licensed vaccine, developed primarily for Vibrio cholerae and which provides moderate protection against enterotoxigenic E. coli (ETEC) (Dukoral(®)), as well as a few additional candidates in advanced stages of development for ETEC and one candidate for Shigella spp. Numerous vaccine candidates in earlier stages of development are discussed.

Antibacterial activity of the roots, stems and leaves of Alchornea floribunda.

ETHNOPHARMACOLOGICAL RELEVANCE: Alchornea floribunda Müll. Arg. is used in traditional medicine across Africa for the treatment of bacterial, fungal, parasitic and inflammatory disorders. AIM OF THE STUDY: To evaluate the antibacterial activity of the crude extracts of different plant parts in order to provide a scientific rationale for the proposed broad efficacy of Alchornea floribunda in the treatment of bacterial infections. MATERIALS AND METHODS: Extracts of roots, stems and leaves were prepared using solvents of various polarities in order to extract a wide range of phytochemicals. The antibacterial activity of these crude extracts was evaluated by micro-dilution assay, against Gram-positive (i.e. Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Staphylococcus saprophyticus) as well as Gram-negative (i.e. Escherichia coli, Klebsiella pneumoniae, Moraxella catarrhalis and Proteus mirabilis) bacteria. RESULTS: Generally, the ethanol (EtOH), methanol (MeOH), ethyl acetate (EtOAc) and chloroform (CHCl3) extracts demonstrated the best activities, with the leaves exhibiting the highest average activity for six of the eight pathogens. Of these, the ethanolic leaf extract was the most active against Staphylococcus aureus with an MIC value of 50µg/mL. Some other notable activity was observed for the ethyl acetate and chloroform root extracts against Staphylococcus aureus (50µg/mL), and for selected stem extracts against Staphylococcus aureus (50µg/mL), Klebsiella pneumoniae (63µg/mL) and Staphylococcus saprophyticus (63µg/mL). CONCLUSION: This study demonstrates the promising antibacterial activity of Alchornea floribunda against both Gram-positive and Gram-negative bacteria responsible for gastrointestinal, skin, respiratory and urinary ailments, and validates its use in the ethnopharmacology of the region.

Ethno-medicinal uses and screening of plants for antibacterial activity from Similipal Biosphere Reserve, Odisha, India.

ETHNOPHARAMCOLOGICAL RELEVANCE: The present study examined the variety and the extent of medicinal plants used in the health care system of tribal inhabitants of Similipal Biosphere Reserve. In addition to this, such plants were also screened for antibacterial properties against common pathogenic bacteria. MATERIAL AND METHODS: Semi-structured interview was carried out with 42 informants (mean age 42, 86% male, 14% female) at 24 locations in and around SBR, regarding the use of plants for the treatment of various human ailments. Antibacterial screening is adopted with the documented ethnomedicinal plants using methanol and aqueous extracts against eight bacterial strains. RESULTS: A total of 187 plant species belonging to 74 families were documented for frequent medicinal uses against common ailments such as stomach problems, fever, skin diseases, diarrhea and dysentery. Although all parts of plant are used, leaves and bark are most common. Tribals used the plant parts both in form of decoction (taken orally as in internal problems) and paste (external use). Out of 187 plant species, 120 plants recorded antibacterial activity against test bacterial strain. CONCLUSIONS: This study revealed that self care using medicinal plants is a common practice by the tribes of SBR. About 64% of the used plants have scientifically proved medicinal values with respect to the antibacterial properties.