Difference In Number Of Colonies Research Articles

Comparison of Antibacterial Properties of Salvia Officinalis with Commercially Available Mouth-Rinse

Dental caries, commonly known as cavities/tooth decay, is a widespread and one of the most prevalent diseases affecting people of all ages. Despite its prevalence, dental caries is largely preventable through good oral hygiene practices and reducing sugar intake. A variety of commercially available mouthwashes are being recommended by dental health professionals because of their antibacterial properties but their side effects limits the use of these mouth washes hence, plant based natural compounds are being evaluated for their antibacterial properties in the current study Salvia Officinalis was evaluated in this regard. Methodology: It was an in-vivo preclinical experimental study conducted from October 2022 to January 2023. The calculated sample size was n =60. The participants were given an envelope for group randomization. The organisms were grown on appropriate media and extract was prepared and diluted. The extract was diluted in distilled water at 1:4 (Extract: Distilled water) concentration. Study participants were instructed to not brush their teeth before sampling. Study participants were divided into three groups (negative control, positive control, Salvia Officinalis extract rinse group) each group had 20 participants. Diluted SO extract was given to the experimental group for rinses, distilled water was given to the negative control group and positive controls were given a standard commercially available mouth rinse. The next sample of plaque was collected after two hours to observe the effects of SO extract on bacterial colonies. Results: The pre and post-samples showed a significant (p-value <0.05) decrease in number of colonies in the positive control group (conventional rinse) and Salvia officinalis extract group. The intra-group comparison of negative and positive control showed a significant difference in number of colonies and the same was observed with the Salvia officinalis rinse. However, the positive control and Salvia officinalis extract comparison was insignificant. Conclusion: The invitro experiment highlighted that the S. officinalis plant extract has equivalent effects against S. mutans and P. gingivalis as that of commercially available mouth rinse. Keywords: Salvia officinalis, S. mutans, P. gingivalis, Moth rinse

Comparison of flax seeds extract and fenugreek seed extract rinse against streptococcus mutans colonies

Objective: To compare flax seeds extract and fenugreek seed extractrinse against streptococcus mutans colonies Study setting: The bench work of this preclinical experimental study was carried out at Baqai Medical and Dental College Karachi., from December 2021 to January 2022. Materials and Methods: It was a preclinical experimental study. Participants were divided into 4 groups randomization was performed in allocating the groups of participants. Film of plaque from labial surface of teeth of study participants was collected on sterile strips before and after the 2 hours of intervention. The extract preparation of both the plants and culture of S. mutantcolonies were performed as per standard protocol. The extract was diluted at 1:4 for rinsing and study participants were given the rinses according to their groups. Data was analyzed by SPSS v. 22 Results: The growth of colonies was calculated on growth media plates in samples prior to rinsing and samples that were taken after the rinsing. The pre and post samples showed significant (p-value <0.05) decrease in number of colonies in positive control group (conventional rinse), flax seed extract group and fenugreek seed extract group as shown in table 1. The intra group comparison of negative and positive control showed significant difference in number of colonies and same was observed with the both the extract rinses. However, the positive control and flax seed extract and fenugreek seed extract comparison was insignificant that highlighted the equal potential of herbal extracts in reducing the streptococcus mutans colonies. Conclusion: Flax seeds extract and fenugreek seed extract mouth rinses reduced the number of colonies equally as that of commercially available mouth rinse. Keywords: Flax seed extract, Fenugreek Seed Extract, Rinse, Antibacterial, streptococcus mutans

Abstract IA13: Combination of DNA methyltransferase and PARP inhibitors as a novel therapy strategy for multiple cancers: Key data in AML and triple negative breast cancer

Abstract Poly (ADP-ribose) polymerase (PARP) inhibitors represent one of the most exciting recent developments in cancer therapy. While substantial efficacy has been shown with clinically available PARP inhibitors (PARPis), to date, in treatment of hereditary deletions of BRCA1/2 in breast and ovarian cancers, the high promise of these drugs has not yet been realized in sporadic cancers. We present here strong preclinical data for a novel, mechanistically based, combinatorial approach to using DNA methyltransferase inhibitors (DNMTis), such as decitabine (DAC) and 5-Azacytidine (5-AZA), with PARP inhibitors (PARPis) as a treatment strategy for acute myelogenous leukemias (AML) and triple negative breast cancer (TNBC). We have previously demonstrated that low doses of 5-AZA and DAC alone show efficacy in AML and TNBC, and propose treatment with PARPis to enhance sensitivity of cancer cells to DNMTis. The mechanistic rationale for our approach is based upon: 1) data from our group and others showing DNMT1 and PARP1 associate in a complex, and this association increases with DNA damage; 2) the fact that 5-AZA and DAC trap DNMTs led us to hypothesize that these drugs might also increase PARP trapping at DNA damage sites; and 3) the cytotoxicity of the most potent PARPis (e.g. BMN 673) appears to correlate with the degree of trapping of PARP1 in chromatin. We first find that in cultured human AML and TNBC cells, the DNMTis (5 to 20 nM DAC or 100 to 200nM 5-AZA) and PARPis (1 to 10 nM BMN 673) alone trap PARP into chromatin, and this effect is enhanced when the drugs are combined. In addition, the PARPi-DNMTi combination treatment of TNBC cell line MDA-MB-231 resulted in significantly enhanced retention of PARP1 and DNMT1 at sites of double strand breaks (DSBs) induced by laser microirradiation. Concomitant with this, the combined doses resulted in significant increases in cytotoxic DSBs, observed 4-24 hours after DSB induction, when compared to single-drug treatments. Homologous recombination (HR) DSB repair activity also appears decreased, as measured by GFP reporter assays. In keeping with these findings, colony survival assays demonstrated that the combination treatment, compared to either drug alone, strongly inhibited colony formation of TNBC cell lines (N=4). Notably non-tumorigenic MCF10A cells showed no significant differences in colony numbers with single or combination drug treatments. Similar to TNBCs, AML cell lines (N=3) as well as primary AML cells (N=8) showed dramatic decreases in colonies in combination vs single agent drug treatments. In the most important translational implications of the preliminary studies, in in vivo therapy TNBC and AML models in immune-deficient mice, our low dose combinations of DNMTis and PARPis provide for potent anti-tumor responses. Mouse xenograft experiments using BRCA mutant TNBC cell line SUM149PT demonstrated that the combination treatment has a significant (p<0.05) survival advantage compared to control (vehicle), AZA (0.5mg/kg) or BMN (0.3 mg/kg) alone. Importantly, mouse MDA-MB-231 xenografts with intact BRCA1 showed significant survival with the combined drug treatment. Likewise in AML xenografts of MOLM14 and MV411 cell lines treated with the drug combination show significantly decrease leukemia burden, as measured by luciferase imaging. Our data suggest a novel use of both DNMTis and PARPis in a compelling therapeutic strategy for TNBCs independent of BRCA mutations and poor prognosis AML; the latter will be investigated in a clinical trial to be based at the University of Maryland and funded by Van Andel-SU2C. Citation Format: Nidal Muvarak, Khadiza Chowdhury, Carine Robert, Xia Limin, Eun Yong Choi, Yi Cai, Marina Bellani, Michael Seidman, Maria R. Baer, Rena Lapidus, Stephen B. Baylin, Feyruz V. Rassool. Combination of DNA methyltransferase and PARP inhibitors as a novel therapy strategy for multiple cancers: Key data in AML and triple negative breast cancer [abstract]. In: Proceedings of the AACR International Conference: New Frontiers in Cancer Research; 2017 Jan 18-22; Cape Town, South Africa. Philadelphia (PA): AACR; Cancer Res 2017;77(22 Suppl):Abstract nr IA13.

Bare below elbows: does this policy affect handwashing efficacy and reduce bacterial colonisation?

UK Department of Health guidelines recommend that clinical staff are 'bare below the elbows'. There is a paucity of evidence to support this policy. One may hypothesise that absence of clothing around wrists facilitates more effective handwashing: this study aims to establish whether dress code affects bacterial colonisation before and after handwashing. Sixty-six clinical staff volunteered to take part in the study, noting whether they were bare below the elbows (BBE) or not bare (NB). Using a standardised technique, imprints of left and right fingers, palms, wrists and forearms were taken onto mini agar plates. Imprints were repeated after handwashing. After incubation, colonies per plate were counted, and subcultures taken. Thirty-eight staff were BBE and 28 were not. A total of 1112 plates were cultured. Before handwashing there was no significant difference in number of colonies between BBE and NB groups (Mann-Whitney, P < 0.05). Handwashing reduced the colony count, with greatest effect on fingers, palms and dominant wrists (t-test, P < 0.05). Comparing the two groups again after handwashing revealed no significant difference (Mann-Whitney, P < 0.05). Subcultures revealed predominantly skin flora. There was a large variation in number of colonies cultured. Handwashing resulted in a statistically significant reduction in colony count on fingers, palms and dominant wrist regardless of clothing. We conclude that handwashing produces a significant reduction in number of bacterial colonies on staff hands, and that clothing that is not BBE does not impede this reduction.

Frequent decreased expression of candidate tumor suppressor gene, DEC1, and its anchorage‐independent growth properties and impact on global gene expression in esophageal carcinoma

Previous studies showed that expression of the novel candidate tumor suppressor gene, DEC1 (Deleted in Esophageal Cancer 1), is reduced in esophageal carcinoma and suppresses cancer cell growth in vitro and tumor growth in vivo in nude mice. This study shows that DEC1 gene expression was downregulated in 100% of 16 esophageal squamous cell carcinoma (ESCC) cell lines and 52 and 45%, respectively, of esophageal tumor specimens from Hong Kong and a high-risk ESCC region of Henan, China. Using epitope tagging, the DEC1 protein was localized to both the cytoplasm and nucleus of the cell. In 3D Matrigel culture, no significant difference in colony numbers formed was observed for DEC1 stable transfectants, as compared to vector-alone transfectant controls. However, significantly smaller colony sizes were observed for the DEC1 transfectants. In in vitro cell migration, invasion and soft agar assays of DEC1 transfectants, only the soft agar assay showed statistically significant differences in colony numbers with the vector-alone controls, indicating that DEC1 may be involved in anchorage-independent cell growth. In addition, the global gene expression affected by DEC1 in tumor-suppressive stable transfectants was investigated using cDNA oligonucleotide microarray hybridization. Three candidate genes, TFPI-2, GDF15 and DUSP6, were identified through this approach; they are downregulated in tumor segregants of DEC1 stable transfectants, ESCC cell lines and esophageal tumors and have a potential role in tumor growth and progression. These studies show that DEC1 is involved in esophageal cancer development and help elucidate its functional role in tumor development.

THE EFFECT OF ETHANOL ON RHESUS MONKEY EMBRYONIC STEM CELLS

Fetal Alcohol Syndrome results from ethanol exposure and can be deleterious to a developing fetus. The earliest stages of fetal development are most sensitive to the effects of ethanol, however that time of development is very difficult to study in vivo. Embryonic stem cells (ESCs) are derived from blastocyst stage embryos making ESCs an appropriate model for early development. Rhesus monkey (Macaca mulatta) ESC lines (ORMES 6 and 7) were treated with 0, 0.01%, 0.1%, or 1.0% ethanol in modified DMEM media for one month. Approximately 30 colonies were cultured per 60mm dish in 5 mL of media. Culture media were changed daily and ESC colonies were passaged twice weekly. Colony number and morphology were observed every 3–4 days and by week 2, morphological differences became apparent. ESCs in the 1.0% ethanol treatment group were either hypotrophic with decreased colony area or showed abnormal growth patterns that may indicate differentiation. No differences in colony numbers or viability were observed. Area measurements indicated a significant decrease (p<0.05) between the control and 1.0% ethanol treatment group for ORMES 6 at fourteen days and for ORMES 7 at twenty-four days and between the control and 0.1% ethanol treatment group at day twenty-eight in ORMES 7. Immunofluorescence staining of key pluripotency markers (OCT4, SSEA1 & 4, TRA-1-60 & 81, and alkaline phosphatase) indicated a loss of pluripotency of the ESC lines used. The morphological changes and immunofluorescence indicates that ethanol affects these early cells of development and may cause uncontrolled differentiation. Further studies are warranted to determine possible mechanisms for the effects observed, especially in the higher ethanol treatment groups. Supported by NIH grants RR00169 and AA014173. (platform)

Number and Proliferative Capacity of Human Mesenchymal Stem Cells Are Modulated Positively in Multiple Trauma Patients and Negatively in Atrophic Nonunions

Mesenchymal stem cells (MSCs) participate in regenerative osteogenesis by generating bone-forming cells. To examine the proliferative capacity of MSC populations from bone marrow and their relationship to trauma severity (multiple trauma, monofracture, atrophic nonunion), we quantified colony properties of human MSCs in vitro. Serum levels of mediators associated with bone formation were also assessed. Fifty-five individuals were enrolled in this study (13 multiple trauma patients, 15 patients with monofracture, 20 patients with atrophic nonunions, 7 healthy volunteers). The colony forming unit-fibroblast (CFU-F) assay was used to quantify total colony number, mean cell density per colony, and mean colony area. MSC phenotype was established using flow cytometry and osteogenic differentiation. MSCs obtained from multiple-trauma patients yielded the highest reservoir. Significant differences in colony numbers of MSCs in female subjects were found between multiple-trauma patients (mean +/- SD 48 +/- 21 CFU-F/culture) and healthy volunteers (18.7 +/- 3.3 CFU-F/culture, P < 0.05), patients with monotrauma (15 +/- 10 CFU-F/culture, P < 0.05), and patients with atrophic nonunions (6.3 +/- 4.1 CFU-F/culture, P < 0.05). In male participants, significant differences were found between patients with nonunions (14 +/- 14 CFU-F/culture) and healthy volunteers (54 +/- 17 CFU-F/culture, P < 0.05) as well as multiple-trauma patients (59 +/- 25 CFU-F/culture, P < 0.05). The highest proliferative capacity (cell density) was seen in multiple-trauma patients. These data suggest that trauma severity and gender affect the reservoir and proliferation capacity of bone marrow-derived MSCs.

Evaluation of abundance of aerobic bacteria in the rhizosphere of transgenic and non-transgenic alfalfa lines

Fourteen genetically modified lines of alfalfa (Medicago sativa) containing the gene Ov from Japanese quail, coding for a methionine-rich protein ovalbumin, were evaluated for nodulation ability and concentration of aerobic bacteria in the rhizosphere. The transgenic lines were derived from a highly regenerable genotype Rg9/I-14-22, selected from cv. Lucia. On selective media, a higher concentration of ammonifying bacteria, bacterial spores, denitrifying and nitrifying bacteria were observed in the rhizosphere of transgenic clonesand, on the other hand, lower concentration of cellulolytic bacteria and Azotobacter spp. compared with the rhizosphere of non-transgenic clone SE/22-GT2. A statistically significant difference in the concentration of all the bacterial types was found between samples taken from two types of substrates (i.e. sterile vs. nonsterile). Higher bacterial concentration (measured as colony forming units per g soil dry mass) were observed for all tested groups of culturable bacteria in the non-sterile substrate. The presence of Azotobacter spp. was found only in the rhizosphere of plants grown in non-sterile soil in which the highest number of fertile soil particles (97 %) was observed in transgenic clones SE/22-9-1-12 and SE/22-11-1-1S.1. Concentration of bacteria involved in the N cycle in the soil was increased in the rhizosphere of transgenic clones and decreased in the rhizosphere of non-transgenic plants compared with the average value. In spite of some differences in colony numbers in samples isolated from the root rhizosphere of transgenic and nontransgenic alfalfa plants, we could not detect any statistically significant difference between individual lines.

In vitro Studies of Haematopoietic Colony-Forming Capacity of Human Fetal Liver Cells at Exposure to Cytotoxic and Immunomodulatory Drugs

Objective: Intrauterine transplantation with haematopoietic stem cells (SC) as a treatment for immunological, haematological and metabolic inherited disorders has not been effective in fetuses with a normal immunological status. Inhibition of the fetal haematopoiesis or immunosuppression might therefore be a therapeutic alternative in fetal SC transplantation. The aim of the present study was to evaluate the effects of drugs that might be considered as therapeutic options in fetal transplantations on the colony-forming capacity of fetal haematopoietic SC. Methods: Fetal liver cells were incubated with doxorubicin, daunorubicin, antithymocyte-globulin (ATG), OKT-3 (Orthoclone) and betamethasone. The effects of these drugs on colony formation by fetal hematopoietic SC were evaluated. Colony-forming capacity assays were cultured during 10 days after drug incubation on day 1 and evaluated for differences in number of colonies compared to unexposed cells. For betamethasone, similar studies were performed with adult bone marrow. Results: A significant reduction in fetal liver haematopoietic colony formation of BFU-E, CFU-GM and CFU- GEMM was detected when 0.1 M doxorubicin (p < 0.05) and 0.5 µM daunorubicin (p < 0.05) were added. OKT-3 (5 µg/ml) and ATG (4 µg/ml) significantly reduced BFU-E, CFU-GM and CFU-GEMM (p < 0.05). A concentration of 0.2 µg/ml betamethasone caused a significant reduction of BFU-E, CFU-GM and CFU-GEMM (p < 0.05). Conclusions: The drugs investigated in this in vitro study were capable to different degrees to decrease the colony-forming capacity of fetal haematopoietic progenitor cells. Whether this strategy will become an alternative in fetal SC transplantations is an open question and further studies are required to elucidate the potential use of these drugs in fetal transplantation.

Predictive value of thymidylate synthase and dihydropyrimidine dehydrogenase

before injection, no differences in colony numbers were shown. These results suggest that different AML progenitor cells can be defined in the whole AML population, dependent on the test system used. Moreover, the findings indicate a hierarchy in AML progenitors, including cells with short-term and longterm repopulating abilities, as has already been defined in the normal counterpart. This expanding knowledge on the biological behaviour of AML progenitors ita vitro may be used in future clinical trials to optimise new therapeutic strategies, especially in the possible elimination of AML progenitor cells from the graft.

Number of Colonies Produced by 22 Isolates of Powdery Mildew on Primary Leaves of 10 Barley Cultivars

AbstractPrimary leaves of 10 barley cultivars were uniformly inoculated with 22 virulent powdery mildew isolates using a jet spore trap. The number of colonies per unit area leaf was counted. Significant differences in number of colonies among cultivars were found, indicating different degrees of susceptibility among cultivars. While cultivars were rather consistent in their rankings for number of colonies produced by most isolates, interactions between some cultivars and isolates were found. In spite of great differences in geographical origin of cultivars and isolates, the interactions were not associated with origin.

Circulating haematopoietic progenitors in myelofibrosis.

We studied circulating erythroid and granulocyte-monocyte progenitors in 18 patients with idiopathic myelofibrosis and in healthy controls, using the methyl cellulose assay. 9 of the patients had been splenectomized prior to the study. The median number of circulating erythroid burst-forming units (BFU-E) was 8 times higher than that of the controls. 12 patients also had CFU-E (colony-forming unit, erythroid) in the blood. 10 patients had spontaneous BFU-E colony formation, and 8 patients had spontaneous CFU-E colony growth. Granulocyte-monocyte progenitors (CFU-GM) were increased 47 times compared to the controls. There were no differences in colony numbers between splenectomized and non-splenectomized patients. We conclude that in myelofibrosis, circulating erythroid and granulocyte-monocyte progenitors are usually markedly increased in number, but erythroid precursors to a lesser extent than granulocyte-monocyte precursors. Many patients, but not all, show spontaneous erythroid colony formation.

Overwintering of Venturia inaequalis, the causal agent of apple scab, on different cultivars

SUMMARYA maturation index summarising pseudothecial development of Venturia inaequalis was devised. The index demonstrated differences in overwintering on 10 apple cultivars and the crab apple Malus toringo in experiments over two winters. At about the green cluster stage, when there is a high proportion of susceptible foliage, there were 50–100 fold differences in the yield of ascospores from these hosts, due mainly to different rates of pseudothecial maturation. These rates were shown to be independent of cultivar differences in colony numbers on leaves in the previous autumn.Cultivar differences in the timing of fruit bud development were independent of differences in the overwintering of V. inaequalis. These findings confirm the importance of considering cultivar identity in apple scab management.

CHANGES IN THE METABOLISM OF WHEAT LEAVES INDUCED BY INFECTION WITH POWDERY MILDEW

CHANGES IN THE METABOLISM OF WHEAT LEAVES INDUCED BY INFECTION WITH POWDERY MILDEW