Effects of salvadora persica extract on the bacterial population in single-species biofilm

The effect of S. persica extract on the population of bacteria in single-species biofilm was investigated. An artificial mouth (NAM) system was used for the development of biofilm. In one experiment, aqueous extract of S. persica at sub-MIC concentration (5 mg/mL) was first pumped into the NAM system to treat the experimental pellicle on glass beads before inoculating it with the bacteria (Strep. mitis, Strep. mutans and Strep. sanguinis). This would resemble using the aqueous extract of S. persica before meals. In another experiment, the aqueous extract of S. persica was pumped into the NAM system after the bacterial inoculation and this would resemble using it after meals. In both experiments, 24 h biofilms were allowed to form. The bacterial population of the biofilms was determined and expressed as colony forming unit per mL (cfu mL-1). For negative control, sterilized distilled water was used whereas for positive control, commercial Listerine. It was found that the population of Strep. mitis, Strep. mutans and Strep. sanguinis in the respective biofilms for both experiments involving treatment with S. persica extract or Listerine was significantly reduced by more than 70% (p<0.05) when compared with the negative control. Comparing the effect of S. persica with Listerine on the bacterial population of the biofilms when used either before or after meal showed that S. persica is slightly less effective towards S. sanguinis before meal and towards S. mutans after meal (p<0.05). This study suggests that the efficiency of the antibacterial effect of S. persica is species-related and by its way of usage

Revisiting the association between candidal infection and carcinoma, particularly oral squamous cell carcinoma

Background: Tobacco and alcohol are risk factors associated with cancer of the upper aerodigestive tract, but increasingly the role of infection and chronic inflammation is recognized as being significant in cancer development. Bacteria, particularly Helicobacter pylori, and viruses such as members of the human papilloma virus family and hepatitis B and C are strongly implicated as etiological factors in certain cancers. There is less evidence for an association between fungi and cancer, although it has been recognized for many years that white patches on the oral mucosa, which are infected with Candida, have a greater likelihood of undergoing malignant transformation than those that are not infected. Objective: This article reviews the association between the development of oral squamous cell carcinoma in potentially malignant oral lesions with chronic candidal infection and describes mechanisms that may be involved in Candida-associated malignant transformation

Health behaviour and its relationship with the risk of getting dengue fever at the district of Temerloh Pahang Darul Makmur; a case control study, 1999

Dengue Fever and Dengue Haemorrhagic Fever infections are common infectious diseases in Malaysia, which caust'S mortality and morbidity and thus losses in teml of economics to the individuals and the country .No satisfactory outcomes has been gained from various prevention and control programmes with multiple strategies. Health behaviour is recognized to be one of the causes for the increase risk of getting the disease. In order to identify the specific health behaviour for future interventions, a matched 1: 1 case control study was conducted at the District of Temerloh Pahang Darul Makmur in June 1999. The factors studied were the knowledge, attitudes and practices regarding prevention and control, treatment, law enforcement and level of awareness of the danger caused by Dengue/Dengue Haemorrhagic Fever. Cases were selected from the registration data available at the Temerloh's Health Office with confinned diagnosis starting from the period 1st of January to 31st of December 1998. Whilst the controls were those who were never diagnosed to have DF/DHF and free from the signs and symptoms of the disease until the 31st. of December 1998. Sixty seven pairs of case and control were surveyed. The univariate analysis showed that agreeable to mosquito net usage (p=0.00), agreeable to blood taking (p=0.00), agreeable to stay longer in hospital if complications arise(p=0.00), allowing sick family members to be warded (p=0.03), letting blood to be tested (p=0.001 ). sending sick family members to hospital (p=0.003), aware of the danger of Dengue fever (p=0.039), agreeable that DF must be known by everybody regardless of their disease status (p=0.027), mosquito coil usage (p=0.02), cleaning water containers (p=0.03) and breeding larvae (p=0.03) were factors significantly related to the risk of getting DF/DHF. The multivariate analysis narrowed down the predictors of getting DF/DHF to only agreeable to blood taking (p=0.02), agreeable to mosquito net usage (p=0.03), agreeable to stay longer in hospital if complications arise (p=0.03) and lastly allowing sick family members to be warded (p=0.01). On conclusion, there is a relationship between bad health behaviour and increase risk of getting dengue fever

Boletín Oficial de la Provincia de Oviedo: Número 252 - 1941 noviembre 12

Oral squamous cell carcinoma (OSCC) can remain undiagnosed until in an advanced, and sometimes lethal, state. It is often preceded by a potentially malignant lesion, which may manifest as a white patch 'leukoplakia' of the oral mucosa. Previous research has implicated an association between the presence of Candida albicans and the progression of leukoplakias to OSCC. Alcohol may contribute to oral cancer via its conversion to acetaldehyde, a known carcinogen, which is also a product of C. albicans metabolism. The reversible conversion of ethanol to acetaldehyde is catalysed by enzymes known as alcohol dehydrogenases (ADHs) and in C. albicans, it is not known which ADH is responsible for acetaldehyde production. Aims of this study: To investigate expression of the CaADH genes in vitro and to identify the C. albicans genes responsible for acetaldehyde production. It is also the aim of this study to detect the expression of the CaADH genes in archival formalin-fixed paraffin-embedded (FFPE) samples from leukoplakia biopsies. Study hypotheses: (i) that production of acetaldehyde by C. albicans via expression of ADH genes can be detected in lesions diagnosed as CHC; (ii) that the presence of C. albicans in CHC lesions is associated with expression of CaADH1 mRNA; (iii) that acetaldehyde production may provide a mechanism for the previously reported putative link between oral infection/colonisation with C. albicans and oral cancer. Methods: The levels of expression for three C. albicans genes, CaADH1, CaADH2 and CaADH3, were measured under various growth conditions using Northern blot analysis and qRT-PCR. The three C. albicans genes were also cloned and expressed in the model yeast Saccharomyces cerevisiae. CaAdhp polypeptide expression was confirmed by Western blot analysis. Ethanol utilisation was assayed in cell extracts of recombinant S. cerevisiae strains expressing CaADH1 or CaADH2. Disruption of the endogenous ScADH2 gene, which is responsible for production of acetaldehyde in S. cerevisiae, was carried out in the recombinant S. cerevisiae strain that expressed CaADH1 to reduce background ScAdhp activity. The presence of Candida was investigated in archival FFPE samples from four sample groups: normal oral mucosa, non-dysplastic leukoplakia, chronic hyperplastic candidosis (CHC) and non-CHC dysplastic leukoplakia. Candida was detected in FFPE samples by histology and immunocytochemistry and C. albicans ADH1 and ADH2 mRNAs were detected by RT-PCR and qRT-PCR. Results: Northern blot analysis showed that CaADH1 mRNA was expressed under a variety of in vitro growth conditions while CaADH2 was only detected during stationary phase in a rich medium. No expression of CaADH3 was detected. Each gene was cloned in S. cerevisiae but CaADH3 again was not expressed. Cell extracts from the Adh1p-expressing S. cerevisiae recombinant, but not the Adh2p-expressing recombinant, or an empty vector control strain, possessed ethanol utilising Adh activity. Expression of C. albicans Adh1p in a recombinant S. cerevisiae strain in which the endogenous ScADH2 gene had been deleted conferred an NAD-dependant ethanol utilising and hence acetaldehyde producing Adh activity. FFPE samples were analysed by immunocytochemistry for C. albicans and by RT-PCR for C. albicans gene expression. C. albicans was detected by both methods in FFPE samples diagnosed as CHC, but not in any other sample group, including the non-dysplastic leukoplakias. RT-PCR confirmed the presence of high levels of C. albicans ADH1 mRNA expression, compared to a house-keeping gene, in CHC biopsies but CaADH2 expression was variable. Conclusions: C. albicans Adh1p was shown to be the major Adh isozyme involved in the production of acetaldehyde by this human commensal yeast. Immunohistopathological evidence was obtained from FFPE samples from patients with previously diagnosed CHC indicating that C. albicans was the predominant species in the lesions. The presence of C. albicans in CHC lesions was associated with a high expression of CaADH1 mRNA. The results of this study provide the first experimental support for the hypothesis of a putative link between acetaldehyde production by oral infection/colonisation with C. albicans and oral cancer despite the lack of malignant transformation in the clinical cases of chronic hyperplastic candidosis studied

Evaluation of Wound Closure Activity of Nigella sativa

Nigella sativa, Melastoma malabathricum, Pluchea indica, and Piper sarmentosum are common Asian traditional medicines to treat minor wounds. This study aimed to investigate the in vitro wound healing properties of aqueous extracts of these plants using human gingival fibroblast (HGF) monolayer as study model. DPPH scavenging activity of the extracts was evaluated and effect on HGF proliferation was determined. Their effect on HGF’s function to synthesize collagen was indicated by the level of hydroxyproline produced and effect on wound healing activity was assessed using an in vitro scratch assay. The influence of the extracts on expression of bFGF and TGF-β was also determined. Results revealed all four extracts to exhibit low free radical scavenging activity. The extract from N. sativa (NSSE) compared to the others showed favourable enhancement of HGF proliferation with EC50 of 22.67±3.06 µg/mL (P<0.05) with accelerated wound closure activity despite its nonsignificant effect on collagen synthesis. In addition to the elevated level of bFGF by up to 15% at 100 µg/mL of NSSE, a slightly better effect was observed on the expression of TGF-β. NSSE thus showed that promising wound healing properties and data obtained may contribute towards validation of its traditional use for the healing of oral wounds

Saliva Sampling of Alcoholic Participants using Three Saliva Collection Methods

The potential of using saliva as a diagnostic fluid is well documented. The aim of this study was to assess the quality and quantity of saliva DNA of alcoholic and non-alcoholic participants using three saliva collection methods; DNA-SalTM (Oasis Diagnostics, USA), Oragene-DNA (DNA Genotek Inc, Ontario, Canada) and whole saliva collection method. Saliva DNA of non-alcoholic (n=30) and alcoholic participants (n=10) age between 25 and 35 years was assessed qualitatively and quantitatively using spectrophotometry. Saliva DNA quantity was the highest for all participants when using the DNA-Sal TM saliva collection kit (p<0.05). The use of a mechanical scraper provided only in the DNA-Sal TM kit may have contributed to the highest DNA yield for all participants. The quantity of saliva DNA when assessed using spectrophotometer was found to be significantly lower (p<0.05) for the alcoholic (16±3.57 ng/μL) than non-alcoholic participants (19.92±6.18 ng/μL). To determine the integrity of the DNA samples, PCR amplification of the Alcohol Dehydrogenase gene, ADH1B was carried out and the PCR was found to be successful. For all participants, the DNA quality of the saliva collected using the three saliva collection methods was found to be in the acceptable range considered as pure DNA. The DNA quality and quantity of saliva collected from the three saliva collection methods were considered suitable for research purposes

The Antiproliferative and Apoptotic Effects of Capsaicin on an Oral Squamous Cancer Cell Line of Asian Origin, ORL-48

Background and Objectives: The antitumor activities of capsaicin on various types of cancer cell lines have been reported but the effect of capsaicin on oral cancer, which is prevalent among Asians, are very limited. Thus, this study aimed to investigate the effects of capsaicin on ORL-48, an oral cancer cell line of Asian origin. Materials and Methods: Morphological changes of the ORL-48 cells treated with capsaicin were analyzed using fluorescence microscopy. The apoptotic-inducing activity of capsaicin was further confirmed by Annexin V-Fluorescein isothiocyanate/Propidium iodide (V-FITC/PI) staining using flow cytometry. In order to establish the pathway of apoptosis triggered by the compound on ORL-48 cells, caspase activity was determined and the mitochondrial pathway was verified by mitochondrial membrane potential (MMP) assay. Cell cycle analysis was also performed to identify the cell cycle phase of ORL-48 cells being inhibited by the capsaicin compound. Results: Fluorescence microscopy exhibited the presence of apoptotic features in capsaicin-treated ORL-48 cells. Apoptosis of capsaicin-treated ORL-48 cells revealed disruption of the mitochondrial-membrane potential, activation of caspase-3,-7 and-9 through an intrinsic apoptotic pathway and subsequently, apoptotic DNA fragmentation. The cell cycle arrest occurred in the G1-phase, confirming antiproliferative effect of capsaicin in a time-dependent manner. Conclusion: This study demonstrated that capsaicin is cytotoxic against ORL-48 cells and induces apoptosis in ORL-48 cells possibly through mitochondria mediated intrinsic pathway resulting in cell cycle arrest. © 2019 by the authors

Acute and sub-acute oral toxicity of Dracaena cinnabari resin methanol extract in rats

Abstract Background Dracaena cinnabari (DC) is a perennial tree that located on the Southern coast of Yemen native to the Socotra Island. This tree produces a deep red resin known as the Dragon’s blood, the Twobrother’s Blood or Damm Alakhwain. The current study performed to evaluate the safety of the DC resin methanol extract after a single or 28 consecutive daily oral administrations. Methods In assessing the safety of DC resin methanol extract, acute and sub-acute oral toxicity tests performed following OECD guidelines 423 and 407, respectively, with slight modifications. In acute oral toxicity test, DC resin methanol extract administered to female Sprague Dawley rats by oral gavage at a single dose of 300 and 2000 mg/kg body weight. Rats observed for toxic signs for 14 days. In sub-acute oral toxicity test, DC resin methanol extract administered to the rats by oral gavage at 500, 1000, and 1500 mg/kg body weight daily up to 28 days to male and female Spradgue Dawley rats. The control and high dose in satellite groups were also maintained and handled as the previous groups to determine the late onset toxicity of DC resin methanol extract. At the end of each test, hematological and biochemical analysis of the collected blood were performed as well as gross and microscopic pathology. Results In acute oral toxicity, no treatment-related death or toxic signs were observed. It revealed that the DC resin methanol extract could be well tolerated up to the dose 2000 mg/kg body weight and could be classified as Category 5. The sub-acute test observations indicated that there are no treatment-related changes up to the high dose level compared to the control. Food consumption, body weight, organ weight, hematological parameters, biochemical parameters and histopathological examination (liver, kidney, heart, spleen and lung) revealed no abnormalities. Water intake was significantly higher in the DC resin methanol extract treated groups compared to the control. Conclusion This study demonstrates tolerability of DC resin methanol extract administered daily for 28 days up to 1500 mg/kg dose

Antitumor Activity of Ficus deltoidea Extract on Oral Cancer: An In Vivo Study

Background. The aim of this study is to evaluate the chemopreventive and chemotherapeutic activities of Ficus deltoidea (FD) in an animal model induced for oral cancer using 4-nitroquinoline-1-oxide (4NQO). Methods. Male Sprague-Dawley (SD) rats were randomized into six groups (n = 7 per group): Group 1 (untreated group); Group 2 (control cancer group) received 4NQO only for 8 weeks in their drinking water; Groups 3 and 4 (chemopreventive) received 4NQO for 8 weeks and were simultaneously treated with FD extract at 250 and 500 mg/kg, respectively, by oral gavage; Groups 5 and 6 (chemotherapeutic) received 4NQO for 8 weeks followed by the administration of FD extract at 250 and 500 mg/kg, respectively, for another 10 weeks. The incidence of oral cancer was microscopically evaluated. Moreover, immunohistochemical expression was analysed in tongue specimens using an image analyser computer system, while the RT2 profiler PCR array method was employed for gene expression analysis. Results. The results of the present study showed a beneficial regression effect of the FD extract on tumor progression. The FD extract significantly reduced the incidence of oral squamous cell carcinoma (OSCC) from 100% to 14.3% in the high-dose groups. The immunohistochemical analysis showed that the FD extract had significantly decreased the expression of the key tumor marker cyclin D1 and had significantly increased the expression of the β-catenin and e-cadherin antibodies that are associated with enhanced cellular adhesion. Based on the gene expression analysis, FD extract had reduced the expression of the TWIST1 and RAC1 genes associated with epithelial-mesenchymal transition (EMT) and had significantly downregulated the COX-2 and EGFR genes associated with cancer angiogenesis, metastasis, and chemoresistance. Our data suggest that the FD extract exerts chemopreventive and chemotherapeutic activities in an animal model induced for oral cancer using 4NQO, thus having the potential to be developed as chemopreventive and chemotherapeutic agents