Resistotyping and extended-spectrum beta-lactamase genes among Escherichia coli from wastewater treatment plants and recipient surface water for reuse in South Africa.

The spread of extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli has increased in different environments. This study aimed to evaluate the patterns of antibiotic resistance and ESBL genes among E. coli isolates collected from wastewater and recipient surface water in South Africa. Fifteen samples containing nine wastewater and six river water samples were collected from a local wastewater treatment plant. The E. coli isolates were detected using standard microbiology methods. Antibiotic susceptibility testing was performed using disc diffusion agar. The occurrence of bla CTX-M, bla SHV and bla TEM ESBL genes was investigated by PCR. Exactly 140 isolates were selected from the primary enumeration plates with a log10 CFU/mL count that ranged from 4.1 to 4.2 (influent), 4.2 to 4.5 (biofilter) and 2.5 to 3.3 (effluent). The wastewater effluent showed an impact on the receiving water environment, as the treatment efficiency was 92% and the downstream log10 CFU/mL count (range, 3.6-3.8 log10 CFU/mL) was higher than the upstream count (range, 3.3-3.6 log10 CFU/mL). Antibiotic testing results showed that 40% to 100% of E. coli isolates were resistant to ampicillin, penicillin, tetracycline and cefotaxime but susceptible to imipenem, meropenem and ciprofloxacin. A total of 40 studied isolates (28.6%) had both the bla TEM and bla CTX-M genes, while no bla SHV was detected. The wastewater treatment plants contributed multidrug-resistant ESBL-producing E. coli isolates that can be potential environmental health risks. Regular monitoring policies are recommended to prevent the spread of antibiotic resistance in the region.

Computer-Based Oral Hygiene Instruction versus Verbal Method in Fixed Orthodontic Patients.

STATEMENT OF PROBLEM: Fixed orthodontic appliances in the oral cavity make tooth cleaning procedures more complicated. OBJECTIVES: This study aimed to compare the efficacy of computerized oral hygiene instruction with verbal technique among fixed orthodontic patients referred to the evening clinic of Orthodontics of Shiraz Dental School. MATERIALS AND METHODS: A single-blind study was performed in Orthodontic Department of Shiraz, Islamic Republic of Iran, from January to May 2015 following the demonstrated exclusion and inclusion criteria. The sample size was considered 60 patients with 30 subjects in each group. Bleeding on probing and plaque indices and dental knowledge were assessed in the subjects to determine pre-intervention status. A questionnaire was designed for dental knowledge evaluation. The patients were randomly assigned into the computerized and verbal groups. Three weeks after the oral hygiene instruction, indices of bleeding on probing and plaque index and the dental knowledge were evaluated to investigate post-intervention outcome. The two groups were compared by chi-square and student t tests. The pre- and post-intervention scores in each group were compared using paired t-test. RESULTS: In the computerized group, the mean score for plaque index and bleeding on probing index was significantly decreased while dental health knowledge was significantly increased after oral hygiene instruction, in contrast to the verbal group. CONCLUSIONS: Within the limitations of the current study, computerized oral hygiene instruction is proposed to be more effective in providing optimal oral health status compared to the conventional method in fixed orthodontic patients.

Analysis of 41 suicide attempts by wrist cutting: a retrospective analysis.

PURPOSE: Self-cutting injuries have a low mortality rate, but this type of injuries has special clinical significance because they have the potential of leading to devastating disability and repeated suicide attempts. The purpose of this study is to analyze the nature and outcomes of wrist-cutting injuries. MATERIAL AND METHOD: A retrospective study was designed in order to investigate 41 suicide attempts by wrist cutting attended to Uludag University Faculty of Medicine Emergency Department between June 2008 and December 2014. The patients were analyzed for age, gender, alcohol intake, psychological state, prior suicide attempts, and clinical features such as injury side, injury pattern, and used tool. RESULTS: It was seen that the severity of wrist-cutting injury variates between gender and age. CONCLUSION: Alcohol or drug consumption and having a diagnosed psychiatric disorder create a higher risk for extensive wrist lacerations. It was seen that skin only lacerations were most likely to repeat the act and therefore are most in need of psychiatric intervention. LEVEL OF EVIDENCE: Level III, retrospective study.

Targeted next-generation sequencing of locally advanced squamous cell carcinomas of the head and neck reveals druggable targets for improving adjuvant chemoradiation.

BACKGROUND: Despite clear differences in clinical presentation and outcome, squamous cell carcinomas of the head and neck (SCCHN) arising from human papilloma virus (HPV) infection or heavy tobacco/alcohol consumption are treated equally. Next-generation sequencing is expected to reveal novel targets for more individualised treatment. PATIENTS AND METHODS: Tumour specimens from 208 patients with locally advanced squamous cell carcinoma of the hypopharynx, oropharynx or oral cavity, all uniformly treated with adjuvant cisplatin-based chemoradiation, were included. A customised panel covering 211 exons from 45 genes frequently altered in SCCHN was used for detection of non-synonymous point and frameshift mutations. Mutations were correlated with HPV status and treatment outcome. RESULTS: Mutational profiles and HPV status were successfully established for 179 cases. HPV- tumours showed an increased frequency of alterations in tumour suppressor genes compared to HPV+ cases (TP53 67% versus 4%, CDKN2A 18% versus 0%). Conversely, HPV+ carcinomas were enriched for activating mutations in driver genes compared to HPV- cases (PIK3CA 30% versus 12%, KRAS 6% versus 1%, and NRAS 4% versus 0%). Hotspot TP53 missense mutations in HPV- carcinomas correlated with an increased risk of locoregional recurrence (hazard ratio [HR] 4.3, 95% confidence interval [CI] 1.5-12.1, P=0.006) and death (HR 2.2, 95% CI 1.1-4.4, P=0.021). In HPV+ SCCHN, driver gene mutations were associated per trend with a higher risk of death (HR 3.9, 95% CI 0.7-21.1, P=0.11). CONCLUSIONS: Distinct mutation profiles in HPV- and HPV+ SCCHN identify subgroups with poor outcome after adjuvant chemoradiation. Mutant p53 and the phosphoinositide 3-kinase pathway were identified as potential druggable targets for subgroup-specific treatment optimisation.

Cellular and molecular properties of (90)Y-labeled cetuximab in combination with radiotherapy on human tumor cells in vitro.

PURPOSE: Anti-EGFR antibody cetuximab (C225) is used in combination with radiotherapy of head and neck squamous cell carcinoma (HNSCC) patients. We investigated whether conjugation of cetuximab with trans-cyclohexyl-diethylene-triamine-pentaacetic acid (CHX-A″-DTPA) and radiolabeling with (90)Yttrium affect the molecular and cellular function of cetuximab and improve its combined effect with external-beam irradiation (EBI). METHODS: The following cell lines were used: HNSCC UT5, SAS, FaDu, as well as A43, Chinese hamster ovary cells (CHO), and human skin fibroblast HSF7. Binding affinity and kinetics, specificity, retention, and the combination of (90)Y-cetuximab with EBI were evaluated. RESULTS: Control cetuximab and CHX-A″-DTPA-cetuximab blocked the proliferation activity of UT5 cells. In combination with EBI, CHX-A″-DTPA-cetuximab increased the radiosensitivity of UT5 to a similar degree as control cetuximab did. In contrast, in SAS and HSF7 cells neither proliferation nor radiosensitivity was affected by either of the antibodies. Binding [(90)Y]Y-CHX-A″-DTPA-cetuximab ((90)Y-cetuximab) to EGFR in HNSCC cells occurred time dependently with a maximum binding at 24 h. Retention of (90)Y-cetuximab was similar in both HNSCC cell lines; 24 h after treatment, approximately 90% of bound activity remained in the cell layer. Competition assays, using cell membranes in the absence of an internalized fraction of cetuximab, showed that the cetuximab affinity is not lost as a result of conjugation with CHX-A″-DTPA. Cetuximab and CHX-A″-DTPA-cetuximab blocked EGF-induced Y1068 phosphorylation of EGFR. The lack of an effect of cetuximab on EGF-induced Akt and ERK1/2 phosphorylation and the inhibition of irradiation (IR)-induced Akt and ERK1/2 phosphorylation by cetuximab were not affected by DTPA conjugation. (90)Y-cetuximab in combination with EBI resulted in a pronounced inhibition of colony formation of HNSCC cells. CONCLUSIONS: Conjugation of CHX-A″-DTPA to cetuximab does not alter the cellular and biological function of cetuximab. (90)Y-labeling of cetuximab in combination with EBI may improve radiotherapy outcome.

Progress in pursuit of therapeutic A2A antagonists: the adenosine A2A receptor selective antagonist KW6002: research and development toward a novel nondopaminergic therapy for Parkinson's disease.

Research and development of the adenosine A2A receptor selective antagonist KW6002 have focused on developing a novel nondopaminergic therapy for Parkinson's disease (PD). Salient pharmacologic features of KW6002 were investigated in several animal models of PD. In rodent and primate models, KW6002 provides symptomatic relief from parkinsonian motor deficits without provoking dyskinesia or exacerbating existing dyskinesias. The major target neurons of the A2A receptor antagonist were identified as GABAergic striatopallidal medium spiny neurons. A possible mechanism of A2A receptor antagonist action in PD has been proposed based on the involvement of striatal and pallidal presynaptic A2A receptors in the "dual" modulation of GABAergic synaptic transmission. Experiments with dopamine D2 receptor knockout mice showed that A2A receptors can function and anti-PD activities of A2A antagonists can occur independent of the dopaminergic system. Clinical studies of KW6002 in patients with advanced PD with L-dopa-related motor complications yielded promising results with regard to motor symptom relief without motor side effects. The development of KW6002 represents the first time that a concept gleaned from A2A biologic research has been applied successfully to "proof of concept" clinical studies. The selective A2A antagonist should provide a novel nondopaminergic approach to PD therapy.

Immunohistochemical examination of NOS and SOD in nasal mucosa.

An immunohistochemical examination was performed to detect the localization of neural-type nitric oxide synthase (nNOS), endothelial-type NOS (eNOS) and inducible-type NOS (iNOS) in the human and mouse nasal mucosa. nNOS-immunoreactive nerve fibers were found in the subepithelial layer and around the seromucous glands of mice. In these fibers, immunodouble staining revealed co-localization of nNOS and superoxide dismutase (SOD). In the human nasal mucosa of normal subjects, strong eNOS immunoreactivity and weak iNOS immunoreactivity were found in the columnar epithelium.

Removal of power line interference in signal-averaged electrocardiography systems.

A method for line interference reduction to be used in signal-averaged electrocardiography (SAECG) systems is proposed and its performance is analyzed. This new method is an adaptation of a previously reported technique for removal of line interference from conventional electrocardiograms. It involves the recording of a line reference signal simultaneous with the lead signals, so that a shifted and scaled version of it can be used to subtract line interference from the leads. It is shown that this line interference subtraction method can reduce line interference effectively and without introducing any additional noise into the ECG signal. It is also shown that Late Potential diagnostic decisions are not altered when this filter is applied. It is recommended that this technique be used in SAECG when line interference is unavoidable.

Emergence of Buruli ulcer disease in the Daloa region of Cote d'Ivoire.

Recent reports have suggested increases in Buruli ulcer (BU), an infection caused by Mycobacterium ulcerans in west Africa. In 1991, we conducted surveillance for BU in a rural area of Cote d'Ivoire and identified 312 cases of active or healed ulceration. A case-control study was then performed to investigate risk factors for this infection. The rate of illness did not appear to differ between males and females (5.2% versus 7.5%; P = 0.11). The highest rate of illness was seen in the 10-14-year-old age group (143 cases per 1,000 population). New cases increased more than three-fold between 1987 and 1991, and local prevalence of BU was as high as 16.3%. Twenty-six percent of persons with healed ulcers had chronic functional disability. Participation in farming activities near the main river in the region was identified in the case-control study as a risk factor for infection (odds ratio [OR] for each 10-min decrease in walking distance between the fields and the river = 1.52, 95% confidence interval [CI] 1.01, 2.28, P = 0.046). Wearing long pants was protective (OR 0.20, 95% CI 0.06, 0.62, P < 0.005). We conclude that the incidence of BU is increasing rapidly in Cote d'Ivoire. Specific causes of this increase were not identified, but wearing protective clothing appeared to decrease the risk of disease.

Subcellular localization and characterization of interleukin-1 alpha produced by rat bladder cancer cells.

A rat bladder cancer cell line BC31ad, previously reported to constitutively release tumor necrosis factor alpha (TNF alpha), was found to produce an interleukin (IL)-1-like factor inducing proliferation of murine thymocytes in a standard costimulation assay (LAF activity). IL-1-like factor was partially purified by DEAE-Sephacel, Sephacryl S-200 and MonoQ-FPLC chromatographies from the serum-free conditioned medium of a BC31 adderived clone, C19, to the specific activity of 3.2 x 10(6) U/mg and characterized as a protein of molecular weight 17 kDa with isoelectric point 5.2. LAF activity of IL-1-like factor was specifically neutralized with anti-rat IL-1 alpha but not with anti-rat IL-1 beta. These findings show that the IL-1-like factor is rat IL-1 alpha, although the precise molecular relationship with monocyte IL-1 alpha is unclear. Furthermore, IL-1 alpha was shown to be primarily located in the cytosol and plasma membrane without spontaneous release. A kinetics study indicated that it was leaked into the medium late in culture resulting from cell destruction, in contrast to TNF alpha which was released as the cell number was increased. We discussed the possible role of IL-1 alpha as an autocrine or paracrine growth regulator.