Correlation between tcdB gene PCR cycle threshold and severe Clostridium difficile disease.

A retrospective study, including all samples tested for Clostridium difficile from 2015 to 2018, was conducted. 222 and 199 patients were respectively classified as having a mild/moderate or severe disease. A CT ≤ 26 was significantly associated with severe disease. Furthermore, low CT values were significantly associated to older patients and leukocytosis.

Invasive Candidiasis in Brescia, Italy: Analysis of Species Distribution and Antifungal Susceptibilities During Seven Years.

The aims of this study were to evaluate the epidemiology of nosocomial candidemia in a large teaching hospital in Brescia, Italy, and the in vitro antifungal susceptibility of isolates. We analyzed 196 isolates causing fungemia in patients admitted in our hospital, between January 2009 and December 2015. Strains were identified by VITEK 2 and MALDI-TOF MS. MICs were determined by Sensititre Yeast OneTM. The resistance was defined by using the revised CLSI breakpoints/epidemiological cutoff values to assign susceptibility or wild type to systemic antifungal agents. Most infections were caused by Candida albicans (60%), Candida parapsilosis (15%), Candida glabrata (12%) and Candida tropicalis (6%). The susceptibility rate for fluconazole was 96.5%. Non-Candida species isolates exhibited full susceptibilities to echinocandins according to CLSI breakpoints. Amphotericin B demonstrated excellent activity against all Candida species. Local epidemiological and antifungal susceptibility studies are necessary in order to improve empirical treatment guidelines.

Emergence of the first levofloxacin-resistant strains of Streptococcus agalactiae isolated in Italy.

Of 901 group B streptococcus strains analyzed, 13 (1.4%) were resistant to levofloxacin (MICs of >32 µg/ml for seven isolates, 2 µg/ml for four isolates, and 1.5 µg/ml for four isolates). Mutations in the quinolone resistance-determining regions (QRDRs) of gyrase and topoisomerase IV were identified. A double mutation involving the Ser-81 change to Leu for gyrA and the Ser-79 change to Phe or to Tyr for parC was linked to a high level of fluoroquinolone resistance. In addition, two other mutational positions in parC were observed, resulting in an Asp-83-to-Tyr substitution and an Asp-83-to-Asn substitution. Different mutations were also observed in gyrB, with unknown significance. Most levofloxacin-resistant GBS strains were of serotype Ib and belonged to sequence type 19 (ST19) and clonal complex 19 (CC-19). Most of them exhibited the epsilon gene.

Lymphatic endothelial cells derived from metastatic and non-metastatic lymph nodes of human colorectal cancer reveal phenotypic differences in culture.

Colorectal cancer is one of the most frequent causes of death in Western countries. Most patients develop metastasis traveling through the lymphatic system, and regional lymph node metastasis is considered a marker for dissemination, increased stage, and worse prognosis. Despite rapid advances in tumor biology, the processes that underpin lymphatic invasion and lymph node metastasis remain poorly understood. The aim of this study was to establish an easy protocol for isolation of pure tumor lymphatic endothelial cells derived from lymph nodes to study differences compared with normal endothelial cells of uninvolved tissue from the same patients. Cells were isolated with very high purity via magnetic cell sorting and express the specific lymphatic markers Prox-1 and Lyve-1. They show differences in expression of adhesion molecules, chemokines, and growth factor secretion, and capability to form capillaries when seeded on basal membrane, thereby, revealing important differences between the two cell type. These cultures may provide a promising platform for the comparative analysis of both cell types at the molecular and biological level and to optimize treatment strategies.

Evaluation of three different manual techniques of sharpening curettes through a scanning electron microscope: a randomized controlled experimental study.

OBJECTIVE: The purpose of this study was to compare the effectiveness of three different techniques for manually sharpening of periodontal curettes (PCs) by examining the blades with the aid of scanning electron microscope (SEM). METHODS: Three groups were considered based on three sharpening methods used: group A (moving a PC over a stationary stone); group B (moving a stone over a stationary PC) and group C (moving a PC over a stone fixed, placed on a 'sharpening horse'). After the sharpening, the blades were examined using SEM. The SEM images were assessed independently by five different independent observers. An evaluation board was used to assign a value to each image. A preliminary pilot study was conducted to establish the number of samples. Pearson's correlation test was used to assess the correlations between measurements. anova test with Bonferroni's post hoc test was used to compare the three groups. RESULTS: Sixty PCs (20 PCs per group) were used in this study. Statistically significant differences emerged between the three groups (P-value = 0.001). Bonferroni's test showed that the difference between groups A and B was not statistically significant (P-value = 0.80), while it was significant for the comparisons between groups A and C (P-value = 0.005) and between groups B and C (P-value = 0.001). CONCLUSIONS: The sharpening technique used in group C, which involved the use of the sharpening horse, proved the most effective.

Experimental assessment of oral hygiene achieved by children wearing rapid palatal expanders, comparing manual and electric toothbrushes.

OBJECTIVES: The aim was to compare the efficacy of the electric versus the manual toothbrush in terms of the oral hygiene achieved by patients wearing rapid palatal expanders (RPEs). METHODS: Forty patients were randomly divided into two groups; one equipped with a manual toothbrush (Group A), the other with an electric toothbrush (Group B). Each child's plaque index (PI) and gingival index (GI) were calculated at banded molar level at times T0 (before banding), T1 (a month later), T2 (3 months later) and T3 (when the expander was removed). At each appointment, the PI and GI were recorded and the patient was remotivated. RESULTS: The level of oral hygiene achieved by the group using an electric toothbrush produced a greater improvement in the two indexes than in the group using the manual toothbrush that showed no statistically significant improvement (PI T0-T3: P = 0.309; GI T0-T3: P = 0.141). Both indexes dropped considerably in both groups from T0 to T2, but more so in the group B. From T2 to T3, although the electric toothbrush continued to be substantially more effective, Group B showed a statistically significant deterioration in the oral hygiene (PI +20%; GI +33%). Other assessments conducted on particular areas of the tooth showed improvements in the PI (-33%) for the vestibular region, and for the GI (-57%) in the palatal region among the patients in Group B, while there were no significant changes in these indexes in Group A. CONCLUSIONS: Our findings show that the electric toothbrush is statistically more efficient in performing an adequate level of oral hygiene in children wearing RPE.

In vitro comparison of the bond strength to the enamel of conventional and self-etching dental fissure sealants.

AIM: Dental caries in pits and fissures of molars is still very common in young people, despite a gradual reduction in their incidence and prevalence. Prevention with the aid of dental fissure sealants can help to reduce the onset of decay. In vitro tests were conducted to compare the bond strength to enamel of self-etching sealants versus those applied using the conventional procedure. METHODS: The lingual surface of 40 extracted, caries-free, mandibular third molars was milled to make them flat. The prepared teeth were randomly divided into two groups of 20 teeth each: those in Group A were treated with Clinpro (3M ESPE, St. Paul, MN, USA); those in Group B with Quick Seal (BJM Laboratories Ltd, Or-Yehuda, Israel). Cylinders of sealant were attached to the enamel of the flat surfaces of the samples using a polymerisation process treating the surfaces involved according to the type of material. All samples underwent load testing by means of a universal test machine. RESULTS: The results of the load testing, measured in MPa, were analysed using the Student's t-test for independent samples and the differences proved significant, indicating that the traditionally- applied sealant (mean strength 21.06 MPa) assured a significantly stronger bond (p <.05) than the self-etching sealant (mean strength 10.43 MPa) under our experimental conditions. CONCLUSION Conventional sealants generally provide a considerably higher bond strength than self-etching sealants.

Phenotypes, genotypes, serotypes and molecular epidemiology of erythromycin-resistant Streptococcus agalactiae in Italy.

The purpose of this investigation was to analyse Streptococcus agalactiae (group B Streptococcus, GBS) isolates collected in Italy from vaginal and urine samples in respect to their clonality, distribution of virulence factors and antimicrobial resistance determinants. Three hundred and eighty-eight GBS were recovered from clinical samples. They were analysed for antibiotic resistance profiling. Erythromycin-resistant strains were further characterised by multilocus sequence typing (MLST), serotyping and the detection of alp genes of the alpha-like protein (Alp) family. GBS isolates represented 40 different sequence types (STs), grouped in five clonal complexes (CCs) and belonged to seven serotypes. Most serotype V strains (81%) possessed alp2-3; serotype Ia carried mainly epsilon, while the serotype III mainly rib. All isolates were susceptible to penicillin, whereas resistance to erythromycin was detected in 15% of isolates. Most erythromycin-resistant GBS strains were of serotype V (56.8%) and belonged to the CC-1 group (50%). Macrolide resistance phenotypes were the cMLS(B) (46.5%) and the M phenotypes (46.5%) due to the presence of ermB and mefA/E genes, respectively. These results provide data which establish a baseline for monitoring erythromycin resistance in this region and also provide an insight into the correlation among clonal types, serotypes, surface protein and resistance genes. The increased prevalence of strains that displayed the M phenotype strengthens the importance of the epidemiological surveillance of macrolide resistance in GBS, which may also represent an important reservoir of resistance genes for other species.

Endovascular radiofrequency renal denervation in treating refractory arterial hypertension: a preliminary experience.

PURPOSE: This study was done to investigate the efficacy and safety of percutaneous renal denervation with the Symplicity catheter for reducing blood pressure in patients with essential hypertension resistant to medical therapy (systolic blood pressure >160 mmHg despite the use of three or more antihypertensive drugs, including a diuretic). MATERIALS AND METHODS: In September 2010, five patients affected by essential hypertension resistant to medical therapy were treated. All patients were studied by computed tomography angiography (CTA) of the renal arteries before the procedure and underwent follow-up at 30 and 60 days with colour Doppler ultrasound (CDUS) with evaluation of resistive index, glomerular filtration rate (GFR), 24-h blood pressure and serum catecholamine concentration. Student's t test was used to assess the effectiveness of the procedure in lowering blood pressure. RESULTS: In treated patients, mean blood pressure at baseline was 171/100 mmHg [standard deviation (SD) ± 8/10]; mean GFR was 91.6 ml/min/1.73 m(2) (SD ± 15). Blood pressure after the procedure was reduced by -18/-5 and -13/-10 mmHg at 30 and 60 days, respectively, with a mean medication reduction of 3.6. No complications occurred during the intra- or periprocedural period or during short-term follow-up. CONCLUSIONS: The Symplicity system proved to be efficacious and without serious adverse events in reducing blood pressure and antihypertensive medication use in patients affected by essential hypertension resistant to medical therapy. Although encouraging, our data are preliminary and need to be validated by larger prospective randomised studies.

Experimental evaluation of two methodologies for the restoration of crown fracture in permanent anterior teeth.

AIM: The purpose of this study is to compare, by means of mechanical tests, the resistance of two types of dental restoration in the uncomplicated crown fracture of human permanent maxillary central incisor: rebonding of the fractured tooth fragment and application of ceramic veneers. MATERIALS AND METHODS: Thirty human maxillary central incisors were divided into three groups of ten teeth each. The teeth of the first group (Group A) were kept intact and used as controls, while the remaining teeth were sectioned, in order to simulate a crown fracture, and restored respectively with either the rebonding of the fragment (Group B) or with the use of ceramic veneers (Group C). All samples underwent mechanical tests by means of a universal test machine. RESULTS: The mean resistance to fracture recorded is respectively 425.2N for Group A, 233.3N for Group B, and 347.3N for Group C. One-way ANOVA analysis demonstrates statistical significance for at least one couple of values (Group B compared to Group A). The group of restorations with rebonding of fractured fragments has a mean value of resistance to fracture that is lower than both intact teeth and teeth restored with ceramic veneers. The resistance of teeth with rebonded restorations is 54.9% of that of, intact teeth, and resistance increases for teeth restored with ceramic veneers (81.8% of that of intact teeth). CONCLUSION: Ceramic veneers showed a higher resistance than rebonding and resulted to be more reliable in case of further trauma.

Genomic characterisation of Escherichia coli isolates co-producing NDM-5 and OXA-1 from hospitalised patients with invasive infections.

OBJECTIVES: Carbapenems are one of the last-report therapeutic choices to treat infections due to multidrug-resistant (MDR) micro-organisms. For this reason, the spread of carbapenemase-producing Enterobacteriaceae represents a serious health-public problem. Here we describe isolates co-producing blaNDM-5 and blaOXA-1. METHODS: Three Escherichia coli isolates obtained from patients with invasive infections were analysed by phenotypic antibiotic susceptibility testing and whole-genome sequencing (WGS). RESULTS: All of the isolates were resistant to carbapenems, most ß-lactam antibiotics, piperacillin/tazobactam, amoxicillin/clavulanic acid and ciprofloxacin, remaining susceptible to amikacin, fosfomycin, colistin and tigecycline. The isolates belonged to sequence types ST44, ST405 and ST167 and co-harboured the blaNDM-5 and blaOXA-1 genes. Two of the isolates also harboured extended-spectrum ß-lactamase (ESBL) genes (blaCTX-M-15 and blaTEM-1b). The blaNDM-5 gene was probably carried chromosomally even if different plasmids were identified. Various virulence genes were also identified. CONCLUSION: Our results highlight that continuous surveillance is essential to monitor the spread of clinically important MDR pathogens.

Persistence of human T cell lymphotropic virus type 1 (HTLV-1) sequences in peripheral blood mononuclear cells from patients with mycosis fungoides.

Mycosis fungoides (MF) is a rare form of cutaneous T cell lymphoma suspected of having a viral etiology. As in adult T cell leukemia, the virus involved may be human T lymphotropic virus type 1 (HTLV-1). We cultured the peripheral blood mononuclear cells (PBMC) of 29 patients with MF HTLV-1 seronegative by enzyme-linked immunosorbent assay and Western blot. The presence of reverse transcriptase (RT) and p24 antigen was investigated in the concentrate supernatant of the culture. The DNA of all studied patients was submitted to polymerase chain reaction and Southern blot analysis using primers and probes recognizing the tax region of HTLV-1/2 and the pol region of HTLV-1. 10 of 29 patients were found positive to HTLV-1, whereas they were always negative to RT and p24. The same results were confirmed in double blind after 6 mo. Our findings suggest HTLV-1 may be involved in the etiology of MF, at least in certain cases.

In vitro inhibitory effect of two commercial probiotics on chromogenic actinomycetes.

PURPOSE: Black extrinsic discoloration is a common clinical and aesthetic problem. This study aims to evaluate the potential in vitro antagonistic activity of two commercial probiotics, Streptococcus salivarius M18 and Lactobacillus reuteri ProDentis, against microorganisms associated with black stains. METHODS: Streptococcus salivarius M18 and Lactobacillus reuteri were tested against Aggregatibacter actinomycetemcomitans and Actinomyces naeslundiiusing their cell-free fermentative broth in a planktonic growth inhibition test. RESULTS: Both probiotic cell-free supernatants showed the ability to reduce the pathogenic bacteria growth in a dose-dependent way. Streptococcus salivarius M18 showed a stronger antimicrobial activity than Lactobacillus reuteri ProDentis against the two indicator strains used. A. naeslundi was less susceptible to the probiotic activity of both S. salivarius and L. reuteri compared to A. actinomycetemcomitans. CONCLUSIONS: The obtained results demonstrate a potent antagonistic ability of probiotics to reduce the growth of microorganisms associated with black tooth stains. Therefore, these strains could be evaluated for a therapeutic use against dental pigmentations.

Building Capacity for Evidence-Informed Priority Setting in the Indian Health System: An International Collaborative Experience.

India's rapid economic growth has been accompanied by slower improvements in population health. Given the need to reconcile the ambitious goal of achieving Universal Coverage with limited resources, a robust priority-setting mechanism is required to ensure that the right trade-offs are made and the impact on health is maximised. Health Technology Assessment (HTA) is endorsed by the World Health Assembly as the gold standard approach to synthesizing evidence systematically for evidence-informed priority setting (EIPS). India is formally committed to institutionalising HTA as an integral component of the EIPS process. The effective conduct and uptake of HTA depends on a well-functioning ecosystem of stakeholders adept at commissioning and generating policy-relevant HTA research, developing and utilising rigorous technical, transparent, and inclusive methods and processes, and a strong multisectoral and transnational appetite for the use of evidence to inform policy. These all require myriad complex and complementary capacities to be built at each level of the health system . In this paper we describe how a framework for targeted and locally-tailored capacity building for EIPS, and specifically HTA, was collaboratively developed and implemented by an international network of priority-setting expertise, and the Government of India.

Detection of Ureaplasma biovars and polymerase chain reaction-based subtyping of Ureaplasma parvum in women with or without symptoms of genital infections.

Ureaplasma parvum colonises human mucosal surfaces, primarily in the urogenital and respiratory tracts, causing a wide spectrum of diseases, from non-gonococcal urethritis to pneumonitis in immunocompromised hosts. Although the basis for these diverse clinical outcomes is not yet understood, it has been suggested that only certain strains of these micro-organisms are disease-associated. The aim of this study was to determine the distribution of Ureaplasma biovars and U. parvum serovars and to estimate their possible association with age, absence of lactobacilli, clinical symptoms and antibiotic resistance. DNA was extracted by endocervical, vaginal and urethral samples obtained from 158 women positive for U. urealyticum by culture and were biotyped by polymerase chain reaction (PCR) targeting the multiple-banded gene. Parvo biovar (biovar 1) was found in 136 (86%) and T960 biovar (biovar 2) in 22 (14%) patients. Among the different serovars of U. parvum, we found that serovar 3/14 was present maximally in the 21-25-year-old age group, while T960 biovar was distributed with quite similar frequency in women of 26-30 and >40 years of age. In this study, U. parvum serovar 3/14 and T960 biovar were found to be significantly associated with symptomatic patients and a loss of lactobacilli, while, on the contrary, U. parvum serovar 6 was significantly correlated with asymptomatic women and normal vaginal flora. The most active antibiotic for the majority of Ureaplasma isolates was tetracycline. These preliminary data show the possibility of distinguishing between the more or less virulent strains of Ureaplasma, with important consequences for therapeutic treatment.

Comparison of the AMPLICOR human papillomavirus test and the hybrid capture 2 assay for detection of high-risk human papillomavirus in women with abnormal PAP smear.

Infection with human papillomavirus (HPV) is a necessary step in the progression to cervical cancer. Many methods for HPV testing are currently available, mostly developed to detect pools of HPV types. Hybrid Capture 2 (HC2) is one of the most widely used. A new PCR-based assay, the Roche AMPLICOR HPV test, has been recently developed. Both assays recognize a group of 13 HR HPV types contemporaneously. This study evaluated the performance of both methods for detecting high-grade cervical lesions as a part of management for abnormal PAP smears. The study population was composed of 213 women, all referred to colposcopy and histologic diagnosis following an abnormal PAP test. Biopsy-confirmed high-grade cervical intraepithelial neoplasia was used as a gold standard. Overall agreement was 84.9% with a kappa value of 0.6. When comparing the ability to detect moderate cervical intraepithelial neoplasia (CIN2+) and high-grade cervical intraepithelial neoplasia (CIN3+/cancer), AMPLICOR proved slightly more sensitive than HC2, a finding that is important when HPV testing is used in a triage of borderline smear results. Genotyping of discordant results showed a prevalence of LR-HPV types in HC2 positive/AMPLICOR negative samples, and a similar prevalence of HR- and LR-HPV types in AMPLICOR positive/HC2 negative samples. In conclusion, the study shows that the AMPLICOR assay is more sensitive than HC2, which makes it a valid alternative for routine clinical use.

Different sequence strains of Streptococcus agalactiae elicit various levels of cytokine production.

Group B streptococcus (GBS) is the most common cause of neonatal and obstetric sepsis and an increasingly important cause of septicemia in elderly subjects and immunocompromised patients. Our aim was to evaluate whether different genotypes of GBS may induce a different production of pro-inflammatory and anti-inflammatory cytokines. We used multilocus sequence typing to identify 71 clones isolated from asymptomatic healthy carriers and symptomatic individuals. All these clinical isolates were used to infect purified human monocytes. TNF-alpha, IL-6, IL-8 and IL-10 secretion was measured. Fifteen allelic sequence types (STs) were identified. The MLST (multilocus sequence typing) analysis grouped the bacteria into four different lineages (clonal cluster) and two of these were closely involved in the infection of symptomatic subjects: CC17 and CC19. Furthermore, CC17 and CC19 stimulated TNF-alpha, IL-6 and IL-8 production significantly more than the other lineages, while CC17 induced a decreased IL-10 production. These results suggest the existence of differences in immune response to infection with particular genotypes of GBS.

Prevalence and distribution of single and multiple HPV infections in cytologically abnormal cervical samples from Italian women.

The prevalence of single and multiple HPV infections was assessed over a cohort of 213 women with cytological abnormalities and its association with cervical neoplasia established. Roche linear array HPV genotyping test was used to identify HPV genotypes. The most prevalent HPV genotypes in cervical cancer samples were HPV16 (61.2%), HPV52 (16.1%), HPV18 (12.9%) and HPV 31 (9.6%). Multiple HR and LR HPV infections, comprising between two and 5+ HPV types, were identified in 49.7% of samples, with a significantly lower number in severe dysplasia and cervical cancer samples (p<0.05). These results seem to indicate that detection of multiple HPV infection with HR-HPV types is not significantly better as a predictor of cervical cancer than single HR-HPV infection, though further longitudinal studies are needed to better clarify the relevance of these infections to the progression of cervical neoplasia.

Comparative evaluation of different techniques of surface preparation for occlusal sealing.

AIM: Aim of this experiment is the efficacy evaluation of three different types of surface preparation techniques by microleakage assessment of two different sealing materials. MATERIALS AND METHODS: Sixty teeth were used for the experiment: half were treated with Admira seal (Voco) and the remaining half with Delton FS+ (Dentsply). Each group was then divided into three subgroups, which underwent three different preparation techniques before sealant placement: mechanical brushing, air abrasion and Intensive bur FG 40D4, respectively. After immersion of the specimens in methylene blue solution, the degree of microleakage was assessed by mean of optic microscope. RESULTS: Results show no microleakage in samples treated with air abrasion, followed by the samples treated with mechanical brushing. The teeth prepared by bur treatment before sealant application show a significant number and degree of marginal infiltration. CONCLUSION: It can be concluded that air abrasion technique is able to guarantee a reliable seal of occlusal surfaces.