Exploring the root canal microbiome in previously treated teeth: A comparative study of diversity and metabolic pathways across two geographical locations.

AIM: To analyse and compare the root canal microbiome present in root-filled teeth of two different geographical populations, and to study their functional potential using a next-generation sequencing approach. METHODOLOGY: Sequencing data obtained from surgical specimens from previously treated teeth with periapical bone loss from Spain and USA were included in the study. Taxa were classified using SILVA v.138 database. Differences in genera abundances among the 10 most abundant genera were evaluated using a Kruskal-Wallis test. Alpha diversity indices were calculated in mothur. The Shannon and Chao1 indices were used. Analyses of similarity (ANOSIM) to determine differences in community composition were done in mothur, with Bonferroni correction for multiple comparisons. p-Values < .05 were considered statistically significant. Identification of enriched bacteria function prediction in the study groups (KEGG pathways) was carried out by linear discriminant analysis effect size (LEfSe) via Python 3.7.6. RESULTS: A greater alpha-diversity (Shannon and Chao1 indices) was observed from samples obtained in Spain (p = .002). Geography showed no significant effects on community composition via an ANOSIM using Bray-Curtis dissimilarities (R = 0.03, p = .21). Bacterial functional analysis prediction obtained by PICRUSt showed that 5.7% KEGG pathways differed between the Spain and US samples. CONCLUSIONS: The taxonomic assessment alone does not fully capture the microbiome's differences from two different geographical locations. Carbohydrate and amino acid metabolism were enriched in samples from Spain, while samples from USA had a higher representation of pathways related to nitrogen, propanoate metabolism, and secretion systems.

Achieving Comprehensive Contraceptive Counseling for Young Women and Improved Contraceptive Uptake Through the Implementation of a Reproductive Health Educator Program.

OBJECTIVES: To evaluate the effect of student-volunteer reproductive health educators (RHEs) on frequency of counseling about long acting reversible contraception (LARC) and uptake of more effective contraception. METHODS: This was a non-randomized intervention study conducted at a resident continuity clinic. Eligible patients were females aged 14-25. Participants met with an RHE during their visit in addition to the standard care team. Patients in the historical group met only with the standard care team. We compared counseling patterns and uptake of more effective contraceptive methods between the baseline historical comparison and intervention groups. RESULTS: Nearly all eligible patients in the intervention group received counseling about LARC during their visit, whereas less than half of patients seen in the baseline period received the same counseling. Approximately two-thirds of patients who had no contraceptive method at the beginning of the visit chose to initiate a short- or long-acting method in the intervention group, compared to less than half of patients who were using no method in the baseline group. CONCLUSIONS FOR PRACTICE: Integration of student-volunteer RHEs increased the frequency with which patients were counseled about LARC and also increased the number of patients who switched to more effective contraception during their visit compared to patients seen in the same clinic before RHEs were introduced. Student-volunteer RHEs are a potentially cost-effective way to provide comprehensive health education in busy clinical settings.

Prospective comparison among three intrarectal anesthetic treatments combined with periprostatic nerve block during transrectal ultrasonography-guided prostate biopsy.

BACKGROUND: Recent trends in prostate biopsy analgesia suggest a combination anesthetic to provide better pain relief than periprostatic nerve block (PPNB) alone. This study aimed to demonstrate the efficacy and safety of three intrarectal local anesthesia (IRLA) combined with PPNB in patients undergoing transrectal ultrasonography (TRUS)-guided prostate biopsy. METHODS: In this prospective, randomized study, 120 prostate biopsy patients were equally divided into four IRLA groups: group 1 (placebo) received simple lubrication; group 2 received 2% lidocaine gel; group 3 received 100 mg indomethacin suppository and group 4 received 5% prilocaine/lidocaine (EMLA) cream. PPNB with 2% lidocaine was applied in all groups. A ten-point visual analog scale evaluated both pain associated with the probe insertion and pain associated with prostate sampling. Adverse effects or complications due to anesthesia during and after the procedure were documented. RESULTS: Compared with group 1, groups 3 and 4 had significantly lower pain scores at both probe insertion and prostate sampling while group 2 showed no significant differences at both pain scores. Moreover, group 4 showed significantly lower pain scores at probe insertion compared to group 3, while no significant difference was observed at prostate sampling. Mild complications were observed in all groups with no significant difference in the incidence of complications between groups. CONCLUSION: Intrarectal application of EMLA cream is a more efficient pain reduction than either 2% lidocaine gel or 100 mg indomethacin suppository when applied combined with PPNB. This combination represents an effective option of pain relief for patients undergoing TRUS-guided prostate biopsy.

Characterization of porcine milk oligosaccharides during early lactation and their relation to the fecal microbiome.

The composition of porcine milk oligosaccharides (PMO) was analyzed during early lactation and their relation to piglet gut microbiome was investigated. Pigs are considered ideal intestinal models to simulate humans because of the striking similarity in intestinal physiopathology to humans. The evolution of PMO was investigated in the milk from 3 healthy sows at prefarrowing, farrowing, and d 7 and 14 postpartum by Nano-LC Chip Quadrupole-Time-of-Flight mass spectrometer (Agilent Technologies, Santa Clara, CA). Previously sequenced metagenome libraries were reanalyzed to examine changes with specific gut bacterial populations. Over 30 oligosaccharides (OS) were identified in the milk, with 3'-sialyllactose, lacto-N-tetraose, α1-3,ß1-4-d-galactotriose, 2'-fucosyllactose, and 6'-sialyllactose being the most abundant species (accounting for ~70% of the total OS). Porcine milk had lower OS diversity (number of unique structures) than human milk, and appeared closer to bovine and caprine milk. In agreement with previous studies, only 3 fucosylated OS were identified. Surprisingly, their contribution to total OS abundance was greater than in bovine milk (9 vs. 1%). Indeed, fucosylated PMO increased during lactation, mirroring a similar trend observed for neutral and type I OS content during early lactation. Taken together, these results suggest that, in terms of abundance, PMO are closer to human milk than other domestic species, such as bovine and caprine milks. Metagenomic sequencing revealed that fucose-consuming bacterial taxa in the gut microbiota of piglets were qualitatively but not quantitatively different between nursing and weaning stages, suggesting that both the composition and structure of dietary glycans may play a critical role in shaping the distal gut microbiome. The similarity of both intestinal physiopathology and milk OS composition in human and porcine species suggests similar effects on gastrointestinal development of early nutrition, reinforcing the use of the pig intestinal model to simulate human intestinal models in the clinical setting.

The stress and workload of virtual reality training: the effects of presence, immersion and flow.

The present investigation evaluated the effects of virtual reality (VR) training on the performance, perceived workload and stress response to a live training exercise in a sample of Soldiers. We also examined the relationship between the perceptions of that same VR as measured by engagement, immersion, presence, flow, perceived utility and ease of use with the performance, workload and stress reported on the live training task. To a degree, these latter relationships were moderated by task performance, as measured by binary (Go/No-Go) ratings. Participants who reported positive VR experiences also tended to experience lower stress and lower workload when performing the live version of the task. Thus, VR training regimens may be efficacious for mitigating the stress and workload associated with criterion tasks, thereby reducing the ultimate likelihood of real-world performance failure. Practitioner Summary: VR provides opportunities for training in artificial worlds comprised of highly realistic features. Our virtual room clearing scenario facilitated the integration of Training and Readiness objectives and satisfied training doctrine obligations in a compelling engaging experience for both novice and experienced trainees.

HBV/HDV co-infection in the Western Brazilian Amazonia: an intriguing mutation among HDV genotype 3 carriers.

HDV infection still remains a serious public health problem in Amazonia. There are few data regarding the biomolecular aspects of HBV/HDV co-infection in this region. We studied 92 patients HBsAg(+) /anti-HDV IgG(+) followed at the Hepatitis Referral Centers of Porto Velho (RO), Rio Branco and Cruzeiro do Sul (AC), Brazil, from March 2006 to March 2007 for whom the HDV and/or the HBV genotype could be determined. The HDV genotype could be determined in 90 patients, while the HBV genotypes could be positively determined in 74. HBV subgenotype F2 is the most prevalent (40.2%), followed by the subgenotypes A1 (15.2%) and D3 (8.7%), while 16.4% were other subgenotypes or genotypes, 4.3% were discordant and 15.2% were unamplifiable. Surprisingly, HDV genotype 3 (HDV-3) was found in all of the HBV/HDV-infected patients that could be genotyped for HDV, confirming that HDV-3 can associate with non-F HBV genotypes. However, a HDV-3 mutant was found in 29.3% of patients and was more frequently associated with non-F HBV genotypes (P < 0.001) than were nonmutant strains, suggesting that the mutation may facilitate association of HDV-3 with non-F HBV genotypes.

Sclerostin serum levels in prostate cancer patients and their relationship with sex steroids.

UNLABELLED: The role of sclerostin on bone metabolism and its relation to sex steroids in patients with prostate cancer (PC) is not well known. We found that sclerostin levels are significantly increased in PC patients, particularly in those with androgen deprivation therapy (ADT), and there is an inverse relationship between sclerostin levels and testosterone. INTRODUCTION: Recent studies have evaluated sclerostin levels in bone diseases as osteoporosis. However, there are few data in PC patients, particularly in patients with hypogonadism related to ADT. The aim of the present study was to compare serum sclerostin levels in ADT/non-ADT-treated PC patients and healthy controls and to evaluate their relationship with sex steroids and bone metabolism. METHODS: We performed a cross-sectional study involving 81 subjects: 25 ADT-treated PC patients, 34 PC patients without ADT treatment, and 22 healthy controls. We measured serum sclerostin levels, bone turnover markers, bone mineral density (BMD) in all individuals, and sex steroids levels in PC patients. RESULTS: Serum sclerostin levels were significantly higher in PC patients compared to those in control subjects. ADT-treated patients had significantly higher sclerostin levels than PC patients without ADT treatment: ADT 64.52 ± 27.21 pmol/L, non-ADT 48.24 ± 15.93 pmol/L, healthy controls 38.48 ± 9.19 pmol/L, p < 0.05. In PC patients, we found a negative relationship between serum sclerostin levels and androgens after age adjustment (total testosterone: r = -0.309, p = 0.029; bioavailable testosterone: r = -0.280, p = 0.049; free testosterone: r = -0.299, p = 0.035). We did not observe any relationship between sclerostin levels and bone turnover markers or BMD in any group. CONCLUSIONS: Circulating sclerostin levels are significantly increased in patients with PC and particularly in those receiving ADT. The inverse relationship between serum sclerostin and testosterone in these patients suggests that androgens are key regulators of bone metabolism in this population.

[Enterocolitis associated with Hirschsprung's Disease. Experience in a pediatric teaching hospital]. / Enterocolitis asociada a enfermedad de Hirschsprung. Experiencia en un Hospital Universitario Pediátrico.

Hirschsprung's associated enterocolitis (HAEC) is the most serious and potentially fatal complication of the disease, with a significant morbidity and mortality. The effect of HAEC varies remarkably among the publications since the lack of consensus and the absence of established diagnostic criteria. Patients diagnosed with Hirschsprung's disease (HD) were selected and treated between January 2009 and December 2012 in Fundación HOMI, finding 41 cases of HD, 6 of them with total aganglionosis. 76% of the patients had HAEC, 18 of them during the neonatal period and 6 of them after the final pull-through; 23% presented recurrent enterocolitis episodes. HAEC was the first clinical manifestation in 25 (61%) of the patients with HD. From HD patients limited to the rectum and sigmoid 67% had enterocolitis unlike TCA (total colonic agangliniosis) cases in which all presented HAEC episodes. In our setting, HAEC remains to be an unknown complication; this revision demonstrates its high incidence and mortality.

La enterocolitis asociada a la enfermedad de Hirschsprung (EAEH) es la complicación más grave y potencialmente fatal de la enfermedad, con una morbilidad y mortalidad significativas. La incidencia de EAEH varía notablemente entre las publicaciones dada la falta de consenso y la ausencia de criterios diagnósticos establecidos. Se seleccionaron los pacientes con diagnóstico de enfermedad de Hirschsprung (EH) tratados entre Enero de 2009 a Diciembre de 2012 en la Fundación Hospital de La Misericordia (HOMI) encontrando 41 casos de EH, 6 de ellos con aganglionosis total. En el 76% de los pacientes se realizó el diagnóstico de EAEH, 18 de ellos en el periodo neonatal y 6, tras el descenso definitivo; 23% presentaron episodios de enterocolitis recurrente. La EAEH fue la primera manifestación clínica en 25 (61%) de los pacientes con EH. De los pacientes con EH limitada al recto y sigmoides, un 67% tuvieron enterocolitis a diferencia de los casos de aganglionosis total del colon (ATC), donde todos presentaron episodios de EAEH.

Serum osteoprotegerin: bone or cardiovascular marker in Type 2 diabetes males?

BACKGROUND: The role of osteoprotegerin (OPG) as a marker of cardiovascular disease (CVD) in Type 2 diabetes (T2DM) is not well established. Moreover, the relationship between OPG, osteoporosis, and vertebral fractures in T2DM remains to be elucidated. AIM: To determine the role of serum OPG in the prediction of CVD and bone disease in T2DM males. SUBJECTS AND METHODS: Cross-sectional study with 68 males, 43 with T2DM and 25 subjects without diabetes. We measured: serum OPG by inmunoassay, the presence of CVD (coronary heart disease, cerebrovascular and peripheral artery disease), surrogate markers of CVD [intima- media thickness (IMT) and aortic calcification] and bone disease (bone mineral density and prevalent vertebral fractures). RESULTS: OPG serum levels (in pmol/l) were significantly higher in T2DM males with abnormal IMT (5.12 ± 1.59 vs 3.76 ± 1.98), carotid plaque (5.46 ± 1.67 vs 4.20 ± 1.81), aortic calcification (5.91 ± 1.39 vs 4.07 ± 1.76), hypertension (5.11 ± 1.86 vs 3.81 ± 1.47), and peripheral artery disease (6.24 ± 1.64 vs 4.21 ± 1.63, p < 0.05 for all comparisons). In the logistic regression analysis (after adjustment for age and main cardiovascular risk factors), serum OPG (per 1 pmol/l increase in OPG) was associated with increased risk of abnormal IMT [odds ratio (OR) 1.84, confidence interval (CI) 1.21-2.79, p = 0.004), carotid plaque (OR 1.71, CI 1.13-2.58, p = 0.012), aortic calcification (OR 2.21, CI 1.27-3.84, p = 0.05) and peripheral artery disease (OR 4.02, CI 1.65-9.8 p = 0.002). However, OPG were not related to bone mass or vertebral fractures. CONCLUSIONS: Our results suggest that in T2DM males OPG serum concentrations constitute a marker of CVD, but not a marker of bone disease.

Cervical human papillomavirus infection in Mexican women with systemic lupus erythematosus or rheumatoid arthritis.

Cervical human papillomavirus (HPV+) infection is associated with an increased risk of cervical dysplasia. Although the frequency of HPV+ in systemic lupus erythematosus (SLE) has been investigated in some races its prevalence in Hispanic women is still unknown. This cross-sectional study evaluated the prevalence of cervical HPV+ in Mexican women with SLE (n = 34) or rheumatoid arthritis (RA) (n = 43) and in healthy controls (n = 146). These women were interviewed about risk factors for sexually transmitted infections and cervical cytology analysis was performed. HPV+ viral types were identified using PCR: HPV+ was observed in 14.7% of SLE, 27.9% of RA and 30.8% of controls. High-risk HPV types were observed in 11.7% of women with SLE, 27.9% of women with RA, and in 26% of the controls. High-risk viral types 58, 35 and 18 were the most frequently identified in SLE. Two women with SLE had a high-grade squamous intraepithelial lesion and one had cervical cancer. An association was observed between methotrexate utilization, longer duration of therapy with prednisone, and HPV+ in RA or SLE. Thus, there is a high prevalence of cervical HPV infection in Mexican women with SLE or RA, and physicians must be vigilant in preventing the development of cervical dysplasia.

Linac stereotactic radiosurgery for the treatment of small arteriovenous malformations: lower doses can be equally effective.

OBJECTIVE: The purpose of this study was to examine the efficacy and toxicity of treating small arteriovenous malformations (AVMs) (≤3 cm in diameter) with a median marginal applied dose of 14 Gy. METHODS: Two hundred and thirteen patients diagnosed with AVMs were treated between January 1991 and December 2005. Seventy-three percent of the patients had hemorrhaged prior to treatment, 13% had had previous surgery and 19.2% had had previous embolization. The median follow-up duration was 48.1 months. RESULTS: The Kaplan-Meier analysis estimated that the 36-month obliteration rate was 65.5% for patients undergoing their first stereotactic radiosurgery (SRS) and 68.3% for those undergoing repeated SRS. The Kaplan-Meier analysis estimated the 60-month AVMs obliteration rate for the entire cohort to be 82.4%. The median time to AVM obliteration was 40 ± 2.8 months. We found a statistically significant relationship between the time of obliteration and the following factors: site of the AVMs (sites other than brainstem), a higher prescribed dose and a positive history of previous hemorrhage. Thirteen patients (7.6%) experienced toxicities. CONCLUSIONS: SRS was an effective and safe treatment for AVMs ≤3 cm in diameter, with acceptable toxicity.

Metastatic involvement of skeletal muscle from gastric adenocarcinoma.

Gastric cancer represents the fifth most common cancer diagnosis worldwide and the third leading cause of cancer-related mortality. In the USA, the overall 5-year survival rate is 31%, with distant disease nearing 5%. The most common sites of metastasis are the liver and peritoneum. Skeletal muscle involvement has been rarely reported. Since clinical and imaging findings overlap with primary sarcomas, a confirmatory biopsy is required for diagnosis. Prognosis remains poor with treatment options including palliative chemotherapy, radiotherapy and surgical resection. We report the case of a 57-year-old female presenting with extensive involvement of skeletal muscle 10 years after achieving remission. In addition to illustrating the refractoriness and poor outcomes associated with muscle involvement, this case and comprehensive review of the literature highlights important characteristics of disease biology and tumor genomics that warrant detailed discussion and exposition to a wider audience.

Carcinoma of unknown primary site: development in a single institution of a prognostic model based on clinical and serum variables.

PURPOSE: To identify clinical and biologic variables with significant impact on survival in patients with carcinomas of an unknown primary site (CUP) and to develop a simple prognostic model. PATIENTS AND METHODS: In this retrospective study, univariate and multivariate prognostic factors analyses were conducted in a population of 100 patients with CUP. Patients with features requiring well defined treatments had previously been excluded. RESULTS: Overall survival (OS) was significantly related to the following pretreatment adverse prognostic clinical factors: a poor performance status (2 or 3), weight loss more than 10% in the last six months, the presence of liver metastases and more than two metastatic sites. Two biological parameters predicted a significantly shorter survival: elevated serum levels of alkaline phosphatase and of lactate dehydrogenase. In the multivariate analysis, only two independent adverse prognostic parameters were retained: a poor performance status and the presence of liver metastases. We developed a prognostic model for OS based on the following subgroups: good prognosis (PS 0 or 1 and absence of liver metastases), intermediate prognosis (PS> or =2 or presence of liver metastases) and poor prognosis (PS> or =2 or presence of liver metastases). Median OS for the three groups was 10.8, 4 and 1.9 months respectively, p<0.0001. CONCLUSION: A simple prognostic model using performance status and presence of liver metastases was developed. It allowed the assignment of patients into three subgroups with different outcomes. Treatment strategies could be adapted for each subgroup. We think that this prognostic model could be useful and should be validated in other patient series.

[Peritoneal mesothelioma: an inusual clinical presentation in a patient without exposure to asbestos]. / Mesotelioma peritoneal: una presentación clínica inusual en un paciente sin exposición al asbesto.

Malignant mesothelioma is an insidious neoplasm arising from the mesotelial surfaces, of the pleural and peritoneal cavities, the tunica vaginalis, or the pericardium. The predominant cause is inhalation exposure to asbestos. We present a rare case of primary malignant mesothelioma of the peritoneum in a 64 year old man without history of inhalation exposure to asbestos. The initial symptoms were constitutional syndrome and right pleural effusion. Positron emission tomography combined with computed tomography (PET/TC) was useful for a supporting diagnosis and to determine the extension. The patient received treatment with systemic palliative chemotherapy, cisplatin-pemetrexed. After three cycles, partial response was observed, but the evolution was fatal due to secondary toxicity of chemotherapy.

[Comorbidity, pluripathology, resource use and prognosis of patients hospitalized in internal medicine areas]. / Comorbilidad, pluripatología, consumo de recursos y pronóstico de pacientes ingresados en una unidad de Medicina Interna.

OBJECTIVE: To compare the concept of patient with pluripathology (PP) with the index of comorbidity of Charlson (IC) respect to the resources use and prognosis of hospitalized patients. PATIENTS AND METHODS: An observational prospective study of 207 consecutively hospitalized patients in an internal medicine unit was conducted. The PP, a variant of PP with three or more criteria (PP3), IC and IC fit to the age (ICE) were determined, and their relation with the consumption of resources (emergency attentions, programmed consultations of specialties and episodes of hospitalization in the last year, length of stay, own procedures, other specialty procedures and consulting) and prognosis (hospital mortality, significant functional deterioration and rehospitalization) were stayed. RESULTS: The frequency of PP and PP3 was 40.6 and 16.9%, respectively. The median of the IC and ICE score were 2 (0-7) and 4 (0-11), respectively. We found the following independent relations: IC of 3 or greater with the rehospitalization and the length of stay; ICE of 5 or greater with mortality; PP3 with mortality, rehospitalization and emergency visits, hospitalizations and consultations in the last year; the categories B and F of PP with hospitalizations in the last year; the category D of PP with consultations in the last year; and the association of categories A, B and C with mortality and hospitalizations in the last year. PP was not related in an independent form with any variable. CONCLUSION: The IC and the concept of PP do not identify the same hospitalized patient population. Modifications of the PP definition, like PP3, or the analysis of the different clinical categories of PP and their associations, could improve the utility of this concept.

Behavioral effects of dopaminergic agonists in transgenic mice overexpressing human wildtype alpha-synuclein.

Overexpression of alpha-synuclein causes familial Parkinson's disease and abnormal aggregates of the protein are present in sporadic cases of the disease. We have examined the behavioral effects of direct and indirect dopaminergic agonists in transgenic mice expressing human alpha-synuclein under the Thy-1 promoter (Thy1-aSyn, alpha-synuclein overexpressor), which exhibit progressive impairments in behavioral tests sensitive to nigrostriatal dopamine dysfunction. Male Thy1-aSyn and wild-type mice received vehicle, benserazide/L-DOPA (25 mg/kg, i.p.), high (2 mg/kg, s.c.) and low doses (0.125, 0.25, 0.5 mg/kg, s.c.) of apomorphine, and amphetamine (5 mg/kg, i.p.), beginning at 3 months of age, and were tested on the challenging beam, spontaneous activity, pole test, and gait. l-DOPA had a paradoxical effect and worsened the deficits in Thy1-aSyn mice compared with controls, whereas the high dose of apomorphine only produced few deficits above those already present in Thy1-aSyn. In contrast to wild-type mice, Thy1-aSyn mice did not show amphetamine-induced stereotypies. The results indicate that chronic overexpression of alpha-synuclein led to abnormal pharmacological responses in mice.

Characterization of goat colostrum oligosaccharides by nano-liquid chromatography on chip quadrupole time-of-flight mass spectrometry and hydrophilic interaction liquid chromatography-quadrupole mass spectrometry.

A detailed qualitative and quantitative characterization of goat colostrum oligosaccharides (GCO) has been carried out for the first time. Defatted and deproteinized colostrum samples, previously treated by size exclusion chromatography (SEC) to remove lactose, were analyzed by nanoflow liquid chromatography-quadrupole-time of flight mass spectrometry (Nano-LC-Chip-Q-TOF MS). Up to 78 oligosaccharides containing hexose, hexosamine, fucose, N-acetylneuraminic acid or N-glycolylneuraminic acid monomeric units were identified in the samples, some of them detected for the first time in goat colostra. As a second step, a hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS) methodology was developed for the separation and quantitation of the main GCO, both acidic and neutral carbohydrates. Among other experimental chromatographic conditions, mobile phase additives and column temperature were evaluated in terms of retention time, resolution, peak width and symmetry of target carbohydrates. Narrow peaks (wh: 0.2-0.6min) and good symmetry (As: 0.8-1.4) were obtained for GCO using an acetonitrile:water gradient with 0.1% ammonium hydroxide at 40°C. These conditions were selected to quantify the main oligosaccharides in goat colostrum samples. Values ranging from 140 to 315mgL(-1) for neutral oligosaccharides and from 83 to 251mgL(-1) for acidic oligosaccharides were found. The combination of both techniques resulted to be useful to achieve a comprehensive characterization of GCO.