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.

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.

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.

Robot-assisted laparoscopic partial nephrectomy with hydrodissection. / Nefrectomía parcial laparoscópica asistida por robot con hidrodisección.

INTRODUCTION: Nephron-sparing surgery is reporting satisfactory oncological results, with improved quality of life. The laparoscopic approach is technically complex and requires advanced skills; however, the use of robotic systems facilitates the transition to this procedure. With improved strategies for diagnosis and follow-up, as well as ablative technologies, the role of partial nephrectomy continuous to evolve. The use of water-jet technology to dissect and resect organs helps create anatomical planes in a relatively blood-free field, which improves vision and does not cause heat damage. MATERIAL AND METHODS: We present the case of a patient with an incidental diagnosis of a complex right renal cyst managed with robot-assisted laparoscopic partial nephrectomy using hydrodissection for tumour resection. RESULTS: The surgery was performed with 2 robotic arms. Four ports were routinely placed, the colon was mobilised, and the tumour was dissected. Tumour delimitation was performed with laparoscopic ultrasound, and the renal artery was clamped. We then performed the tumour resection assisted by a laparoscopic water-jet applicator, operated by the assistant through an auxiliary port. The renal parenchyma was reconstructed in 2 planes, without knots, and the arterial clamp was withdrawn at 23minutes. The resection margins were negative. CONCLUSIONS: The use of water-jet technology to assist with the renal tumour resection during minimally invasive nephron-sparing surgery facilitates the dissection, making it more fluid and atraumatic. Simultaneously, the technique provides a clean surgical field, which improves visibility. This procedure can help decrease ischaemia times, thereby positively affecting the patient's evolution without compromising the oncological results.

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.

Surgical technique for en bloc transurethral resection of bladder tumour with a Hybrid Knife(®). / Técnica quirúrgica de la resección transuretral de tumor vesical en bloque con Hybrid Knife(®).

BACKGROUND: Bladder cancer is the second most common malignancy of the urinary tract and the 9th worldwide. Latin American has an incidence of 5.6 per 100,000 inhabitants per year. Seventy-five percent of newly diagnosed cases are nonmuscle invasive bladder cancer, and 25% of cases present as muscle invasive. The mainstay of treatment for nonmuscle invasive bladder cancer is loop transurethral resection. In 2013, the group led by Dr Mundhenk of the University Hospital of Tübingen, Germany, was the first to describe the Hybrid Knife(®) equipment for performing en bloc bladder tumour resection, with favourable functional and oncological results. OBJECTIVE: To describe the surgical technique of en bloc bladder tumour resection with a Hybrid Knife(®) as an alternative treatment for nonmuscle invasive bladder tumours. MATERIAL AND METHODS: A male patient was diagnosed by urotomography and urethrocystoscopy with a bladder tumour measuring 2×1cm on the floor. En bloc transurethral resection of the bladder tumour was performed with a Hybrid Knife(®). RESULTS: Surgery was performed for 35min, with 70 watts for cutting and 50 watts for coagulation, resecting and evacuating en bloc the bladder tumour, which macroscopically included the muscle layer of the bladder. There were no complications. CONCLUSION: The technique of en bloc bladder tumour resection with Hybrid Knife(®) is an effective alternative to bipolar loop transurethral resection. Resection with a Hybrid Knife(®) is a procedure with little bleeding and good surgical vision and minimises the risk of bladder perforation and tumour implants. The procedure facilitates determining the positivity of the neoplastic process, vascular infiltration and bladder muscle invasion in the histopathology study.

[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.

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.

[D-repeat polymorphism in the ASPN gene in knee osteoarthritis in females in Torreón, Coahuila. Case-control study]. / Polimorfismo del D repetido en gen ASPN en osteoartritis de rodilla en mujeres de Torreón, Coahuila. Estudio de casos y controles.

BACKGROUND: Knee osteoarthritis (OA) is one of the most common and disabling disorders of the musculoskeletal system. It may affect any ethnic group and causes variable degrees of disability. Various risk factors have been associated with the development and progression of this condition, such as: age, genetic and occupational factors, trauma, menopause, diabetes mellitus, obesity, and gender, among others. Distinguishing these factors, whether individually or altogether, is important to prevent or diagnose and treat the disease early on. METHODS: A case-control study was conducted in 260 females in Torre6n, Coahuila, to analyze the relationship between primary knee osteoarthritis and the D-repeat polymorphism in the ASPN gene (asporin). 130 females with knee osteoarthritis and 130 healthy female controls were included. RESULTS: In this study, menopause and the D16 allele variant were found to be significant risk factors for knee osteoarthritis (p = 0.002, OR 2.656, CI 95% 1.412-4.998; p = 0.026, OR 2.418, CI 95% 1.111-5.263, respectively). The D12 variant was found to be a significant protective allele. CONCLUSIONS: As far as we know, this is the first case-control study in Mexican women that suggests that menopause and the D-repeat polymorphism in the ASPN gene are associated with knee OA.

Identification of the NF-κB inhibitor A20 as a key regulator for human adipogenesis.

The zinc-finger protein A20 is a key player in the negative feedback regulation of the nuclear factor kappa-light-chain-enhancer of activated B-cell (NF-κB) pathway in response to multiple stimuli. Tumor necrosis factor alpha (TNFα), a cytokine with pleiotropic effects on cellular proliferation and differentiation, dramatically increases A20 expression in all tissues. As TNFα inhibits adipocyte differentiation, we have determined the contribution of A20 to the adipogenic capacity of human mesenchymal stromal cells (MSCs). Here we show that A20 is constitutively expressed in MSCs, which previously has been observed only in cells that are either tumor or immune cells (T/B lymphocytes). TNFα stimulation induced a rapid degradation of A20 protein mediated exclusively by the proteasome in MSCs and not by caspases. This degradation is concomitant to the induction of its own mRNA, which suggests that a tight regulation of NF-κB signaling in MSCs is fundamental. On one hand, we demonstrate that the knockdown of A20-mediated transcript dramatically decreases the adipogenic capacity of MSCs, which correlates with the phenotype observed in the presence of TNFα. On the other hand, A20 overexpression blocks NF-κB activation and drives to increased adipogenesis, even in the presence of TNFα treatment. In conclusion, our data demonstrate that the presence of A20 allows MSCs to differentiate into adipocytes by maintaining NF-κB signaling at a basal state.

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.

Gangliosides and sialic acid effects upon newborn pathogenic bacteria adhesion: an in vitro study.

The effect of the main gangliosides (GM(1), GM(3), GD(3)) and free sialic acid (Neu5Ac) upon the adhesion of pathogenic bacteria implicated in infant diarrhoea is assessed in vitro using the Caco-2 cell line. Concentrations of the bioactive compounds found in the bioaccessible (soluble) fraction of infant formula and human milk are employed. Bacterial adhesion behaviour included enterotoxigenic Escherichia coli (ETEC), enteropathogenic E.coli (EPEC), Listeria monocytogenes, Salmonella entericaserovartyphi, Shigella sonnei, Campylobacter jejuni and Helicobacter pylori. Three different approaches were assayed: pre-incubation of bacteria and compounds before addition to cells (competition); pre-incubation of the cells with compounds (exclusion); and pre-incubation of cells with bacteria (displacement). Furthermore, the spatial localization of the most abundant gangliosides, GM(3) and GD(3), in Caco-2 cells has been determined using confocal microscopy. Results show that GM(3), GD(3), GM(1) and Neu5Ac at the assayed concentrations are able to interfere with the adhesion of several pathogenic bacteria involved in neonatal diseases-the greatest effect corresponding to Neu5Ac, followed by GD(3), GM(1) and GM(3). Gangliosides GM(3) and GD(3) are located in the apical and basolateral membranes of the Caco-2 cells.

Posible síndrome serotoninérgico relacionado con la administración de venlafaxina y tramadol / Possible serotonin syndrome associated with administration of venlafaxine and tramadol

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