Single-cell proteomics reveals decreased abundance of proteostasis and meiosis proteins in advanced maternal age oocytes.

Advanced maternal age is associated with a decline in oocyte quality, which often leads to reproductive failure in humans. However, the mechanisms behind this age-related decline remain unclear. To gain insights into this phenomenon, we applied plexDIA, a multiplexed, single-cell mass spectrometry method, to analyze the proteome of oocytes from both young women and women of advanced maternal age. Our findings primarily revealed distinct proteomic profiles between immature fully grown germinal vesicle and mature metaphase II oocytes. Importantly, we further show that a woman's age is associated with changes in her oocyte proteome. Specifically, when compared to oocytes obtained from young women, advanced maternal age oocytes exhibited lower levels of the proteasome and TRiC complex, as well as other key regulators of proteostasis and meiosis. This suggests that aging adversely affects the proteostasis and meiosis networks in human oocytes. The proteins identified in this study hold potential as targets for improving oocyte quality and may guide future studies into the molecular processes underlying oocyte aging.

Bioinspired gelatin/bioceramic composites loaded with bone morphogenetic protein-2 (BMP-2) promote osteoporotic bone repair.

There are currently several commercialized products approved by the Food and Drug Administration and the European Medicines Agency based on the use of recombinant human BMP-2 for the treatment of non-unions long fractures and spinal fusion. However, the adverse effects recorded with the use of BMPs suggest the need for drug delivery carriers that allow reducing the required doses and improve their cost-effectiveness. Herein, we have developed a new osteoconductive scaffold that reduces the required doses of BMP-2 for promoting bone regeneration in an osteoporotic defect model. The composite is, in brief, a gelatin-based 3D scaffold reinforced with either calcium sulfate or hydroxyapatite as an inorganic osteoconductive biomaterial. To this end, the organic/inorganic composite systems showed high hydration capacity and good in vitro degradability. The incorporation of 7.5% (m/v) ceramic compounds resulted in scaffolds with stiffer Young modulus (179 and 75 kPa for CaSO4_7 and HA_7, respectively) than bare gelatin hydrogels (48 kPa). Studies with human bone-marrow derived mesenchymal stem cells (hBM-MSCs) revealed that the 3D scaffolds promote cell adhesion and proliferation along with osteogenic differentiation capabilities. Specifically, downregulation of stemness (Nanog, Oct4) genes and upregulation of osteogenic markers (ALP, Col1a1, Fmod) by two fold were observed over 10 days under basal culture conditions. Promisingly, the sustained in vitro release of BMP-2 observed from the porous reinforced scaffolds allowed us to address the critical-sized osteoporotic mice calvarial defects with a relatively low growth factor doses (600 ng BMP-2/scaffold) compared to conventional doses at 2-15 micrograms. Overall, this study demonstrates the promising potential of osteoconductive gelatin/calcium bioceramics composites as osteogenic growth factors delivery carriers for bone-regeneration via ultra-low growth factor doses.

A microfluidic sperm-sorting device reduces the proportion of sperm with double-stranded DNA fragmentation.

Sperm DNA fragmentation can be produced in one (ssSDF) or both (dsSDF) DNA strands, linked to difficulties in naturally achieving a pregnancy and recurrent miscarriages, respectively. The techniques more frequently used to select sperm require centrifugation, which may induce sperm DNA fragmentation (SDF). The objective of this study was to assess whether the microfluidic-based device FertileChip® (now ZyMot®ICSI) can diminish the proportion of sperm with dsSDF. First, in a blinded split pilot study, the semen of nine patients diagnosed with ≥60% dsSDF, was divided into three aliquots: not processed, processed with FertileChip®, and processed with swim up. The three aliquots were all analyzed using neutral COMET for the detection of dsSDF, resulting in a reduction of 46% (P < 0.001) with FertileChip® (dsSDF: 34.9%) compared with the ejaculate and the swim up (dsSDF: 65%). Thereafter, the FertileChip® was introduced into clinical practice and a cohort of 163 consecutive ICSI cycles of patients diagnosed with ≥60% dsSDF was analyzed. Fertilization rate was 75.41%. Pregnancy rates after the first embryo transfer were 53.2% (biochemical), 37.8% (clinical), 34% (ongoing) and the live birth rate was 28.8%. Cumulative pregnancy rates after one (65.4% of patients), two (27.6% of patients) or three (6.4% of patients) transfers were 66% (biochemical), 56.4% (clinical), 53.4% (ongoing) and the live birth rate was 42%. The selection of spermatozoa using Fertile Chip® significantly diminishes the percentage of dsSDF, compared with either the fresh ejaculate or after swim up. Its applicability in ICSI cycles of patients with high dsSDF resulted in good laboratory and clinical outcomes.

Thyroid as a target of adjuvant autoimmunity/inflammatory syndrome due to mRNA-based SARS-CoV2 vaccination: from Graves' disease to silent thyroiditis.

BACKGROUND: As COVID-19 became a pandemic, the urgent need to find an effective treatment vaccine has been a major objective. Vaccines contain adjuvants which are not exempt from adverse effects and can trigger the autoimmune/inflammatory syndrome induced by adjuvants (ASIA). There is very little information about autoimmune endocrine disease and the ASIA after the use of mRNA-based SARS-CoV2 vaccination. CASE SERIES: We report three cases and also review the literature showing that the thyroid gland can be involved in the ASIA induced by the mRNA-based SARS-CoV2 vaccination. We present the first case to date of silent thyroiditis described in the context of SARS-CoV2 vaccination with Pfizer/BioNTech. Also, we discuss the first subacute thyroiditis in the context of SARS-CoV2 vaccination with the Moderna's vaccine. Finally, we provide another case to be added to existing evidence on Graves' disease occurring post-vaccination with the Pfizer/BioNTech vaccine. DISCUSSION: Adjuvants play an important role in vaccines. Their ability to increase the immunogenicity of the active ingredient is necessary to achieve the desired immune response. Both the Moderna and the Pfizer/BioNTech vaccines use mRNA coding for the SARS-CoV2 S protein enhanced by adjuvants. In addition, the cross-reactivity between SARS-CoV2 and thyroid antigens has been reported. This would explain, at least, some of the autoimmune/inflammatory reactions produced during and after SARS-CoV2 infection and vaccination. CONCLUSION: The autoimmune/inflammatory syndrome induced by adjuvants involving the thyroid could be an adverse effect of SARS-CoV2 vaccination and could be underdiagnosed.

Predictive value of transcriptional expression of Krüppel-like factor-6 (KLF6) in head and neck carcinoma patients treated with radiotherapy

Purpose To analyse the relationship between the transcriptional expression of Krüppel-like factor-6 (KLF6) and local response to treatment with radiotherapy in patients with head and neck squamous cell carcinoma (HNSCC). Methods We determined the transcriptional expression of KLF6 in tumour biopsies obtained before treatment with radiotherapy in 83 HNSCC patients. The KLF6 expression was categorized according to the local control of the disease with a recursive partitioning analysis. Results During the follow-up period, 27 patients (32.5%) had a local recurrence of the tumour. Patients with local recurrence had significantly higher levels of KLF6 expression than patients in which radiotherapy achieved local control of the disease (P = 0.029). Five-year local recurrence-free survival for patients with a high transcriptional expression of KLF6 (n = 46) was 51.1% (95% CI 36.4–66.2%), and for patients with low expression it was 85.6% (95% CI 73.9–97.3%) (P = 0.0001). The results of a multivariate analysis showed that patients with a high KLF6 expression had a 3.8 times higher risk of local recurrence after treatment with radiotherapy (95% CI 1.4–10.5, P = 0.008). Conclusion Transcriptional expression of KLF6 was significantly related to local control in HNSCC patients treated with radiotherapy. Patients with high levels of KLF6 expression had a significantly higher risk of local recurrence after treatment (AU)

Predictive value of transcriptional expression of Krüppel-like factor-6 (KLF6) in head and neck carcinoma patients treated with radiotherapy.

PURPOSE: To analyse the relationship between the transcriptional expression of Krüppel-like factor-6 (KLF6) and local response to treatment with radiotherapy in patients with head and neck squamous cell carcinoma (HNSCC). METHODS: We determined the transcriptional expression of KLF6 in tumour biopsies obtained before treatment with radiotherapy in 83 HNSCC patients. The KLF6 expression was categorized according to the local control of the disease with a recursive partitioning analysis. RESULTS: During the follow-up period, 27 patients (32.5%) had a local recurrence of the tumour. Patients with local recurrence had significantly higher levels of KLF6 expression than patients in which radiotherapy achieved local control of the disease (P = 0.029). Five-year local recurrence-free survival for patients with a high transcriptional expression of KLF6 (n = 46) was 51.1% (95% CI 36.4-66.2%), and for patients with low expression it was 85.6% (95% CI 73.9-97.3%) (P = 0.0001). The results of a multivariate analysis showed that patients with a high KLF6 expression had a 3.8 times higher risk of local recurrence after treatment with radiotherapy (95% CI 1.4-10.5, P = 0.008). CONCLUSION: Transcriptional expression of KLF6 was significantly related to local control in HNSCC patients treated with radiotherapy. Patients with high levels of KLF6 expression had a significantly higher risk of local recurrence after treatment.

Phenotypic correlations in a large single-center cohort of patients with BSCL2 nerve disorders: a clinical, neurophysiological and muscle magnetic resonance imaging study.

BACKGROUND AND PURPOSE: BSCL2 heterozygote mutations are a common cause of distal hereditary motor neuropathies (dHMNs). A series of BSCL2 patients is presented and clinical, neurophysiological and muscle magnetic resonance imaging (MRI) findings are correlated. METHODS: Twenty-six patients from five families carrying the p.N88S mutation were identified. Age of onset, clinical phenotype (dHMN, Charcot-Marie-Tooth, spastic paraplegia), physical examination, disability measured as a modified Rankin Scale score and neurophysiological findings were collected. A whole body muscle MRI had been performed in 18 patients. The pattern of muscle involvement on T1-weighted and short time inversion recovery sequences was analysed. Hierarchical analysis using heatmaps and an MRI Composite Score were generated. Statistical analysis was carried out with STATA SE v.15 (TX, USA). RESULTS: The mean age was 51.54 ± 19.94 years and 14 patients were men. dHMN was the most common phenotype (50%) and five patients (19.23%) showed no findings on examination. Disease onset was commonly in childhood and disability was low (modified Rankin Scale score 1.34 ± 1.13) although median time since onset of disease was 32 years (range 10-47). Charcot-Marie-Tooth-like patients were more disabled and disability correlated with age. On muscle MRI, thenar eminence, soleus and tibialis anterior were most frequently involved, irrespective of clinical phenotype. MRI Composite Score was strongly correlated with disability. CONCLUSION: Patients with the p.N88S BSCL2 gene mutation are phenotypically variable, although dHMN is most frequent and generally slowly progressive. Muscle MRI pattern is consistent regardless of phenotype and correlates with disease severity, probably serving as a reliable outcome measure for future clinical trials.

Dexmedetomidine implementation in pain management in the immediate postoperative period of cardiac surgery. / Implementación de la dexmedetomidina en el manejo del dolor en el postoperatorio inmediato de cirugía cardíaca.

BACKGROUND: This study arose from the need to improve all tasks related to monitoring pain in post cardiac surgery patients. OBJECTIVES: Checking and quantifying the pain suffered by patients undergoing cardiac surgery in the first 24hours of their stay in the intensive care unit (ICU), treated with Dexmedetomidine (Dex) as analgesic adjuvant, as well as their degree of sedation and the need for opiates such as rescue analgesia. MATERIAL AND METHODS: Unicentric study, observational, descriptive, from April 2016 to September 2017. Both genders, all adult, undergoing cardiac surgery, operating theatre-extubated and Dex continuous infusion carriers. Pain degree level was evaluated by the Visual analogue Scale of Pain (VAS), from immediately post-surgery until 24hours from ICU entry and sedation degree, by the Richmond Sedation Agitation Scale (RASS), only while the Dex infusion lasted. RESULTS: 109 patients were included. The results obtained showed that the average pain suffered by patients during the first 24hours was .47 with standard deviation (SD) of 1.25; the average maximum pain experienced was 3.58 with a range of 0 to 8 and the RASS average was -.68 (SD: 0,80). In addition, 44.04% of the patients needed rescue opiates, although only 7.32% showed severe pain greater than 6. CONCLUSIONS: The Dex infusion patients showed mild levels of pain, however, a small percentage, who must be taken into account, suffered severe pain.

Comparison of two different oocyte vitrification methods: a prospective, paired study on the same genetic background and stimulation protocol.

STUDY QUESTION: Can two different methods for oocyte vitrification, one using an open tool and the other a closed tool, result in similar oocyte survival rates? SUMMARY ANSWER: The oocyte survival rate was found to be higher in the closed method. WHAT IS KNOWN ALREADY: Open vitrification is performed routinely in oocyte donation cycles. Closed oocyte vitrification may result in slower cooling rates and thus it is less used, even though it has been recommended in order to avoid the risk of cross-contamination between material from different patients. STUDY DESIGN, SIZE, DURATION: This is a prospective cohort study with sibling oocytes carried out in a fertility center between July 2014 and January 2016. The study included 83 oocyte donors each providing a minimum of 12 mature oocytes (metaphase II: MII) at oocyte retrieval. Oocyte survival rate and fertilization rate, as well as reproductive outcomes (biochemical, clinical, ongoing pregnancy and live birth rates) per embryo transfer and also cumulatively between the two methods were compared by Chi2 tests. PARTICIPANTS/MATERIALS, SETTING, METHODS: Donor oocytes were denuded and six MII oocytes from each donor were vitrified using an open method and later assigned to one recipient, while another six MII oocytes were vitrified using a closed method and assigned to a different recipient (paired analysis). ICSI was used in all cases and embryo transfer was performed on Day 2-3 in all cases. MAIN RESULTS AND THE ROLE OF CHANCE: Oocyte donors were 24.8 years old on average (SD 4.7). Recipient age (average 41.2 years, SD 4.7) and BMI (mean 23.8 kg/m2, SD 4.0) were similar between recipient groups. Oocytes vitrified using the closed method had higher survival rate (94.5% versus 88.9%, P = 0.002), but lower fertilization rate (57.1% versus 69.8%, P < 0.001) compared to the open method. The number of fresh embryos transferred in the two groups was 1.8 on average (SD 0.4). Biochemical (45% closed versus 50% open), clinical (40% versus 50%) and ongoing (37.5% versus 42.5%) pregnancy rates were not different between groups (P > 0.05) and neither were live birth rates (37.5% versus 42.5%, P > 0.05). Cumulative reproductive results (obtained after the transfer of all the embryos) were also similar between groups. LIMITATIONS, REASONS FOR CAUTION: The participants of this study were oocyte donors, i.e. young women in good health, and care should be exerted in extending our results to other populations such as infertility patients, oncofertility patients and women freezing oocytes to delay childbearing. WIDER IMPLICATIONS OF THE FINDINGS: Our results suggest that, in spite of different survival and fertilization rates, closed and open oocyte vitrification methods should offer similar reproductive outcomes up to cumulative live birth rates. STUDY FUNDING/COMPETING INTEREST(S): The authors report no conflict of interest. Vitrolife provided the media and the closed method tool needed for the study at no cost.

Arytenoid asymmetry in opera singers.

INTRODUCTION: Otolaryngologists commonly observe asymmetrical movements of the arytenoid cartilages, but few authors have described the clinical implications of this asymmetry, especially in singers. OBJECTIVES: The aim of this study was to determine the epidemiological and clinical characteristics of arytenoid asymmetry in adduction in a group of healthy opera singers and to evaluate the impact of this asymmetry on the voice. PATIENTS AND METHODS: The medical charts and laryngeal video recordings of 245 healthy opera singers were retrospectively reviewed. Arytenoid asymmetry was defined in relation to the position of the corniculate cartilages, cuneiform cartilages and aryepiglottic angle. RESULTS: The subjects had a mean age of 38.54years (range: 18 to 85 years) and presented a male-to-female sex ratio of 1.02. About 5% of subjects had a history of smoking. The most common vocal symptoms were occasional dysphonia (4%), followed by vocal fatigue (2%) and pharyngeal dryness (2%). Arytenoid asymmetry was slightly more common in males (50.6%) and predominantly affected the right side (64.5%). The most common form of asymmetry was cuneiform asymmetry (37.5%), followed by aryepiglottic angle asymmetry in 33.1% of cases and corniculate asymmetry in 29.4% of cases. Mild asymmetry was observed in the majority of cases. No correlation was observed between arytenoid asymmetry and vocal symptoms. CONCLUSION: Almost one out of two singers was likely to present arytenoid asymmetry in adduction. The presence of this asymmetry does not appear to be correlated with any vocal symptoms.

Effect of kaolin silver complex on the control of populations of Brettanomyces and acetic acid bacteria in wine.

In this work, the effects of kaolin silver complex (KAgC) have been evaluated to replace the use of SO2 for the control of spoilage microorganisms in the winemaking process. The results showed that KAgC at a dose of 1 g/L provided effective control against the development of B. bruxellensis and acetic acid bacteria. In wines artificially contaminated with an initial population of B. bruxellensis at 104 CFU/mL, a concentration proven to produce off flavors in wine, only residual populations of the contaminating yeast remained after 24 days of contact with the additive. Populations of acetic bacteria inoculated into wine at concentrations of 102 and 104 CFU/mL were reduced to negligible levels after 72 h of treatment with KAgC. The antimicrobial effect of KAgC against B. bruxellensis and acetic bacteria was also demonstrated in a wine naturally contaminated by these microorganisms, decreasing their population in a similar way to a chitosan treatment. Related to this effect, wines with KAgC showed lower concentrations of acetic acid and 4-ethyl phenol than wines without KAgC. The silver concentration from KAgC that remained in the finished wines was below the legal limits. These results demonstrated the effectiveness of KAgC to reduce spoilage microorganisms in winemaking.

Is there a relation between the time to ICSI and the reproductive outcomes?

STUDY QUESTION: Does time to ICSI affect reproductive outcomes? SUMMARY ANSWER: Biochemical and clinical pregnancy diminish progressively as time between oocyte pick up (OPU) and ICSI increases after fresh embryo transfer. WHAT IS KNOWN ALREADY: Appropriate oocyte cytoplasmic and nuclear maturation are of paramount importance to ensure an optimal embryonic developmental competence. While nuclear maturation is usually attained by the time an oocyte reaches OPU, cytoplasmic maturation cannot be readily assessed and might be incomplete. On the other hand, excessive in vitro culture of mature human oocytes can affect their ultrastructural characteristics and, in mice, induces alterations in gene expression and changes of chromatin and histone modification patterns. STUDY DESIGN, SIZE, DURATION: Retrospective consecutive cohort study including 1468 ICSI cycles carried out in a single center between December 2012 and September 2015. All cycles were with patient's own oocytes and fresh embryo transfer (ET). A radiofrequency-based system was used to record exact culture times, namely, OPU-denudation (DN); DN-ICSI and OPU-ICSI. We analyzed the effect of total and partial time intervals between procedures, from OPU to ICSI, on fertilization rate and biochemical, clinical, ongoing pregnancy and live birth rates. PARTICIPANTS/MATERIALS, SETTING, METHODS: Differences in laboratory times between positive and negative biochemical, clinical, ongoing pregnancies and birth results were tested by Mann-Whitney U test. The likelihood of positive clinical outcomes was further modeled by locally weighted scatterplot smoothing (LOWESS) regression and logistic regression, adjusting for woman's age and BMI, number of transferred embryos; mean embryo morphological score, sperm origin and status, and number of mature oocytes obtained at OPU. Effect of time on fertilization rate was modeled by Generalized Linear Modeling (GLM) and LOWESS regression. MAIN RESULTS AND THE ROLE OF CHANCE: The mean woman's age was 38.4 years (SD 4.6). Biochemical, clinical, ongoing pregnancy and live birth rates after the fresh ET were: 39.6, 33.1, 25.7 and 20.8%, respectively. Cumulative values for biochemical pregnancy and live birth were 46.4 and 26.3%, respectively. Mean times in hours for OPU-DN, DN-ICSI and OPU-ICSI were: 1.00 (SD 0.20); 3.86 (SD 1.93) and 4.87 (SD 1.96), respectively, and were not different for pregnant and non-pregnant patients. However, multivariate analyses showed that on average (anti-log transformed), each 1-h increase in the OPU-ICSI time reduced the likelihood of biochemical pregnancy by 7.3% (95% CI: 0.7-13.5%) and of clinical pregnancy by 7.7% (95% CI 0.8-14.1%), after the fresh ET. No effect of time was observed for ongoing pregnancy or live birth rates. Increasing OPU-ICSI time increased the fertilization rate (B = 0.052, 95% CI: 0.022, 0.082). LIMITATIONS, REASONS FOR CAUTION: The lack of relationship between incubation time of oocytes and live birth rates might be due to uncontrolled variables. Given the population analyzed, these results should not be extended to other ART protocols such as in vitro maturation of oocytes or classical IVF fertilization. WIDER IMPLICATIONS OF THE FINDINGS: This study indicates that in vitro ageing of mature oocytes significantly affects the chances to become pregnant. Effect on live birth rates, although not evident in this study, cannot be excluded. Limiting incubation time of mature oocytes in the embryology laboratory should improve reproductive results for patients using their own oocytes and with a transfer of fresh embryos. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: NA.

Utilidad de la técnica ROLL en la exéresis de lesiones no palpables de mama / Use of the ROLL technique for lumpectomy in non-palpable breast lesions

Objetivo: Evaluar la eficacia de la localización radioguiada de lesiones no palpables de mama (LNPM) respecto al arpón quirúrgico. Método: Se han estudiado prospectivamente 161 mujeres con LNPM, 80 marcadas con arpón (grupo 1) y 81 con inyección intratumoral de 99mTc nanocoloide (grupo 2). Las lesiones se localizaron por ecografía o estereotaxia. Las tumorectomías se realizaron, en el grupo 1 siguiendo la dirección del arpón y en el grupo 2 con la ayuda de una sonda gammadetectora. Posteriormente se comprobaron los márgenes quirúrgicos, determinando la necesidad de ampliación si el margen era menor a 5 mm en el estudio intraoperatorio y menor a 2mm en el estudio diferido. Se recogieron datos de porcentaje de detección quirúrgica, afectación de márgenes quirúrgicos, número de ampliaciones, presencia de lesión residual en la ampliación, número de reintervenciones, volumen de la tumorectomía y volumen total extraído, ratio volumen/tumor y complicaciones. Resultados: No hubo diferencias significativas entre ambos grupos en porcentaje de detección, afectación de márgenes, número de ampliaciones, presencia de lesión residual en la ampliación, reintervenciones, volumen de la tumorectomía, volumen total extraído, ratio volumen/tumor y complicaciones. El análisis multivariante mostró que los factores condicionantes del volumen extraído son la técnica de marcaje radiológico y el cirujano. Conclusiones: La técnica de localización radioguiada de lesiones ocultas permite la detección y exeresis de las LNPM con la misma eficacia que el arpón y añade la posibilidad de detección simultánea del ganglio centinela. Los condicionantes del volumen extraído son la técnica de marcaje radiológico y el cirujano (AU)

Objective: To evaluate the efficiency of radioguided occult lesion localising in non-palpable breast lesions (NPBL) compared to the surgical wire technique. Method: A prospective study was conducted on 161 women with NPBL, of whom 80 marked with the wire (group 1), whereas 81 women were marked with an intratumour injection of 99mTc-nanocoloid (group 2). The NPBL were located by ultrasound or stereotactic guidance. The lumpectomies were performed following the wire direction in group 1, and with the aid of a gamma-probe in group 2. Surgical margins were then checked, determining the need of extension if the margin was less than 5mm in the intra-surgical study, and less than 2mm in the deferred study. Data were collected on the mean number detected by surgery, surgical margins, number of extensions, presence of residual tumour in the extension, second surgeries, lumpectomy volume, as well as total resected volume, volume/tumour ratio, and complications. Results: No significant differences were observed between the two groups in the mean number detected, surgical margins, number of extensions, presence of residual tumour in the extension, second surgeries, lumpectomy volume, total resected volume, volume/tumour ratio or complications. The multivariate analysis showed the determining factors of the resected volume were the radiological guidance technique, as well as the surgeon. Conclusions: The radioguided occult lesion localising technique helps in the detection and resection of NPBL with the same efficiency as the surgical wire, and adds the possibility of sentinel node detection in the same surgery. The determining factors of the resected volume were the radiological guidance technique and the surgeon (AU)

Use of the ROLL technique for lumpectomy in non-palpable breast lesions. / Utilidad de la técnica ROLL en la exéresis de lesiones no palpables de mama.

OBJECTIVE: To evaluate the efficiency of radioguided occult lesion localising in non-palpable breast lesions (NPBL) compared to the surgical wire technique. METHOD: A prospective study was conducted on 161 women with NPBL, of whom 80 marked with the wire (group 1), whereas 81 women were marked with an intratumour injection of 99mTc-nanocoloid (group 2). The NPBL were located by ultrasound or stereotactic guidance. The lumpectomies were performed following the wire direction in group 1, and with the aid of a gamma-probe in group 2. Surgical margins were then checked, determining the need of extension if the margin was less than 5mm in the intra-surgical study, and less than 2mm in the deferred study. Data were collected on the mean number detected by surgery, surgical margins, number of extensions, presence of residual tumour in the extension, second surgeries, lumpectomy volume, as well as total resected volume, volume/tumour ratio, and complications. RESULTS: No significant differences were observed between the two groups in the mean number detected, surgical margins, number of extensions, presence of residual tumour in the extension, second surgeries, lumpectomy volume, total resected volume, volume/tumour ratio or complications. The multivariate analysis showed the determining factors of the resected volume were the radiological guidance technique, as well as the surgeon. CONCLUSIONS: The radioguided occult lesion localising technique helps in the detection and resection of NPBL with the same efficiency as the surgical wire, and adds the possibility of sentinel node detection in the same surgery. The determining factors of the resected volume were the radiological guidance technique and the surgeon.

De novo loss of function mutations in KIAA2022 are associated with epilepsy and neurodevelopmental delay in females.

Intellectual disability (ID) affects about 3% of the population and has a male gender bias. Of at least 700 genes currently linked to ID, more than 100 have been identified on the X chromosome, including KIAA2022. KIAA2022 is located on Xq13.3 and is expressed in the developing brain. The protein product of KIAA2022, X­linked Intellectual Disability Protein Related to Neurite Extension (XPN), is developmentally regulated and is involved in neuronal migration and cell adhesion. The clinical manifestations of loss­of­function KIAA2022 mutations have been described previously in 15 males, born from unaffected carrier mothers, but few females. Using whole­exome sequencing, we identified a cohort of five unrelated female patients with de novo probably gene damaging variants in KIAA2022 and core phenotypic features of ID, developmental delay, epilepsy refractory to treatment, and impaired language, of similar severity as reported for male counterparts. This study supports KIAA2022 as a novel cause of X­linked dominant ID, and broadens the phenotype for KIAA2022 mutations.

Trends in incidence and survival analysis in non-melanoma skin cancer from 1994 to 2012 in Girona, Spain: A population-based study.

INTRODUCTION: We present an epidemiological study focused on Non-melanoma skin cancers (NMSC), including squamous cell carcinoma (SCC), basal cell carcinoma (BCC), Merkel cell carcinoma (MCC), dermatofibrosarcoma protuberans (DFS) and adnexal and skin appendages neoplasm (ASAN), a neoplasm understudied in cancer registries. MATERIAL AND METHODS: We analyze trends of incidence and survival of NMSC registered with the Cancer Registry of Girona, Spain. RESULTS: We found 14389 cases of NMSC, accounting 3,474 SCC, 10729 BCC, 33 MCC, 61 DFSP and 71 ASAN. Incidence increased significantly in SCC and BCC with annual percentage of change of 1.6 and 1.5, respectively, but not in MCC, DFS or ASAN. Five-year relative survival for both sexes was 90.1% in SCC, 99.8% in BCC, 44.2% in MCC, 93.7% in DFS and 84% in ASAN. CONCLUSIONS: Our study confirms the increasing incidence and good survival of SCC and BCC and enhances knowledge on the epidemiology of the less incidental NMSC.

Ruminococcus gnavus E1 modulates mucin expression and intestinal glycosylation.

AIMS: The molecular cross-talk between commensal bacteria and the gut play an important role in the maintenance of the intestinal homeostasis and general health. Here, we studied the impact of a major Gram-positive anaerobic bacterium of the human gut microbiota, that is, Ruminococcus gnavus on the glycosylation pattern and the production of intestinal mucus by the goblet cells. METHODS AND RESULTS: Our results showed that R. gnavus E1 specifically increases the expression and the glycosylation level of the intestinal glyco-conjugates by goblet cells in the colonic mucosa of mono-associated mice with R. gnavus E1 as well as in human HT29-MTX cells. Such an effect was mediated through induction of the level of mRNA encoding for the major intestinal gel-forming mucin such as MUC2 and various glycosyltransferase enzymes. CONCLUSIONS: This study demonstrates for the first time that R. gnavus E1 possess the ability to modulate the glycosylation profile of the glyco-conjugate molecules and mucus in goblet cells. SIGNIFICANCE AND IMPACT OF THE STUDY: Furthermore, we demonstrated that R. gnavus E1 modified specifically the glycosylation pattern and MUC2 expression by means of a small soluble factor of peptidic nature (<3 kDa) and heat stable in the HT29-MTX cell.

Effect of ribonucleic acid (RNA) isolation methods on putative reference genes messenger RNA abundance in human spermatozoa.

Although the male gamete participates in a significant proportion of infertility cases, there are currently no proven molecular markers of sperm quality. The search for significant gene expression markers is partially hindered by the lack of a recognized set of reference genes (RGs) to normalize reverse transcription quantitative PCR (RT-qPCR) data across studies. The aim of this study is to define a set of RGs in assisted reproduction patients undergoing different sample collection and RNA isolation methods. Twenty-two normozoospermic men were included in the study. From each man, semen was either cryopreserved by slow freezing or analyzed fresh, and, for each, RNA was extracted with either phenol-free or phenol-based methods. In two cases, both methods were used to isolate RNA. Twenty putative RGs were analyzed and their mRNA abundance across samples was estimated by RT-qPCR. To determine the genes whose steady-state mRNA abundance remains unchanged, three different algorithms (geNorm, BestKeeper and NormFinder) were applied to the qPCR data. We found that RGs such as GAPDH or ACTB, useful in other biological contexts, cannot be used as reference for human spermatozoa. It is possible to compare gene expression from fresh and cryopreserved sperm samples using the same isolation method, while the mRNA abundance of expressed genes becomes different depending on the RNA isolation technique employed. In our conditions, the most appropriate RGs for RT-qPCR analysis were RPLP1, RPL13A, and RPLP2. Published discrepancies in gene expression studies in human spermatozoa may be due in part to inappropriate RGs selection, suggesting a possible different interpretation of PCR data in several reports, which were normalized using unstable RGs.

Análisis comparativo de los conocimientos en ventilación mecánica no invasiva de profesionales de cuidados intensivos / Intensive care unit profesionals's knowledge about non invasive ventilation comparative analysis

Objetivo: La bibliografía destaca la falta de protocolos de ventilación mecánica no invasiva (VNI) y la variabilidad de conocimientos entre unidades y hospitales, por lo que se pretende comparar los conocimientos en VNI en enfermeras de 4 UCI polivalentes y una quirúrgica. Métodos: Estudio descriptivo multicéntrico en hospitales universitarios de nivel 3. Se diseñó una encuesta ad-hoc, con fiabilidad interobservador Kappa = 0,9. Contestar la encuesta era indicación de consentimiento informado. Análisis con Chi cuadrado. Resultados: Respondieron 117 (65%) enfermeras, con 11 ± 9,7 años de experiencia en UCI y 9,2 ± 7,2 en uso de VNI. Una de las UCI polivalentes había iniciado la VNI una media de 6 años más tarde que las otras (IC 95% [3,3 a 8,6], P <0,001).Solo el 23,1% de las enfermeras colocaría una mascarilla sin puerto espiratorio en respirador convencional, el resto cualquier mascarilla buconasal. El 12,7% cree que la mascarilla debe permitir el paso de 2 dedos mientras que el 29% apretaría la máscara a la cara del paciente y taparían el puerto espiratorio para facilitar la sincronización con el respirador. En la UCI quirúrgica se identifica mayoritariamente la agitación como una complicación de la VNI comparado con las polivalentes (31,6 vs. 1,8%, P < 0,001).El 56,4% de las enfermeras no consideran la fisioterapia respiratoria como un cuidado enfermero, sin diferencia entre unidades. Conclusiones: El conocimiento en tipos de interfase es muy dependiente del material de la unidad. Falta formación para detectar complicaciones de VNI como la agitación y el manejo de secreciones

Aims: The literature highlights the lack of noninvasive vntilation (NIV) protocols and the variability of the knowledge of NIV between intensive care units (ICU) and hospitals, so we want to compare NIV nurses's Knowledge from 4 multipurpose ICU and one surgical ICU. Methods: Multicenter, crosscutting, descriptive study in three university hospitals. The survey instrument was validated in a pilot test, and the calculated Kappa index was 0.9. Returning a completed survey is an indication of informed consent. Analysis by Chi square test. Results: 117 responded (65%) nurses, 11 ± 9.7 years of experience in ICU and 9.2 ± 7.2 in use of NIV. One of the multipurpose ICU, was initiated NIV an average of 6 years later than the others (95% CI [3.3 to 8.6], P < .001). Only 23.1% of nurses would place a non-vented mask (with no exhalation port) by conventional ventilator, the rest any kind of face mask. 12.7% believed that the mask must be adjusted to the '2-finger' fit while 29% would seal the mask to the patient's face and cover the mask opening where air escapes to facilitate patient/ventilator synchronization. In the surgical ICU agitation identifies mostly as a complication of NIV compared with multipurpose UCIs (31.6% vs 1.8%, P < .001). 56.4% of nurses do not consider respiratory physiotherapy as nursing care, with no difference between units. Conclusions: Knowledge about types of interface is very dependent on the material of the unit. More training for complications of NIV as agitation and handling secretions it is necessary

[Intensive care unit profesionals's knowledge about non invasive ventilation comparative analysis]. / Análisis comparativo de los conocimientos en ventilación mecánica no invasiva de profesionales de cuidados intensivos.

AIMS: The literature highlights the lack of noninvasive ventilation (NIV) protocols and the variability of the knowledge of NIV between intensive care units (ICU) and hospitals, so we want to compare NIV nurses's Knowledge from 4 multipurpose ICU and one surgical ICU. METHODS: Multicenter, crosscutting, descriptive study in three university hospitals. The survey instrument was validated in a pilot test, and the calculated Kappa index was 0.9. Returning a completed survey is an indication of informed consent. Analysis by Chi square test. RESULTS: 117 responded (65%) nurses, 11±9.7 years of experience in ICU and 9.2±7.2 in use of NIV. One of the multipurpose ICU, was initiated NIV an average of 6 years later than the others (95% CI [3.3 to 8.6], P<.001). Only 23.1% of nurses would place a non-vented mask (with no exhalation port) by conventional ventilator, the rest any kind of face mask. 12.7% believed that the mask must be adjusted to the "2-finger" fit while 29% would seal the mask to the patient's face and cover the mask opening where air escapes to facilitate patient/ventilator synchronization. In the surgical ICU agitation identifies mostly as a complication of NIV compared with multipurpose UCIs (31.6% vs 1.8%, P<.001). 56.4% of nurses do not consider respiratory physiotherapy as nursing care, with no difference between units. CONCLUSIONS: Knowledge about types of interface is very dependent on the material of the unit. More training for complications of NIV as agitation and handling secretions it is necessary.