Protamine folds DNA into flowers and loop stacks.

DNA in sperm undergoes an extreme compaction to almost crystalline packing levels. To produce this dense packing, DNA is dramatically reorganized in minutes by protamine proteins. Protamines are positively charged proteins that coat negatively charged DNA and fold it into a series of toroids. The exact mechanism for forming these ∼50-kbp toroids is unknown. Our goal is to study toroid formation by starting at the "bottom" with folding of short lengths of DNA that form loops and working "up" to more folded structures that occur on longer length scales. We previously measured folding of 200-300 bp of DNA into a loop. Here, we look at folding of intermediate DNA lengths (L = 639-3003 bp) that are 2-10 loops long. We observe two folded structures besides loops that we hypothesize are early intermediates in the toroid formation pathway. At low protamine concentrations (∼0.2 µM), we see that the DNA folds into flowers (structures with multiple loops that are positioned so they look like the petals of a flower). Folding at these concentrations condenses the DNA to 25% of its original length, takes seconds, and is made up of many small bending steps. At higher protamine concentrations (≥2 µM), we observe a second folded structure-the loop stack-where loops are stacked vertically one on top of another. These results lead us to propose a two-step process for folding at this length scale: 1) protamine binds to DNA, bending it into loops and flowers, and 2) flowers collapse into loop stacks. These results highlight how protamine uses a bind-and-bend mechanism to rapidly fold DNA, which may be why protamine can fold the entire sperm genome in minutes.

Extracellular Vesicles Secreted by Pre-Hatching Bovine Embryos Produced In Vitro and In Vivo Alter the Expression of IFNtau-Stimulated Genes in Bovine Endometrial Cells.

The embryo-maternal interaction occurs during the early stages of embryo development and is essential for the implantation and full-term development of the embryo. In bovines, the secretion of interferon Tau (IFNT) during elongation is the main signal for pregnancy recognition, but its expression starts around the blastocyst stage. Embryos release extracellular vesicles (EVs) as an alternative mechanism of embryo-maternal communication. The aim of the study was to determine whether EVs secreted by bovine embryos during blastulation (D5-D7) could induce transcriptomic modifications, activating IFNT signaling in endometrial cells. Additionally, it aims to assess whether the EVs secreted by embryos produced in vivo (EVs-IVV) or in vitro (EVs-IVP) have different effects on the transcriptomic profiles of the endometrial cells. In vitro- and in vivo-produced bovine morulae were selected and individually cultured for 48 h to collect embryonic EVs (E-EVs) secreted during blastulation. E-EVs stained with PKH67 were added to in vitro-cultured bovine endometrial cells to assess EV internalization. The effect of EVs on the transcriptomic profile of endometrial cells was determined by RNA sequencing. EVs from both types of embryos induced several classical and non-classical IFNT-stimulated genes (ISGs) and other pathways related to endometrial function in epithelial endometrial cells. Higher numbers of differentially expressed genes (3552) were induced by EVs released by IVP embryos compared to EVs from IVV (1838). Gene ontology analysis showed that EVs-IVP/IVV induced the upregulation of the extracellular exosome pathway, the cellular response to stimulus, and the protein modification processes. This work provides evidence regarding the effect of embryo origin (in vivo or in vitro) on the early embryo-maternal interaction mediated by extracellular vesicles.

Evaluation of extracellular vesicles and gDNA from culture medium as a possible indicator of developmental competence in human embryos.

Human embryos generated in vitro have a high incidence of chromosomal abnormalities that negatively affect pregnancy rate. Embryos generated in vitro secrete extracellular vesicles (EVs) into the culture medium that could be used potentially as indicators of embryo competence. This research aimed to evaluate the concentration and size of EVs and their gDNA content as an indicator of developmental competence in human embryos. Human embryos generated by intracytoplasmic sperm injection (ICSI) were classified morphologically as of either TOP, FAIR or POOR quality. Culture medium and developmentally arrested embryos (which were not able to be used for embryo transfer) were collected. Microvesicles, exosomes (MV/Exo) and apoptotic bodies (ABs) were isolated from culture medium. Nanoparticle tracking analysis (NTA) and array comparative genomic hybridization (aCGH) analysis were performed to evaluate EVs and their gDNA content. From NTA, the diameter (mean) of MVs/Exo from TOP quality embryos was higher (112.17 nm) compared with that of FAIR (108.02) and POOR quality embryos (102.78 nm) (P < 0.05). aCGH analysis indicated that MVs/Exo and ABs carried gDNA with the presence of 23 chromosome pairs. However, when arrested embryos were compared with their respective MVs/Exo and ABs, the latter had an increased rate of chromosomal abnormalities (24.9%) compared with embryos (8.7%) (P < 0.05). In conclusion, the size of EVs from culture medium might be an alternative for evaluating competence of human embryos, however more studies are needed to validate the use of gDNA from EVs as an indicator of embryo competence.

Extracellular vesicles secreted during blastulation show viability of bovine embryos.

Extracellular vesicles (EVs) secreted by blastocysts may be clinically relevant, as indicator of embryo viability on in vitro fertilization. We tested if the characteristics of EVs secreted during blastulation are related to embryo viability. Morulae were individually cultured in SOF media depleted of EVs until day 7.5 post IVF. Viable embryos were determined by a system of extended in vitro culture of bovine embryos until day 11 (post-hatching development). Afterward, a retrospective classification of blastocyst and culture media was performed based on blastulation time (early blastulation (EB) or late blastulation (LB)) and post-hatching development at day 11 (viable (V) or non-viable embryo (NV)). A total of 254 blastocysts and their culture media were classified in four groups (V-EB, NV-EB, V-LB, NV-LB). Group V-EB had a larger blastocyst diameter (170.8 µm), higher proportion of good-quality blastocysts (77%) and larger mean size of population of EVs (122.9 nm), although the highest concentration of EVs (5.75 × 109 particles/mL) were in group NV-EB. Furthermore, small RNA sequencing detected two biotypes, miRNA (86-91%) and snoRNA (9-14%), with a total of 182 and 32 respectively. In differential expression analysis of miRNAs between V versus NV blastocysts, there were 12 miRNAs upregulated and 15 miRNAs downregulated. Binary logistic regression was used to construct a non-invasive novel model to select viable embryos, based on a combination of variables of blastocyst morphokinetics and EVs characteristics, the ROC-AUC was 0.853. We concluded that characteristics of EVs secreted during blastulation vary depending on embryo quality.

Effect of BMP15 and/or AMH during in vitro maturation of oocytes from involuntarily culled dairy cows.

The high metabolic activity to which the dairy cattle are exposed to maintain milk production altered steroid metabolism that affects reproductive physiology and reduce oocyte competence. Our aims were (a) to characterize the competence of immature oocytes collected from dairy cattle based on the expression of genes in cumulus cells (CCs) and (b) to improve oocyte competence to support preimplantation embryo development by the supplementation of maturation medium with bone morphogenetic protein 15 (BMP15) and/or anti-mullerian hormone (AMH). Oocyte donors were identified at the moment of ovary collection and grouped by involuntarily culled dairy cows (Holstein breed) or beef cattle. The embryo development speed to blastocyst of the cull dairy cattle versus beef cattle (control group) was lower. Besides, <10% of oocytes (with CC biopsies) derived from dairy cattle were able to develop to the blastocyst stage. In addition, a higher level of expression and a positive correlation were observed in the expression of most of the genes evaluated (LUM, KRT18, KRT8, CLIC3, BMPR1B, and SLC38A3) in the cumulus-oocyte complexes that produced blastocysts versus those which did not develop correctly (arrested development). Further, use of BMP15 in the maturation of oocytes from dairy cattle seems to increase competence, modulating the expression of OCT4, SOX2, CDX2, GATA6, and TP1 in resulting blastocysts.

Splitting of IVP bovine blastocyst affects morphology and gene expression of resulting demi-embryos during in vitro culture and in vivo elongation.

Embryo splitting might be used to increase offspring yield and for molecular analysis of embryo competence. How splitting affects developmental potential of embryos is unknown. This research aimed to study the effect of bovine blastocyst splitting on morphological and gene expression homogeneity of demi-embryos and on embryo competence during elongation. Grade I bovine blastocyst produced in vitro were split into halves and distributed in nine groups (3 × 3 setting according to age and stage before splitting; age: days 7-9; stage: early, expanded and hatched blastocysts). Homogeneity and survival rate in vitro after splitting (12 h, days 10 and 13) and the effect of splitting on embryo development at elongation after embryo transfer (day 17) were assessed morphologically and by RT-qPCR. The genes analysed were OCT4, SOX2, NANOG, CDX2, TP1, TKDP1, EOMES, and BAX. Approximately 90% of split embryos had a well conserved defined inner cell mass (ICM), 70% of the halves had similar size with no differences in gene expression 12 h after splitting. Split embryos cultured further conserved normal and comparable morphology at day 10 of development; this situation changes at day 13 when embryo morphology and gene expression differed markedly among demi-embryos. Split and non-split blastocysts were transferred to recipient cows and were recovered at day 17. Fifty per cent of non-split embryos were larger than 100 mm (33% for split embryos). OCT4, SOX2, TP1 and EOMES levels were down-regulated in elongated embryos derived from split blastocysts. In conclusion, splitting day-8 blastocysts yields homogenous demi-embryos in terms of developmental capability and gene expression, but the initiation of the filamentous stage seems to be affected by the splitting.

Nanoparticles from culture media are internalized by in vitro-produced bovine embryos and its depletion affect expression of pluripotency genes.

Extracellular vesicles are nanoparticles secreted by cell and have been proposed as suitable markers to identify competent embryos produced in vitro. Characterizing EVs secreted by individual embryos is challenging because culture medium itself contributes to the pool of nanoparticles that are co-isolated. To avoid this, culture medium must be depleted of nanoparticles that are present in natural protein source. The aim of this study was to evaluate if the culture medium subjected to nanoparticle depletion can support the proper in vitro development of bovine embryos. Zygotes were cultured in groups on depleted or control medium for 8 days. Nanoparticles from the medium were characterized by their morphology, size and expression of EVs surface markers. Isolated nanoparticles were labelled and added to depleted medium containing embryos at different developmental stages and evaluated after 24 hours at 2, 8-16 cells, morula and blastocyst stages. There were no statistical differences on blastocyst rate at day 7 and 8, total cell count neither blastocyst diameter between groups. However, morphological quality was better in blastocysts cultured in non-depleted medium and the expression of SOX2 was significantly lower whereas NANOG expression was significantly higher. Few nanoparticles from medium had a typical morphology of EVs but were positive to specific surface markers. Punctuated green fluorescence near the nuclei of embryonic cells was observed in embryos from all developmental stages. In summary, nanoparticles from culture medium are internalized by in vitro cultured bovine embryos and their depletion affects the capacity of medium to support the proper embryo development.

DMSO supplementation during in vitro maturation of bovine oocytes improves blastocyst rate and quality.

The molecule Dimethyl sulfoxide is widely used as drug solvent. However, its antioxidant property was poorly explored. In this study, we evaluated the effect of DMSO supplementation during oocyte in vitro maturation (IVM) on embryo development and quality. Bovine oocytes were matured with different DMSO concentrations (0, 0.1, 0.25, 0.5, 0.75, 1 and 10% v:v) followed by in vitro fertilization. Subsequently, quality indicators such as gene expression of SOX2, OCT4, CDX2, SOD1, oocyte and embryo redox status and DNA damage were evaluated. Polar body extrusion and blastocyst rates increased with 0.5% v:v DMSO. Moreover, first polar body extrusion and blastocyst rates did not increase with 1%, and 10% of DMSO reduced polar body extrusion and did not produce blastocyst. Optimal concentration of DMSO for the use on the maturation was estimated at around 0.45% v:v. Supplementation with 0.5% v:v DMSO has not affected mRNA abundance of genes key in blastocyst, however 0.75% increased gene expression of OCT4 and SOX2. Oocytes matured with 0.5% v:v DMSO and blastocyst from DMSO group showed reduced lipid peroxidation respect control. Total Glutathione concentrations increased in blastocyst stage in DMSO group. DMSO increased the total cell number of blastocysts but not TUNEL positive cells. In conclusion, our results suggest that low DMSO concentrations used during bovine oocytes in vitro maturation increases the maturation, as well as the blastocyst rate and its quality, without demonstrating deleterious effect on embryo cells.

[Sarcomatoid carcinoma of larynx. A histological challenge?] / Carcinoma sarcomatoide de laringe. ¿Un reto histológico?

INTRODUCTION: Sarcomatoid carcinoma can occur in any part of the body; in the head and neck it occurs most frequently in the major salivary glands, with only about 1% of cases found in the larynx. As it has both epithelial and mesenchymal components, there are many theories concerning its origin and it can prove a diagnostic challenge. CASE REPORT: A 76 year old male smoker presented with dysphonia. Vocal cord injury was found on examination but no lymphadenopathy or metastases were present. Laryngeal microsurgery was performed with complete excision of the lesion. Histopathology showed it to be a carcinosarcoma which showed intense and diffuse positivity for vimentin and focal positivity for AE1-AE3, CK5 and p63. The patient underwent radiotherapy as complementary treatment. DISCUSSION: Sarcomatoid carcinoma usually presents with obstructive symptoms such as dysphonia. Prognosis depends on the stage and the presence or not of metastases. Both epithelial markers EMA, cytokeratin (AE1-AE3), epithelial membrane antigen, Ki 67 and mesenchymal markers such as vimentin, desmin, S-100 may be positive in these tumours. Recommended treatment for T2-T1 stages is an excisional biopsy which can be followed by adjuvant radiotherapy; radiotherapy alone has also been successful. T3-T4 stages can be treated with local excision, partial laryngectomy or total laryngectomy with subsequent ganglion emptying and concomitant radio and chemotherapy.

Factores que dificultan el diagnóstico del síndrome de taquicardia ortostática postural / Factors that hinder the diagnosis of postural orthostatic tachycardia syndrome

Las disautonomías son el resultado de un mal funcionamiento del sistema nervioso autónomo, entre las cuales se encuentra el síndrome de taquicardia ortostática postural, un problema de salud variable y complejo que tiene una prevalencia considerable, principalmente en adolescentes y mujeres. Este produce una amplia variedad de signos y síntomas que son similares a los de otras patologías, lo que, sumado a la falta de pruebas diagnósticas específicas, muchas veces retrasa el diagnóstico. A pesar de la existencia de criterios estandarizados para determinar la presencia del síndrome, existe aún una brecha en el conocimiento acerca del mismo. Esto dificulta el abordaje del padecimiento y, por consiguiente, su tratamiento oportuno. Conocer más sobre este síndrome y los factores que dificultan su diagnóstico temprano permitiría mejorar la atención de los pacientes y su calidad de vida

Dysautonomias are the result of a malfunction of the autonomic nervous system, among which is the postural orthostatic tachycardia syndrome, a variable and complex health problem that has a considerable prevalence, mainly in adolescents and women. It produces a wide variety of signs and symptoms that are similar to those of other pathologies, which, added to the lack of specific diagnostic tests, often delays diagnosis. Despite the existence of standardized criteria to determine the presence of the syndrome, there is still a gap in knowledge about it. This hinders the approach to the condition and, consequently, its timely treatment. Understanding more about this syndrome and the factors that hinder its early diagnosis would improve patient care and quality of life

FSH stimulation of anestrous cats improves oocyte quality and development of parthenogenetic embryos.

In the domestic cat, the efficiency of in vitro embryo production systems is negatively affected during the nonbreeding season. The objective of this research was to evaluate the effect of FSH stimulation in anestrous cats, on quality of cumulus-oocyte complexes (COCs) and in vitro developmental competence after parthenogenetic activation. To accomplish this purpose, anestrous cats were grouped into: (1) FSH treated (serial doses of 5 mg of porcine FSH each, every 24 hours, for 4 days) and (2) untreated control. The COCs were classified morphologically and a proportion of grade I and II COCs was used for expression analysis of FSHR, LHCGR, EGFR, PTGS2, EGR1, GDF9, and GATM by RT-qPCR. In addition, another proportion of grade I and II COCs was matured in vitro and used for parthenogenetic activation. After 8 days in culture, blastocyst and hatching blastocyst rates were assessed, and the expression of OCT4, SOX2, NANOG, CDX2, and GATA6 was evaluated. The COCs in the FSH group had an enhanced quality, a higher expression of LHCGR and a lower expression of GATM than did COCs from the control group (P < 0.05). Furthermore, embryos in the FSH group had increased blastocyst and hatching blastocyst rates, and those embryos had a higher expression of OCT4 and GATA than their counterparts from the control group (P < 0.05). In conclusion, ovarian stimulation of anestrous cats with FSH improved quality and increased the expression of LHCGR in COCs. The enhanced in vitro developmental competence, after parthenogenetic activation of oocytes from FSH-treated cats, coincided with an increased expression of OCT4 and GATA6 in blastocysts and hatching blastocysts.

Endometritis and In Vitro PGE2 Challenge Modify Properties of Cattle Endometrial Mesenchymal Stem Cells and Their Transcriptomic Profile.

Mesenchymal stem cells (MSCs) were isolated and characterized from postpartum bovine endometrium of animals with subclinical (n = 5) and clinical endometritis (n = 3) and healthy puerperal females (n = 5). Cells isolated displayed mean morphological features of MSCs and underwent osteogenic, chondrogenic, and adipogenic differentiation after induction (healthy and subclinical). Cells from cows with clinical endometritis did not undergo adipogenic differentiation. All cells expressed mRNAs for selected MSC markers. Endometrial MSCs were challenged in vitro with PGE2 at concentrations of 0, 1, 3, and 10 µM, and their global transcriptomic profile was studied. Overall, 1127 genes were differentially expressed between unchallenged cells and cells treated with PGE2 at all concentrations (763 up- and 364 downregulated, fold change > 2, and P < 0.05). The pathways affected the most by the PGE2 challenge were immune response, angiogenesis, and cell proliferation. In conclusion, we demonstrated that healthy puerperal bovine endometrium contains MSCs and that endometritis modifies and limits some functional characteristics of these cells, such as their ability to proceed to adipogenic differentiation. Also, PGE2, an inflammatory mediator of endometritis, modifies the transcriptomic profile of endometrial MSCs. A similar situation may occur during inflammation associated with endometritis, therefore affecting the main properties of endometrial MSCs.

Identification and characteristics of extracellular vesicles from bovine blastocysts produced in vitro.

Extracellular vesicles (EVs) have been identified within different body fluids and cell culture media. However, there is very little information on the secretion of these vesicles during early embryonic development. The aims of this work were first to demonstrate the secretion of extracellular vesicles by pre-implantation bovine embryos and second to identify and characterize the population of EVs secreted by bovine blastocysts during the period from day seven to nine of embryo culture and its correlation with further embryo development up to day 11. Bovine embryos were produced by in vitro fertilization (IVF) or parthenogenetic activation (PA) and cultured until blastocyst stage. Blastocyst selection was performed at day 7 post IVF/PA considering two variables: stage of development and quality of embryos. Selected blastocysts were cultured in vitro for 48 hours in groups (exp. 1) or individually (exp. 2) in SOF media depleted of exosomes. At day 9 post IVF/PA the media was collected and EVs isolated by ultracentrifugation. Transmission electron microscopy revealed the presence of heterogeneous vesicles of different sizes and population: microvesicles (MVs) and exosomes (EXs) of rounded shape, enclosed by a lipid bi-layer and ranging from 30 to 385 nm of diameter. Flow cytometry analysis allowed identifying CD63 and CD9 proteins as exosome markers. Nanoparticle tracking analysis generated a large number of variables, which required the use of multivariate statistics. The results indicated that the concentration of vesicles is higher in those blastocysts with arrested development from day 9 up to day 11 of in vitro development (6.7 x 108 particles/ml) derived from IVF (p <0.05), compared to PA blastocysts (4.7 x 108 particles/ml). Likewise, the profile (concentration and diameter) of particles secreted by embryos derived from IVF were different from those secreted by PA embryos. In conclusion, we demonstrated that bovine blastocysts secrete MVs/EXs to the culture media. Data suggest that characteristics of the population of EVs vary depending on embryo competence.

Carotid blowout syndrome in patients treated by larynx cancer.

INTRODUCTION: Carotid blowout syndrome is an uncommon complication for patient treated by head and neck tumors, and related to a high mortality rate. OBJECTIVE: The aim of this study was to study the risk of carotid blowout in a large cohort of patients treated only by larynx cancer. METHODS: Retrospective analysis of patients older than 18 years, treated by larynx cancer who developed a carotid blowout syndrome in a tertiary academic centre. RESULTS: 197 patients met the inclusion criteria, 192 (98.4%) were male and 5 (1.6%) were female. 6 (3%) patients developed a carotid blowout syndrome, 4 patients had a carotid blowout syndrome located in the internal carotid artery and 2 in the common carotid artery. According to the type of rupture, 3 patients suffer a type I, 2 patients a type III and 1 patient a type II. Five of those patients had previously undergone radiotherapy and all patients underwent total laryngectomy. We found a statistical correlation between open surgical procedures (p=0.004) and radiotherapy (p=0.023) and the development of a carotid blowout syndrome. CONCLUSION: Carotid blowout syndrome is an uncommon complication in patients treated by larynx tumours. According to our results, patient underwent radiotherapy and patients treated with open surgical procedures with pharyngeal opening have a major risk to develop this kind of complication.

Complications in CO2 Laser Transoral Microsurgery for Larynx Carcinomas.

Introduction Transoral laser microsurgery (TLM) has established itself as an effective option in the management of malignant tumors of the glottis, supraglottis, and hypopharynx. Nonetheless, TLM is not a harmless technique. Complications such as bleeding, dyspnea, or ignition of the air may appear in this type of surgery. Objective The aim of this study is to describe the complications that occurred in a group of patients treated for glottic and supraglottic carcinomas in all stages by TLM. Methods This study is a retrospective analysis of patients diagnosed with squamous cell carcinoma of the glottis and supraglottis for all stages (T1, T2, T3, T4), N -/ + , M -/+ treated with TLM between January 2009 and March 2012 in a tertiary hospital. Results Ninety-eight patients met the inclusion criteria, which had undergone a total of 131 interventions. Ninety-four (95.9%) patients were male and 4 (4.1%) were female. The mean age was 64.2 years (± 10.7 years = min 45; max 88). The presence of intraoperative complications was low, affecting only 2% of patients. Immediate postoperative complications occurred in 6.1%, whereas delayed complications affected 13.2% of patients, without any of them being fatal. Conclusion TLM has shown good oncologic results and low complication rate compared with traditional open surgery during intervention, in the immediate and delayed postoperative period and in the long-term with respect to radiotherapy.

Transoral CO2 Laser Microsurgery Outcomes for Early Glottic Carcinomas T1-T2.

INTRODUCTION: Transoral laser microsurgery (TLM) has won territory in larynx oncology, establishing itself as an effective option in treatment of glottic, supraglottic, and hypopharynx tumors. Its advantages include limited resections, a reduction in number of tracheostomies, and the use of nasogastric tubes. Moreover, its oncological benefits are similar to those from open surgery in patients with early or advanced stages, when correctly selected. OBJECTIVE: The objective of this study is to review oncologic outcomes obtained with the treatment of a series of glottic tumors, treated by TLM. METHODS: Retrospective analysis of patients previously untreated, diagnosed with squamous cell carcinoma of the glottis (T1a, T1b, T2) in a tertiary university hospital. Endpoints for analysis were local control, overall and disease-specific survival, and larynx preservation rate. RESULTS: The study group included 58 patients that met the inclusion criteria: 57 (98.3%) men and 1 (1.7%) woman. Mean age was 65.5 ± 10.7 years (Min: 46/Max: 88). The tumor stages of the patients included were 30 T1a, 11 (19%) T1b, and 17 (29.3%) T2. Three-year overall survival rate was 89.7% (Fig. 1), and three-year disease-specific survival rate was 96.5%, three-year local control rate was 98.3%, and three-year organ preservation rate was 98.3%. CONCLUSION: TLM is a safe and effective option in the treatment of glottis carcinomas, associated with less morbidity and a high percentage of local control, overall survival, specific survival, and organ preservation.

The risk of venous thromboembolism in ENT and head & neck surgery.

OBJECTIVES: Venous thromboembolic disease (VTD) includes deep vein thrombosis (DVT) and pulmonary embolism (PE), thus is one of the most feared postoperative complications developed by patients at any surgical department, because of high morbidity and mortality associated with it. MATERIALS AND METHODS: We performed a retrospective study including all patients operated on at the Otolaryngology Head and Neck Department (tertiary hospital) between January 2009 and December 2013. RESULTS: A total of 9007 surgical procedures were performed, including 7150 elective surgeries under general anesthesia, with 2127 on children and 5023 on adults. A total of 1989 patients had oncological head and neck surgery, eight cases had VTE complications, which represents 0.08% of patients. All of those complicated cases had head and neck cancer (8/1989 = 0.4%) and belonged to the group of scheduled surgeries under general anesthesia (8/7150 = 0.1%). CONCLUSION: The incidence of DVT and PE in ENT and head and neck surgery appears to be lower than in other surgical specialties. Oncological surgery of the head and neck, usually associated with other risk factors, appears to increase the risk of VTD.

Compensatory hypertrophy of the contralateral inferior turbinate in patients with unilateral nasal septal deviation. A computed tomography study.

INTRODUCTION: The compensatory hypertrophy of the inferior turbinate in patients with septal deviation to one of the nostrils is considered to protect the airways from the excess of air that could enter through the nostril and its potential negative effects such as dryness, alteration of air filtration, mucociliary flow, or lung involvement. MATERIALS AND METHODS: A prospective, longitudinal, non-randomized study. Patients were divided in two groups: 10 consecutive patients, with nasal septal deviation and compensatory hypertrophy of the inferior turbinate in the contralateral nasal cavity (10 non-hypertrophied turbinates as control and 10 contralateral hypertrophied turbinates as study cases), and the second group with 5 patients without any nasal pathology (10 turbinates without any obvious pathology). In both groups CT scans of the nasal region were performed. A comparison of patients with nasal septal deviation with compensatory hypertrophy of the inferior turbinate in the contralateral nasal cavity and with non-pathological inferior turbinate was carried out. RESULTS: When analyzing the groups of patients with septal deviation, the contralateral hypertrophied turbinate and the non-hypertrophied turbinate side, we found a significant hypertrophy in the anterior portion of the inferior turbinate, at the level of the medial mucosa (P = 0.002) and bone (P = 0.001) in the group of patients with contralateral hypertrophied turbinate. However, when we compared the contralateral hypertrophic turbinate with the turbinate of patients without septal deviation, we found a significant difference in all volumes of the medial and lateral mucosa and the bone portion (P = 0.001, P = 0.005). CONCLUSION: Surgical correction of the nasal septum and lateralization or reduction of the volume of the inferior turbinate (which may include the medial mucosa, head or part of the bone) is necessary in order to improve air passage into the nasal valve.

Constitutive expression of the embryonic stem cell marker OCT4 in bovine somatic donor cells influences blastocysts rate and quality after nucleus transfer.

Nuclear transfer (NT) is associated with epigenetic reprogramming of donor cells. Expression of certain genes in these cells might facilitate their expression in the NT embryo. This research was aimed to investigate the effect of constitutive expression of OCT4 in bovine somatic cells used for NT on the developmental potential of derived cloned embryos as well as in the expression of pluripotency markers in the Day-7 resulting embryos. Cloned blastocysts were generated from five cell lines that expressed OCT4. Pools of blastocysts were screened to detect OCT4, SOX2, and NANOG by qPCR. In vitro-fertilized time-matched blastocysts were used as controls. The development potential was assessed on the basis of blastocysts rate; grading and total cell counts at Day 7. OCT4 expression in the cell lines positively correlates with blastocysts rate (r = 0.92; p = 0.02), number of grade I blastocysts (r = 0.96; p = 0.01), and total cell number (r = 0.98; p = 0.002). The high expression of OCT4 in the cell line did not improve the final outcome of cloning. Somatic expression of OCT4 lead to increased expression of OCT4 and SOX2 in cloned grade I blastocysts; however, there was a bigger variability in OCT4 and SOX2 (p = 0.03; p = 0.02) expression in the embryos generated from cells expressing highest levels of OCT4. Probably the higher variability in OCT4 expression in cloned embryos is due to incorrect reprogramming and incapability of the oocyte to correct for higher OCT4 levels. For that reason, we concluded that OCT4 expression in somatic cells is not a good prognosis marker for selecting cell lines.

Carotid blowout syndrome in patients treated by larynx cancer / Síndrome da ruptura da artéria carótida em pacientes tratados para câncer de laringe

Abstract Introduction: Carotid blowout syndrome is an uncommon complication for patient treated by head and neck tumors, and related to a high mortality rate. Objective: The aim of this study was to study the risk of carotid blowout in a large cohort of patients treated only by larynx cancer. Methods: Retrospective analysis of patients older than 18 years, treated by larynx cancer who developed a carotid blowout syndrome in a tertiary academic centre. Results: 197 patients met the inclusion criteria, 192 (98.4%) were male and 5 (1.6%) were female. 6 (3%) patients developed a carotid blowout syndrome, 4 patients had a carotid blowout syndrome located in the internal carotid artery and 2 in the common carotid artery. According to the type of rupture, 3 patients suffer a type I, 2 patients a type III and 1 patient a type II. Five of those patients had previously undergone radiotherapy and all patients underwent total laryngectomy. We found a statistical correlation between open surgical procedures (p = 0.004) and radiotherapy (p = 0.023) and the development of a carotid blowout syndrome. Conclusion: Carotid blowout syndrome is an uncommon complication in patients treated by larynx tumours. According to our results, patient underwent radiotherapy and patients treated with open surgical procedures with pharyngeal opening have a major risk to develop this kind of complication.

Resumo Introdução: A síndrome da ruptura da carótida é uma complicação incomum no paciente em tratamento para tumores de cabeça e pescoço, relacionada com uma alta taxa de mortalidade. Objetivo: O objetivo deste estudo foi estudar o risco de ruptura da carótida em uma grande coorte de pacientes tratados isoladamente por um câncer de laringe. Método: Análise retrospectiva de pacientes com mais de 18 anos, tratados por câncer de laringe em um centro de assistência terciária, que desenvolveram a síndrome da ruptura da carótida. Resultados: Ao todo, 197 pacientes atenderam aos critérios de inclusão, 192 (98,4%) eram do sexo masculino e 5 (1,6%) eram do sexo feminino. 6 (3%) desenvolveram síndrome da ruptura da carótida, 4 tiveram síndrome da ruptura da carótida localizada na artéria carótida interna e 2 na artéria carótida comum. De acordo com o tipo de ruptura, 3 pacientes apresentaram síndrome da ruptura da carótida tipo I, 2 pacientes, síndrome da ruptura da carótida Tipo III e um tipo II. Cinco desses pacientes haviam sido previamente tratados com radioterapia e todos os pacientes foram submetidos a laringectomia total. Encontrou-se uma correlação estatística entre procedimentos cirúrgicos abertos (p = 0,004) e radioterapia (p = 0,023) e o desenvolvimento de síndrome da ruptura da carótida. Conclusão: A síndrome de ruptura da carótida é uma complicação rara em pacientes tratados para tumores de laringe. De acordo com nossos resultados, pacientes submetidos a radioterapia e pacientes tratados com procedimentos cirúrgicos abertos com abertura da faringe apresentam um risco maior de desenvolver essa complicação.