Clinical and molecular characterization of Chilean patients with X-linked hypophosphatemia.

We report the most comprehensive clinical and molecular characterization of XLH patients performed in Chile. We show high prevalence of musculoskeletal burden and pain, associated with significantly impaired physical capacity and quality of life, with many relevant complications presenting more frequently than previously reported in cohorts from developed countries. INTRODUCTION: Our current understanding of the clinical presentation and natural history of X-linked hypophosphatemia (XLH) comes mainly from cohorts from developed countries, with limited data on the clinical and genetic abnormalities of XLH patients in South America. OBJECTIVE: To describe the clinical, biochemical, and molecular presentation of patients with XLH in Chile. METHODS: Patients with XLH referred by endocrinologist throughout Chile were included. Demographic data and clinical presentation were obtained from a clinical interview. Surveys were applied for quality of life (QoL), pain, and functionality. FGF23 was measured by ELISA, and genetic testing was performed. Imaging studies were conducted to assess skeletal and renal involvement. RESULTS: We included 26 patients, aged 2-64 years, from 17 unrelated Chilean families. All pediatric patients but only 40% of adults were receiving conventional therapy, while 65% of all patients had elevated alkaline phosphatase. All patients had mutations in PHEX, including 5 novel variants. Radiographic skeletal events (RSE) and enthesopathies in adults were frequent (34% and 85%, respectively). The duration of treatment was associated with fewer RSE (p < 0.05). Most adults reported pain and impaired QoL, and 50% had impaired physical capacity. The number of enthesopathies was associated with worse pain and stiffness scores (p < 0.05). CONCLUSION: Chilean patients with XLH have a high prevalence of musculoskeletal burden associated with pain and impaired physical capacity and QoL, especially in adults who were generally undertreated. These data identify a significant unmet need, inform our understanding of the current status of patients, and can guide care for XLH patients in similarly socioeconomically defined countries.

[Clarifying concepts on euthanasia]. / Eutanasia: aclarando conceptos.

Euthanasia is a subject of permanent discussion everywhere, mainly about its ethical appropriateness and about its legalization in different countries. This debate includes many ethical, legal, social, political, spiritual, religious, and public health issues. A relevant problem is the frequent misunderstanding that many have about the meaning of the terms and concepts used in different ethical and clinical end of life settings. Many views and even texts are affected by such mistakes, rendering a constructive discussion almost impossible. The goal of this article is to contribute to an improvement of the social and legislative discussion about euthanasia through an elucidation of the main terms and notions linked to this topic. We present definitions and explanations for euthanasia, assisted suicide, withdrawal or foregoing of therapy, treatment rejection, palliative sedation, dignified death and others.

Novel Way to Search for Light Dark Matter in Lepton Beam-Dump Experiments.

A novel mechanism to produce and detect light dark matter in experiments making use of GeV electrons (and positrons) impinging on a thick target (beam dump) is proposed. The positron-rich environment produced by the electromagnetic shower allows us to produce an A^{'} via nonresonant (e^{+}+e^{-}→γ+A^{'}) and resonant (e^{+}+e^{-}→A^{'}) annihilation on atomic electrons. The latter mechanism, for some selected kinematics, results in a larger sensitivity with respect to limits derived by the commonly used A^{'}-strahlung. This idea, applied to beam-dump experiments and active beam-dump experiments, pushes down the current limits by an order of magnitude.

High sodium intake is associated with increased glucocorticoid production, insulin resistance and metabolic syndrome.

OBJECTIVE: High sodium (HS) diet is associated with hypertension (HT) and insulin resistance (IR). We evaluated whether HS diet was associated with a dysregulation of cortisol production and metabolic syndrome (MetS). PATIENTS AND MEASUREMENTS: We recruited 370 adults (18-85 years, BMI 29·3 ± 4·4 kg/m(2) , 70% women, 72% HT, 61% MetS). HS diet (urinary sodium >150 mEq/day) was observed in 70% of subjects. We measured plasma hormones, lipid profile, urinary free cortisol (UFC) and cortisol tetrahydrometabolites (THM). RESULTS: Urinary sodium was correlated with UFC (r = +0·45, P < 0·001), cortisol THM (r = +0·41, P < 0·001) and inversely with adiponectin, HDL and aldosterone, after adjusting by age, gender and BMI. Subjects with high, compared with adequate sodium intake (50-149 mEq/day) had higher UFC (P < 0·001), THM (P < 0·001), HOMA-IR (P = 0·04), HT (81% vs 50%, P < 0·001), MetS (69% vs 41%, P < 0·001) and lower adiponectin (P = 0·003). A multivariate predictive model adjusted by confounders showed a high discriminative capacity for MetS (ROC curve 0·878) using four clinical variables: HS intake [OR = 5·6 (CI 2·3-15·3)], HOMA-IR [OR 1·7 (1·3-2·2)] cortisol THM [OR 1·2 (1·1-1·4)] and adiponectin [OR = 0·9 (0·8-0·9)], the latter had a protective effect. CONCLUSIONS: High sodium diet was associated with increased urinary cortisol and its metabolites. Also, HS diet was associated with HT, insulin resistance, dyslipidaemia and hypoadiponectinaemia, even when adjusting by confounding variables. Further, we observed that high salt intake, IR and higher cortisol metabolites, alone or combined in a clinical simple model, accurately predicted MetS status, suggesting an additive mechanism in obesity-related metabolic disorders.

The estimation of visceral adipose tissue with a body composition monitor predicts the metabolic syndrome.

BACKGROUND: Central obesity has a higher risk of the metabolic syndrome (MetS) and cardiovascular diseases. It is estimated by measuring waist circumference (WC) and waist-to-hip ratio (WHR), which are operator-dependent. The present study aimed to validate a body composition monitor (BCM) as a tool for estimating visceral adipose tissue (VAT), as well as to assess its capacity to predict the MetS and its correlation with anthropometric parameters. METHODS: We measured WC, WHR and body mass index (BMI) in 60 recruited subjects. BCM estimated VAT (1-30 points). Body composition and resting energy expenditure (REE) were compared with bioelectrical impedance analysis (BIA) and indirect calorimetry, respectively. VAT was estimated by BCM (range 1-30 points), We evaluated the capability of VAT, WC, BMI and WHR to predict the MetS by ATP-III criteria. RESULTS: The mean (SD) age of subjects was 36.8 (12.9) years, 80% were female, and 47% had the MetS. Body composition and REE estimated by BCM had a significant correlation with BIA (r = 0.85-0.91, P< 0.001) and REE (r = 0.86, P < 0.001), respectively, even after adjusting by sex. VAT estimation by BCM was positively correlated with WC (r = 0.75, P< 0.001) and WHR (r = 0.61, P < 0.001). The area under the receiver operator characteristic curves to predict the MetS was 0.93 for VAT, 0.81 for WC, 0.76 for WHR and 0.74 for BMI. VAT ≥10 points had a sensitivity of 100% and a specificity of 82% for predicting the MetS. CONCLUSIONS: VAT estimation by BCM efficiently predicts the MetS and correlates with anthropometric parameters of central obesity. Its routine use could facilitate cardiovascular risk estimation and follow-up in overweight and obese patients in ambulatory practice.

Plasma levels of potassium and magnessium after modified ultrafiltration in pediatric cardiac surgery with cardiopulmonary bypass.

OBJECTIVE: Modified ultrafiltration (MUF) reduces some of the complications associated with cardiopulmonary bypass (CPB) in pediatric cardiac surgery. However, we have observed hypokalemia and hypomagnesemia in children when MUF is used. Such alterations may elicit severe arrhythmias in the postoperative period. To date, no studies have focused on the effects MUF may have in plasma levels of potassium (K) and magnesium (Mg). The objective of our study was to determine if there is any variation in plasma levels of K (plK) and Mg (plMg) after MUF in children undergoing cardiac surgery with CPB. PATIENTS: Sixteen children scheduled for cardiac surgery with CBP and MUF were prospectively studied. Anesthetic, CPB and MUF management were standardized for all patients, the latter lasting for 10 minutes. RESULTS: Plasma K average levels before and after MUF were 4.16 mmol/L and 3.58 mmol/L, respectively. The average plasma Mg levels before and after MUF were 4.82 mmol/L and 4.81 mmol/L, respectively. CONCLUSIONS: The plasma level of K is reduced by 13.7% after MUF (p<0.0001). The reduction in Mg at the same period of time was not statistically significant (p<0.970).

[Eugenic abortion could explain the lower infant mortality in Cuba compared to that in Chile]. / El aborto eugenésico podría explicar la menor mortalidad infantil existente en Cuba comparada con la de Chile.

BACKGROUND: Cuba and Chile have the lower infant mortality rates of Latin America. Infant mortality rate in Cuba is similar to that of developed countries. Chilean infant mortality rate is slightly higher than that of Cuba. AIM: To investigate if the lower infant mortality rate in Cuba, compared to Chile, could be explained by eugenic abortion, considering that abortion is legal in Cuba but not in Chile. MATERIAL AND METHODS: We compared total and congenital abnormalities related infant mortality in Cuba and Chile during 2008, based on vital statistics of both countries. RESULTS: In 2008, infant mortality rates in Chile were significantly higher than those of Cuba (7.8 vs. 4.7 per 1,000 live born respectively, odds ratio (OR) 1.67; 95% confidence intervals (Cl) 1.52-1.83). Congenital abnormalities accounted for 33.8 and 19.2% of infant deaths in Chile and Cuba, respectively. Discarding infant deaths related to congenital abnormalities, infant mortality rate continued to be higher in Chile than in Cuba (5.19 vs. 3.82 per 1000 live born respectively, OR 1.36; 95%CI 1.221.52). CONCLUSIONS: Considering that antenatal diagnosis is widely available in both countries, but abortion is legal in Cuba but not in Chile, we conclude that eugenic abortion may partially explain the lower infant mortality rate observed in Cuba compared to that observed in Chile.

[The change in the epidemiological profile of maternal mortality in Chile will hinder the fulfillment of the Millennium 5th goal]. / El cambio del perfil epidemiológico de la mortalidad materna en Chile dificultará el cumplimiento del 5° objetivo del Milenio.

BACKGROUND: The death of women during pregnancy, childbirth or puerperium, remains a serious public health problem worldwide. Chile is committed to comply with the Millennium 5th Goal of reducing maternal mortality to 9.9/100,000 live births in 2015. AIM: To analyze trends in maternal mortality in Chile during 2000-2009. MATERIAL AND METHODS: A descriptive population analysis using raw data obtained from the yearbooks of the National Institute of Statistics of Chile. Maternal mortality, causes of death and age of the dead mothers were evaluated. The causes of maternal death were classified according to the tenth revision of International Classification of Diseases. Trend studies were performed using Pearson correlation analysis. RESULTS: In the studied period there were no significant changes in maternal mortality and fertility. The five major causes of maternal death were concurrent diseases, hypertension, abortion, obstetric embolism and postpartum hemorrhage. Mortality associated with concurrent illness showed a significant upward trend (r = 0.656, p = 0.035). Abortion associated mortality had a significant downward trend (r = -0.712, p = 0.023). The group of women 40 years and older significantly increased its birth rate (r = 0.930, p < 0.001), this group showed the highest maternal mortality, especially in association with concurrent diseases. CONCLUSIONS: The increased birth rate occurring in women over 40 years old and its larger maternal mortality rate, probably will hinder the fulfillment of the Millennium 5th goal in Chile.

The effect of the chemical composition on the sensory characterization of Ecuadorian coffee.

The study aims to test if Ecuadorean coffee's symbolic and material contents agree with the instrumental analysis and grading protocols. We studied the relationship between the chemical composition and the organoleptic characteristics of eight non-specialty and six specialty coffee samples. Firstly, the study addresses the grading following the Specialty Coffee American Association (SCAA) method. The second stage focuses on the qualitative composition of the coffee brews employing GC-MS and caffeine concentrations using HPLC. Then, we employed statistical tools such as Cohen's concordance coefficients, dissimilarity dendrograms, and linear correlations between the chemical compounds in the beverage and the attributes' scores. The grading panel consisted of 6 semi-trained-testers who would assess if the primary cultural capital can provide a criterion to identify specialty coffee. The variety of compounds allowed the evaluators to distinguish between commercial and specialty coffees. However, the composition analysis identified molecules that would imply greater gradation in the tasting, a prevision that was not reflected in the results. Finally, we confirmed that basic training could create cultural capital to distinguish non-specialties from specialty coffees through their chemistry and organoleptic attributes.

A de novo unequal cross-over mutation between CYP11B1 and CYP11B2 genes causes familial hyperaldosteronism type I.

UNLABELLED: Familial hyperaldosteronism type I (FH-I) is an autosomal dominant disorder caused by an unequal cross-over of the gene encoding steroid 11ß-hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2), giving rise to a chimeric CYP11B1/CYP11B2 gene that displays aldosterone synthase activity regulated by ACTH instead of angiotensin II. AIM: To report an unprecedented case of a de novo unequal crossover mutation between CYP11B1 and CYP11B2 genes causing FH-I. PATIENTS AND METHODS: The index case is a 45-yr-old Chilean male diagnosed with primary aldosteronism (PA). All family members were also studied: his biological parents, 1 brother, 6 sisters, 2 daughters, and 1 son. Plasma renin activity, serum aldosterone, and its ratio were measured in all patients. Genetic analyses were performed using long-extension PCR (XL-PCR), DNA sequencing and Southern blot methods. RESULTS: PA was diagnosed for the index case, 1 of his daughters, his son but not for his parents or siblings. XLPCR and Southern blotting demonstrated the presence of the chimeric CYP11B1/CYP11B2 gene solely in PA-affected subjects, suggesting a case of a de novo mutation. Sequence analysis showed the unequal cross-over CYP11B1/CYP11B2 at intron 2 (c.2600-273 CYP11B2). We also identified a polymorphism at the same intron (c.2600-145C>A CYP11B2) in the genome of the index case's father. CONCLUSION: We describe an unprecedented case of unequal cross-over mutation for the chimeric CYP11B1/CYP11B2 gene causing FH-I, which may be linked to a polymorphism in the index case's father germ line.

Harnessing of geothermal energy for a greenhouse in Ecuador employing a heat pump: design, construction, and feasibility assessment.

Globally, the greenhouses' farming area comprises 500 000 ha, and they efficiently produce more than half of the vegetables consumed around the world. Nevertheless, high-yield crops tend to be incredibly energy-intensive. This study proposes designing and building a coupled geothermal heat pump for a 470 m2 greenhouse in the Andean zone conditions addressing a requirement of 15 °C at night and 30 °C during the day. Firstly, the study determined the energy potential of the solar and geothermal sources employing actual measurements and contrasting the results with theoretical models. Then, it developed an energy balance in the greenhouse to size the geothermal heat pump using the vapor compression cycle. Finally, the comprehensive system was built and evaluated through the Leveled Cost of Heat (LCOH). The operation requires a potential of 29.56 and 65.76 kW for heating and cooling; this is technically feasible when running the system with a heating flow driven by an optimized temperature ramp of 1.64 °C h-1. Also, the capacity factor (CF) shows that a lifespan between 12 to 14 years is required to reach acceptable LCOH when CF is as low as 0.45. Financially, it is necessary to foster customs exemptions to make it competitive versus more traditional sources such as electricity and LPG since the main components of the heat pump and the geothermal exchanger are not produced locally and represent nearly 70 % of the upfront costs.

Landmark versus ultrasound-guided insertion of femoral venous catheters in the pediatric intensive care unit: An efficacy and safety comparison study.

BACKGROUND: Central venous cannulation (CVC) is common and necessary in pediatric intensive care. However, this procedure is not without risks or complications. Although CVCs have classically been placed following anatomical landmarks, the use of ultrasound guidance has largely replaced the latter, given its better profile of efficacy and safety, demonstrated at least in adult populations. OBJECTIVES: To compare the effectiveness and safety in the insertion of femoral central venous catheters guided by ultrasound (US) versus the anatomical method (LM) in critical care pediatric patients. METHODS: 100 patients were randomized: 50 were assigned to the US group and 49 to the LM group. In the LM group the traditional method consisted in palpating the femoral artery pulse as a; in the US group the CVC was inserted using a real time technique. Success at the first attempt, overall success in cannulation, number of attempts and arterial puncture were the variables studied in both groups. RESULTS: Success at the first attempt and overall success in cannulation were significantly higher in the US group versus the LM (US 42% vs. LM 18%, p 0.011, US 84% vs. LM 51% p <0.001, respectively). The incidence of puncture of the femoral artery was lower in the US group (LM 12 vs. US 5, p 0.056) without achieving statistical significance. CONCLUSIONS: According to our results, the placement of central venous access via the femoral approach should be preferably performed under ultrasound guidance, however, further studies in larger populations are needed to confirm this findings.

Energy Return on Investment (EROI) and Life Cycle Analysis (LCA) of biofuels in Ecuador.

In Ecuador, the net energy contribution of biofuels is unknown or unnoticed. To address this issue, we determined the Energy Return on Investment (EROI) for bioethanol and biodiesel. The selection of raw materials relied on their productive capacity, export and import records, and historical yields. Consequently, the scope included three raw materials for ethanol (sugar cane, corn, and forest residues) and four for biodiesel (African palm, pinion, bovine fat, and swine fat). Using a method based on the Life Cycle Analysis (LCA) of each biofuel, we assessed the entire production chain through statistical processing of primary and secondary information. Then we calculated the calorific values in the laboratory, compared energy inputs/outputs, and finally obtained the energetic returns. EROIs for bioethanol were: 1.797 for sugarcane, 1.040 for corn, and 0.739 for wood. The results for biodiesel were: 3.052 for African palm, 2.743 for pinion, 2.187 for bovine fat, and 2.891 for swine fat. These values suggest feasibility only for sugarcane in the case of ethanol. In contrast, biodiesel has better prospects because all the feedstocks analyzed had EROIs higher than two. Nevertheless, biodiesel is not available for trading in Ecuador because energy policy has overlooked systems based on higher energy return. Future studies should consider more comprehensive variables such as climate change, land use, and water management.

Primary aldosteronism can alter peripheral levels of transforming growth factor beta and tumor necrosis factor alpha.

UNLABELLED: Primary aldosteronism (PA) is the most common secondary cause of hypertension that has recently been implicated in alterations of the immune system and progression of cardiovascular disease. OBJECTIVE: To study the cytokines transforming growth factor beta1 (TGF-beta1), tumor necrosis factor alpha (TNF-alpha), and interleukin 10 (IL-10) in patients with PA and essential hypertensives (EH) and evaluate its association with the renin-angiotensin-aldosterone system. PATIENTS AND METHODS: We studied 26 PA and 52 EH patients as controls, adjusted by their blood pressure, body mass index, age, and gender. In both groups, PA and EH, we measured serum aldosterone (SA), plasma renin activity (PRA), and cytokines TGF- beta1, TNF-alpha, and IL-10. In addition, 17 PA patients were treated for 6 months with spironolactone, a mineralocorticoid receptor (MR) antagonist. RESULTS: PA patients had lower levels of TGF-beta1 (17.6+/-4.1 vs 34.5+/-20.5 pg/ml, p<0.001) and TNF-alpha (17.0+/-4.4 vs 35.6+/-21.7 pg/ml, p<0.001) and similar IL-10 levels (99.7+/-18.7 vs 89.4+/-49.5 pg/ml, p: ns), as compared with EH controls. TGF-beta1 and TNF-alpha levels showed a remarkable correlation with SA/PRA ratio in the total group (PA+EH). The treatment of PA patients with spironolactone increased the TGF-beta1 levels (18.3+/-5.9 to 28.4+/-6.3 pg/ml, p<0.001), while TNF-alpha, and IL-10 remained unchanged. CONCLUSION: Our results showed that PA patients have lower TGF-beta1 and TNF-alpha cytokine serum levels than EH. TGF-beta1 levels were restored with spironolactone, showing a MR-dependent regulation. In this way, the chronic aldosterone excess modifies the TGF-beta1 levels, which could produce an imbalance in the immune system homeostasis that may promote an early proinflammatory cardiovascular phenotype.

The impact of joint bleeding and synovitis on physical ability and joint function in a murine model of haemophilic synovitis.

Haemophilia is a congenital disorder that commonly results in musculoskeletal bleeding and orthopaedic complications. After an acute joint haemorrhage, an increase in intra-articular pressure and inflammation cause pain, swelling and limited motion. Blood in the joint space provokes a proliferative disorder known as haemophilic synovitis. Overgrowth of the synovial membrane causes mechanical dysfunction. Eventually, there is destruction of the articular surface and underlying bone. The aim of this project was to test the hypothesis that a minimum number of haemarthroses negatively impacts on joint function and that this would be reflected by decreased physical performance of experimental animals. Mice deficient in factor VIII coagulant activity were trained to ambulate on a rotating rod then injured three times at weekly intervals. Their ability to walk was then compared to a group of uninjured mice. Cohorts of mice were killed after 1, 2 or 3 months and the knee joints examined by gross and histological methods. The results supported the following conclusions: (i) haemophilic mice can be trained to ambulate on a rotating rod; (ii) acute hemarthrosis temporarily impairs their ability to ambulate and (iii) following recovery from acute injury, mice developing synovitis demonstrated inferior physical ability compared to mice not developing synovitis. This is the first description of a quantitative assay to monitor joint function in experimental animals and should be useful to evaluate the efficacy of new therapies developed to prevent and treat bleeding and to test strategies to counter the devastating effects of synovitis.

Manejo de embarazos únicos con diagnóstico antenatal de vasa previa / Management of singleton pregnancies with prenatal diagnosis of vasa previa

Introducción: vasa previa (VP) corresponde al paso de los vasos umbilicales por las membranas amnióticas, sin protección de gelatina de Wharton o placenta, antes de la presentación fetal, sobre el orificio cervical interno. Pese a su baja incidencia, el diagnóstico prenatal es relevante por las graves consecuencias que puede tener esta patología en caso de no ser diagnosticada. El objetivo de esta revisión es presentar la evidencia disponible para el manejo de embarazadas con diagnóstico antenatal de VP. Materiales y métodos: analizamos todos los estudios publicados (prospectivos, retrospectivos y reporte de casos) entre los años 1999 y 2023, con diagnóstico VP en embarazo único, reportando la edad gestacional de interrupción y el resultado neonatal. Resultados: incluimos 19 investigaciones (18 en la búsqueda primera y una adicional por relevancia). Las pacientes con manejo intrahospitalario desde las 34 semanas tuvieron mayor latencia al parto, mejores resultados neonatales y menor tasa de cesárea de urgencia que las pacientes con manejo ambulatorio. La edad gestacional de interrupción es variable entre los estudios, sin embargo, no se evidenció beneficio de interrupción a las 34 semanas comparado con manejo expectante hasta las 37 semanas de edad gestacional. Conclusión: existiría beneficio de hospitalización entre las 32-34 semanas en mujeres con diagnóstico de VP, siendo razonable la interrupción cercana a las 37 semanas por cesárea electiva.

Introduction: vasa previa (VP) corresponds to the passage of the umbilical vessels through the amniotic membranes, without the protection of Wharton's gelatin or placenta, in front of the fetal presentation, over the internal cervical os. Despite its low incidence, prenatal diagnosis is relevant due to the severe consequences of this pathology if the diagnosis is missed. This review presents the available evidence for pregnant women's management with an antenatal diagnosis of VP. Materials and methods: we analyzed all the studies published (prospective, retrospective, and case reports) between 1999 and 2023, with a diagnosis of VP in a single pregnancy, reporting gestational age at delivery and neonatal outcome. Results: We included 19 investigations (18 in the first search and another for relevance). Patients with in-hospital management from 34 weeks had a more extended latency period until delivery, better neonatal outcomes, and a lower rate of emergency cesarean section than patients with outpatient management. The gestational age at birth is variable between the studies; however, no benefit of delivery at 34 weeks was evidenced compared with expectant management until 37 weeks of gestational age. Conclusion: there would be a benefit of hospitalization between 32-34 weeks in women diagnosed with VP, being reasonable to schedule the delivery close to 37 weeks by elective cesarean section.

Positional uncertainty of vaginal cuff and feasibility of implementing portable bladder scanner in postoperative cervical cancer patients.

PURPOSE: To propose a geometrical margin for definition of the vaginal cuff PTV using only CT images of the full bladder (CTfull) in postoperative cervical cancer patients. METHODS: Twenty-nine operated cervical cancer patients underwent volumetric arc therapy with a bladder filling protocol. This study assessed bladder filling using a portable bladder scanner and cone-beam computed tomography (CBCT) during the entire treatment period. The measured bladder volumes with a BladderScan® were compared with the delineated volume on CBCT. Titanium clips in the vaginal cuff were analysed to assess geometrical uncertainty and the influence of rectal and bladder volume changes. RESULTS: BladderScan® showed good agreement with the delineated volume (R = 0.80). The volume changes in the bladder have a greater influence on the clip displacements than in the rectum. The 95th percentile of uncertainty of the clips in reference to CTfull in the right-left (RL), the superoinferior (SI), and the anteroposterior (AP) was 0.32, 0.65, and 1.15 cm, respectively. From this result and intra-fractional movements of the vaginal cuff reported by Haripotepornkul, a new geometrical margin was proposed for definition of the vaginal cuff planning target volume (PTV): 0.5, 0.9, and 1.4 cm in the RL, SI, and AP directions, respectively. CONCLUSIONS: A new geometrical margin was proposed for definition of the vaginal cuff PTV based on CTfull, which will be needless of empty bladder at the planning CT scan. This method allows patients to reduce the burden and efficient routine CT scans can be improved.

New splicing mutation of MEN1 gene affecting the translocation of menin to the nucleus.

UNLABELLED: Multiple endocrine neoplasia type 1 (MEN1) is a syndrome inherited in an autosomal dominant trait caused by the inactivation of the tumor suppressor gene MEN1. OBJECTIVE: To communicate a family with a new heterozygous germ line mutation in the intronic region of MEN1 gene and to study its influence in the menin expression. PATIENTS AND METHODS: We studied 5 members of a family with symptomatic hyperparathyroidism (HPT). One of them had also a neuroendocrine pancreatic tumor, and 2 had non-functional multinodular cortical adrenal hyperplasia compatible with the diagnosis of MEN1. After the mutation was identified, HSP92II restriction enzyme was used to determine both zygosity and segregation of the mutation. RT-PCR from leukocyte's isolated mRNA and western blot from pancreatic tumor tissue were performed. In vitro studies were done in Chinese hamster ovary (CHO) cells transfected with reporter minigenes carrying the coding regions spanning exon (EX)-intron 9 and EX10 with the mutant and the wild type sequences. RESULTS: We identified a heterozygous G-to-T substitution in the intron-EX junction (IVS9-1 G>T) of MEN1 gene in the index case and the family members. The mRNA from patient's leukocytes was larger (934 bp) in comparison to the normal transcript (717 bp). Immunoblot analysis demonstrated that wild type (67 kDa) and two additional mutant proteins (approximately 55 and approximately 90 kDa) were expressed in the pancreatic tissue. The in vitro study showed a 45% nuclear localization of the mutated menin signal and a 95% in the wild type protein. CONCLUSIONS: We identified a new intronic heterozygous germ line mutation (IVS9-1G>T) of MEN1 gene in a family affected by MEN1 syndrome. This mutation alters the splice acceptor site of intron 9 that promotes an incorrect splicing, generating aberrant proteins without the nuclear localization signals necessary for the normal menin translocation to the nucleus.

M. tuberculosis DNA detection in nasopharyngeal mucosa can precede tuberculosis development in contacts.

BACKGROUND: The nasopharynx is a known gateway for some mycobacterial species such as Mycobacterium bovis and M. leprae. M. tuberculosis can cross lymphoepithelial barriers in vitro, but its ability to colonise the nasopharyngeal mucosa in vivo has not been established. OBJECTIVE: To determine if M. tuberculosis can be transiently detected in nasopharyngeal mucosa of tuberculosis (TB) contacts as a preliminary step in the development of tuberculous infection. DESIGN: Exploratory study conducted among asymptomatic household contacts of pulmonary TB cases. A chest X-ray, QuantiFERON(®) TB-Gold or tuberculin skin test and a bilateral nasopharyngeal swab for Xpert(®) MTB/RIF and mycobacterial culture were performed at baseline and repeated 8-12 weeks later. RESULTS: Eighty-nine contacts were enrolled a median of 9 days after the diagnosis of the index case. At baseline, 29.9% were positive for latent tuberculous infection and one subject (1.1%) had a positive Xpert in the nasopharyngeal swab with a normal chest X-ray, negative QuantiFERON and negative induced sputum. After 12 weeks' follow-up, this subject developed a new cough and upper lobe infiltrates and M. tuberculosis grew in sputum. No other cases of active TB were detected at follow-up. CONCLUSION: The detection of M. tuberculosis DNA in the nasopharyngeal mucosa of contacts is an infrequent event that in this instance preceded the development of pulmonary TB. Its pathogenic role requires further investigation.