Effects of glucose variability on hematopoietic stem/progenitor cells in patients with type 1 diabetes.

BACKGROUND AND PURPOSE: Diabetes reduces the levels of hematopoietic stem/progenitor cells (HSPCs), which can contribute to organ and tissue homeostasis. Among patients with diabetes, lower HSPC levels predict the development or worsening of micro- and macro-angiopathy. High glucose variability is also associated with diabetic complications and we have previously shown that acute hypoglycaemia can stimulate stem/progenitor cells. Thus, we evaluated the relationship between glucose variability or time in hypoglycaemia and HSPCs in patients with type 1 diabetes (T1D). METHODS: Patients with T1D were compared to healthy subjects. HSPCs (CD34+, CD133+, CD34+CD133+, CD34 + CD45dim) were quantified by flow cytometry. Using flash glucose monitoring system for 90 days, we calculated several measures of glucose variability and time in hypoglycaemia. RESULTS: Forty-four patients with T1D and 44 healthy subjects were enrolled. Compared to healthy controls, T1D patients had significantly lower levels of HSPCs and duration of diabetes was inversely correlated with HSPC levels. Significant direct correlations were found between HSPC levels and the coefficient of variation of glucose levels or time in hypoglycaemia, which were stronger in patients with short-term than in those with long-standing diabetes. CONCLUSION: This study confirms the pauperization of HSPCs in T1D patients and demonstrates a potential HSPC-stimulatory effect of hypoglycaemia, which mitigates with long-lasting diabetes. These data are consistent with a model whereby disease chronicity progressively blunts the release of HSPCs in response to adrenergic triggers, like hypoglycaemic events.

Variable rescue of microtubule and physiological phenotypes in mdx muscle expressing different miniaturized dystrophins.

Delivery of miniaturized dystrophin genes via adeno-associated viral vectors is one leading approach in development to treat Duchenne muscular dystrophy. Here we directly compared the functionality of five mini- and micro-dystrophins via skeletal muscle-specific transgenic expression in dystrophin-deficient mdx mice. We evaluated their ability to rescue defects in the microtubule network, passive stiffness and contractility of skeletal muscle. Transgenic mdx mice expressing the short dystrophin isoform Dp116 served as a negative control. All mini- and micro-dystrophins restored elevated detyrosinated α-tubulin and microtubule density of mdx muscle to values not different from C57BL/10, however, only mini-dystrophins restored the transverse component of the microtubule lattice back to C57BL/10. Passive stiffness values in mdx muscles expressing mini- or micro-dystrophins were not different from C57BL/10. While all mini- and micro-dystrophins conferred significant protection from eccentric contraction-induced force loss in vivo and ex vivo compared to mdx, removal of repeats two and three resulted in less protection from force drop caused by eccentric contraction ex vivo. Our data reveal subtle yet significant differences in the relative functionalities for different therapeutic constructs of miniaturized dystrophin in terms of protection from ex vivo eccentric contraction-induced force loss and restoration of an organized microtubule lattice.

Mouse models of two missense mutations in actin-binding domain 1 of dystrophin associated with Duchenne or Becker muscular dystrophy.

Missense mutations in the dystrophin protein can cause Duchenne muscular dystrophy (DMD) or Becker muscular dystrophy (BMD) through an undefined pathomechanism. In vitro studies suggest that missense mutations in the N-terminal actin-binding domain (ABD1) cause protein instability, and cultured myoblast studies reveal decreased expression levels that can be restored to wild-type with proteasome inhibitors. To further elucidate the pathophysiology of missense dystrophin in vivo, we generated two transgenic mdx mouse lines expressing L54R or L172H mutant dystrophin, which correspond to missense mutations identified in human patients with DMD or BMD, respectively. Our biochemical, histologic and physiologic analysis of the L54R and L172H mice show decreased levels of dystrophin which are proportional to the phenotypic severity. Proteasome inhibitors were ineffective in both the L54R and L172H mice, yet mice homozygous for the L172H transgene were able to express even higher levels of dystrophin which caused further improvements in muscle histology and physiology. Given that missense dystrophin is likely being degraded by the proteasome but whole body proteasome inhibition was not possible, we screened for ubiquitin-conjugating enzymes involved in targeting dystrophin to the proteasome. A myoblast cell line expressing L54R mutant dystrophin was screened with an siRNA library targeting E1, E2 and E3 ligases which identified Amn1, FBXO33, Zfand5 and Trim75. Our study establishes new mouse models of dystrophinopathy and identifies candidate E3 ligases that may specifically regulate dystrophin protein turnover in vivo.

Independent variability of microtubule perturbations associated with dystrophinopathy.

Absence of the protein dystrophin causes Duchenne muscular dystrophy. Dystrophin directly binds to microtubules in vitro, and its absence in vivo correlates with disorganization of the subsarcolemmal microtubule lattice, increased detyrosination of α-tubulin, and altered redox signaling. We previously demonstrated that the dystrophin homologue utrophin neither binds microtubules in vitro nor rescues microtubule lattice organization when overexpressed in muscles of dystrophin-deficient mdx mice. Here, we fine-mapped the dystrophin domain necessary for microtubule binding to spectrin-like repeats 20­22. We show that transgenic mdx mice expressing a full-length dystrophin/utrophin chimera completely lacking microtubule binding activity are surprisingly rescued for all measured dystrophic phenotypes, including full restoration of microtubule lattice organization. Conversely, despite the presence of dystrophin at the sarcolemma, ß-sarcoglycan-deficient skeletal muscle presents with a disorganized and densified microtubule lattice. Finally, we show that the levels of α-tubulin detyrosination remain significantly elevated to that of mdx levels in transgenic mdx mice expressing nearly full-length dystrophin. Our results demonstrate that the microtubule-associated perturbations of mdx muscle are distinct, separable, and can vary independently from other parameters previously ascribed to dystrophin deficiency.

Water vapor thermal therapy: Technical variations among spanish hospitals and efficacy at 2-year follow-up.

AIM: The aim of this work is to evaluate the initial experience with water vapor thermal therapy (WVTT) for benign prostatic hyperplasia (BPH) in Spanish university hospitals, as well as to describe the differences in technique and follow-up between centers. MATERIALS AND METHODS: This retrospective observational multicenter study collected baseline characteristics, surgical, postoperative and follow-up data at 1, 3, 6, 12 and 24 months, including validated questionnaires, flowmetric variations, complications, and the need for pharmacological or surgical treatment following the procedure. Possible triggers for postoperative acute urinary retention (AUR) were also analyzed. RESULTS: A total of 105 patients were included. No differences were observed between the groups with and without AUR with respect to catheterization time (5 and 4.3 days respectively, P=.178), or prostate volume (47.9g and 41.4g respectively, P=.147). The mean improvement at 3, 6, 12 and 24 months in terms of peak flow was 5.3, 5.2, 4.2 and 3.8ml/s, respectively. As for ejaculation, an improvement was observed after 3 months of follow-up and was maintained over time. CONCLUSIONS: Minimally invasive treatment for BPH with WVTT shows good functional outcomes at 24 months follow-up, without significant impairment of sexual function and a low incidence of complications. There are minor inter-hospital variations, mainly in the immediate postoperative period.

[Trend of the asbestos related diseases mortality in workers of a company in the province Cremona which manufactured asbestos products]. / Andamento delle patologie e della mortalità asbesto correlate in lavoratori di un'azienda della provincia di Cremona che produceva manufatti in amianto.

The study analyzes the trend of asbestos-related diseases and mortality in workers of a company in the province of Cremona which manufactured asbestos products. It is confirmed that the exposure to a high concentration of asbestos fibers (estimated to more than 20 fibers/cc) strictly correlates with the onset of pathologies from asbestos. In the studied population were found 19 cases of neoplastic diseases (12 mesotheliomas and 7 bronchopulmonary carcinomas). This figure, compared to the company working population, which over the years has been an average of 80 units, while not enabling to calculate an incidence rate due to the lack of reliable data on population, is indicative of a very significant cause-effect relationship since these are neoplastic diseases that can still arise. So it is necessary to continue the health monitoring of formerly exposed workers and appropriate to try to extend it to all workers of the asbestos compartment.

An Advanced Surgical Dressing for High-risk Patients Undergoing Breast Cancer Surgery: a Case-control Study.

Oncological breast surgeries, classified as breast conserving surgery, oncoplastic surgery, and mastectomies (standard or with tissue sparing and reconstruction), are burdened with an overall complication rate up to 33%. Aquacel Ag Surgical is a combined hydrofiber-hydrocolloids dressing. The aim of this study is to evaluate the incidence of surgical site complications in patients presenting with three or more risk factors (or two, of which at least one classified as "high risk"), undergoing breast cancer surgery with/without reconstruction, comparing advanced (Aquacel Ag Surgical) with traditional dressing. METHODS: This is a retrospective, monocentric, case-control study based at the breast unit of the Città della Salute e della Scienza Hospital of Turin, Italy. Forty-two patients who underwent breast surgeries and met the inclusion criteria were enrolled, from February 1 to July 31, 2018. The primary endpoint was comparing the incidence of surgical site complications (skin alterations, infection, and wound dehiscence) in the two groups. The secondary endpoints were evaluating patient's quality of life, aesthetic outcomes, and compliance to the dressings. RESULTS: The distribution of risk factors at the baseline between the two groups was balanced, without statistically significant differences. Wound complications' incidence at 1 week was lower in the advanced dressing group (P = 0.015). On the bivariate descriptive analysis, advanced dressing proved to be easier to remove for the operator (P = 0.026). The aesthetic outcomes vouched for better scores in the advanced dressing group. CONCLUSION: In the presented study Aquacel Ag Surgical dressing reduces surgical site complications in the first week after surgery in patients affected by three or more risk factors (or two with at least one classified as "high risk").

Rapid, redox-mediated mechanical susceptibility of the cortical microtubule lattice in skeletal muscle.

The highly ordered cortical microtubule lattice of skeletal muscle is disorganized in dystrophin-deficient mdx mice. Implicated mechanisms include loss of dystrophin binding, altered α-tubulin posttranslational modification, expression of a ß-tubulin involved in regeneration, and reactive oxygen species (ROS). Here we show that the transverse microtubules in mdx muscle expressing miniaturized dystrophins are rapidly lost after eccentric contraction. Analysis of mdx lines expressing different dystrophin constructs demonstrate that spectrin-like repeats R4-15 and R20-23 were required for mechanically stable microtubules. Microtubule loss was prevented by the non-specific antioxidant N-acetylcysteine while inhibition of NADPH oxidase 2 had only a partial effect, suggesting that ROS from multiple sources mediate the rapid loss of transverse microtubules after eccentric contraction. Finally, ablation of α-dystrobrevin, ß- or γ-cytoplasmic actin phenocopied the transverse microtubule instability of miniaturized dystrophins. Our data demonstrate that multiple dystrophin domains, α-dystrobrevin and cytoplasmic actins are necessary for mechanically stable microtubules.

Office stent placement under local anesthesia is a safe and efficient procedure for the management of multiple ureteral disorders. / La colocación de catéteres doble J con anestesia local en régimen ambulatorio es un procedimiento seguro y eficiente para el manejo de distintas patologías ureterales.

OBJECTIVE: To assess the outcomes of ureteral stent placement under local anesthesia for the management of multiple ureteral disorders. METHODS: Retrospective study of 45 consecutive ureteral stents placed under local anesthesia from January 2015 to July 2016. Inclusion criteria were hemodynamically stable patients with urinary obstruction, urinary fistula or for prophylactic ureteral localization during surgery. Five minutes before the procedure, 10ml of lidocaine gel and 50ml of lidocaine solution were instilled in the bladder. A 4.8Fr ureteral stent was placed using a 15.5Fr flexible cystoscope under fluoroscopic control. Characteristics of procedures and outcomes were analysed. RESULTS: A total of 45 procedures (33 placement, 12 replacements) were attempted in 37 patients, of which 40 (89%) were successful. There were 10 male (27%) and 27 female patients (73%) with a mean age of 58.6 years (±17.5). Main indications for stent placement were stones (37.8%), extrinsic ureteral compression (28.9%) and surgery ureteral localization (22.2%). The reasons for failing to complete a procedure were the inability to pass the guidewire/stent in 4 cases (8.8%) or to identify the ureteral orifice in 1 (2.2%). Postoperative complications occurred in 8 patients (17.8%) (7 Clavien I, 1 Clavien IIIa). No procedure was prematurely terminated due to pain. Statistical analysis did not find significant successful predictors. The outpatient setting provided a fourfold cost decrease. CONCLUSIONS: Ureteral stent placement can be safely and effectively performed under local anesthesia in the office cystoscopy room. This procedure could free operating room time, reduce costs and minimize side effects of general anesthesia.

Impaired muscle relaxation and mitochondrial fission associated with genetic ablation of cytoplasmic actin isoforms.

While α-actin isoforms predominate in adult striated muscle, skeletal muscle-specific knockouts (KOs) of nonmuscle cytoplasmic ßcyto - or γcyto -actin each cause a mild, but progressive myopathy effected by an unknown mechanism. Using transmission electron microscopy, we identified morphological abnormalities in both the mitochondria and the sarcoplasmic reticulum (SR) in aged muscle-specific ßcyto - and γcyto -actin KO mice. We found ßcyto - and γcyto -actin proteins to be enriched in isolated mitochondrial-associated membrane preparations, which represent the interface between mitochondria and sarco-endoplasmic reticulum important in signaling and mitochondrial dynamics. We also measured significantly elongated and interconnected mitochondrial morphologies associated with a significant decrease in mitochondrial fission events in primary mouse embryonic fibroblasts lacking ßcyto - and/or γcyto -actin. Interestingly, mitochondrial respiration in muscle was not measurably affected as oxygen consumption was similar in skeletal muscle fibers from 12 month-old muscle-specific ßcyto - and γcyto -actin KO mice. Instead, we found that the maximal rate of relaxation after isometric contraction was significantly slowed in muscles of 12-month-old ßcyto - and γcyto -actin muscle-specific KO mice. Our data suggest that impaired Ca2+ re-uptake may presage development of the observed SR morphological changes in aged mice while providing a potential pathological mechanism for the observed myopathy.

Uterine sarcomas. A retrospective study of 17 cases.

Seventeen documented cases of uterine sarcoma were studied in an effort to establish the relationship between prognostic factors and patients' survival. The analysis of prognostic factors showed that there was a significant difference in survival between patients with stages I and II tumors and those with more advanced lesions. It has been also noted that prognosis was worse in patients with history of previous abortions and in patients who had abdominal or pelvic pain.

Terapia termal con vapor de agua: variaciones técnicas entre hospitales universitarios españoles y eficacia a 2 años de seguimiento / Water vapor thermal therapy: technical variations among spanish hospitals and efficacy at 2-year follow-up

Objetivo El objetivo de este trabajo es conocer experiencia inicial con la terapia térmica con vapor de agua (TTVA) para hiperplasia benigna de próstata (HBP) en los hospitales universitarios españoles, así como describir las diferencias en cuanto a técnica y seguimiento observadas entre los centros. Materiales y métodos Este estudio multicéntrico observacional retrospectivo recogió características basales, datos quirúrgicos, posoperatorios y seguimiento a los uno, tres, seis, 12 y 24 meses, incluyendo cuestionarios validados, variaciones flujométricas, complicaciones y la necesidad de tratamiento farmacológico y quirúrgico tras el procedimiento. También se analizaron los posibles desencadenantes de retención aguda de orina (RAO) en el posoperatorio. Resultados Se incluyeron un total de 105 pacientes. No se observaron diferencias entre los grupos con y sin RAO con respecto a tiempo de sondaje (cinco y 4,3 días respectivamente, p = 0,178), ni volumen prostático (47,9 y 41,4 g, respectivamente, p = 0,147). La mejoría media a los tres, seis, 12 y 24 meses en cuanto de flujo máximo fue de 5,3, 5,2, 4,2 y 3,8 mL/s, respectivamente. En cuanto a la eyaculación, se observa una mejoría en la misma a partir de los tres meses de seguimiento que se mantiene en el tiempo. Conclusiones El tratamiento mínimamente invasivo de HBP con TTVA presenta buenos resultados funcionales a 24 meses de seguimiento, sin afectación significativa de la función sexual y con una baja incidencia de complicaciones. Existen pequeñas variaciones principalmente en cuanto al posoperatorio inmediato entre los hospitales participantes en el estudio. (AU)

Aim The aim of this work is to evaluate the initial experience with water vapor thermal therapy (WVTT) for benign prostatic hyperplasia (BPH) in Spanish university hospitals, as well as to describe the differences in technique and follow-up between centers. Materials and Methods This retrospective observational multicenter study collected baseline characteristics, surgical, postoperative and follow-up data at 1, 3, 6, 12 and 24 months, including validated questionnaires, flowmetric variations, complications, and the need for pharmacological or surgical treatment following the procedure. Possible triggers for postoperative acute urinary retention (AUR) were also analyzed. Results A total of 105 patients were included. No differences were observed between the groups with and without AUR with respect to catheterization time (5 and 4.3 days respectively, p = 0.178), or prostate volume (47.9 gr and 41.4 gr respectively, p = 0.147). The mean improvement at 3, 6, 12 and 24 months in terms of peak flow was 5.3, 5.2, 4.2 and 3.8 ml/s, respectively. As for ejaculation, an improvement was observed after 3 months of follow-up and was maintained over time. Conclusions Minimally invasive treatment for BPH with WVTT shows good functional outcomes at 24 months follow-up, without significant impairment of sexual function and a low incidence of complications. There are minor inter-hospital variations, mainly in the immediate postoperative period. (AU)

La colocación de catéteres doble J con anestesia local en régimen ambulatorio es un procedimiento seguro y eficiente para el manejo de distintas patologías ureterales / Office stent placement under local anesthesia is a safe and efficient procedure for the management of multiple ureteral disorders

Objetivo: Evaluar los resultados de la colocación de catéteres doble J con anestesia local para el manejo de distintas patologías ureterales. Métodos: Estudio retrospectivo de 45 cateterismos ureterales consecutivos con anestesia local desde enero de 2015 hasta julio de 2016. Se incluyeron pacientes hemodinámicamente estables con una obstrucción o fístula urinaria o para la identificación ureteral durante una cirugía abdominopélvica. Cinco minutos antes del procedimiento se instilaron 10 ml de gel-lidocaína y 50 ml de suero-lidocaína en la vejiga. Se colocaron catéteres 4,8 Fr mediante un cistoscopio flexible de 15,5 Fr y escopia. Se analizaron las características y resultados de los procedimientos. Resultados: Se realizaron 45 procedimientos (33 colocaciones, 12 recambios) en 37 pacientes, de los cuales 40 (89%) fueron exitosos. Con una edad media de 58,6 años (±17,5), se intervinieron 10 hombres (27%) y 27 mujeres (73%). Las principales indicaciones fueron litiasis (37,8%), compresiones extrínsecas del uréter (28,9%) y la localización intraoperatoria ureteral (22,2%). Los intentos infructuosos fueron debidos a la incapacidad para ascender la guía/catéter en 4 casos (8,8%) o para identificar el meato ureteral en uno (2,2%). Ocho pacientes (17,8%) presentaron alguna complicación postoperatoria (7 Clavien I, uno Clavien IIIa). Ningún procedimiento se interrumpió por dolor. El análisis estadístico no encontró ningún factor predictor de éxito. El régimen ambulatorio fue 4 veces más barato. Conclusiones: La colocación de catéteres ureterales se puede realizar de forma eficaz y segura bajo anestesia local en el gabinete de cistoscopias. Este procedimiento podría ahorrar tiempo operatorio, reducir costes y minimizar los efectos secundarios de la anestesia general

Objective: To assess the outcomes of ureteral stent placement under local anesthesia for the management of multiple ureteral disorders. Methods: Retrospective study of 45 consecutive ureteral stents placed under local anesthesia from January 2015 to July 2016. Inclusion criteria were hemodynamically stable patients with urinary obstruction, urinary fistula or for prophylactic ureteral localization during surgery. Five minutes before the procedure, 10 ml of lidocaine gel and 50 ml of lidocaine solution were instilled in the bladder. A 4.8 Fr ureteral stent was placed using a 15.5 Fr flexible cystoscope under fluoroscopic control. Characteristics of procedures and outcomes were analysed. Results: A total of 45 procedures (33 placement, 12 replacements) were attempted in 37 patients, of which 40 (89%) were successful. There were 10 male (27%) and 27 female patients (73%) with a mean age of 58.6 years (±17.5). Main indications for stent placement were stones (37.8%), extrinsic ureteral compression (28.9%) and surgery ureteral localization (22.2%). The reasons for failing to complete a procedure were the inability to pass the guidewire/stent in 4 cases (8.8%) or to identify the ureteral orifice in 1 (2.2%). Postoperative complications occurred in 8 patients (17.8%) (7 Clavien I, 1 Clavien IIIa). No procedure was prematurely terminated due to pain. Statistical analysis did not find significant successful predictors. The outpatient setting provided a fourfold cost decrease. Conclusions: Ureteral stent placement can be safely and effectively performed under local anesthesia in the office cystoscopy room. This procedure could free operating room time, reduce costs and minimize side effects of general anesthesia

An extremely potent anilinoacridine inhibitor of aldehyde oxidase.

An acridine antitumor agent, 4'-(9-acridinylamino)methanesulfon-m-anisidide, has been found to be an extremely potent competitive inhibitor of aldehyde oxidase (EC 1.2.3.1). The inhibitor constant (Ki) was determined to be 0.06 microM. The degree of enzyme inhibition was quite sensitive to small changes in the structure of the inhibitor's anisidide moiety. Drug inhibition was specific for aldehyde oxidase and inhibition was not detected with the other mammalian molybdenum iron-sulfur flavoenzyme, xanthine oxidase (EC 1.2.3.2). Members of the 4'-(9-acridinylamine)methanesulfonanilide series might be useful probes in the study of the structure and function of aldehyde oxidase.

Hysteroscopic treatment of uterine septum.

Authors report their 4 years experience in the treatment of uterine septum by hysteroscopic metroplasty. 35 patients underwent procedure; no complications occurred. Postoperative reproductive outcome was evaluated in 29 women with follow-up longer than 6 months. Patients were divided into 2 groups according to presence or absence of associated factors compromising fertility. Group A composed of 19 women in which uterine septum was the only cause of pregnancy wastage. Preoperatively they had 40 pregnancies all ending in spontaneous abortion. Postoperatively 15 (79%) patients conceived and 13 (68%) had a live baby. Totally they had 18 pregnancies, 2 (12%) ended in abortion, 1 in molar pregnancy, 1 in premature delivery, 11 delivered at term and 3 are currently beyond 20 weeks pregnant, for a live birth rate of 78%. Life table analysis showed an estimated pregnancy rate of 82% at 12 months, monthly fecundability was 0.13. Group B composed of 10 women in which other factors compromising fertility were present. Preoperatively only 5 experienced pregnancy. Totally they had 8 pregnancies 7 (86%) of which ended in abortions and one in extrauterine pregnancy. Postoperatively only 3 (30%) had pregnancy and all had a live baby. Totally they had 4 pregnancies, 1 ended in abortion and 3 at term for a live birth rate of 75%. Life table analysis showed an estimated pregnancy rate of 11% at 12 months, monthly fecundability rate was 0.01. Hysteroscopic metroplasty proved to be safe and effective for solving pregnancy wastage caused by uterine septum. If other factors compromising fertility were present metroplasty did not increase fecundability, but improved live birth rate.(ABSTRACT TRUNCATED AT 250 WORDS)

[Peritoneal cystic mesothelioma]. / Mesotelioma cistico del peritoneo.

Cystic mesothelioma of peritoneum is a rare neoplasia that was identified as a individual entity by ultrastructural and immunoistochemical studies. It is more frequent in young woman and often it tends to recur. Because of its rarity and difficulty in differential diagnosis, the Authors describe a case of cystic mesothelioma observed.