The Large Uterus Classification System: a prospective observational study.

OBJECTIVE: To investigate the Large Uterus Classification System (LUCS) ability to predict surgical outcomes and complications in total laparoscopic hysterectomies (TLHs) for large uteri. DESIGN: Prospective observational study. SETTING: Two referral centres. POPULATION OR SAMPLE: Three hundred and ninety-two women who underwent TLH for a large uterus (uterine fundus at or over the transverse umbilical line). METHODS: Between 2004 and 2019, the intraoperative LUCS was estimated in all patients. The LUCS considers the uterine and adnexal vascular pedicles displacement. Type 1 is without vascular pedicles displacement. Type 2 has the cephalad displacement of adnexal vascular pedicles. The uterine vessels displacement regardless of adnexal pedicles defines Type 3. MAIN OUTCOME MEASURES: Patients' characteristics with perioperative outcomes were prospectively collected and compared between the three types of large uteri. RESULTS: Two hundred and fifty-one (64%), 82 (20.9%) and 59 (15.1%) women had Type 1, Type 2 and Type 3 uteri, respectively. Women with Type 1 uteri had a lower uterine weight, shorter operative time, less blood loss and lower complication rates than women with Types 2 and 3. The conversion rate to laparotomy in Type 1 was similar to that in Type 2 (odds ratio [OR] 0.98; 95% CI 0.32-3.56) but lower than Type 3 (OR 0.35; 95% CI 0.14-0.97); in Type 2 it was lower than Type 3, although without the conventional statistical significance (OR 0.36; 95% CI 0.13-1.13; P = 0.07). Multivariable analysis showed that the uterine Type (1 versus 2-3) was independently associated with the total complications rate (OR 2.00; 95% CI 1.09-3.68; P = 0.02). CONCLUSIONS: The LUCS appears associated with surgical outcomes and complications, potentially stratifying the surgical risk and guiding the surgical technique in TLHs for large uteri. TWEETABLE ABSTRACT: The Large Uterus Classification System may predict outcomes in total laparoscopic hysterectomy of large uteri.

Evaluation Through Transcutaneous Tissue Oximetry (Near Infrared Spectroscopy) of Spinal Cord Stimulation in Patients With Complex Regional Pain Syndrome Type I.

OBJECTIVE: One of the physiopathological hypothesis behind complex regional pain syndrome (CRPS) type I involves the deep-tissue hypoxia of the affected areas. Spinal cord stimulation (SCS) appears to be effective in the treatment of these patients. We evaluated whether ESCS modifies tissue oxygen saturation (StO2 ) measured with near-infrared spectroscopy (NIRS) in the affected limbs in patients diagnosed with CRPS type I. MATERIALS AND METHODS: Nonrandomized, cross-sectional study that evaluated 16 patients with CRPS type I who were receiving SCS applied to the posterior cords. NIRS was used to evaluate baseline StO2 (primary outcome) and variations in StO2 (secondary outcome) during an ischemia-reperfusion test performed using a vascular occlusion test, comparing the hands of limbs unilaterally affected by CRPS type I with the unaffected contralateral hands. We also determined whether the variations in StO2 were related to a modification in the percentage of subjective pain improvement and in the visual analog scale score. RESULTS: The baseline StO2 of the affected hands was significantly higher than that of the unaffected hands (mean 4.7%; 95% confidence interval: 1.41, 6.7; p = 0.005). Variations in StO2 during the ischemia-reperfusion test revealed no differences between affected and unaffected hands. No significant correlations were detected between baseline StO2 values or variations in StO2 during the vascular occlusion test and the pain measurements. CONCLUSIONS: Baseline StO2 evaluated by NIRS was greater in the affected hands of patients with CRPS type I treated with SCS than in the unaffected, contralateral hands.

Almost 2% of Spanish breast cancer families are associated to germline pathogenic mutations in the ATM gene.

PURPOSE: There is still a considerable percentage of hereditary breast and ovarian cancer (HBOC) cases not explained by BRCA1 and BRCA2 genes. In this report, next-generation sequencing (NGS) techniques were applied to identify novel variants and/or genes involved in HBOC susceptibility. METHODS: Using whole exome sequencing, we identified a novel germline mutation in the moderate-risk gene ATM (c.5441delT; p.Leu1814Trpfs*14) in a family negative for mutations in BRCA1/2 (BRCAX). A case-control association study was performed to establish its prevalence in Spanish population, in a series of 1477 BRCAX families and 589 controls further screened, and NGS panels were used for ATM mutational screening in a cohort of 392 HBOC Spanish BRCAX families and 350 patients affected with diseases not related to breast cancer. RESULTS: Although the interrogated mutation was not prevalent in case-control association study, a comprehensive mutational analysis of the ATM gene revealed 1.78% prevalence of mutations in the ATM gene in HBOC and 1.94% in breast cancer-only BRCAX families in Spanish population, where data about ATM mutations were very limited. CONCLUSION: ATM mutation prevalence in Spanish population highlights the importance of considering ATM pathogenic variants linked to breast cancer susceptibility.

Persistent hepatitis C virus (HCV) infection impairs HCV-specific cytotoxic T cell reactivity through Mcl-1/Bim imbalance due to CD127 down-regulation.

In persistent hepatitis C virus (HCV) infection, HCV-specific cytotoxic T lymphocyte (CTL) reactivity is impaired and this affects HCV control. Interleukin-7 receptor (CD127) expression on these cells could regulate CTL reactivity through Mcl-1/Bim balance modulation. Bim is a pro-apoptotic molecule blocked by the action of Mcl-1. Mcl-1/Bim expression and T cell reactivity on HCV-specific CTLs were compared according to CD127 phenotype. Peripheral blood lymphocytes (PBL) from HLA-A2(+) HCV(+) patients were obtained. HCV-specific CTLs were visualized by staining PBL with anti-CD8 and HLA-A2/peptide pentameric complexes (pentamer). Mcl-1/Bim/CD127 phenotype of HCV-specific CTLs was tested by staining detectable CD8(+)/pentamer(+) cells with anti-Mcl-1/Bim/CD127 antibodies. HCV-specific CTL proliferation ability after specific in vitro challenge was tested in the presence and absence of pancaspase inhibitor z-VAD-fmk. All stained cells were analysed by flow cytometry. CD127(low)-expressing HCV-specific CTLs associated with high HCV viraemia, while CD127(high) correlated with undetectable viral loads (P < 0.001). Directly ex vivo, pentamer(+) cell frequency was similar according to CD127 expression level. Nevertheless, CD127(low) pentamer(+) cell proliferation after specific in vitro challenge was impaired (P < 0.05), although this was corrected by z-VAD-fmk treatment (P < 0.05). Mcl-1 expression was low directly ex vivo (P < 0.01), and Bim was up-regulated after antigen encounter (P < 0.05) of CD127(low) pentamer(+) cells. The ex vivo difference between Mcl-1 and Bim expression on pentamer(+) cells correlated positively with CD127 expression level (P < 0.001) and with pentamer(+) cell reactivity (P < 0.05). In summary, a low ex vivo Mcl-1 expression and Bim up-regulation after antigen encounter are involved in CD127(low) HCV-specific CTL hyporeactivity during chronic infection, but it can be overcome by apoptosis blockade.

Cold-induced sweating syndrome type 1, with a CRLF1 level mutation, previously associated with Crisponi syndrome.

INTRODUCTION: Cold-induced sweating syndrome type 1 (CISS1) is a rare autosomal recessive genodermatosis caused by mutations in the CRLF1 gene, characterized by profuse sweating when the ambient temperature is below 22°C and morphological alterations. CRLF1 mutations also cause Crisponi syndrome (CS), which presents neonatal muscle contractions, morphological disorders and alterations in the autonomous nervous system. CASE REPORT: A 30-year-old man sought treatment for profuse sweating. His medical record included neonatal admission for generalized hypertonicity. Clinical examination revealed morphological alterations. A genetic study was requested, detecting a c.713dupC mutation in homozygosity in the CRLF1 gene. CONCLUSIONS: We report the case of a male with clinical and genetic diagnosis of CISS1 who in childhood presented clinical characteristics of CS. The mutation detected in CRLF1 has not been described in patients with CISS1, but in one with CS. These data seem to support the theory that CS and CISS1 are variants of the same disorder.

Gestational diabetes mellitus: Prevention, diagnosis and treatment. A fresh look to a busy corner.

BACKGROUND: Gestational diabetes mellitus (GDM) is a common pregnancy complication characterized by hyperglycaemia with onset or first recognition during pregnancy. Risk factors include family history of diabetes, previous GDM, genetic predisposition for GDM/type 2 diabetes, insulin resistance conditions such as overweight, obesity and ethnicity. Women with GDM are at high risk for fetal macrosomia, small for gestational age, neonatal hypoglycaemia, operative delivery and caesarean delivery. The aim of this narrative review is to summarize the most recent findings of diagnosis and treatment of GDM in order to underline the importance to promote adequate prevention of this disease, especially through lifestyle interventions such as diet and physical activity. METHODS: The research was conducted using the following electronic databases, MEDLINE, EMBASE, Web of Science, Scopus, ClinicalTrial.gov, OVID and Cochrane Library, including all published randomized and non-randomized studies as well as narrative and systematic reviews. RESULTS: The lack of universally accepted criteria makes the definition of diagnosis and prognosis of this condition difficult. Early diagnosis and glucose blood level control may improve maternal and fetal short and long-term outcomes. Treatment strategies include nutritional interventions and exercise. Medical treatment can be necessary if these strategies are not effective. Moreover, novel non-pharmacologic agents such as myo-inositol seem to be effective and safe both in the prevention and the treatment of GDM. CONCLUSIONS: It is important to promote adequate prevention of GDM. Further studies are needed in order to better define the most appropriate strategies for the clinical management of women affected by GDM.

Management of diabetic ketoacidosis in a teaching hospital.

The aim of our study was to evaluate clinical management of diabetic ketoacidosis (DKA) in a teaching hospital. We followed all the patients hospitalised for DKA over six years (1995-2000), and we recorded clinical data, laboratory finding at entrance and clinical management. We compared the data to the standards set in guidelines. Our study showed an important delay of initiation of intravenous fluid (70% of cases), an under-replacement with intravenous fluid (69% of cases) and with potassium therapy (80% of cases), and an excessive use of alkali therapy. In conclusion, suboptimal management of DKA occurred in a large percentage of patients.

Mutational analysis of the HIV-1 Vpu protein.

The human immunodeficiency virus type-1 (HIV-1) encoded Vpu protein facilitates the release of budding virions from the surface of infected cells and delays the rate of syncytium formation of the virus. Furthermore, Vpu induces rapid degradation of nascent CD4 molecules that are retained in the endoplasmic reticulum by the formation of gp160-CD4 complexes. Currently, little is known of the precise mechanism(s) by which Vpu function. Whether or not these different events are related remain unclear. In this report, we describe our recent structure/function studies on vpu suggesting that the protein may have independent biological activities during the HIV-1 infection.

Histochemical and immunochemical study of the fate of Candida albicans inside human neutrophil phagolysosomes.

To further define the ultrastructural events associated with the killing of Candida albicans by human neutrophils, four methods were used: (1) the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of vicinal-glycol-containing complex carbohydrates; (2) the localization of thermostable immunodeterminants of the yeast cell wall, mannans or mannoproteins, using monospecific antibodies and a protein A-gold complex (monAb-gold); (3) the localization of mannose residues with concanavalin A labeled with gold particles (Con A-gold); (4) the localization of chitin oligomers using wheat germ agglutinin and ovomucoid labeled with gold particles (WGA-gold). The mannan-rich cell wall layers were progressively lost as shown by altered PA-TCH-SP reactivity and a diffuse pattern of staining with Con A-gold and monAb-gold. The de novo appearance of conspicuous amounts of glycogen-like particles near the plasmalemma and in the cell wall was interpreted as evidence of a reparative process of the yeast cell wall. Chitin was seemingly unaltered and readily demonstrated by the WGA-gold in the wall remnants of ghost cells.

Pegylated interferon and ribavirin for the recurrence of chronic hepatitis C genotype 1 in transplant patients.

The efficacy of pegylated interferon (p-IFN) and ribavirin (RB) in transplant patients is not well known. Chronic hepatitis C evolves in a more aggressive form after transplantation, causing a worse survival. Twenty-one naïve patients with recurrent chronic hepatitis C demonstrated by biopsy were treated for 48 weeks with p-IFN alpha2b (1.5 microg/kg/wk) and RB (>10.6 mg/kg/d). Quantification of RNA was performed (Amplicor Cobas 2.0 Roche) at baseline, 4, 12, 24, 48, and 72 weeks. A qualitative technique was used when quantitative levels were undetectable. At more than 1 year since liver transplantation we did not detect coinfection with human immunodeficiency virus or use steroid treatment. Among the cohort there were 16 men (76.2%). The mean overall age was 52 +/- 12 years. Time from liver transplant to treatment was 1637 +/- 1030 days. They were all infected with genotype 1. Eight patients received cyclosporine and the others tacrolimus. One patient was coinfected with hepatitis B virus and was receiving lamivudine. The mean initial histological activity index was 6.9 +/- 1.5 and fibrosis, 2.52 +/- 1.8 (Ishak). Two patients needed spleen embolization before the treatment. Two patients had to stop the treatment: one due to clinical intolerance, and the other one due to a cholangitis. In 14%, p-IFN doses were adjusted. In 32% RB was adjusted. Five (23.8%) did not respond at 24 weeks. Fourteen (66.7%) showed end-treatment responses but four relapsed at 72 weeks. A sustained viral response was achieved in 9 (42.8%). One patient died due to arterial thrombosis just after completing the treatment.

Depth of bacterial invasion in resected intestinal tissue predicts mortality in surgical necrotizing enterocolitis.

OBJECTIVE: Up to a third of all infants who develop necrotizing enterocolitis (NEC) require surgical resection of necrotic bowel. We hypothesized that the histopathological findings in surgically resected bowel can predict the clinical outcome of these infants. STUDY DESIGN: We reviewed the medical records and archived pathology specimens from all patients who underwent bowel resection/autopsy for NEC at a regional referral center over a 10-year period. Pathology specimens were graded for the depth and severity of necrosis, inflammation, bacteria invasion and pneumatosis, and histopathological findings were correlated with clinical outcomes. RESULT: We performed clinico-pathological analysis on 33 infants with confirmed NEC, of which 18 (54.5%) died. Depth of bacterial invasion in resected intestinal tissue predicted death from NEC (odds ratio 5.39 per unit change in the depth of bacterial invasion, 95% confidence interval 1.33 to 21.73). The presence of transmural necrosis and bacteria in the surgical margins of resected bowel was also associated with increased mortality. CONCLUSION: Depth of bacterial invasion in resected intestinal tissue predicts mortality in surgical NEC.

The effect of vpu on HIV-1-induced syncytia formation.

To investigate the role of vpu in the cytopathicity of human immunodeficiency type 1 (HIV-1), the MT4 CD4+ T-cell line was infected with viruses that were isogenic except for their ability to produce the vpu protein. The experiments described here demonstrate that expression of vpu reduces HIV-1 cytopathic effects by decreasing the rate of syncytia formation. By reducing the concentration of gp 120 at the cell surface, vpu limits cell killing by syncytia formation.

Protein G-gold complex: comparative evaluation with protein A-gold for high-resolution immunocytochemistry.

We combined the protein G-gold complex with several polyclonal and monoclonal antibodies for localization of various antigenic sites. The labelings were compared with those obtained using the protein A-gold complex. The results from either the immunodot experiment or immunoelectron microscopy have demonstrated that, for rabbit and guinea pig antibodies, both protein G-gold and protein A-gold complexes label several different specific antibodies with similar efficiency. However, with antibodies raised in goats or in mice, and particularly with mouse monoclonal antibodies, protein G-gold yielded intense and specific labeling, whereas protein A-gold yielded intense and specific labeling, whereas protein A-gold was very variable; it either gave weaker signals or failed to reveal any specific site or, as with one monoclonal, both protein G and protein A gave similar results. The higher affinity and versatility of protein G over protein A, established by the immunochemical approach, was confirmed by immunocytochemistry. Because of its enhanced reactivity with monoclonal antibodies and its broader affinity for polyclonal antibodies, protein G-gold complex appears to be a better and more versatile probe for high-resolution immunocytochemistry.

Establishment of persistent reovirus infection in SC1 cells: absence of protein synthesis inhibition and increased level of double-stranded RNA-activated protein kinase.

In the present study we report the establishment and characterization of an SC1 cell line persistently infected by reovirus. We observed that a significant percentage of SC1 cells was resistant to cell lysis upon infection with non-defective reovirus stocks. The apparent resistance of SC1 cells to the virus-induced inhibition of protein synthesis is probably an important factor favoring the establishment of such a persistence. The remaining cells, obtained following reovirus infection at a high multiplicity of infection, were kept as a continuous cell line and shown to have normal growth rate. They also released a high titer of virus that did not appear to differ from the original stock in neither infectivity nor genomic pattern. Electron microscopic examination further confirmed the presence of well-developed viral inclusions in the persistently infected cells. These cells were resistant to viral superinfection and exhibited a high constitutive level of the double-stranded RNA-activated protein kinase that might be involved in this resistance. We suggest that this cell line might be an interesting, and possibly more natural system than most previously used cell lines, for the continuing study of virus-host cell interactions during establishment of viral persistence using the much-studied model of reovirus infection.

Early infarct artery collateral flow does not improve long-term survival following thrombolytic therapy for acute myocardial infarction.

It is known that acutely developed collaterals can prevent the onset of acute myocardial infarction (AMI) in the presence of a total coronary occlusion. However, there still is controversy concerning long-term follow-up of coronary collateral circulation to the infarct-related artery. In this study we analyze the prognostic role of collateral flow (degrees 0 to 3) as well as anterograde flow (degrees 0 to 3) in patients with AMI treated with thrombolytic therapy. Four hundred twenty-two patients (median age 57 years, 355 men) with AMI were treated with intravenous streptokinase and followed prospectively for up to 8 years. At the end of the study period, patients with collateral coronary flow 3 (n = 30) and those with flow <3 (n = 392) at in-hospital coronary arteriography had survival rates of 66% and 85%, respectively (p <0.12). Meanwhile, patients with coronary anterograde flow 3 (n = 189) and those with flow <3 (n = 233) had survival rates of 89% and 80%, respectively (p <0.04). By censored regression analysis, a negative correlation was found between coronary collateral flow degree and survival (p = 0.0498) and, inversely, a positive correlation was found between coronary anterograde flow degree and survival (p = 0.0053). By Cox multivariate analysis, the following variables showed significant correlations with long-term survival: global left ventricular ejection fraction (p = 0.0003), anterograde flow degree (p = 0.0006), collateral flow degree (negative correlation, p = 0.0179), and medical treatment (negative correlation, p = 0.0464). Thus, patients treated with intravenous streptokinase during AMI and with adequate coronary collateral circulation had a worse prognosis than those who developed adequate anterograde flow, probably because of residual myocardial ischemia. Such patients may benefit from coronary revascularization (angioplasty or surgery) to restore anterograde blood flow and minimize myocardium at risk.

Diltiazem improves left ventricular systolic function following acute myocardial infarction treated with streptokinase. The Calcium Antagonist in Reperfusion Study (CARES) Group.

The role of diltiazem on left ventricular systolic function was analyzed in 101 patients with acute myocardial infarction treated with streptokinase, being obtained, for the total of the population, higher LV global ejection fraction (p = 0.022), LV regional shortening (p = 0.046) and LV global shortening (p = 0.064) for the treated group, relative to the placebo group; the p values were, respectively, 0.005, 0.009, and 0.012, for patients that achieved TIMI-3 antegrade coronary flow. It is concluded that diltiazem is useful as adjuvant to streptokinase, especially when antegrade coronary blood flow TIMI-3 is obtained.

Surgical revascularization after fibrinolysis in acute myocardial infarction. Long-term follow-up.

One hundred twenty-eight patients with myocardial infarction who underwent operation for myocardial revascularization and 147 patients who received medical therapy were followed up for up to 6 years: all patients had received treatment with intravenous streptokinase. In the surgical group, 91.5% of the patients had the region related to the infarction revascularized, and in 82.8% of them the mammary artery was used. Statistically significant differences were not detected between the groups according to infarct size, clinical features, and left ventricular ejection fraction. However, there was a higher risk in the surgical group, as compared with that in the medical group, in terms of anatomic characteristics: 99.2% versus 77.1% of the patients showed more than 70% residual obstruction at the "culprit" coronary artery (p < 0.001, 95% confidence interval 14.1% to 30.1%) and 76.8% versus 40.7% showed multivessel coronary disease (p < 0.001, 95% confidence interval 23.7% to 48.5%). In-hospital survival was 95.3% in the surgical group and 89.1% in the medical group (p = 0.096, 95% confidence interval -0.2% to 12.6%). Significantly higher survivals were obtained for the surgical group both during the first (93% +/- 2.3% versus 80.3% +/- 3.3%, p = 0.005) and the sixth (86.4% +/- 3.4% versus 68.4% +/- 4.3%, p = 0.003) year of follow-up. Statistically significant differences were also obtained when in-hospital deaths were excluded. A Cox regression model with 13 variables showed that only age (p = 0.0422) and medical treatment (p = 0.0194) correlated independently with mortality. It is concluded that in this nonrandomized study, operation led to a significantly higher survival both on a medium- and long-term basis, when compared with that obtained for patients receiving medical therapy.

Prognosis of acute myocardial infarction in the thrombolytic era: medical evaluation is still valuable.

BACKGROUND: Modern and sophisticated technology for the management of myocardial infarction has progressively devalued medical evaluation. HYPOTHESIS: This study was undertaken to assess the importance of the findings of medical evaluation at hospital presentation, in patients with acute myocardial infarction. METHODS: Data from 590 thrombolytic-treated myocardial infarction patients were analyzed. The patients were grouped according to their clinical status on arrival at hospital. A modified Forrester classification--subset II was divided according to the absence (IIa) or presence (IIb) of symptoms--was applied. Short- (14 days) and long-term (up to 10 years) survival was analyzed and 19 independent variables were included in the multivariate models. RESULTS: Short-term survival was 95.6% for subset I, 83.3% for subset IIa, 60% for subset IIb, 54.6% for subset III, and 34.8% for subset IV (P<0.001). By multiple regression analysis, lower clinical subsets (P<0.001), fewer coronary arteries with disease (P=0.006), younger age (P=0.014), absence of reinfarction (P=0.034), longer interval between streptokinase infusion and coronary arteriography (P=0.016), and higher left ventricular ejection fraction (P=0.037) demonstrated significant and independent correlation with short-term survival. Long-term survival for the total population was 71+/-3.6% for subset I, 54.4+/-8.5% for subset IIa, 20.8+/-9.4% for subset IIb, 54.5+/-15% for subset III, and 0% for subset IV (P<0.001). Using Cox regression analysis, lower clinical subsets (P<0.001), younger age (P<0.001), higher global left ventricular ejection fraction (P<0.001), and fewer coronary arteries with disease (P=0.021) correlated independently and significantly with long-term survival. When excluding data from patients who died before the short-term follow-up (n=532), lower clinical subsets remained an important predictor of long-term survival (P<0.001). CONCLUSION: Clinical classification at hospital presentation is a powerful predictor of short- and long-term survival post-myocardial infarction.

Ultrastructural localization of viral antigens using the protein A-gold technique.

HSV and DNV viral antigens have been localized by electron microscopy using the protein A-gold technique. The labelling of HSV antigens was detected over the (naked and enveloped) viral particles as well as on the cytoplasm and the nucleoplasm. In contrast, DNV antigens were revealed only over clusters of viral particles in the nucleus. The high sensitivity of the technique and the good ultrastructural preservation allowed a very fine identification of the labelled structures. Thus, the protein A-gold technique can be applied generally for the ultrastructural detection and identification of viral antigens and might be useful for diagnostic purposes.