Pulmonary Ultrasound and Diaphragmatic Shortening Fraction Combined Analysis for Extubation-Failure-Prediction in Critical Care Patients

Introduction

Invasive respiratory support is a cornerstone of Critical Care Medicine, however, protocols for withdrawal of mechanical ventilation are still far from perfect. Failure to extubation occurs in up to 20% of patients, despite a successful spontaneous breathing trial (SBT).

Search for Neuro-Endocrine Markers (Chromogranin A,Synaptophysin and VGF) in Breast Cancers. An integrated Approach Using Immunohistochemistry and Gene Expression Profiling

Discordant data are reported in the literature on the definition, incidence and clinical features of neuroendocrine (NE) carcinomas of the breast. This tumour entity is currently assessed by immunohistochemistry (IHC) detecting “general” NE markers such as chromogranin A (CHGA) and synaptophysin (SYP), but other markers have been considered as well. In the present study, in addition to CHGA and SYP, we investigated the expression of VGF, a neurotrophin-inducible gene, which is emerging as a new specific NE marker. In order to evaluate the differential expression of these neuro-endocrine markers in breast cancers, we conducted parallel immunohistochemical and gene expression analyses, using PCR, gene array and real-time quantitative PCR procedures. Data obtained in 28 cases were further validated with a meta-analysis of published datasets of 103 breast cancer cases. The value of IHC positivity (irrespective of the percentage of positive cells) was confirmed by over-expression of the related gene. However, the genetic approach emerged as more sensitive, showing over-expression of NE markers in a subset of IHC-negative carcinomas. In conclusion, the present study confirms, by a novel approach, the occurrence of NE differentiation in breast cancers. Over-expression of one or more NE marker (CHGA and/or SYP and/or VGF) characterizes a significant fraction (approximately 10 %) of infiltrative breast cancers.     

Trypanosoma cruzi nucleoside triphosphate diphosphohydrolase 1 (TcNTPDase-1) biochemical characterization,immunolocalization and possible role in host cell adhesion

Previous work has suggested that Trypanosoma cruzi diphosphohydrolase 1 (TcNTPDase-1) may be involved in the infection of mammalian cells and serve as a potential target for rational drug design. In this work, we produced recombinant TcNTPDase-1 and evaluated its nucleotidase activity, cellular localization and role in parasite adhesion to mammalian host cells. TcNTPDase-1 was able to utilize a broad range of triphosphate and diphosphate nucleosides. The enzyme's K_m for ATP (0.096 mM) suggested a capability to influence the host's ATP-dependent purinergic signaling. The use of specific polyclonal antibodies allowed us to confirm the presence of TcNTPDase-1 at the surface of parasites by confocal and electron microscopy. In addition, electron microscopy revealed that TcNTPDase-1 was also found in the flagellum, flagellum insertion region, kinetoplast, nucleus and intracellular vesicles. The presence of this enzyme in the flagellum insertion region and vesicles suggests that it may have a role in nutrient acquisition, and the widespread distribution of TcNTPDase-1 within the parasite suggests that it may be involved in other biological process. Adhesion assays using anti-TcNTPDase-1 polyclonal antibodies as a blocker or purified recombinant TcNTPDase-1 as a competitor revealed that the enzyme has a role in parasite–host cell adhesion. These data open new frontiers to future studies on this specific parasite–host interaction and other unknown functions of TcNTPDase-1 related to its ubiquitous localization.     

Functional mechanism of tracheal relaxation,antiasthmatic, and toxicological studies of 6-hydroxyflavone

Previously, we described tracheal rat rings relaxation by several flavonoids, being 6-hydroxyflavone (6-HOF) the most active derivative of the series. Thus, its mechanism of action was determined in an ex vivo tracheal rat ring bioassay. The anti-asthmatic effect was assayed in in vivo OVAlbumin (OVA)-sensitized guinea pigs. Finally, the toxicological profile of 6-HOF was studied based on Organization of Economic Cooperation and Development guidelines with modifications. 6-HOF–induced relaxation appears to be related with receptor-operated calcium channel and voltage-operated calcium channel blockade as the main mechanism of action, and also through the production of relaxant second messengers NO and cGMP. Molecular docking supports that 6-HOF acts as calcium channel blocker and by activation of nitric oxide synthase. In addition, the in vivo anti-asthmatic experiments demonstrate the dose-dependent significant anti-allergic effect of 6-HOF induced by OVA, with best activity at 50 /kg. Finally, toxicological studies determined a LD50 > 2,000 mg/kg and, after 28 day of treatment with 6-HOF (50 mg/kg) by intragastric route, mice did not exhibit evidence of any significant toxicity. In conclusion, experiments showed that 6-HOF exerts significant relaxant activity through calcium channel blockade, and possibly, by NO/cGMP-system stimulation on rat trachea, which interferes with the contraction mechanism of smooth muscle cells in the airways. In addition, the flavonoid shows potential anti-asthmatic properties in an anti-allergic pathway. Furthermore, because the pharmacological and safety evidence, we propose this flavonoid as lead for the development of a novel therapeutic agent for the treatment of asthma and related respiratory diseases.     

Lean mass and insulin resistance in women with polycystic ovary syndrome

Insulin resistance is common in women with polycystic ovary syndrome (PCOS). Muscle is the major tissue utilizing glucose while excess adipose tissue relates to insulin resistance. Thus, body composition is likely to be an important regulator of insulin sensitivity. Thirty-nine PCOS patients (age: 29.9 ± 1.0 years; BMI: 33.8 ± 1.2 kg/m²) participated in a cross sectional study. Body composition was measured by dual energy x-ray absorptiometry (DEXA). Insulin resistance and secretion were assessed using oral glucose tolerance test (OGTT) and frequently sampled intravenous glucose tolerance test (FS-IVGTT). In contrast with the conventional expectations, lean mass correlated directly (P < .05) with the insulin resistance measure HOMA (r = 0.440); and inversely with the insulin sensitivity index QUICKI (r = ?0.522) independent of fat mass. In 11 pairs of subjects matched for fat mass (35.6 ± 2.2 and 35.6 ± 2.4 kg) but with discordant lean mass (52.8 ± 1.8 vs 44.4 ± 1.6 kg), those with higher lean mass had a higher glucose response during OGTT (AUC_Glucose; P = .034). In contrast, 17 pairs matched for lean mass (48.7 ± 1.7 and 48.9 ± 1.6 kg) but discordant for fat mass (43.3 ± 2.6 vs 30.3 ± 8.9 kg) showed no differences in insulin resistance parameters. These novel findings indicate that lean mass relates directly to insulin resistance in PCOS.     

One year postoperative hard and soft tissue volumetric changes after a BSSO mandibular advancement

In this study, cone beam computed tomography (CBCT) and three dimensional (3D) stereophotogrammetry are used to compare the 3D skeletal and soft tissue changes caused by a bilateral sagittal split osteotomy (BSSO) 1 year after a mandibular advancement. Eighteen consecutive patients with a hypoplastic mandible were treated with a BSSO according to the Hunsuck modification. Preoperatively and 1 year postoperatively, a CBCT scan was acquired and a 3D photograph. The pre- and postoperative CBCT scans were matched using voxel based registration. After registration, the mandible could be segmented in the pre- and postoperative scans. The preoperative scan was subtracted from the postoperative scan, resulting in the hard tissue difference. To investigate the soft tissue changes, the pre- and postoperative 3D photographs were registered using surface based registration. After registration the preoperative surface could be subtracted from the postoperative surface, resulting in the overall volumetric difference. As expected, a correlation between mandibular advancent and volumetric changes of the hard tissues was found. The correlation between advancement and soft tissues was weak. The labial mental fold stretched after surgery. This study proved that using 3D imaging techniques it is possible to document volumetric surgical changes accurately and objectively.     

Influence of Er,Cr:YSGG laser irradiation on enamel caries prevention

The objective of this study was to evaluate the effects of chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation on the acid resistance of dental enamel. Forty human enamel samples were divided into four groups. They were manually irradiated with an Er,Cr:YSGG laser device (λ?=?2.78 µm, 20 Hz, 20 s), in a scanning mode, with and without water cooling, according to the following parameters: Group 1: 0.25 W, 62.5 J/cm², no water cooling; group 2: 0.25 W, 62.5 J/cm², 5.0 ml/min; group 3: 0.5 W, 125 J/cm², no water cooling; group 4: 0.5 W, 125 J/cm², 5.0 ml/min. No airflow was used. Afterwards, the samples were submitted to an acid challenge and assessed by cross-sectional Knoop microhardness at different depths (20, 40, 60, 80, and 100 µm) from the outer enamel surface. Average values were obtained for both irradiated and control areas in each sample and they were compared to obtain a percentage of microhardness increase. Data were analyzed by analysis of variance and Fisher’s exact test (α?=?5%). The percentage of microhardness increase observed in group 1 (+23.58%) was similar to group 3 (+19.12%), but higher than groups 2 (+3.61%) and 4 (10.9%) (p?<?0.05). The comparison of the depths showed that the Er,Cr:YSGG laser acted in the superficial layers of the dental enamel. The findings of the present study suggest that the energy densities of 62.5 and 125 J/cm² were capable of increasing the acid resistance of human enamel. The presence of water during irradiation makes it difficult to obtain an enamel surface more resistant to acids.