Synergistic Effects of BAY 60‐4552 and Vardenafil on Relaxation of Corpus Cavernosum Tissue of Patients with Erectile Dysfunction and Clinical Phosphodiesterase Type 5 Inhibitor Failure

IntroductionOverall efficacy rates of phosphodiesterase type 5 inhibitors (PDE5‐i) for erectile dysfunction (ED) are 60–70%. PDE5‐i treatment failures currently have to resort to invasive treatment options for restoration of erectile function.AimsTo assess changes in the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP)/protein kinase (PKG) pathway in human corpus cavernosum (HCC) of PDE5‐i nonresponders compared with healthy controls. To evaluate the effects of BAY 60‐4552, a stimulator of soluble guanylate cyclase (sGC), and vardenafil on relaxation of HCC strips from PDE5‐i nonresponders.Main Outcome MeasuresmRNA expression, morphological localization of the NO/cGMP/PKG pathway, and relaxant capacity of both compounds alone or combined. Analysis of variance, t‐test or Mann–Whitney test based upon number of groups and normality of data.MethodsHCC tissues were harvested after consent from individuals undergoing penile prosthesis implantation (patients) and potent patients undergoing transurethral surgery (healthy controls, needle biopsy). HCC tissues of patients were compared with those of healthy controls for the expression of mRNA coding for PDE5A, eNOS, PKGα1, PKG2, sGCα1, sGCα2, sGCβ1, sGCβ2, α‐smooth muscle actin (aSMA) and β‐actin by quantitative polymerase chain reaction (qPCR). The respective proteins were localized using immunofluorescence. Tissue strips of patients were precontracted with phenylepinephrine followed by incubation with 1 μM of either vardenafil or BAY 60‐4552, or both simultaneously.ResultsThe main targets in the NO/cGMP/sGC pathway were downregulated in PDE5‐i nonresponders. The pathway was morphologically located to HCC smooth muscle, of which the overall content was preserved in ED patients based on aSMA expression. BAY 60‐4552 and vardenafil have synergistic effects on relaxation of HCC of PDE5‐i nonresponders. The main limitation is the small amount of control tissue precluding functional testing on these samples.ConclusionDespite downregulation of the NO/cGMP/PKG pathway, combining BAY 60‐4552 with vardenafil significantly enhanced relaxation HCC strips of PDE5‐i nonresponders.     

Calcium Sulfate with Stearic Acid as an Encouraging Carrier for Reindeer Bone Protein Extract

Various bone proteins and growth factors in specific concentrations are required for bone formation. If the body cannot produce sufficient quantities of these factors, bone trauma can be healed with an implant that includes the required factors in a carrier. This study was designed to evaluate various calcium salt candidates that can be used as carrier with reindeer bone protein extract to induce ectopic bone formation in the muscle pouch model of mouse. The bone protein extract was either impregnated into the disc form of carrier or mixed with carrier powder before implantation. The radiographic analysis indicated increased bone formation in all of the active groups containing the bone protein extract compared to the controls within 21 days follow-up. The highest bone formation was seen in the group with calcium sulfate with stearic acid where new bone and calcified cartilage were clearly visible. The greatest bone formation occurred in the groups that had bone protein extract readily available. This indicates that the bone forming factors in sufficient concentrations are required at the early stage of bone formation. The calcium sulfate with stearic acid was the most suitable and effective carrier for reindeer bone protein extract.     

Digital analysis yields more reliable and accurate measures of dermal and epidermal thickness in histologically processed specimens compared to traditional methods

Changes in the thickness of the dermis and epidermis have been described in the scenario of tissue expansion as well as inflammatory skin processes (psoriasis, contact hypersensitivity and so on). These changes have previously been quantified using ocular micrometers to obtain and then average a limited number of spot measurements, leading to suboptimal accuracy. We describe a rapid method of using freely available ImageJ software to analyze digitized images of fixed skin specimens. By determining the cross‐sectional area and surface length of a skin layer, a simple calculation produces more accurate and reproducible measurements of its thickness compared to historical methods, with excellent inter‐rater reliability.     

Unowocześnienie diagnostyki nocnej napadowej hemoglobinurii – wstępne wyniki badań przy użyciu odczynnika FLAER

Blood samples of 50 patients diagnosed for paroxysmal nocturnal haemoglobinuria (PNH) were examined using FLAER screening test for the detection of PNH clone. The results were compared with routine testing using monoclonal antibodies detecting CD59 and CD66b antigens on neutrophils and CD59 on erythrocytes. Assay sensitivity (≥ 0.1%) and PNH clone size analysed using FLAER and anti-CD66b antibodies was fully consistent (R² = 0.9991) and higher than with anti-CD59 antibodies. Preliminary results indicate that for diagnosis of PNH the FLAER based test is quick, reliable and cost-effective. It might seem that the conventional screening methods could therefore be abandoned. It was found however, that in some cases of abnormal granulocyte population, additional studies should be performed using multiparameter flow cytometry blood cell analysis.     

Root cause analysis of internal hernia and Roux limb compression after laparoscopic Roux-en-Y gastric bypass using observational clinical human reliability assessment

BackgroundInternal hernia (IH) and Roux limb compression (RC) are recognized complications after retrocolic laparoscopic Roux-en-Y gastric bypass for obesity. The aim of the present study was to systematically identify the surgical technical errors leading to these complications.MethodsAn observational clinical human reliability assessment approach was used to analyze the operating videos of 3 groups: an IH group (n = 12), a Roux compression group (n = 13), and a control group (no complications, n = 21). Two investigators, unaware of the outcomes, reviewed all videos, using special rating software. All errors were categorized using the external error mode system and further described if a direct consequential error (e.g., bleeding) was found.ResultsAn analysis of data showed that, on average, more errors occurred in the complication groups than in the control group (IH 5.85, Roux compression 3.54, control .90, P < .001). The strongest differences were found for missing intermesenteric stitches on both sides of the Roux limb. Logistic regression analysis showed that a missed stitch between the mesentery of the Roux limb and the transverse mesocolon was an independent predictor for IH (B = 1.727, P = .025). No technical or consequential errors could be identified as responsible for RC.ConclusionThe observational clinical human reliability analysis is a useful method to identify operative failure. For retrocolic, retrogastric laparoscopic Roux-en-Y gastric bypass, a systematic approach for the closure of the transverse mesenteric window might prevent IH complications.