Role of p21‐activated kinase in cell polarity and directional mesendoderm migration in the Xenopus gastrula

The p21 activated kinases (Paks) are prominently involved in the regulation of cell motility. Using a kinase‐dead mutant of xPak1, we show that during Xenopus gastrulation, the kinase activity of Pak1 is required upstream of Cdc42 for the establishment of cell polarity in the migrating mesendoderm. Overactivation of Pak1 function by the expression of constitutively active xPak1 compromises the maintenance of cell polarity, by indirectly inhibiting RhoA function. Inhibition of cell polarization does not affect the migration of single mesendoderm cells. However, Pak1 inhibition interferes with the guidance of mesendoderm migration by directional cues residing in the extracellular matrix of the blastocoel roof, and with mesendoderm translocation in the embryo. Developmental Dynamics 238:1709–1726, 2009. © 2009 Wiley‐Liss, Inc.     

Enhancing memory for lists by grouped presentation and rehearsal: A pilot study in healthy subjects with unexpected results

List learning is probably the most established paradigm for the psychometric evaluation of episodic memory deficits in different neuropsychiatric conditions including epilepsy. Strategies which are capable of increasing the test performance might be promising candidates for a therapeutic improvement of daily memory performance. Based on the classical ‘temporal grouping effect’ we wanted to evaluate the memory-enhancing potential of disentangling perceiving, rehearsing and encoding by temporally grouped presentation and group-wise reproduction during acquisition. According to the ethical principle of subsidiary the study was performed in healthy adolescents (N = 126) before setting-up a patient study. Subjects had to learn a list of 12 semantically unrelated nouns and a list of 12 figures during two acquisition trials under one of four experimental conditions defined by the size of presented item groups (GS): GS = 1 (single items, i.e., 12 × 1 item), GS = 3 (4 × 3 items), GS = 6 (2 × 6 items), and GS = 12 (standard presentation mode, i.e., 1 × 12 items). Repeated measures MANOVA confirmed a positive effect of smaller GS on acquisition performance but the grouping condition obtained no effect on immediate and delayed free recall or on yes/no recognition. For verbal retention, GS = 12 even showed a tendency toward an advantage as compared to GS = 3. Although appearing reasonable and promising, facilitating acquisition during list learning by temporal grouping and grouped overt rehearsal turned out to be ineffective with regard to long-term memory encoding and retrieval. A strategy however which fails in healthy subjects is unlikely to obtain a therapeutic potential in patients with memory deficits.     

Thoracic surgery in solving enormous elevation of the left hemidiaphragm

BACKGROUND: Acquired elevation of the diaphragm is mostly the result of phrenic nerve paralysis, some of thoracic and abdominal patological states, and also some of neuromuscular diseases. Surgical treatment is rarely performed and is indicated when lung compression produces disabilitating dyspnea, and includes plication of diaphragm. The goal of this case report has been to show completely documented diagnostic procedures and surgical treatment one of rare pathological condition. CASE REPORT. A 62-year-old patient was admitted to our clinic because of surgical treatment of the enormous elevation of the left hemidiaphragm. After thoracotomy and plication of the bulging diaphragm, lung compression did not exist any more and mediastinum went back in the normal position. CONCLUSION: Elevation of the diaphragm rarely demands surgical correction. When it is complicated with lung compression and disabilitating dyspnea, surgical treatment has extremely useful functional effect.     

Divergent Squamous Differentiation in Upper Urothelial Carcinoma—Comparative Clinicopathological and Molecular Study

Upper urothelial carcinoma (UUC) has a plasticity to demonstrate divergent differentiation with squamous metaplastic elements. There was no previous study exploring profiling of molecular markers in metaplastic squamous upper urothelial carcinoma (SUUC) and conventional upper urothelial carcinoma (CUUC). The aims of this study was to compare expression of the phenotypic characteristics of tumors and molecular markers (p53, p16, cyclin D1, E-cadherin, HER-2, Ki-67, Bcl-2, Bax) in SUUC and CUUC. SUUC was detected in 20% of 44 patients. There was significant difference between SUUC and CUUC in the pathological stage, grade, growth and presence of lymphovasular invasion (p < 0.05; 0.05; 0.05; 0.01 respectively). The mean Ki-67 and p53 labeling index was significantly higher in SUUC than in CUUC (p < 0.05; 0.05). There was no significant difference in the expression of p16, cyclin D1, E-cadherin, HER-2, Bcl-2 and Bax between SUUC and CUUC. Univariant model showed that SUUC was significantly associated with lymphovascular invasion (p = 0.007), Ki-67 activity (p = 0.016) and growth (p = 0.026). Exploration of UUC with squamous divergent differentiation showed changes in phenotypic characteristics and Ki-67, as well as similar molecular profile with CUUC.     

Inactivation of harmful tumour-associated proteolysis by nanoparticulate system

The primary aim in cancer therapy is to deliver anti-cancer drugs to their specific molecular targets in the tumour. Here we present a system composed of poly(d,l-lactide-co-glycolide) nanoparticles, cytokeratin specific monoclonal antibody and cystatin, a potent protease inhibitor, that can neutralize the excessive proteolytic activity associated with the invasive and metastatic potential of breast tumour cells. The antibody provides specific targeting of the delivery system to invasive breast epithelial cells and, additionally, prevents the generation of plasmin, a central extracellular protease involved in malignant progression. Polymeric nanoparticles rapidly enter the targeted cells and release the inhibitor cargo within the endosomes/lysosomes. The inhibitor is capable to inactivate lysosomal cysteine proteases, in particular cathepsin B, which is involved in the degradation of extracellular matrix inside the tumour cells. Our approach, which combines nanoparticulate delivery system with the inhibitory potential against extracellular and intracellular proteases, may improve the efficacy of therapy in patients with breast tumours compared to the application of individual protease inhibitors.     

Is bridging to transplantation with a left ventricular assist device a risk factor for transplant coronary artery disease?

BACKGROUND: Left ventricular assist device (LVAD) support is associated with coagulopathy, bleeding, increased blood transfusion, and increased anti-HLA antibody production. Increased anti-HLA antibody production is associated with early transplant rejection, transplant coronary artery disease (CAD), and decreased post-transplant survival rates. We asked whether bridging to transplantation with an LVAD increases the risk of transplant CAD. METHODS: We reviewed data for all adults (>18 years old) who underwent heart transplantation at our institution between 1988 and 2000. After exclusion of transplant recipients who survived <3 years, we divided the remaining cohort into 2 groups: those bridged to transplantation with LVADs (mean duration of support, 149 +/- 107 days, n = 29) and those in United Network for Organ Sharing Status 1 bridged to transplantation without LVADs (controls, n = 86). We compared groups in terms of disease cause, age, sex, donor age, panel-reactive antibody testing, crossmatching, pre- and post-transplant cholesterol concentrations, diagnosis of diabetes mellitus or treated hypertension, infections, calcium channel blocker use, transplant rejection, ischemic time, cytomegalovirus infection, pre-transplant transfusion, and incidence of transplant CAD (defined as any coronary lesion identified by coronary angiography). We considered p < 0.05 to be significant. RESULTS: The bridged and control groups were similar in all respects except mean ischemic time (217 +/- 58 minutes vs 179 +/- 67 minutes, p = 0.007), post-transplant cholesterol concentration (212 +/- 55 mg/dl vs 171 +/- 66 mg/dl, p = 0.007), and pre-transplant transfusion incidence (100% vs 22%, p < 0.001). The incidence of transplant CAD was similar in both groups during a 3-year follow-up period (28% vs 17%, p = 0.238) and during total follow-up (34% vs 35%, p = 0.969). Multivariate logistic regression analysis identified cholesterol concentration at 1 year after transplantation as a significant predictor of CAD at 3 years after heart transplantation (p = 0.0029, odds ratio = 0.984). CONCLUSIONS: Bridging to transplantation with an LVAD does not increase the risk of transplant CAD. Nevertheless, aggressive prophylactic therapy to minimize potential risk factors for transplant CAD, such as increased cholesterol concentration, is warranted in all transplant recipients.     

Growth hormone replacement normalizes impaired fibrinolysis: New insights into endothelial dysfunction in patients with hypopituitarism and growth hormone deficiency

BackgroundCardiovascular morbidity in adult patients with growth hormone deficiency (GHD) and hypopituitarism is increased. Clustering of cardiovascular risk factors leading to endothelial dysfunction and impaired fibrinolysis has also been reported and may account for progression to overt vascular changes in these patients. However, effect of long lasting GH replacement therapy on fibrinolytic capacity in GH deficient patients has not been investigated so far.ObjectiveTo investigate fibrinolysis before and after challenge with venous occlusion in GHD patients with hypopituitarism before and during one year of growth hormone replacement.DesignHospital based, interventional, prospective study.Investigated subjectsTwenty one patient with GHD and fourteen healthy control subjects matched for age, sex and body mass index (BMI).MethodsAnthropometric, metabolic and fibrinolytic parameters were measured at the start and after three, six and twelve months of treatment with human recombinant GH.ResultsAt baseline GHD patients had significantly impaired fibrinolysis compared to healthy persons. During treatment with GH, significant changes were observed in insulin like growth factor 1(IGF-1) [from baseline 6.9(2.4–13.5) to 22.0(9.0–33.0) nmol/l after one month of treatment; p < 0.01] and fibrinolysis. Improvement in fibrinolysis was mostly attributed to improvement of stimulated endothelial tissue plasminogen activator (t-PA) release in response to venous occlusion [from baseline 1.1(0.4–2.6) to 1.9(0.5–8.8) after one year of treatment; p < 0.01].ConclusionGrowth hormone replacement therapy has favorable effects on t-PA release from endothelium and net fibrinolytic capacity in GHD adults, which may contribute to decrease their risk of vascular complications.