Polo‐like‐kinase 1 (PLK1) as a molecular target to overcome SYK‐mediated resistance of B‐lineage acute lymphoblastic leukaemia cells to oxidative stress

SYK tyrosine kinase has emerged as a master regulator of cellular resistance to oxidative stress (OS) by mediating the activation of the anti‐apoptotic nuclear factor κB and phosphatidylinositol‐3 kinase/AKT pathways after OS exposure. Here, we present unprecedented experimental evidence that polo‐like kinase 1 (PLK1) is the upstream regulator of SYK in B‐lineage acute lymphoblastic leukaemia (ALL) cells. Selective inhibition of PLK‐1 with the leflunomide metabolite analogue α‐cyano‐β‐hydroxy‐β‐methyl‐N‐[4‐(trifluoromethoxy) phenyl]‐propenamide/LFM‐A12 abolished the resistance of B‐lineage ALL cells to OS by preventing the activation of the anti‐apoptotic SYK signal transduction pathway. Notably, LFM‐A12 treatments at non‐cytotoxic concentrations resulted in marked augmentation of clonogenic death in resistant human B‐lineage ALL cell lines challenged with OS. Further, LFM‐A12 augmented OS‐induced apoptosis of chemotherapy‐resistant primary leukaemic cells from relapsed B‐lineage ALL patients in vitro and markedly potentiated the in vivo anti‐leukaemic activity of total body irradiation (TBI) against leukaemia‐initiating cells in severe combined immunodeficient mouse xenograft models of B‐lineage ALL. This study is the first to identify PLK1 as a regulator of SYK tyrosine kinase and a molecular target to overcome SYK‐mediated resistance of B‐lineage ALL cells to OS.     

Acute hemiscrotum due to inguinoscrotal Littre hernia: a case report

Meckel diverticulum (MD) is the most common congenital anomaly of the gastrointestinal tract that is generally asymptomatic and only manifests in a specific way when complications exist. An unusual complication of MD is known as Littre hernia. It comprises less than 1% of all MD. Littre hernia is the protrusion of an MD through a potential abdominal opening. Usual sites of Littre hernia are right inguinal (50% of cases), umbilical hernia (20%), and femoral hernia (20%). We report a case of Littre hernia in a boy who presented with acute scrotal pain and swelling.     

Dilution of rocuronium to 0.5 mg/mL with 0.9% NaCl eliminates the pain during intravenous injection in awake patients

In a randomized, double-blinded, controlled study, we evaluated the effect of diluting rocuronium 10 mg/mL to 1 or 0.5 mg/mL with 0.9% NaCl on the pain associated with IV administration of rocuronium with small doses given before succinylcholine or nondepolarizing agent administration. One hundred fifty patients undergoing surgical procedures that required general anesthesia were randomized into three groups. Group 1 received rocuronium 10 mg/mL. Groups 2 and 3 received 1 and 0.5 mg/mL of rocuronium, respectively. Patient demographics, pain scores, osmolality, and the pH of the solutions were recorded. Group 1 had the most intense and frequent pain response. Eighty percent of patients in this group reported pain on injection. In Group 2, the incidence and intensity of pain were significantly less when compared with those of Group 1. In this group, 38% of patients reported pain during injection. In Group 3, none of the patients experienced pain on injection. The pH values and osmolalities of study solutions were not significantly different among groups. In conclusion, in awake patients, dilution of rocuronium 10 mg/mL at small doses given before succinylcholine or nondepolarizing agent administration of 0.06 mg/kg to 0.5 mg/mL with 0.9% NaCl is a simple and cost-effective strategy for preventing pain during IV rocuronium injection.     

Semen characteristics after overnight shipping: preservation of sperm concentrations, HspA2 ratios, CK activity, cytoplasmic retention, chromatin maturity, DNA integrity, and sperm shape

We tested several approaches that can be used to preserve sperm attributes and the objective biochemical markers of sperm maturity and function for assessment in a remote centralized laboratory after overnight shipping of semen samples. Addition of phenyl-methyl-sulfonyl-fluoride (PMSF) to a final concentration of 20 microg/mL semen at 4 degrees C has preserved sperm concentrations and HspA2 isoform ratios, even at room temperature, simulating a shipping delay in moderate ambient temperatures. Regarding the attributes of individual spermatozoa, the patterns of CK-immunocytochemistry (demonstrates cytoplasmic retention in diminished-maturity spermatozoa); aniline blue staining pattern (tests chromatin maturity); sperm shape assessed by both Kruger strict morphology and computer assisted morphometry; and sperm DNA integrity, as tested by DNA nick translation, all remained unchanged. Thus, the PMSF-4 degrees C conditions preserved sperm concentrations and the cytoplasmic and nuclear biomarkers of sperm cellular maturity and function for next-day analysis. This shipping method will facilitate the early detection of subtle changes in semen quality that can affect sperm function, even when there has been no decline in sperm concentrations to signal possible toxic effects. Furthermore, sample preservation will enable investigators to evaluate semen for toxicology studies and for diagnosis of male infertility from remote locations. Home collection of semen should enhance study participation, and semen assessment in centralized laboratories will address concerns regarding interlaboratory variations and quality control.     

Retinal pigment epithelium cell culture on surface modified poly(hydroxybutyrate-co-hydroxyvalerate) thin films

There is currently no effective treatment for the retinal disorders caused by retinal pigment epithelium (RPE) degeneration. Transplantation of allografts is the main strategy towards correction of this malady. Tissue engineering could offer hope and involve the use of biodegradable polymeric templates to replace diseased or lost RPE. In this study PHBV8 film was chosen as a temporary substrate for growing retinal pigment epithelium cells as an organized monolayer before their subretinal transplantation. The surface of the PHBV8 film was rendered hydrophilic by oxygen plasma treatment to increase the reattachment of D407 cells on the film surface. Power and duration was changed, from 50 W, 10 min to 100 W, 20 min during plasma treatment. The effect of these two parameters on surface hydrophilicity, morphology, topography, surface composition of PHBV8 thin films was studied using AFM, SEM, and phase contrast microscopy. The effect of changes in surface characteristics on cell reattachment, spreading and cell growth rate was investigated. It was found that as the treatment level was increased the surface hydrophilicity increased and roughness was decreased probably due to ablation. The PHBV8 film treated with 100 W 10 min was found to be the most suitable for 24 h reattachment of D407 cells. The cells were also grown to confluency as an organized monolayer suggesting PHBV8 film as a potential temporary substrate for subretinal transplantation to replace diseased or damaged retinal pigment epithelium.     

In acute organophosphate poisoning, the efficacy of hemoperfusion on clinical status and mortality

The aims of this study were to report experience in patients with organophosphate poisoning (OPP) and to discuss the potential role for hemoperfusion (HP) in the management of severe OPP. At the emergency service of a university hospital, 52 patients with acute OPP were included in this retrospective study. The patients were divided into 2 groups (Group 1, severe poisoning, n = 25; and Group 2, mild poisoning, n = 27). All patients with mild OPP survived. Seven patients (28%) of the 25 with severe OPP died. This study supports previous data documenting that HP is unnecessary in the management of mild OPP. Although there was not a control group (severe poisoning without HP treatment) in this study, experience suggests that HP can be useful in severe cases. Reports from centers having experience with severe OPP can help clarify this controversial issue. Randomized controlled (prospective) studies investigating the possible beneficial effects of HP on patient survival in patients with severe OPP with control group are needed.     

The restorative effect of a selective cyclooxygenase-2 inhibitor on urothelial cell-cell interactions after partial bladder outlet obstruction in rats

OBJECTIVE: To determine the changes in cyclooxygenase-2 (COX-2), E-cadherin and alpha-catenin expression after partial bladder outlet obstruction (PBOO), and whether a selective COX-2 inhibitor (celecoxib) might inhibit COX-2 expression and have beneficial effects on urothelial cell-to-cell interactions in rats subjected to PBOO. MATERIALS AND METHODS: Thirty-six male rats were divided into six equal groups; celecoxib was administered after creating PBOO for 1 and 4 weeks in groups 1 and 2, respectively. Two further obstructed groups (3 and 4, PBOO for 1 and 4 weeks, respectively) received no treatment. Sham-operated animals served as controls (group 5 and 6, assessed at 1 and 4 weeks, respectively). After 1 and 4 weeks of PBOO or a sham procedure the bladder weight was recorded before sampling the bladder for Western blotting and immunohistological analysis, to assess the expressions of COX-2 and adherens proteins, E-cadherin and alpha-catenin. Urothelial cell-to-cell interactions were evaluated using electron microscopy. RESULTS: The bladder mass increased rapidly during the first 7 days after PBOO in groups 1-4 compared with 5 and 6 (P < 0.05). While the bladder mass then continued to increase for the next 21 days in group 4, it was constant in group 2 (P < 0.001). Immunohistochemical staining and Western blotting analyses showed that E-cadherin and alpha-catenin expression were reversibly decreased in rats with PBOO, while COX-2 protein expression was up-regulated. After giving celecoxib there was a significant decrease in COX-2 expression and a restoration of intercellular adherens junctions and desmosomes, as assessed on electron microscopy and expression of adherens proteins combined. CONCLUSION: The increase in COX-2 expression attributable to hypoxia and the tensile strength of bladder wall was attenuated by celecoxib. Selective COX-2 inhibitors have important restorative effects on intercellular adherens junctions and desmosomes in PBOO.     

Conversion disorder and its subtypes: a need for a reclassification

Four subtypes of conversion disorder were described in DSM-IV. There are few publications concerning studies aimed at separating the subtypes of the conversion disorder. Usually, pseudoseizures are in focus and attempts are made to differentiate these seizures from other disorders. The aim of the present study has been to investigate differences between the four subtypes of the conversion disorder and to discuss the possibilities for a reclassification. Ninety-five patients were seen by two researchers and diagnosed as conversion disorders. The subtypes were determined according to DSM-IV criteria. All completed the Patients Information Form, developed by the researchers, and the Dissociative Experience Scale (DES). Twenty-four (25.2%) of the patients had motor symptoms or deficits (Type 1), 5 (5.2%) sensory symptoms or deficits (Type 2), 23 (24.2%) seizures or convulsions (Type 3) and 43 (47.3%) had mixed presentations (Type 4). There were statistically significant differences between the subtypes as concerns occupation, family history of psychiatric disorders, hospitalizations and place of settlement. Furthermore, the DES scores were statistically different between the groups of patients with different subtypes of conversion disorder.