Frequency-dependent recruitment of inhibition mediated by stellate cells in the rat cerebellar cortex.
In the cerebellum, dendritic inhibition of Purkinje cells (PCs) is mediated by stellate cells (SCs). These inhibitory interneurons are critically involved in the cerebellar network; they control the timing and firing frequency of PCs, the only output cells of the cerebellar cortex. However, the underlying properties of parallel fiber (PF) to SC excitatory synapses have not been fully determined. To characterize the conditions favoring the recruitment of SCs in the cerebellum, we analyzed evoked and spontaneous excitatory postsynaptic currents (EPSCs) recorded from SCs of rat cerebellar slices. We found that SC EPSCs evoked with single suprathreshold-intensity stimulations were mostly unitary, with a large amplitude and variable latencies, and failed with a high rate. Increasing the frequency of stimulation above 60 Hz significantly reduced failures, whereas mean SC EPSC amplitude was increased by less than 20%. Decreasing failures at PF-SC synapses experimentally enhanced the number of asynchronous SC EPSCs per stimulation but, again, moderately increased the mean SC EPSC amplitude. Finally, brief presynaptic bursts transiently depressed synaptic transmission. This depression resulted from the release of endocannabinoids and might act as a negative-feedback mechanism. Thus, we conclude that SC EPSCs evoked with single suprathreshold-intensity stimulations are mostly unitary and that PF-SC synapse efficacy is highly regulated by the presynaptic temporal pattern of activity and the frequency of afferent inputs. Such synaptic properties may control the responsiveness of SC synapses to the frequency of PF stimulations, which may control the spatial extent and duration of the recruitment of inhibition in the cerebellar cortex.


Comprehensive Review on Custodiol-N (HTK-N) and its Molecular Side of Action for Organ Preservation.
The later fate of a graft is highly dependent on its initial quality. Aside from that the three predominant phases during transplantation (Tx) organ retrieval, cold static preservation and reperfusion cause in a direct and indirect manner graft injury. There is complex ischemia reperfusion injury (IRI) triggered during the whole process of Tx which contributes to further damage of the graft. The today's organ preservation with commercially available solutions that all have toxic potential per se is still imperfect. Thus, improved preservation solutions are desperately needed to be able to even safely preserve grafts from extended criteria donors which are more susceptible to especially IRI. Most recently, a modified less toxic histidine-tryptophan-ketoglutarate (HTK; Custodiol®) the so called histidine-tryptophan-ketoglutarate-N (HTK-N) for both better cardioplegia and organ preservation for Tx has been developed. It is characterized as an electrolyte balanced, iron chelatorsupplemented, and amino acid-fortified organ preservation solution with replaced buffer ameliorating resistance to injury during the cold static preservation with subsequent IRI. Numerous in vitro and in vivo experiments have shown the superiority of the HTK-N solution in ROS generation, microcirculation, and subsequent inflammatory response compared with HTK. According to data available to date, HTK-N has both lower cytotoxicity and higher protective potential than HTK. First clinical studies on both HTK-N for cardioplegia in cardiac surgery and for organ preservation for transplantation have been performed or are ongoing. In this review, the novelty and composition of HTK-N and studies investigating the potential of this new solution are focussed, after summarizing the most relevant molecular mechanisms of IRI.


Therapeutic effect of transcranial direct current stimulation on neuropsychological symptoms of an elderly patient: A case report.
Although growing evidence points to the potential therapeutic effects of transcranial Direct Current Stimulation (tDCS), there is still no consensus on the most appropriate protocol to be used in specific neurological and neuropsychological symptoms. This case report evaluated the neuromodulatory therapeutic effects of two 15-day courses of tDCS on an elderly female patient, aged 78 years with mild neurocognitive disorder, chronic pain and depression-related symptoms. Results indicated an overall significant improvement of cognitive and executive functions, as well as reduction in both depression and chronic pain symptoms. These results highlight the potential of tDCS as a safe and useful neuromodulatory clinical tool in the rehabilitation of elderly patients.


Iatrogenic renal disease.
We studied iatrogenic problems in nephrology by classifying all patients for nephrology consultation into nine presenting syndromes and seven etiologic groups. One hundred (2.2%) of all admissions were seen in nephrology consultation. Acute renal failure was the most common presenting syndrome, accounting for 59% of the consultations. Forty-one of the 100 consultations (1% of all admissions) had a renewal syndrome of iatrogenic origin. Of these 41 patients, 38 had acute renal failure and three had fluid and electrolyte problems. Twenty of the 41 patients had drug-induced problems. Eighteen of these patients were dehydrated, and in three patients, acute renal failure occurred after surgery. Of the 20 patients with iatrogenic renal problems caused by drugs, seven problems were antibiotic related, five were due to diuretics, four were due to nonsteroidal anti-inflammatory drugs, three were due to angiotensin-converting enzyme inhibitors, and one was from the use of contrast medium. The 41 patients with iatrogenic-related renal disease were older than the other 59 patients (61.8 vs 49.3 years). Iatrogenic renal disease developed in 1% of all patients admitted to a tertiary care hospital, and 12% of these patients died. The most common renal syndrome is acute renal failure, most often caused by nephrotoxic drugs. The incidence can probably be decreased by better monitoring of body weight and fluid balance to prevent dehydration and by the avoidance of nephrotoxic drugs.


Racial/Ethnic Variations in Acne: A Practical Algorithm for Treatment and Maintenance, Including Skincare Recommendations for Skin of Color Patients With Acne.
Racial/ethnic differences in the clinical presentation, sequelae, and desired treatment outcomes for acne have been reported. Post-inflammatory hyperpigmentation (PIH) frequently occurs in patients with richly pigmented skin complexions and can frequently be the most bothersome aspect of acne in this population. The project used a modified Delphi hybrid process comprising face-to-face discussions followed by an online follow-up. A structured literature search was conducted to identify publications on racial/ethnic differences in the clinical presentation, sequelae, and desired treatment outcomes for skin of color (SOC) patients with acne . The advisors subsequently convened to review the results and draft an algorithm for the treatment and maintenance, including skincare recommendations, for SOC patients with acne. Online, the panel reviewed and adopted the algorithm using published evidence coupled with the panel's expert opinion and clinical experience. Studies suggest that strategies for improving outcomes in patients with acne who have SOC include: the early initiation and maintenance of treatment regimens; careful consideration of the tolerability of active ingredients, vehicle formulations, and dosing; and the use of skin care (eg, pH balanced, non-irritating cleansers, and non-comedogenic moisturizers) to minimize irritation or dryness. Acne treatment in patients with SOC involves unique therapeutic considerations, including management of PIH through efficacious longitudinal acne treatment, prevention of irritation, and potential active treatment of PIH. Skincare products are recommended as an adjunct to prescription therapy to maximize tolerability and may also play a role in maintenance therapy. J Drugs Dermatol. 2022;21:11(Suppl 2):s3-14.


Prolonged haematologic toxicity in CAR-T-cell therapy: A review.
Chimeric antigen receptor-T-cell (CAR-T-cell) therapy is a novel immunotherapy with encouraging results for treatment of relapsed/refractory haematologic malignancies. With increasing use, our understanding of immune-mediated side effects such as cytokine release syndrome and neurotoxicity has improved; nevertheless, prolonged haematologic toxicity (PHT), with a high incidence rate, remains underrecognized. Owing to heterogeneity in populations, the CAR-T cells used and diseases treated as well as differences in the definition of PHT, its rate, risk factors and management vary across studies. In this review, we provide a narrative of PHT occurring in patients following CAR-T-cell therapy; evidence of PHT treatment strategies is also presented, with the aim of contributing to systematic understanding of PHT.


"Trapped in an Empty Waiting Room"-The Existential Human Core of Loneliness in Old Age: A Meta-Synthesis.
Loneliness in old age has a negative influence on quality of life, health, and survival. To understand the phenomenon of loneliness in old age, the voices of lonely older adults should be heard. Therefore, the purpose of this meta-synthesis was to synthesize scientific studies of older adults' experiences of loneliness. Eleven qualitative articles that met the inclusion criteria were analyzed and synthesized according to Noblit and Hare's meta-ethnographic approach. The analysis revealed the overriding meaning of the existential human core of loneliness in old age expressed through the metaphor "trapped in an empty waiting room." Four interwoven themes were found: (a) the negative emotions of loneliness, (b) the loss of meaningful interpersonal relationships, (c) the influence of loneliness on self-perception, and (d) the older adults' endeavors to deal with loneliness. The joint contribution of family members, health care providers, and volunteers is necessary to break the vicious circle of loneliness.


Hygiene measures as primary prevention of toxoplasmosis during pregnancy: A systematic review.
Background Hygiene measures are recommended to prevent toxoplasmosis during pregnancy, although screening for seroconversion in pregnant women currently are debated and practices vary among countries. Objectives The purpose of this systematic literature review was to assess the effectiveness of hygiene measures during pregnancy to prevent toxoplasmosis infection. Search Strategy We followed the standard MOOSE and PRISMA criteria when conducting this systematic review and reporting the results. Selection criteria A systematic literature search was conducted for studies focused on congenital toxoplasmosis prevention, toxoplasmosis prevention during pregnancy, toxoplasmosis prevention and hygiene measures, which were published between 1970 and August 2020, using the databases of PubMed, Scope Med, EMBASE, and the Cochrane library. Data collection and analysis Our literature search identified 3964 articles, 3757 were excluded after review of title or abstract and 67 studies were considered relevant to the subject. We reviewed risk factors for toxoplasmosis infection during pregnancy and for congenital toxoplasmosis, preventive measures for toxoplasmosis during pregnancy, including: dietary recommendations, pet care measures, environmental measures, knowledge of risk factors and ways to control toxoplasmosis infection, knowledge of risk factors for infection by health professionals, knowledge of primary prevention measures by pregnant women. Conclusion: Hygiene measures are effective and applicable primary prevention to reduce toxoplasmosis and avoid congenital toxoplasmosis and its consequences. Funding No.


Does Short Message Service Increase Adherence to Smoking Cessation Clinic Appointments and Quitting Smoking?
Using innovative and scientific methods increases the rate of quitting in smokers. Short message service (SMS) is a communication tool widely used and well integrated in many people's daily lives. To increase adherence to appointments in smoking cessation clinics (SCC), it is thought that increased compliance could be achieved by falling outside the traditional methods. SMS has been shown to increase the compliance of patients with SCC appointments. In this study, we aimed to evaluate the effect of SMS in the compliance of patients with SCC follow-up visits and smoking cessation success. Case-control study. Our study was a controlled, open, prospective study. We enrolled 436 cases applied to SCC of Yedikule Training and Research Hospital between 01.10.2013-30.06.2014 and agreed to follow-up with SMS. SMS was sent to the patients to remind them of appointments at the SCC and to query their smoking state. Two hundred-and-eighty seven (65.8%) of the patients were male and 149 (34.2%) were female. The mean age was 45±12 years. In this study, 296 (67.9%) patients had graduated from primary school. Our patients' smoking state was queried by telephone at the 6-month follow-up and we contacted 348 patients. According to this, 88 (25.3%) patients were not smoking, and 260 (74.7%) patients were smokers. Therefore, the smoking cessation rate was 24% (n=60) in patients who did not respond to SMS reminders at all, and 28.6% (n=28) in patients answering any SMS at least once (p=0.377). Smoking cessation rate of the patients invited by SMS but who did not attend any control visits was 19.1%, and it was 34.5% in patients coming to a control visit at least once. This difference was statistically significant (p=0.001). In our study, there was increased success of smoking cessation in patients coming to control visits. We think that this may result from the possibly increased compliance to SCC appointments following reminders by SMS, and that this may also increase smoking cessation success.


Selective cytotoxic and anti-metastatic activity in DU-145 prostate cancer cells induced by Annona muricata L. bark extract and phytochemical, annonacin.
Annona muricata L. was identified as a popular medicinal plant in treatment regimens among cancer patients in Jamaica by a previously conducted structured questionnaire. Ethnomedically used plant parts, were examined in this study against human prostate cancer cells for the first time and mechanisms of action elucidated for the most potent of them, along with the active phytochemical, annonacin. Nine extracts of varying polarity from the leaves and bark of A. muricata were assessed initially for cytotoxicity using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on PC-3 prostate cancer cells and the ethyl acetate bark (EAB) extract was identified as the most potent. EAB extract was then standardized for annonacin content using High-performance Liquid Chromatography - Mass Spectrometry (HPLC-MS) and shown to be effective against a second prostate cancer cell line (DU-145) also. The mode of cell death in DU-145 cells were assessed via several apoptotic assays including induction of increased reactive oxygen species (ROS) production, reduction of mitochondrial membrane potential, activation of caspases and annexin V externalization combined with morphological observations using confocal microscopy. In addition, the potential to prevent metastasis was examined via inhibition of cell migration, vascular endothelial growth factor (VEGF) and angiogenesis using the chorioallantoic membrane assay (CAM). Annonacin and EAB extract displayed selective and potent cytotoxicity against the DU-145 prostate carcinoma cells with IC50 values of 0.1 ± 0.07 μM and 55.501 ± 0.55 μg/mL respectively, without impacting RWPE-1 normal prostate cells, in stark contrast to chemotherapeutic docetaxel which lacked such selectivity. Docetaxel's impact on the cancerous DU-145 was improved by 50% when used in combination with EAB extract. Insignificant levels of intracellular ROS content, depolarization of mitochondrial membrane, Caspase 3/7 activation, annexin V content, along with stained morphological evaluations, pointed to a non-apoptotic mode of cell death. The extract at 50 μg/mL deterred cell migration in the wound-healing assay, while inhibition of angiogenesis was displayed in the CAM and VEGF inhibition assays for both EAB (100 μg /mL) and annonacin (0.5 μM). Taken together, the standardized EAB extract and annonacin appear to induce selective and potent cell death via a necrotic pathway in DU-145 cells, while also preventing cell migration and angiogenesis, which warrant further examinations for mechanistic insights and validity in-vivo.


Collaborative use of the peer assist model in large transplant programs in the United States.
The basic assumption of the peer assist model is that nearly every transplant center is already doing something that other centers could adopt in order to streamline or enhance their own operations. To describe how the Transplant Growth and Management Collaborative used the peer assist model with 2 large transplant centers in the United States and to identify best practices and outcomes. The University of California, San Francisco, Medical Center. Transplant health care providers (surgeons, nurse transplant coordinators) and administrative staff (program directors, financial advisers, quality representatives). The peer assist model is a mechanism by which a transplant center receives feedback about a specific topic from another transplant center that encountered similar issues and developed effective action plans for overcoming barriers to success; it is a reciprocal model in which knowledge is shared. The peer assist model benefited the preparation for accelerated growth in 2 of the largest transplant centers in the United States. The collaboration during peer assist visits is an opportune time for transplant centers to learn from one another.


[Is there a surgical indication for fracture of the os naviculare pedis?].
In the period from 1986 through 1991 121 patients with injuries of the os naviculare pedis were treated at the Graz Hospital for Accident Surgery. The paper describes our experiences with the diagnosis and surgical management of such lesions. Only 7% of all fractures received surgical treatment. In the overwhelming majority of these cases drill wire osteosynthesis with subsequent plaster cast immobilization proved to be the method of choice, splintered fractures with a tendency to shortening of the limb had to be treated by spongiosa osteoplasty. Follow-up examinations (on average 42.6 weeks after primary management) using the score according to Gainor et al. showed very good and good results in 7 cases and only one negative result. Rotational X-ray examination is indispensable for establishing the diagnosis. Whenever possible, the exact extent of the injury should also be determined by CT.


Immunologically induced sympathectomy of preganglionic nerves by antibodies against acetylcholinesterase: increased levels of peptides and their messenger RNAs in rat adrenal chromaffin cells.
Systemic administration of murine monoclonal acetylcholinesterase antibodies to rats has been shown to cause selective degeneration of sympathetic preganglionic neurons. In the present study rats were subjected to a single i.v. injection of these acetylcholinesterase antibodies, or to normal IgG or saline for control. Exophthalmos, piloerection and eyelid-drooping (ptosis) were observed within 1 h after administration of the antibodies. Rats were killed at different time-points after antibody administration, and the adrenal glands were analysed by means of indirect immunohistochemistry and in situ hybridization histochemistry. As soon as 3 h after the antibody treatment, a marked increase in the number of chromaffin cells expressing mRNA encoding, respectively, enkephalin, calcitonin gene-related peptide, galanin, neurotensin and substance P was seen. At 12 h the peptide mRNA levels were still elevated and there was a concomitant increase in the number of peptide-immunoreactive cells. All peptide levels remained high for at least 48 h; however, 77 days after the antibody treatment only enkephalin-immunoreactive cells could be encountered. A disappearance of acetylcholinesterase- and enkephalin-immunoreactive cells could be encountered. A disappearance of acetylcholinesterase- and enkephalin-positive fibers was already seen 3 h after the antibody treatment, and after 24 h no fibers were encountered. In contrast, up until 48 h there was no apparent change in the number or intensity of immunofluorescent fibers expressing calcitonin gene-related peptide, galanin, neurotensin or substance P. However, 77 days after the antibody treatment the number of calcitonin gene-related peptide- and substance P-immunoreactive fibers was increased as compared to controls. In addition, reappearance of acetylcholinesterase- and enkephalin-immunoreactive fibers was seen 77 days after antibody administration, although their number was still low as compared to controls. Double-labeling immunohistochemistry revealed that the chromaffin cells expressing peptides after the antibody treatment preferentially were adrenaline storing cells (noradrenaline-negative). The majority of these cells expressed only one peptide. Both surgical transection of the splanchnic nerve as well as treatment with acetylcholine receptor antagonists mimicked the effects seen after the acetylcholinesterase-antibody treatment, although changes were less pronounced. The present results show that interruption of splanchnic transmission induces fast, marked, and selective increases in peptide expression in rat adrenal chromaffin cells.


Incidence of Atrial Fibrillation in African Americans post Atrial Flutter Ablation.
African Americans have a lower incidence of atrial arrhythmias both atrial fibrillation (AF) and atrial flutter (AFL) despite having a greater number of traditional risk factors. The incidence of atrial fibrillation after cavotricuspid isthmus (CTI) ablation in patients with typical AFL is markedly increased approaching 40% in some studies. It is unknown if African Americans specifically have a similar rate of increased AF after CTI ablation for typical AFL.We sought to identify differences in development of AF after CTI ablation for typical AFL with regards to race, between African American and non- African Americans. The hypothesis is that African Americans will have a lower incidence of AF after CTI ablation for typical AFL. The electronic medical records of first time, successful CTI ablations performed for typical (AFL) were retrospectively reviewed over a 48 month period. Clinical variables were retrieved from the EMR. AFL was documented on ECG, Holter/ event monitor or device interrogation at clinical follow up visits. Follow up was obtained over a 1 year period. Patients were self-identified as African American, White, Hispanic or Other. The records of a total of 201 patients - 51 African Americans (25.4%) and 150 non-African Americans (74.6%) - who underwent CTI ablation for typical AFL were examined. Average age was similar (67.8 vs 66.3, p=NS) with a preponderance of men (77.6% vs 23.5%). There was a significantly lower rate of development of AF post CTI ablation for typical AFL in African Americans vs non-African Americans (22.2% vs 46.6%, p=0.002). Additionally, women were more likely to develop AF (53.1% vs 36.2%, p=0.026). African American men were the least likely to develop AF post AFL ablation (p=0.0062). We observed a significantly lower incidence of AF among African American patients after CTI ablation for typical AFL.


Cost effectiveness of immune checkpoint inhibitors for treatment of non-small cell lung cancer: A systematic review.
Immune checkpoint inhibitors (ICIs) for treatment of non-small cell lung cancer (NSCLC) have been rapidly evolving. ICIs are likely to be more effective but also lead to escalating healthcare costs. The aim of this study was to evaluate the cost effectiveness of immune checkpoint inhibitors (ICIs) for treatment of non-small cell lung cancer (NSCLC). We searched the PubMed, Web of Science, and Cochrane Library for studies comparing the cost effectiveness of ICIs for NSCLC. Potential studies identified were independently checked for eligibility by two authors, with disagreement resolved by a third reviewer. Quality of the included studies was evaluated using Consolidated Health Economic Evaluation Reporting Standards checklists. A total of 22 economic studies were included. Overall reporting of the identified studies largely met CHEERS recommendations. In the first-line setting, for advanced or metastatic NSCLC patients with PD-L1 ≥ 50%, pembrolizumab appeared cost-effective compared with platinum-based chemotherapy in the US and Hong Kong (China), but not in the UK and China. The cost-effectiveness of pembrolizumab versus chemotherapy for first-line treatment of NSCLC in PD-L1 ≥ 1% patients remained obscure. Regardless of PD-L1 expression status, pembrolizumab in combination with chemotherapy could be a cost-effective first-line therapy in the US. On the contrary, addition of atezolizumab to the combination of bevacizumab and chemotherapy was not cost-effective for patients with metastatic non-squamous NSCLC from the US payer perspective. In the second-line setting compared with docetaxel, pembrolizumab was cost-effective; though nivolumab was not cost-effective in the base case, it could be by increased PD-L1 threshold. Results of the cost-effectiveness of atezolizumab second-line treatment remained inconsistent. In addition, the adoption of durvalumab consolidation therapy after chemoradiotherapy could be cost-effective versus no consolidation therapy for patients with stage III NSCLC. Immunotherapy can be a cost-effective option for treatment of NSCLC in several scenarios. A discount of the agents or the use of PD-L1 expression as a biomarker improves the cost-effectiveness of immunotherapy.


Acute lethal toxicity following passive immunization for treatment of murine cryptococcosis.
Passive immunization with monoclonal antibodies (MAbs) specific for the major capsular polysaccharide of Cryptococcus neoformans alters the course of murine cryptococcosis. During studies of passive immunization for treatment of murine cryptococcosis, we noted the occurrence of an acute, lethal toxicity. Toxicity was characterized by scratching, lethargy, respiratory distress, collapse, and death within 20 to 60 min after injection of antibody. The toxic effect was observed only in mice with a cryptococcal infection and was reduced or absent in the early and late stages of disease. The clinical course and histopathology were consistent with those for shock. There was considerable variation between mouse strains in susceptibility to toxicity. Swiss Webster mice from the Charles River colony were most susceptible, followed by C3H/He, BALB/c, and C57BL/6 mice. DBA/2 mice and Swiss Webster mice from the Simonsen colony were resistant. Acute toxicity was mimicked by injection of preformed complexes of MAb and purified polysaccharide. The toxic effect was also produced by injection of MAbs into mice that were preloaded with polysaccharide. The toxic effect was not blocked by treatment of mice with chloropheniramine or anti-tumor necrosis factor alpha antibodies or by depletion of complement components via pretreatment with cobra venom factor. Toxicity was reduced by treatment of mice with high doses of epinephrine, dexamethasone, or chlorpromazine. Finally, the toxic effect was completely blocked by treatment of mice with the platelet-activating factor antagonist WEB 2170 BS or by pretreatment of mice with the liposome-encapsulated drug dichloromethylene diphosphonate, a procedure which depletes macrophages from the spleen and liver.


Teaching non-specialist health care professionals how to identify the atypical mole syndrome phenotype: a multinational study.
The atypical mole syndrome (AMS) phenotype is the strongest known risk factor for cutaneous melanoma but recognition of the phenotype has been claimed to be problematic and to require specialist assessment. This study determined the ability of previously unskilled doctors and nurses in five countries to recognize the phenotype after brief training. The system used was the AMS scoring system. This incorporates melanocytic naevus counts, clinical atypia of naevi and distribution of naevi. The agreement in scoring between the dermatologist and trained personnel was determined in 986 patients; overall agreement in diagnosis was 94.5% (kappa 0.70, P < 0.0001). The kappa scores in different countries ranged from 0.65 to 0.77 for individual naevus characteristics, indicative of good agreement. Accurate diagnosis of the atypical mole syndrome phenotype is possible by non-specialists. This has implications for collaborative studies of naevi, for screening and for both primary and secondary prevention of melanoma.


Survey and Performance Analysis of Deep Learning Based Object Detection in Challenging Environments.
Recent progress in deep learning has led to accurate and efficient generic object detection networks. Training of highly reliable models depends on large datasets with highly textured and rich images. However, in real-world scenarios, the performance of the generic object detection system decreases when (i) occlusions hide the objects, (ii) objects are present in low-light images, or (iii) they are merged with background information. In this paper, we refer to all these situations as challenging environments. With the recent rapid development in generic object detection algorithms, notable progress has been observed in the field of deep learning-based object detection in challenging environments. However, there is no consolidated reference to cover the state of the art in this domain. To the best of our knowledge, this paper presents the first comprehensive overview, covering recent approaches that have tackled the problem of object detection in challenging environments. Furthermore, we present a quantitative and qualitative performance analysis of these approaches and discuss the currently available challenging datasets. Moreover, this paper investigates the performance of current state-of-the-art generic object detection algorithms by benchmarking results on the three well-known challenging datasets. Finally, we highlight several current shortcomings and outline future directions.


Targeting the expression of functional murine CMP-sialic acid transporter to the E. coli inner membrane.
The heterologous expression of functional mammalian integral membrane proteins still represents a significant hurdle towards their crystallization and structure elucidation. We have therefore explored the use of the OmpA signal sequence to deliberately target the expression of the murine CMP-sialic acid transporter, a Golgi-resident protein with 10 putative transmembrane domains, to the Escherichia coli inner membrane. Here, we show that the expression of an OmpA signal sequence-FLAG-CMP-sialic acid transporter fusion protein in E. coli results in the targeting and insertion of recombinant protein within the inner membrane. Significantly, functionality was confirmed by the ability of spheroplasted E. coli and mixed phosphatidylcholine-E. coli inner membrane proteoliposomes incorporating recombinant CMP-sialic acid transporter to accumulate CMP-sialic acid in vitro.


Neuropathic pain inhibitor, RAP-103, is a potent inhibitor of microglial CCL1/CCR8.
Chemokine signaling is important in neuropathic pain, with microglial cells expressing chemokine (C-C motif) receptor CCR2, CCR5 and CCR8, all playing key roles. In the previous report (Padi et al., 2012), oral administration of a short peptide, RAP-103, for 7 days fully prevents mechanical allodynia and inhibits the development of thermal hyperalgesia after partial ligation of the sciatic nerve in rodents. As for the mechanism of the inhibiting effect of RAP-103, it was speculated to be due to dual blockade of CCR2 and CCR5. We report here that RAP-103 exhibits stronger antagonism for CCR8 (half maximal inhibitory concentration [IC50] 7.7 fM) compared to CCR5 (IC50 < 100 pM) in chemotaxis using primary cultured mouse microglia. In addition, RAP-103 at a concentration of 0.1 pM completely inhibits membrane ruffling and phagocytosis induced by chemokine (C-C motif) ligand 1 (CCL1), an agonist for CCR8. It has been shown that CCL1/CCR8 signaling is important in tactile allodynia induced by nerve ligation. Therefore, CCR8, among other chemokine receptors such as CCR2/CCR5, could be the most potent target for RAP-103. Inhibitory effects of RAP-103 on plural chemokine receptors may play important roles for broad clinical use in neuropathic pain treatment.


SMARCB1/INI1-Deficient Poorly Differentiated Carcinoma of the Colon With Rhabdoid Features-A Rare Tumor With Serrated Phenotype: Case Report and Review of Literature.
Poorly differentiated colonic carcinoma with rhabdoid features is a rarely described entity. Our knowledge regarding the molecular phenotype of the tumor is evolving. We herein report a similar tumor with rhabdoid differentiation identified in the splenic flexure, which on histological examination showed a poorly differentiated phenotype with epithelioid to spindled morphology, tumor giant cells, and rhabdoid differentiation. The tumor was mismatch repair-proficient, deficient of INI1/SMARCB1, KRAS mutated (A146×), BRAFV600E mutated (c.1799T > A), and NRAS wild-type, indicating serrated differentiation in the tumor. The patient died after 3.5 months post-surgery. INI1-deficient poorly differentiated carcinoma of the colon is a rare, aggressive colonic malignancy showing a serrated phenotype. Routine identification and subtyping are important keeping in mind the distinct tumor phenotype, resistance to conventional chemotherapy, and dismal prognosis.


Accuracy and responses of genomic selection on key traits in apple breeding.
The application of genomic selection in fruit tree crops is expected to enhance breeding efficiency by increasing prediction accuracy, increasing selection intensity and decreasing generation interval. The objectives of this study were to assess the accuracy of prediction and selection response in commercial apple breeding programmes for key traits. The training population comprised 977 individuals derived from 20 pedigreed full-sib families. Historic phenotypic data were available on 10 traits related to productivity and fruit external appearance and genotypic data for 7829 SNPs obtained with an Illumina 20K SNP array. From these data, a genome-wide prediction model was built and subsequently used to calculate genomic breeding values of five application full-sib families. The application families had genotypes at 364 SNPs from a dedicated 512 SNP array, and these genotypic data were extended to the high-density level by imputation. These five families were phenotyped for 1 year and their phenotypes were compared to the predicted breeding values. Accuracy of genomic prediction across the 10 traits reached a maximum value of 0.5 and had a median value of 0.19. The accuracies were strongly affected by the phenotypic distribution and heritability of traits. In the largest family, significant selection response was observed for traits with high heritability and symmetric phenotypic distribution. Traits that showed non-significant response often had reduced and skewed phenotypic variation or low heritability. Among the five application families the accuracies were uncorrelated to the degree of relatedness to the training population. The results underline the potential of genomic prediction to accelerate breeding progress in outbred fruit tree crops that still need to overcome long generation intervals and extensive phenotyping costs.


First Report of Sydowia polyspora on Aleppo Pine (Pinus halepensis) in Italy.
Pinus halepensis Mill. is a pine native to the Mediterranean region that is generally located from sea level up to an altitude of 200 meters. In July 2012, extensive leaf yellowing and scorching were observed on the foliage of two specimens of P. halepensis in a public park of Alassio municipality, Liguria region (northern Italy). Diseased needles showed chlorotic and reddish brown colored areas that were randomly distributed among healthy needles. In order to isolate the potential pathogen, diseased needle tissues were surface sterilized for 1 min in 1% NaOCl and plated onto potato dextrose agar (PDA) amended with 100 ppm of streptomycin sulfate. After 7 to 10 days, a brown to dark green mycelium slowly developed on the PDA. From the mycelium, single cell conidia averaging 7.0 (4.7 to 12.2) × 2.9 (2.4 to 5.4) μm (n = 50) were produced. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS1f/ITS4 and sequenced. A BLAST (1) search yielded 100% of maximum identity with Sydowia polyspora (Bref. & Tav.) E. Müller for ITS1f and ITS4. Pathogenicity was confirmed by inoculating 15 3-year-old plants (approximately 5 months after bud break) of P. halepensis grown singly in 8-cm diameter pots and maintained in greenhouse. Twelve of 15 plants were wounded by gently rubbing needles together. Five non-inoculated plants, with wounded and non-wounded branches, were kept as controls. Inoculations were carried out at 16.5 to 18.5°C. A suspension of S. polyspora conidia was made by adding 1 to 2 ml sterile water to the spore mass on 1-month-old PDA cultures in 9-cm petri dishes. Inoculum (107 spores/ml) was applied on the needles with a soft paint brush. After inoculation plants were covered with polythene bags for 5 days, kept at temperatures ranging from 5.5 to 26.4°C (average 14.5°C), and watered as needed. The inoculation trial was repeated once. All inoculated plants, both wounded and non-wounded, developed leaf yellowing and browning since the fifth day after inoculation in both inoculation tests, while control plants remained symptomless. After 15 days from first symptom appearance, S. polyspora was re-isolated from the needles of inoculated plants according to the procedure already described. S. polyspora is associated with current season needle necrosis (CSNN) on various conifer species. P. halepensis was reported in Spain to be a susceptible host (2) and S. polyspora has been isolated from infected tissues of fir (Abies spp.) (3). To our knowledge, this is the first report of S. polyspora on P. halepensis in Italy. Control of the pathogen with fungicides was ineffective on fir species, while application of very high rates of calcium chloride during shoot elongation was able to reduce the severity of CSNN (3). In forest areas, municipality gardens, and parks, effective management strategies have not yet been developed. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) L. Botella et al. Fungal Diversity 40:1, 2010. (3) V. Talgø et al. Fungal Biol. 114:545, 2010.


Proteolytic conversion of insulin-like growth factors to an acidic form(s).
The relative amounts of the various forms of bioassayable insulin-like growth factors (IGF) isolated from human serum or serum fraction Cohn IV-1 depend on the purification procedure. With acid gel filtration or acid/ethanol extraction as the initial step, IGF-II (pI approximately 6.5) was the most abundant (40-70%) followed by somatomedin A (pI approximately 7.4; 15-23%), an acidic form of insulin-like activity (ILA pI 4.8) (13-21%) and IGF-I (pI approximately 8.5; 5-27%). If, however, pH 5.5 ion-exchange chromatography on SP-Sephadex was used prior to acid gel filtration, the acidic pI 4.8 form was the major (greater than 90%) species recovered and was accompanied by a quantitative loss of the other IGF species. This suggested a possible conversion of IGF-I, somatomedin A and/or IGF-II to the acidic ILA pI 4.8 form(s) during the SP-Sephadex procedure. Further experiments indicated that differences in the yields of ILA pI 4.8 were not due simply to differences in the initial pH conditions of the various methods (i.e. acid versus neutral), although exposure to pH 9.7 (a pH experienced during elution of IGF activity from the SP-Sephadex) did appear to play a role. The involvement of the carrier protein in the conversion process was tested by subjecting carrier-free IGF-I and IGF-II to the SP-Sephadex procedure. No conversion of the free forms to ILA pI 4.8 occurred. To examine the possible role of proteinase in the conversion of IGFs to ILA pI 4.8, SP-Sephadex chromatography was performed in the presence of a broad spectrum proteinase inhibitor. The IGF distribution pattern obtained closely resembled the 'normal' pattern seen with acid gel filtration, indicating that proteinase inactivation had prevented conversion to ILA pI 4.8. These data suggest that proteolytic conversion of IGF-I, somatomedin A and IGF-II to more acidic ILA pI 4.8 form(s) (i) occurs during SP-Sephadex chromatography, (ii) is not prevented simply by prior acid exposure, and (iii) takes place only when IGF-I and -II are in their high-Mr carrier-bound forms. Since IGF-I and IGF-II, although homologous, have unique amino acid sequences, the conversion of both IGFs implies that at least two acidic ILA forms exist. Nevertheless, because ILA pI 4.8 retains the full spectrum of IGF bioactivities in vitro, and significant quantities are present in normal human serum (21%), it would suggest that proteolytic conversion of IGF-I, somatomedin A and IGF-II to ILA pI 4.8 in vivo may be a physiologically significant event.


INFLUENCE OF PREGNANCY ON NEUROMYELITIS OPTICA FROM AN OPHTHALMOLOGICAL POINT OF VIEW. A CASE REPORT.
Neuromyelitis Optica (NMO, Devics disease) is a rare demyelinating disease of the central nervous system, leading to optic neuritis and transverse myelitis. The clinical course of the disease and ocular changes in pregnancy are still not well known. Here we present the subclinical ophthalmological changes which were evaluated by spectral domain optical coherence tomography (SD-OCT) during the 39 weeks gestation and postpartum period in a pregnant woman with a diagnosis of NMO. In addition, we present the obstetric and neurological course of our patient and review the literature. A 30-year-old female with a history of NMO was ophthalmologically examined and SD-OCT was performed periodically every trimester to observe the effects of pregnancy on the disease course. No ophthalmological changes were observed during the pregnancy and postpartum period. Caesarean delivery was preferred due to obstetric indication. The patient was discharged on the second day of the postpartum period.


Sexual and Affectionate Behaviors in Asexual and Allosexual Adults.
Prior experience with physical behaviors - both sexual and affectionate - is common among adults in romantic relationships. However, less is known about differences in physical behaviors for asexual and allosexual adults, and these differences may explain how asexual adults navigate sexuality in romantic relationships. In this study we used sexual script theory to examine asexual (n = 109; 59% women; Mage = 27.7) and allosexual adults' (n = 403; 58% women; Mage = 29.0) prior experiences with and future likelihood to engage in sexual and affectionate behaviors in romantic relationships, and how these differences vary by gender. Asexual adults had less prior experience with and were less likely to intend to engage in future physical behaviors than allosexual adults. There were some gender differences in past experience and future likelihood. Prior experience with physical behaviors was associated with expected likelihood to engage in these behaviors for allosexual and asexual adults, although the association was stronger for asexual adults. These results have implications for how asexual adults engage in physical behaviors, and how differing interest in physical behaviors may cause conflict within asexual-allosexual couples.


The influence of self-absorption on the photoluminescence of thin film CdS demonstrated by two-photon absorption.
By means of two-photon excited photoluminescence, we demonstrate the influence of self-absorption on the emission properties of thin (1.5 microm) film CdS formed by laser ablation. The excitation of the sample is performed with 200 fs pulses at 804 nm (1.54 eV). The photoluminescence spectrum takes the form of a single peak centered at 510 nm (2.43 eV) at 300 K. The spectrum is shifted about 45 meV to lower energies with respect to the photoluminescence excited by one photon absorption. By fitting the photoluminescence spectra with the Roosbroeck-Shockley relation and Urbach's rule, it is shown by Beer's law that the shift is caused by self-absorption. The results further provide evidence of low impurity concentration and excellent surface quality. They also confirm the outstanding optical properties of thin film CdS formed by pulsed-laser deposition and suggest the application of the films for effective up-conversation materials in ultra-fast experiments.


Integrating the science of team training: guidelines for continuing education.
The provision of high-quality, efficient care results from the coordinated, cooperative efforts of multiple technically competent health care providers working in concert over time, spanning disciplinary and professional boundaries. Accordingly, the role of medical education must include the development of providers who are both expert clinicians and expert team members. However, the competencies underlying effective teamwork are only just beginning to be integrated into medical school curricula and residency programs. Therefore, continuing education (CE) is a vital mechanism for practitioners already in the field to develop the attitudes, behaviors (skills), and cognitive knowledge necessary for highly reliable and effective team performance.The present article provides an overview of more than 30 years of evidence regarding team performance and team training in order to guide, shape, and build CE activities that focus on developing team competencies. Recognizing that even the most comprehensive and well-designed team-oriented CE programs will fail unless they are supported by an organizational and professional culture that values collaborative behavior, ten evidence-based lessons for practice are offered in order to facilitate the use of the science of team-training in efforts to foster continuous quality improvement and enhance patient safety.


Video-assisted interventional bronchoscopy. The Hong Kong experience.
Major airway obstruction due to benign or malignant etiology is not uncommon and is always distressing. Intraluminal stenting has been shown to be a safe and effective approach for symptomatic relief in selected patients based on the European and North American experience. We reviewed our experience in Hong Kong on airway stenting over a 19-month period. From February 1994 to August 1995, 33 silicone stents (Dumon stent, Cometh, Marseille, France) were placed in 23 patients (20 males, three females with mean age 61.4 years, range from 26 to 81). Eighteen stents were placed in the trachea, nine in the left main stem, five in the right main stem, and one Y-stent over the carina. Twelve patients had esophageal carcinoma involving the airway, seven had bronchial carcinoma, one had metastatic carcinoma, and three had benign strictures (of which two were due to tuberculosis). There was no procedural related mortality. Stent migration occurred in four patients (17%) and required stent change. Symptoms were improved in all patients as documented by the visual analogue scale. Our experience represents the "stentable" diseases seen in Hong Kong, where carcinoma of the esophagus (and tuberculosis) remains prevalent. We conclude that intraluminal stenting remains a safe and effective approach in selected patients with critical airway stenosis. Complications, however, do exist and should be realized by the operator, the patients, and their families.


Inter-Individual Variability in Metabolic Syndrome Severity Score and VO2max Changes Following Personalized, Community-Based Exercise Programming.
This study sought to examine the effectiveness of a personalized, community-based exercise program at reducing MetS severity and consequently Type 2 diabetes mellitus (T2DM) and cardiovascular disease (CVD) risk. One-hundred and fifty physically inactive participants (aged 18-83 years) were randomized to a non-exercise control group (n = 75; instructed to continue their usual lifestyle habits) or treatment group (n = 75). Participants randomized to the treatment group completed a 12 week personalized exercise training program based on the American Council on Exercise (ACE) Integrated Fitness Training (IFT) model guidelines. Z-scores were derived from levels of metabolic syndrome risk factors to determine the severity of MetS (MetS z-score). After 12 weeks, the treatment group showed a significant favorable change in MetS z-score, whereas the control group demonstrated increased severity of the syndrome (between-group difference, p < 0.05). The proportion of MetS z-score responders (Δ > -0.48) was greater following the exercise intervention (71%, 50/70) compared to control (10%, 7/72) (between group difference, p < 0.001). The inter-individual variability in VO2max change also showed a similar trend. These findings provide critical translational evidence demonstrating that personalized exercise programming based upon the ACE IFT model guidelines can be successfully implemented within the community setting to reduce T2DM and CVD risk.


TARGETING THE NS5A PROTEIN OF HCV: AN EMERGING OPTION.
Hepatitis C virus (HCV) infects more than 3% of the world's population, leading to an increased risk of cirrhosis and hepatocellular carcinoma. The current standard of care, a combination of pegylated interferon alfa and ribavirin, is poorly tolerated and often ineffective against the most prevalent genotype of the virus, genotype 1. The very recent approval of boceprevir and telaprevir, two HCV protease inhibitors, promises to significantly improve treatment options and outcomes. In addition to the viral protease NS3 and the viral polymerase NS5B, direct-acting antivirals are now in development against NS5A. A multifunctional phosphoprotein, NS5A is essential to HCV genome replication, but has no known enzymatic function. Here we report how the design of small-molecule inhibitors against NS5A has evolved from promising monomers to highly potent dimeric compounds effective against many HCV genotypes. We also highlight recent clinical data and how the inhibitors may bind to NS5A, itself capable of forming dimers.


Heart rate variability.
Reduced heart rate variability carries an adverse prognosis in patients who have survived an acute myocardial infarction. This article reviews the physiology, technical problems of assessment, and clinical relevance of heart rate variability. The sympathovagal influence and the clinical assessment of heart rate variability are discussed. Methods measuring heart rate variability are classified into four groups, and the advantages and disadvantages of each group are described. Concentration is on risk stratification of postmyocardial infarction patients. The evidence suggests that heart rate variability is the single most important predictor of those patients who are at high risk of sudden death or serious ventricular arrhythmias.


[Current clinical aspects in hemophilia treatment].
The introduction of factor VIII and IX concentrates in the early 1960s brought a significant change in the hemophiliac's life. In consequence hemophilia treatment has been improving rapidly since, and today most life-threatening hemorrhages are controlled by replacement therapy. Hemophilic arthropathy through recurrent joint and muscle bleedings occurs later in life and is often limited to one joint only. Major surgery in hemophiliacs involves little more risk than in non-hemophilic patients, provided of course there is close teamwork between surgeon and hematologist. The most frequent causes of death are no longer hemorrhages but blood-product-associated AIDS and hepatic failure. Fortunately these side effects have been overcome by the use of virus-inactivated concentrates which in Switzerland have been generally administered since 1986. Factor VIII and IX concentrates must contain a precisely declared quantity of factor VIII and IX activity respectively, with a high specific activity. High-purity concentrates should be preferred because of the hazardous effect of foreign proteins administered intravenously in large quantities over a long period. Activation of fibrinolysis with consequent failure of hemostasis or even worsening of hemorrhage may be a clinically relevant side-effect of DDAVP therapy. When DDAVP is used for prophylactic treatment before surgery, an interval of one hour between the intravenous administration of DDAVP and surgery ensures the latter is performed at the time of highest factor VIII and von Willebrand factor level but with already decreased t-PA and fibrinolytic activity. If DDAVP is used in case of hemorrhage or postoperatively, however, the whole fibrinolytic potential must be taken into account. In these cases subcutaneous administration is advantageous due to more protracted t-PA release and the subsequent lower fibrinolytic activity, which can more easily be neutralized by tranexamic acid. To prevent hemophilic arthropathy, correct replacement therapy in hemarthroses is essential: it should be performed as early as possible, preferably in a home therapy program; adequate levels of factor VIII or IX should be achieved and maintained over a sufficient length of time. Hemophiliacs who did not receive replacement therapy during childhood often need major surgery because of severely destructed joints. Joint replacement by total knee and hip prostheses has proved very successful if certain special conditions are fulfilled. Surgical indications should, however, be carefully considered and the possibilities and limits of replacement therapy should be well known. Blood-product-associated hepatitis will be of prognostic relevance in many hemophiliacs treated formerly with non-virus-inactivated concentrates.(ABSTRACT TRUNCATED AT 400 WORDS)


Sympathetic and parasympathetic innervation in cancer: therapeutic implications.
The autonomic nervous system, consisting of sympathetic and parasympathetic/vagal nerves, is known to control the functions of any organ, maintaining whole-body homeostasis under physiological conditions. Recently, there has been increasing evidence linking sympathetic and parasympathetic/vagal nerves to cancers. The present review aimed to summarize recent developments from studies addressing the relationship between sympathetic and parasympathetic/vagal nerves and cancer behavior. Literature review. Human and animal studies have revealed that sympathetic and parasympathetic/vagal nerves innervate the cancer microenvironment and alter cancer behavior. The sympathetic nerves have cancer-promoting effects on prostate cancer, breast cancer, and melanoma. On the other hand, while the parasympathetic/vagal nerves have cancer-promoting effects on prostate, gastric, and colorectal cancers, they have cancer-suppressing effects on breast and pancreatic cancers. These neural effects may be mediated by β-adrenergic or muscarinic receptors and can be explained by changes in cancer cell behavior, angiogenesis, tumor-associated macrophages, and adaptive antitumor immunity. Sympathetic nerves innervating the tumor microenvironment promote cancer progression and are related to stress-induced cancer behavior. The parasympathetic/vagal nerves have variable (promoting or suppressing) effects on different cancer types. Approaches directed toward the sympathetic and parasympathetic/vagal nerves can be developed as a new cancer therapy. In addition to existing pharmacological, surgical, and electrical approaches, a recently developed virus vector-based genetic local neuroengineering technology is a powerful approach that selectively manipulates specific types of nerve fibers innervating the cancer microenvironment and leads to the suppression of cancer progression. This technology will enable the creation of "cancer neural therapy" individually tailored to different cancer types.


A vector boundary matching technique for efficient and accurate determination of photonic bandgaps in photonic bandgap fibers.
A vector boundary matching technique has been proposed and demonstrated for finding photonic bandgaps in photonic bandgap fibers with circular nodes. Much improved accuracy, comparing to earlier works, comes mostly from using more accurate cell boundaries for each mode at the upper and lower edges of the band of modes. It is recognized that the unit cell boundary used for finding each mode at band edges of the 2D cladding lattice is not only dependent on whether it is a mode at upper or lower band edge, but also on the azimuthal mode number and lattice arrangements. Unit cell boundaries for these modes are determined by mode symmetries which are governed by the azimuthal mode number as well as lattice arrangement due to mostly geometrical constrains. Unit cell boundaries are determined for modes at both upper and lower edges of bands of modes dominated by m = 1 and m = 2 terms in their longitudinal field Fourier-Bessel expansion series, equivalent to LP0s and LP1s modes in the approximate LP mode representations, for hexagonal lattice to illustrate the technique. The novel technique is also implemented in vector form and incorporates a transfer matrix algorithm for the consideration of nodes with arbitrary refractive index profiles. Both are desired new capabilities for further explorations of advanced new designs of photonic bandgap fibers.


Accuracy of pulse oximetry screening for detecting critical congenital heart disease in the newborns in rural hospital of Central India.
Congenital cardiovascular malformations are the most common category of birth defects and responsible for mortality in the first twelve months of life. Critical congenital heart disease (CCHD) will be present in approximately one quarter of these children, which requires catheter or surgery intervention in the first year of life. To determine the accuracy of pulse oximetry for detecting clinically unrecognized CCHD in the newborns. Pulse oximetry was performed on clinically normal newborns within first 4 hours of life. If screening oxygen saturation (SpO2) was below 90%, echocardiography was then performed. All newborns who were admitted in postnatal ward & NICU. Out born babies and babies with a prenatal diagnosis of duct dependent circulation. Pulse-oximetric screening was performed on 2110 clinically normal newborns. Low SpO2 (<90%) was found in eight babies seven of them had CCHD, including three neonates with TGA, two with tricuspid atresia, one with tetralogy of fallot and one with transposition of great vessels. A pulse oximetry cut-off value of below 90% for detecting CCHD showed 100% sensitivity, 99.95% specificity, 87.50% positive predictive value, 100% negative predictive value. Pulse oximetry is safe, feasible and noninvasive, can be used as screening tool for detecting CCHD in clinically normal newborn. If SpO2 is below 90% in clinically normal newborns within 4 hours of birth, urgent echocardiography is suggested to rule out CCHD.


Osteoid osteoma in a cervical pedicle. Resection via transpillar approach.
Lesions in the pedicles of the cervical spine are both a diagnostic and technical challenge. An osteoid osteoma of the pedicle of the fourth cervical vertebra occurred in a 17-year-old girl. Resection of the tumor adjacent to the dura medially and the vertebral artery anterolaterally was accomplished via a transpillar approach. The transosseus approach preserved the facet joints and posterior elements and obviated the need for an interbody fusion.


Understanding the process of patient satisfaction with nurse-led chronic disease management in general practice.
To investigate the process of patient satisfaction with nurse-led chronic disease management in Australian general practice. Nurses working in the primary care context of general practice, referred to as practice nurses, are expanding their role in chronic disease management; this is relatively new to Australia. Therefore, determining patient satisfaction with this trend is pragmatically and ethically important. However, the concept of patient satisfaction is not well understood particularly in relation to care provided by practice nurses. A grounded theory study underpinned by a relativist ontological position and a relativist epistemology. Grounded theory was used to develop a theory from data collected through in-depth interviews with 38 participants between November 2007-April 2009. Participants were drawn from a larger project that trialled a practice nurse-led, collaborative model of chronic disease management in three Australian general practices. Theoretical sampling, data collection, and analysis were conducted concurrently consistent with grounded theory methods. Patients undergo a cyclical process of Navigating Care involving three stages, Determining Care Needs, Forming Relationship, and Having Confidence. The latter two processes are inter-related and a feedback loop from them informs subsequent cycles of Determining Care Needs. If any of these steps fails to develop adequately, patients are likely to opt out of nurse-led care. Navigating Care explains how and why time, communication, continuity, and trust in general practitioners and nurses are important to patient satisfaction. It can be used in identifying suitable patients for practice nurse-led care and to inform the practice and organization of practice nurse-led care to enhance patient satisfaction.


H3K14ac is linked to methylation of H3K9 by the triple Tudor domain of SETDB1.
SETDB1 is an essential H3K9 methyltransferase involved in silencing of retroviruses and gene regulation. We show here that its triple Tudor domain (3TD) specifically binds to doubly modified histone H3 containing K14 acetylation and K9 methylation. Crystal structures of 3TD in complex with H3K14ac/K9me peptides reveal that peptide binding and K14ac recognition occurs at the interface between Tudor domains (TD) TD2 and TD3. Structural and biochemical data demonstrate a pocket switch mechanism in histone code reading, because K9me1 or K9me2 is preferentially recognized by the aromatic cage of TD3, while K9me3 selectively binds to TD2. Mutations in the K14ac/K9me binding sites change the sub-nuclear localization of 3TD. ChIP-seq analyses show that SETDB1 is enriched at H3K9me3 regions and K9me3/K14ac is enriched at SETDB1 binding sites overlapping with LINE elements, suggesting that recruitment of the SETDB1 complex to K14ac/K9me regions has a role in silencing of active genomic regions.


Semiautomatic quantification of left ventricular function by two-dimensional feature tracking imaging echocardiography. A comparison study with cardiac magnetic resonance imaging.
To evaluate the accuracy of a semiautomatic quantification of left ventricular (LV) volumes and ejection fraction (EF) using two-dimensional (2D) feature tracking imaging (FTI). Thirty-four consecutive subjects (11 patients with dilated cardiomyopathy, 13 with hypertrophic cardiomyopathy, and 10 subjects with no cardiac disease) underwent, on the same day, trans-thoracic echocardiography (TTE) examination, FTI, and cardiac magnetic resonance imaging (MRI), as gold standard, in order to quantify LV volumes and EF. The echocardiographic quantification of LV volumes and EF was determined from four- and two-chamber views using both standard TTE Biplane Simpson's method and a semiautomatic border detection based on FTI. Furthermore, the time for data analysis for each method was measured. The time required for semiautomatic analysis of volumes and EF was significantly lower (P < 0.0001) by FTI (71 seconds) in comparison with standard biplane Simpson's method (93 seconds). LV volumes obtained by FTI were significant underestimated (P < 0.001) in comparison with MRI. Bland-Altman analysis of EDV and ESV using FTI and cardiac MRI showed a low level of agreement for EDV (mean difference = 40.8; SD = 39) and ESV (mean difference = 38.1; SD = 42). On the contrary, no significant difference between FTI and MRI in assessing the LVEF was found; furthermore, a very low bias (2 ± 12) by Bland-Altman analysis was found between FTI and cardiac MRI for the quantification of EF. Semiautomatic quantification of LV volumes using FTI allows an accurate, rapid, easy and reliable assessment of LV EF and a rough estimation of LV volumes.


Validation of a single point flow cytometric assay for determining P-glycoprotein activity in multidrug resistant cell lines.
P-glycoprotein, a transmembrane protein which acts as an energy dependent efflux pump, has been implicated as one mechanism of multidrug resistance (MDR) in human tumours. Commonly employed assays measure P-glycoprotein immunohistochemically or mdr1 messenger RNA. In this study we compared a single point flow cytometric assay for determining activity of P-glycoprotein with cellular expression of P-glycoprotein determined by Western blot. Five cell lines, with varying levels of multiple drug resistance, were incubated with daunorubicin (DNR) in the presence (treated) and absence (control) of cyclosporine or verapamil, agents known to inhibit the activity of P-glycoprotein. The treated cell lines, along with non-treated controls were examined for intracellular concentrations of DNR measured by fluorescence intensity using a flow cytometer. The ratio of fluorescence intensity expressed in the treated/control was used as an index of functional activity of P-glycoprotein. Functional activity of the P-glycoprotein as determined by flow cytometry correlates highly with cellular content of P-glycoprotein measured by western blot (correlation coefficients of r = 0.90-0.98 for the various cell line combinations). This method represents a rapid single point flow cytometric assay which may be suitable for screening clinical samples for P-glycoprotein activity.


Confocal endomicroscopy: instrumentation and medical applications.
Advances in fiber optic technology and miniaturized optics and mechanics have propelled confocal endomicroscopy into the clinical realm. This high resolution, non-invasive imaging technology provides the ability to microscopically evaluate cellular and sub-cellular features in tissue in vivo by optical sectioning. Because many cancers originate in epithelial tissues accessible by endoscopes, confocal endomicroscopy has been explored to detect regions of possible neoplasia at an earlier stage by imaging morphological features in vivo that are significant in histopathologic evaluation. This technique allows real-time assessment of tissue which may improve diagnostic yield by guiding biopsy. Research and development continues to reduce the overall size of the imaging probe, increase the image acquisition speed, and improve resolution and field of view of confocal endomicroscopes. Technical advances will continue to enable application to less accessible organs and more complex systems in the body. Lateral and axial resolutions down to 0.5 and 3 μm, respectively, field of view as large as 800 × 450 μm, and objective lens and total probe outer diameters down to 0.35 and 1.25 mm, respectively, have been achieved. We provide a review of the historical developments of confocal imaging in vivo, the evolution of endomicroscope instrumentation, and the medical applications of confocal endomicroscopy.


The relationships between neuroticism and depressive symptoms.
A structural equation model designed to estimate the reciprocal associations which may exist between mental state and personality trait variables is presented. This model is applied to measures of neuroticism and depression obtained for a sample of over 1000 women studied over a two year period. The fitted model suggests that while measures of neuroticism are contaminated by the effects of short term mental state on the reporting of personality, there is still a fairly substantial relationship between trait neuroticism (corrected for the effects of mental state contamination) and reports of depressive symptoms.


Mapping of single-base differences between two DNA strands in a single molecule using holliday junction nanomechanics.
The aim of this work is to demonstrate a novel single-molecule DNA sequence comparison assay that is purely based on DNA mechanics. A molecular construct that contained the two homologous but non-identical DNA sequences that were to be compared was prepared such that a four-way (Holliday) junction could be formed by the formation of heteroduplexes through the inter-recombination of the strands. Magnetic tweezers were used to manipulate the force and the winding applied to this construct for inducing both the formation and the migration of a Holliday junction. The end-to-end distance of the construct was measured as a function of the winding and was used to monitor the behavior of the Holliday junction in different regions of the intra-molecular recombination. In the appropriate buffer, the magnet rotation induces the migration of the Holliday junction in the regions where there is no sequence difference between the recombining sequences. In contrast, even a single-base difference between the recombining sequences leads to a long-lasting blockage of the migration in the same buffer; this effect was obtained when the junction was positioned near this locus (the site of the single-base difference) and forced toward the formation of heteroduplexes that comprise the locus. The migration blockages were detected through the identification of the formation of plectonemes. The detection of the presence of sequence differences and their respective mappings were obtained from the series of blockages that were detected. This work presents a novel single-molecule sequence comparison assay that is based on the use of a Holliday junction as an ultra-sensitive nanomechanism; the mismatches act as blocking grains of sand in the Holliday "DNA gearbox". This approach will potentially have future applications in biotechnology.


Holoprosencephaly: clinical evaluation on audiological and brainstem electrophysiological profiles.
This study evaluated audiological and electrophysiological profiles in 13 patients with holoprosencephaly. All patients had imaging evaluation by magnetic resonance imaging and molecular screening for the genes SHH, GLI2, and SIX3. Each patient underwent clinical (otological and vestibular antecedents, otoscopy) and instrumental (tympanometry, auditory brainstem response--ABR) evaluation to compare hearing and the electrophysiological profile possibly occurring in patients with these mutations. To our knowledge there are no systematic studies correlating molecular/imaging and evoked potentials in patients with HPE. Here, we discuss the audiological and electrophysiological profiles of patients and the possible role of the genes studied on the overall findings.


Risk prediction in acute coronary syndrome from serial in-hospital measurements of N-terminal pro-B-type natriuretic peptide.
There is limited information about the in-hospital plasma profile of N-terminal pro-B-type natriuretic peptide (NT-proBNP) in patients with non-ST-elevation acute coronary syndrome (NSTACS) and furthermore, the prognostic influence of the timing of NT-proBNP measurements in NSTACS is unsettled. These subject matters are elucidated in this study composed of 455 patients with NSTACS (symptoms <24 h). NT-proBNP was measured at 0, 6, 12, 24, 36, 48, 72 and 96 h following admission. Any death was registered at follow-up (median: 2.3 years). The study demonstrated a monophasic profile of the plasma NT-proBNP values, reaching a maximum at 6 hours, and it showed an independent prognostic significance of NT-proBNP irrespective of the sampling time. Risk prediction by NT-proBNP was improved by combining the baseline measurement and one value taken between 24 and 96 h (at 48 h, P<0.001). No additional prognostic information was provided by including more than one late in-hospital NT-proBNP value. The in-hospital NT-proBNP measurements exhibit a monophasic profile in patients with NSTACS and these values provide independent prognostic information as regards mortality irrespective of the sampling time. Moreover, risk prediction of NT-proBNP is strengthened by combining the admission measurement with an additional value during the hospitalization.


Suppressed neurogenesis without cognitive deficits: effects of fast neutron irradiation in mice.
This study assessed the effects of combined low-dose neutron and γ-ray irradiation on hippocampal neurogenesis and hippocampal-dependent memory. Neural progenitor cell division and survival were evaluated in brain sections and whole hippocampal preparations following head irradiation at a dose of 0.34 Gy for neutron radiation and 0.36 Gy for γ-ray radiation. Hippocampal-dependent memory formation was tested in a contextual fear conditioning task following irradiation at doses of 0.4 Gy for neutron radiation and 0.42 Gy for γ-ray radiation. Cell division was suppressed consistently along the entire dorsoventral axis of the hippocampus 24 h after the irradiation, but quiescent stem cells remained unaffected. The control and irradiated mice showed no differences in terms of exploratory behavior or anxiety 6 weeks after the irradiation. The ability to form hippocampus-dependent memory was also unaffected. The data may be indicative of a negligible effect of the low-dose of fast neutron irradiation and the neurogenesis suppression on animal behavior at 6 weeks after irradiation.


c-erbB-2 promoter-specific DNA-binding protein isolated from human breast cancer tissues displays mitogenic activity.
Amplification and overexpression of the c-erbB-2 gene appears to play a role in the pathogenesis of human breast and other cancers. Frequent amplification (20-30%) of the c-erbB-2 gene was observed in human adenocarcinomas of kidney, pancreas, lung, ovarian, and breast cancer. The gene product is a 185-kDa glycoprotein that has intrinsic tyrosine kinase activity and is believed to be a receptor. Several candidate ligands have been described. In the present study we have identified and purified a novel DNA-binding protein from malignant human breast tissues. The protein binds to a core element (-22 to +9, +1 being the transcription start site) of the c-erbB-2 promoter region in a sequence-specific manner. The affinity-purified protein has the ability to induce DNA synthesis in quiescent NIH/3T3 cells, suggesting that the factor has mitogenic activity. The purified protein induces c-erbB-2 expression on the surface of microinjected NIH/3T3 cells. This DNA-binding protein is a sequence-specific cellular factor that is associated with high level expression of the c-erbB-2 gene and appears to play a role in cell transformation. Understanding the control and expression of this DNA-binding protein may shed light on the mechanism(s) of c-erbB-2 gene regulation and its potential role in the pathogenesis of human adenocarcinomas.


[ANSA analysis. III. Synthesis of aminonaphthalinesulfonamide chromogenic substrates for protease analysis].
A review of synthetic methods of peptide substrates containing aminonaphthalenesulphonamide (ANSA) as the detected leaving group is presented. Variations of aminoacylic and peptide ANSA derivatives using ANSA as the C-protect group at all stages of the peptide synthesis, condensations of the ANSA with the N-protected peptide fragment obtained preliminary, the application of aminoacyl-ANSA as syntones are discussed. The synthesis scheme used while determining optimal ANSA substrates that involves reactions of aminoacyl derivatives of aminonaphthalenesulfonylchlorides with amines is shown. The application of di-tert-butylpyrocarbonate, DCC, chlorodimethylformiminium chloride, alkylchloroformate as condensing agents is described. The protection of amino groups was carried out by using Boc- and Cbz- groups.


Feasibility, uptake and impact of a hospital-wide tobacco addiction treatment pathway: Results from the CURE project pilot.
Providing comprehensive tobacco addiction treatment to smokers admitted to acute care settings represents an opportunity to realise major health resource savings and population health improvements. The CURE project is a hospital-wide tobacco addiction treatment service piloted in Wythenshawe Hospital, Manchester, UK. The core components of the project are electronic screening of all patients to identify smokers; the provision of brief advice and pharmacotherapy by frontline staff; opt-out referral of smokers to a specialist team for inpatient behavioural interventions; and continued support after discharge. From 01 October 2018 to 31 March 2019, 92% (13,515/14,690) of adult admissions were screened for smoking status, identifying 2,393 current smokers. Of these, 96% were given brief advice to quit by the admitting team. Through the automated 'opt-out' referral process, 61% patients completed inpatient behavioural interventions with a specialist cessation practitioner (69% within the first 48 hours of admission). Overall, 66% of smokers were prescribed pharmacotherapy. Over one in five of all smokers admitted during this pilot reported that they were abstinent from smoking 12 weeks after discharge (22%) at a cost £183 per quit. National implementation of this cost-effective programme would be likely to generate substantial benefits to public health.


Severe idiopathic thymic hemorrhage in two littermate dogs.
Two 6-month-old littermate Shetland Sheepdogs were found to have severe thymic hemorrhage unassociated with trauma or poisoning. One dog survived the insult with supportive care, and the other died and was necropsied. Antemortem laboratory data and radiographic findings for both dogs were consistent with acute blood loss into the mediastinum. Hemorrhage was restricted to the thymus and mediastinum in both dogs.


Patterns of symptoms and prognosis in occlusive thromboaortopathy (Takayasu's disease).
Ninety-five Japanese patients with Takayasu's disease were classified according to four patterns of severity of symptoms in the period from the onset of symptoms to diagnosis. Included were pattern A in 34 patients with a plateau course after the insidious onset of symptoms, pattern B in 18 patients with a decrescendo course after the sudden onset of severe symptoms, pattern C in 5 patients who had no severe symptoms during the years between the early and late periods of severe symptoms and pattern D in 38 patients with a crescendo course after the onset of symptoms and the following period of years of no severe symptoms. Fifteen patients died during the prospective follow-up period of 8.6 +/- 6 years (mean +/- SD) after admission, but 12 had had patterns C and D and the remaining 3 had had patterns A and B. The cardinal signs and symptoms are given in detail, particularly those at the onset. These data should aid in early diagnosis and prediction of the prognosis in Takayasu's disease.


Hypertension-causing cullin 3 mutations disrupt COP9 signalosome binding.
The discovery of new genetic mutations that cause hypertension has illuminated previously unrecognized physiological pathways. One such regulatory pathway was identified when mutations in with no lysine kinase (WNK)4, Kelch-like 3 (KLHL3), and cullin 3 (CUL3) were shown to cause the disease familial hyperkalemic hypertension (FHHt). Mutations in all three genes upregulate the NaCl cotransporter (NCC) due to an impaired ability to degrade WNK protein through the cullin-RING-ligase (CRL) ubiquitin-proteasome system. The CUL3 FHHt mutations cause the most severe phenotype, yet the precise mechanism by which these mutations cause the disease has not been established and current proposed models are controversial. New data have identified a possible novel mechanism involving dysregulation of CUL3 activity by the COP9 signalosome (CSN). The CSN interaction with mutant CUL3 is diminished, causing hyperneddylation of the CRL. Recent work has shown that direct renal CSN impairment mimics some aspects of the CUL3 mutation, including lower KLHL3 abundance and activation of the WNK-NCC pathway. Furthermore, in vitro and in vivo studies of CSN inhibition have shown selective degradation of CRL substrate adaptors via auto-ubiquitination, allowing substrate accumulation. In this review, we will focus on recent research that highlights the role of the CSN role in CUL3 mutations that cause FHHt. We will also highlight how these results inform other recent studies of CSN dysfunction.


Interaction of tubulin with bifunctional colchicine analogues: an equilibrium study.
The interaction of tubulin with simple analogues of colchicine that contain both its tropolone and trimethoxyphenyl rings has been characterized, and the results were analyzed in terms of the simple bifunctional ligand model developed for the binding of colchicine [ Andreu , J. M., & Timasheff , S. N. (1982) Biochemistry 21, 534-543] on the basis of interactions of tubulin with single-ring analogues. The compound 2-methoxy-5-(2,3,4-trimethoxyphenyl)-2,4,6- cycloheptatrien -1-one has been found to bind reversibly to 0.86 +/- 0.06 site of purified calf brain tubulin with an equilibrium constant of (4.9 +/- 0.3) X 10(5) M-1 (25 degrees C), delta H degrees app = -1.6 +/- 0.7 kcal mol-1, and delta S degrees app = 20.5 +/- 2.5 eu. The binding appears specific for the colchicine site. The closely related compound 2-methoxy-5-[[3-(3,4,5-trimethoxyphenyl)-propionyl]amino] -2,4,6- cycloheptatrien -1-one interacts weakly with tubulin. Binding of the first analogue is accompanied by ligand fluorescence appearance, quenching of protein fluorescence, perturbation of the far-ultraviolet circular dichroism of tubulin, and induction of the tubulin GTPase activity, similarly to colchicine binding. Substoichiometric concentrations of the analogue inhibit microtubule assembly in vitro. Excess analogue concentration under microtubule-promoting conditions induces an abnormal cooperative polymerization of tubulin, similar to that of the tubulin-colchicine complex.


Opportunities to Increase Access to HIV Prevention: Evaluating the Implementation of Pharmacist-Initiated Pre-exposure Prophylaxis in California.
Pharmacies are a promising setting through which to expand access to human immunodeficiency virus (HIV) prevention, including pre-exposure and post-exposure prophylaxis (PrEP and PEP, respectively). We aimed to evaluate and inform the implementation of California's Senate Bill 159 (2019), allowing pharmacists to independently prescribe PrEP and PEP. From October through December 2022, we conducted a cross-sectional study of 919 California pharmacists and pharmacy students, primarily recruited via the email listservs of professional organizations. Participants completed an online survey assessing the implementation of pharmacist-initiated PrEP/PEP, including knowledge, attitudes, practices, perceived barriers, and implementation preferences elicited through a discrete choice experiment. Among 919 participants (84% practicing pharmacists, 43% in community pharmacies), 11% and 13% reported that pharmacists at their pharmacy initiate PrEP and PEP, respectively. Most believed that pharmacist-initiated PrEP/PEP is important (96%) and were willing to provide PrEP (81%); fewer (27%) had PrEP/PEP training. Common implementation barriers were lack of staff/time and payment for pharmacist services. Participants preferred PrEP implementation models with in-pharmacy rapid oral HIV testing and pharmacists specifically hired to provide PrEP services. Despite pharmacists' supportive attitudes, Senate Bill 159 implementation in California pharmacies remains limited, in part due to policy-level and organizational-level barriers. Ensuring PrEP/PEP-related payment for services and sufficient workforce capacity is key to leveraging pharmacists' role in HIV prevention.


HPV E6/E7, hTERT, and Ki67 mRNA RT-qPCR Assay for Detecting High-Grade Cervical Lesion with Microscope Slides.
After breast and colon cancer, cervical cancer is the third most common cancer of women worldwide. Since human papillomavirus (HPV) infection is known to be the predominant cause of cervical cancer, molecular HPV screening is currently used along with cytological and histological examination methods for precancer diagnosis. Nevertheless, the sensitivity of the current HPV test is less than 80%; thus, many cervical cancer cases are not able to be diagnosed by HPV screening alone, and likewise, patients with cervical cancer are often determined to be HPV-negative by the current screening methods. Therefore, human telomerase reverse transcriptase (hTERT) and Ki67 previously identified as cancer markers were attempted. And cervical exfoliated cells of high-grade squamous intraepithelial lesion (HSIL), the most severe precancerous lesion of cancer, were used in the study. However, it takes a long time to collect enough specimens to conduct statistical analysis. Therefore, in the present study, microscope slides, cervical exfoliated cells on glass slides, were attempted. The results of the analysis demonstrated that hTERT and Ki67 expression levels were useful in distinguishing between cancerous and normal specimens, exhibiting a higher sensitivity and specificity than conventional HPV E6/E7 testing. And the study suggests clinical slide cell samples could be effectively used in the context of retrospective studies to identify novel biomarkers.


Temporal processing impairment in children with attention-deficit-hyperactivity disorder.
The current study aimed to investigate temporal processing in Chinese children with Attention-Deficit-Hyperactivity Disorder(ADHD) using time production, time reproduction paradigm and duration discrimination tasks. A battery of tests specifically designed to measure temporal processing was administered to 94 children with ADHD and 100 demographically matched healthy children. A multivariate analysis of variance (MANOVA) and a repeated measure MANOVA indicated that children with ADHD were impaired in time processing functions. The results of pairwise comparisons showed that the probands with a family history of ADHD performed significantly worse than those without family history in the time production tasks and the time reproduction task. Logistic regression analysis showed duration discrimination had a significant role in predicting whether the children were suffering from ADHD or not, while temporal processing had a significant role in predicting whether the ADHD children had a family history or not. This study provides further support for the existence of a generic temporal processing impairment in ADHD children and suggests that abnormalities in time processing and ADHD share some common genetic factors.


Regulation of phosphoinositide metabolism, Akt phosphorylation, and glucose transport by PTEN (phosphatase and tensin homolog deleted on chromosome 10) in 3T3-L1 adipocytes.
To investigate the roles of PTEN (phosphatase and tensin homolog deleted on chromosome 10) in the regulation of 3-position phosphorylated phosphoinositide metabolism as well as insulin-induced Akt phosphorylation and glucose metabolism, wild-type PTEN and its phosphatase-dead mutant (C124S) with or without an N-terminal myristoylation tag were overexpressed in Sf-9 cells and 3T3-L1 adipocytes using baculovirus and adenovirus systems, respectively. When expressed in Sf-9 cells together with the p110alpha catalytic subunit of phosphoinositide 3-kinase, myristoylated PTEN markedly reduced the accumulations of both phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate induced by p110alpha. In contrast, overexpression of the C124S mutants apparently increased these accumulations. In 3T3-L1 adipocytes, insulin-induced accumulations of phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate were markedly suppressed by overexpression of wild-type PTEN with the N-terminal myristoylation tag, but not by that without the tag. On the contrary, the C124S mutants of PTEN enhanced insulin-induced accumulations of phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate. Interestingly, the phosphorylation level of Akt at Thr308 (Akt2 at Thr309), but not at Ser473 (Akt2 at Ser474), was revealed to correlate well with the accumulation of phosphatidylinositol 3,4,5-trisphosphate modified by overexpression of these PTEN proteins. Finally, insulin-induced increases in glucose transport activity were significantly inhibited by the overexpression of myristoylated wild-type PTEN, but were not enhanced by expression of the C124S mutant of PTEN. Therefore, in conclusion, 1) PTEN dephosphorylates both phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate in vivo, and the C124S mutants interrupt endogenous PTEN activity in a dominant-negative manner. 2) The membrane targeting process of PTEN may be important for exerting its function. 3) Phosphorylations of Thr309 and Ser474 of Akt2 are regulated differently, and the former is regulated very sensitively by the function of PTEN. 4) The phosphorylation level of Ser474, but not that of Thr309, in Akt2 correlates well with insulin-stimulated glucose transport activity in 3T3-L1 adipocytes. 5) The activity of endogenous PTEN may not play a major role in the regulation of glucose transport activity in 3T3-L1 adipocytes.


Assessing the impact of national antibiotic campaigns in Europe.
Because of evidence of causal association between antibiotic use and bacterial resistance, the implementation of national policies has emerged as a interesting tool for controlling and reversing bacterial resistance. The aim of this study was to assess the impact of public policies on antibiotic use in Europe using a differences-in-differences approach. Comparable data on systemic antibiotics administered in 21 European countries are available for a 11-year period between 1997 and 2007. Data on national campaigns are drawn from the public health literature. We estimate an econometric model of antibiotic consumption with country fixed effects and control for the main socioeconomic and epidemiological factors. Lagged values and the instrumental variables approach are applied to address endogeneity aspects of the prevalence of infections and the adoption of national campaigns. We find evidence that public campaigns significantly reduce the use of antimicrobials in the community by 1.3-5.6 defined daily doses per 1,000 inhabitants yearly. This represents an impact of roughly 6.5-28.3 % on the mean level of antibiotic use in Europe between 1997 and 2007. The effect is robust across different measurement methods. Further research is needed to investigate the effectiveness of policy interventions targeting different social groups such as general practitioners or patients.


Collection of Wet-Origin Footwear Impressions on Various Surfaces Using an Electrostatic Dust Print Lifter.
Electrostatic dust print lift method is known to be able to recover only dry-origin footwear impression. However, the wet-origin footwear impression could also be recovered using this method. As the amount of dust accumulated before deposition of the wet-origin footwear impression increased, the intensity of the footwear impression lifted with this method became stronger. If the footwear impression is not affected by moisture after it is made, the 28-h old wet-origin footwear impression could be recovered using this method. The intensity of the lifted footwear impression did not decrease significantly even when the number of sequential steps increased as long as the shoe sole is wet. However, when the moisture on the shoe sole depleted, the intensity of the footwear impression decreased sharply. This method has the advantage of being able to enhance the footwear impression without being affected by the footwear impressions deposited in the past.


Linkage analysis of multiple endocrine neoplasia type 2A (MEN-2A) and three DNA markers on chromosome 20: evidence against synteny.
Members of four families in which multiple endocrine neoplasia type 2A (MEN-2A) is segregating were genotyped for three chromosome #20 DNA markers. Close linkage is excluded for the genetic locus of MEN-2A (MEN2A) with each marker locus tested. Using multipoint analysis, the MEN2A locus was excluded from a region 32.5 cM in length encompassing D20S5 and D20S6, two loci that previously had been mapped to the chromosome 20p12.2 region containing the putative deletion associated with MEN-2. In total, we excluded approximately 90.5 cM of chromosome #20 as a possible location of the MEN2A locus. The data from this study do not support increased recombination on the short arms of chromosome #20 and were insufficient to support heterogeneity of MEN-2A as possible explanations for failure to detect linkage between MEN2A and D20S5 or D20S6.


Predictors of immunological failure of antiretroviral therapy among HIV infected patients in Ethiopia: a matched case-control study.
In resource constrained settings, immunological assessment through CD4 count is used to assess response to first line Highly Active Antiretroviral Therapy (HAART). In this study, we aim to investigate factors associated with immunological treatment failure. A matched case-control study design was used. Cases were subjects who already experienced immunological treatment failure and controls were those without immunological failure after an exactly or approximately equivalent duration of first line treatment with cases. Data were analyzed using SPSS v16.0. Conditional logistic regression was carried out. A total of 134 cases and 134 controls were included in the study. At baseline, the mean age ± 1 SD of cases was 37.5 ± 9.7 years whereas it was 36.9 ± 9.2 years among controls. The median baseline CD4 counts of cases and controls were 121.0 cells/µl (IQR: 47-183 cells/µl) and 122.0 cells/µl (IQR: 80.0-189.8 cells/µl), respectively. The median rate of CD4 cells increase was comparable for the two groups in the first six months of commencing HAART (P = 0.442). However, the median rate of CD4 increase was significantly different for the two groups in the next 6 months period (M6 to M12). The rate of increment was 8.8 (IQR: 0.5, 14.6) and 1.8 (IQR: 8.8, 11.3) cells/µl/month for controls and cases, respectively (Mann-Whitney U test, P = 0.003). In conditional logistic regressions grouped baseline CD4 count (P = 0.028), old age group and higher educational status (P<0.001) were significant predictors of immunological treatment failure. Subjects with immunological treatment failure have an optimal rate of immunological recovery in the first 6 months of treatment with first line HAART, but relative to the non-failing group the rate declines at a later period, notably between 6 and 12 months. Low baseline CD4 count, old age and higher educational status were associated with immunological treatment failure.


Hypertension among newly diagnosed diabetic patients at Mulago National Referral Hospital in Uganda: a cross sectional study.
The prevalence of hypertension in patients with diabetes is approximately two-fold higher than in age-matched subjects without the disease and, conversely, individuals with hypertension are at increased risk of developing diabetes compared with normotensive persons. Up to 75% of cases of cardiovascular disease (CVD) in patients with diabetes are attributed to hypertension. Diabetics who have hypertension are more likely to develop complications and die, and appropriate blood pressure control in these individuals reduces the risk. This study sought to determine the prevalence and factors associated with hypertension among newly diagnosed adult diabetic patients in a national referral hospital in Uganda. In this cross-sectional study, conducted between June 2014 and January 2015, we recruited 201 newly diagnosed adult diabetic patients. Information on patients' socio-demographics was obtained using a pre-tested questionnaire, while biophysical profile, blood pressure measurement, biochemical testing and echocardiographic findings were obtained by the research team for all the participants. Bivariate and multivariate logistic regression analyses were used to investigate the association of several factors with hypertension. Of the 201 patients recruited, 102 were male (50.8%) and the mean age was 46 ± 15 years. The majority of patients (159) had type 2 diabetes mellitus (DM) (79.1%) with a mean HbA1c level of 13.9 ± 5.3%. The prevalence of hypertension was 61.9% (95% CI: 54.8-68.6%). Knowledge of hypertension status was at 56 (27.7%) patients, 24 (44.4%) hypertensives were on treatment, and 19 (33.9%) were using ACE inhibitors/angiotensin receptor blockers. The independent factors associated with hypertension were being employed (OR 0.37, 95% CI: 0.16-0.90, p = 0.029) and being overweight or obese (OR 11.6, 95% CI: 4.29-31.2, p < 0.0001). The prevalence of hypertension was high in this population of newly diagnosed diabetics, few patients had knowledge of their hypertension status and few were on appropriate treatment. Both modifiable and non-modifiable risk factors were associated with hypertension in this group. Therefore routine assessment, treatment and control of hypertension among diabetics is necessary to prevent cardiovascular complications and death. There is also a need to address the modifiable risk factors.


Multilocus sequence typing of Streptococcus pneumoniae by use of mass spectrometry.
Multilocus sequence typing (MLST) is an important tool for the global surveillance of bacterial pathogens that is performed by comparing the sequences of designated housekeeping genes. We developed and tested a novel mass spectrometry-based method for MLST of Streptococcus pneumoniae. PCR amplicons were subjected to in vitro transcription and base-specific cleavage, followed by analysis of the resultant fragments by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Comparison of the cleavage fragment peak patterns to a reference sequence set permitted automated identification of alleles. Validation experiments using 29 isolates of S. pneumoniae revealed that the results of MALDI-TOF MS MLST matched those obtained by traditional sequence-based MLST for 99% of alleles and that the MALDI-TOF MS method accurately identified two single-nucleotide variations. The MADLI-TOF MS method was then used for MLST analysis of 43 S. pneumoniae isolates from Papua New Guinean children. The majority of the isolates present in this population were not clonal and contained seven new alleles and 30 previously unreported sequence types.