Distribution and neuronal expression of phosphatidylinositol phosphate kinase IIgamma in the mouse brain.
The role of cellular phosphatidylinositol 5-phosphate (PtdIns5P), as a signalling molecule or as a substrate for the production of small, compartmentalized pools of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)], may be dependent on cell type and subcellular localization. PtdIns5P levels are primarily regulated by the PtdIns5P 4-kinases (type II PIP kinases or PIP4Ks), and we have investigated the expression and localization in the brain of the least-studied PIP4K isoform, PIP4Kgamma. In situ hybridization and immunohistochemistry, using antisense oligonucleotide probes and a PIP4Kgamma-specific antibody, revealed that this isoform has a restricted CNS expression profile. The use of antibodies to different cell markers showed that this expression is limited to neurons, particularly the cerebellar Purkinje cells, pyramidal cells of the hippocampus, large neuronal cell types in the cerebral cortex including pyramidal cells, and mitral cells in the olfactory bulb and is not expressed in cerebellar, hippocampal formation, or olfactory bulb granule cells. In neurons expressing this enzyme, PIP4Kgamma has a vesicular distribution and shows partial colocalization with markers of cellular compartments of the endomembrane trafficking pathway. The PIP4Kgamma isoform expression is established after day 7 of postnatal development. Overall, our data suggest that PIP4Kgamma may have a role in neuron function, specifically in the regulation of vesicular transport, in specific regions of the developed brain.


The role of necroptosis in the treatment of diseases.
Necroptosis is an emerging form of programmed cell death occurring via active and well-regulated necrosis, distinct from apoptosis morphologically, and biochemically. Necroptosis is mainly unmasked when apoptosis is compromised in response to tumor necrosis factor alpha. Unlike apoptotic cells, which are cleared by macrophages or neighboring cells, necrotic cells release danger signals, triggering inflammation, and exacerbating tissue damage. Evidence increasingly suggests that programmed necrosis is not only associated with pathophysiology of disease, but also induces innate immune response to viral infection. Therefore, necroptotic cell death plays both physiological and pathological roles. Physiologically, necroptosis induce an innate immune response as well as premature assembly of viral particles in cells infected with virus that abrogates host apoptotic machinery. On the other hand, necroptosis per se is detrimental, causing various diseases such as sepsis, neurodegenerative diseases and ischemic reperfusion injury. This review discusses the signaling pathways leading to necroptosis, associated necroptotic proteins with target-specific inhibitors and diseases involved. Several studies currently focus on protective approaches to inhibiting necroptotic cell death. In cancer biology, however, anticancer drug resistance severely hampers the efficacy of chemotherapy based on apoptosis. Pharmacological switch of cell death finds therapeutic application in drug- resistant cancers. Therefore, the possible clinical role of necroptosis in cancer control will be discussed in brief. [BMB Reports 2018; 51(5): 219-224].


Effects of acute systemic administration of cannabidiol on sleep-wake cycle in rats.
Cannabidiol (CBD) is one of the main components of Cannabis sativa and has a wide spectrum of action, including effects in the sleep-wake cycle. The objective of this paper is to assess the effects on sleep of acute systemic administration of CBD. Adult male Wistar rats were randomly distributed into four groups that received intraperitoneal injections of CBD 2.5 mg/kg, CBD 10 mg/kg, CBD 40 mg/kg or vehicle (n=seven animals/group). Sleep recordings were made during light and dark periods for four days: two days of baseline recording, one day of drug administration (test), and one day after drug (post-test). During the light period of the test day, the total percentage of sleep significantly increased in the groups treated with 10 and 40 mg/kg of CBD compared to placebo. REM sleep latency increased in the group injected with CBD 40 mg/kg and was significantly decreased with the dose of 10 mg/kg on the post-test day. There was an increase in the time of SWS in the group treated with CBD 40 mg/kg, although this result did not reach statistical significance. The systemic acute administration of CBD appears to increase total sleep time, in addition to increasing sleep latency in the light period of the day of administration.


A field trial of the Clinicult system for detection of asymptomatic gonorrhea in women.
Clinicult, a selective medium for culturing Neisseria gonorrhoeae, was field-tested in a gonorrhea screening program in Seattle, Wash., in 1973. The results with this medium and with the Transgrow and Thayer-Martin culture systems were compared as to sensitivity and specificity. A total of 5,141 women from three patient groups were included in the study. Group 1 consisted of 720 female patients of the venereal disease clinic of the Seattle-King County Health Department, who served as the control group. When this group was screened with the Clinicult and Thayer-Martin culture media, the Thayer-Martin medium proved superior in identifying positive carriers. Group 2 was composed of approximately 2,000 patients from five different facilities, including family planning clinics and hospital out patient services. No statistical difference in accuracy was found between the two culture systems used for this group-Clinicult and Transgrow. Group 3 was comprised of approximately 2,500 female patients who were screened with the Clinicult and Transgrow cultures by their own private physician or his staff. The Clinicult system proved significantly less effective than the Transgrow cultures in identifying infected females in group 3. The physicians varied greatly in their ability to use the Clinicult system successfully. Possible reasons for their errors may have been (a) lack of motivation and of care by their office personnel in conducting the necessary additional procedures required with Clinicult, (b) the inhibitory nature of the medium, and (c) the failure of the medium to produce colonies of adequate size. The staffs of communitywide screening programs for gonorrhea need to be highly selective in choosing the medical facilities in which to use the Clinicult culture system. When laboratory facilities are available for the full utilization of the Thayer-Martin medium, this system is preferable. When, however, standard culture procedures are not readily available, Clinicult, properly used, can reduce the central laboratory load by eliminating the need for processing negative cultures.


Evaluating the perceptual and pathophysiological consequences of auditory deprivation in early postnatal life: a comparison of basic and clinical studies.
Decades of clinical and basic research in visual system development have shown that degraded or imbalanced visual inputs can induce a long-lasting visual impairment called amblyopia. In the auditory domain, it is well established that inducing a conductive hearing loss (CHL) in young laboratory animals is associated with a panoply of central auditory system irregularities, ranging from cellular morphology to behavior. Human auditory deprivation, in the form of otitis media (OM), is tremendously common in young children, yet the evidence linking a history of OM to long-lasting auditory processing impairments has been equivocal for decades. Here, we review the apparent discrepancies in the clinical and basic auditory literature and provide a meta-analysis to show that the evidence for human amblyaudia, the auditory analog of amblyopia, is considerably more compelling than is generally believed. We argue that a major cause for this discrepancy is the fact that most clinical studies attempt to link central auditory deficits to a history of middle ear pathology, when the primary risk factor for brain-based developmental impairments such as amblyopia and amblyaudia is whether the afferent sensory signal is degraded during critical periods of brain development. Accordingly, clinical studies that target the subset of children with a history of OM that is also accompanied by elevated hearing thresholds consistently identify perceptual and physiological deficits that can endure for years after peripheral hearing is audiometrically normal, in keeping with the animal studies on CHL. These studies suggest that infants with OM severe enough to cause degraded afferent signal transmission (e.g., CHL) are particularly at risk to develop lasting central auditory impairments. We propose some practical guidelines to identify at-risk infants and test for the positive expression of amblyaudia in older children.


Perspectives from the frontlines: palliative care providers' expectations of Canada's compassionate care benefit programme.
Recognising their valuable role as key informants, this study examines the perspectives of front-line palliative care providers (FLPCP) regarding a social benefit programme in Canada designed to support family caregivers at end-of-life, namely the Compassionate Care Benefit (CCB). The CCB's purpose is to provide income assistance and job security to family caregivers who take temporary leave from employment to care for a dying family member. Contributing to an evaluative study that aims to provide policy-relevant recommendations about the CCB, this analysis draws on semi-structured interviews undertaken in 2007/2008 with FLPCPs (n = 50) from across Canada. Although participants were not explicitly asked during interviews about their expectations of the CCB, thematic content analysis revealed 'expectations' as a key finding. Through participants' discussions of their knowledge of and familiarity with the CCB, specific expectations were identified and grouped into four categories: (1) temporal; (2) financial; (3) informational; and (4) administrative. Findings demonstrate that participants expect the CCB to provide: (1) an adequate length of leave time from work, which is reflective of the uncertain nature of caregiving at end-of-life; (2) adequate financial support; (3) information on the programme to be disseminated to FLPCPs so that they may share it with others; and (4) a simple, clear, and quick application process. FLPCPs hold unique expertise, and ultimately the power to shape uptake of the CCB. As such, their expectations of the CCB contribute valuable knowledge from which relevant policy recommendations can be made to better meet the needs of family caregivers and FLPCPs alike.


Suppressive role of the metastasis-related nm23-H1 gene in human ovarian carcinomas: association of high messenger RNA expression with lack of lymph node metastasis.
The nm23-H1 gene has been proposed as a metastasis suppressor gene. It is located on the long arm of chromosome 17, which is frequently deleted in ovarian cancer, and shows altered expression and structure in some advanced neoplasms. To evaluate the role of nm23-H1 in ovarian carcinogenesis, we have analyzed this gene in 66 primary human ovarian carcinomas at both the DNA and RNA levels. Despite the high frequency (76%) of nm23-H1 loss of heterozygosity (LOH), the complete absence of gene mutations in the coding portions of the retained allele clearly indicated that, in ovarian carcinomas, this gene does not function in the same way as do classic oncosuppressor genes. The relationship of clinicopathological parameters with nm23-H1 gene deletions and expression levels was also investigated. LOHs were more common in the serous and endometrioid histotypes (85 and 93%, respectively), and the highest LOH frequency was detected in poorly differentiated tumors (89%). A significant relationship between nm23-H1 mRNA expression and lymph node metastasis was observed in high-grade tumors, which are intrinsically more invasive than are low-grade tumors. In particular, among the poorly differentiated tumors showing areas of undifferentiated solid carcinoma (classified as G3/G4), lymph node-negative tumors displayed expression levels that were significantly higher than those of lymph node-positive tumors (P < 0.001). In conclusion, our data suggest that the nm23-H1 gene product may exert an inhibitory effect on the lymphatic dissemination of human ovarian tumors. However, several other factors, biological or time and patient dependent, influence the complex metastatic progression of ovarian tumors and may cooperate with nm23-H1 in the promotion or inhibition of this process.


Global and Regional Estimates for Subtype-Specific Therapeutic and Prophylactic HIV-1 Vaccines: A Modeling Study.
Global HIV-1 genetic diversity forms a major obstacle to the development of an HIV vaccine. It may be necessary to employ subtype-specific HIV-1 vaccines in individual countries according to their HIV-1 subtype distribution. We estimated the global and regional need for subtype-specific HIV-1 vaccines. We took into account the proportions of different HIV-1 variants circulating in each country, the genetic composition of HIV-1 recombinants, and the different genome segments (gag, pol, env) that may be incorporated into vaccines. We modeled different scenarios according to whether countries would employ subtype-specific HIV-1 vaccines against (1) the most common subtype; (2) subtypes contributing more than 5% of HIV infections; or (3) all circulating subtypes. For therapeutic vaccines targeting the most common HIV-1 subtype in each country, 16.5 million doses of subtype C vaccine were estimated globally, followed by subtypes A (14.3 million) and B (4.2 million). A vaccine based on env required 2.6 million subtype E doses, and a vaccine based on pol required 4.8 million subtype G doses. For prophylactic vaccines targeting the most common HIV-1 subtype in each country, 1.9 billion doses of subtype A vaccine were estimated globally, followed by subtype C (1.1 billion) and subtype B (1.0 billion). A vaccine based on env required 1.2 billion subtype E doses, and a vaccine based on pol required 0.3 billion subtype G doses. If subtype-specific HIV-1 vaccines are also directed against less common subtypes in each country, vaccines targeting subtypes D, F, H, and K are also needed and would require up to five times more vaccine doses in total. We conclude that to provide global coverage, subtype-specific HIV-1 vaccines need to be directed against subtypes A, B, and C. Vaccines targeting env also need to include subtype E and those targeting pol need to include subtype G.


Synovial cells responding to a 65-kDa mycobacterial heat shock protein have a high proportion of a TcR gamma delta subtype uncommon in peripheral blood.
We have analysed the ability of T cells from synovial fluid mononuclear cells (SFMC) and from peripheral blood mononuclear cells (PBMC) of inflammatory arthritic diseases to proliferate in response to mycobacterial antigens (65-kDa heat shock protein [hsp] of BCG, whole BCG) and to rat collagen type II. The SFMC demonstrated a significantly greater ability to respond to 65-kDa hsp of BCG, and to whole BCG, compared with PBMC from the same patients. With collagen type II, only a small proportion of the patients showed a proliferative response, although with this antigen also SFMC responded better than PBMC. There was no difference between SFMC and PBMC in the response to control antigen (tetanus toxoid), phytohaemagglutinin (PHA), or interleukin 2 (IL-2). A high proportion of cells in SFMC-derived short-term T-cell lines were of TcR gamma delta type, often exceeding the number of TcR gamma beta type. There was a significantly higher proportion of TcR gamma delta cells in the SFMC lines compared with the PBMC lines, and a large part of the TcR gamma delta cells in the SFMC cultures was CD8+. The SFMC lines had a high proportion of delta-TCS-1+ cells (V delta 1) among their TcR gamma delta cells, always exceeding the percentages of Ti gamma A+(V gamma 9) and BB3+ (V delta 2). In the PBMC lines, the distribution of TcR gamma delta subtypes was markedly different, with a Ti gamma A+/BB3+ population in the majority. These data argue for a different subpopulation distribution of TcR gamma delta cells in synovial fluid compared with peripheral blood of patients with inflammatory arthritic diseases.


Human induced pluripotent stem cell-derived B lymphocytes express sIgM and can be generated via a hemogenic endothelium intermediate.
The differentiation of human pluripotent stem cells to the B-cell lymphoid lineage has important clinical applications that include in vitro modeling of developmental lymphogenesis in health and disease. Here, we first demonstrate the capacity of human induced pluripotent stem cells (hiPSCs) to differentiate into CD144(+)CD73(-)CD43/CD235a(-) cells, characterized as hemogenic endothelium, and show that this population is capable of differentiating to CD10(+)CD19(+) B lymphocytes. We also demonstrate that B lymphocytes generated from hiPSCs are able to undergo full VDJ rearrangement and express surface IgM (sIgM(+)), thus representing an immature B-cell subset. Efficiency of sIgM expression on the hiPSC-derived B lymphocytes (∼ 5% of CD19(+) cells) was comparable with B lymphocytes generated from human umbilical cord blood (UCB) hematopoietic progenitor cells. Importantly, when assessed by global transcriptional profiling, hiPSC-derived B-cells show a very high level of similarity when compared with their UCB-derived counterparts, such that from more than 47,000 different transcripts, only 45 were significantly different (with a criteria adjusted P value P<0.05, log FC >1.5 or 2.8-fold). This represents a unique in vitro model to delineate critical events during lymphogeneisis in development and lymphoid diseases such as acute lymphocytic leukemia.


Inter-Subject Variability of Skull Conductivity and Thickness in Calibrated Realistic Head Models.
Skull conductivity has a substantial influence on EEG and combined EEG and MEG source analysis as well as on optimized transcranial electric stimulation. To overcome the use of standard literature values, we propose a non-invasive two-level calibration procedure to estimate skull conductivity individually in a group study with twenty healthy adults. Our procedure requires only an additional run of combined somatosensory evoked potential and field data, which can be easily integrated in EEG/MEG experiments. The calibration procedure uses the P20/N20 topographies and subject-specific realistic head models from MRI. We investigate the inter-subject variability of skull conductivity and relate it to skull thickness, age and gender of the subjects, to the individual scalp P20/N20 surface distance between the P20 potential peak and the N20 potential trough as well as to the individual source depth of the P20/N20 source. We found a considerable inter-subject variability for (calibrated) skull conductivity (8.44 ± 4.84 mS/m) and skull thickness (5.97 ± 1.19 mm) with a statistically significant correlation between them (rho = 0.52). Age showed a statistically significant negative correlation with skull conductivity (rho = -0.5). Furthermore, P20/N20 surface distance and source depth showed large inter-subject variability of 12.08 ± 3.21 cm and 15.45 ± 4.54 mm, respectively, but there was no significant correlation between them. We also found no significant differences among gender subgroups for the investigated measures. It is thus important to take the inter-subject variability of skull conductivity and thickness into account by means of using subject-specific calibrated realistic head modeling.


[Mortality trends in cancer attributable to tobacco in Mexico].
To describe the mortality trends of cancer attributable to tobacco smoking, particularly lung cancer, for the 1980-1997 period in Mexico. Mortality trends were analyzed for each type of cancer associated to tobacco smoking, according to the International Classification of Diseases (ICD). Crude and adjusted mortality rates were estimated for the period between 1980 and 1997, by age, gender, basic death cause, and year of death. The gender ratio and the relative proportion were estimated for cases in the 35-64 age group and for the entire study population. Age population projections by Consejo Nacional de Poblacion (National Population Council), were used as denominators (1970-2010). The gender ratio for mortality rates for lung, esophageal, oral cavity and pharyngeal cancer was 2.10:1.00 (male:female). The gender ratio for laryngeal cancer was striking: 4.21:1.00, probably due to the higher prevalence of male tobacco smokers. The estimated relative proportion, using the total mortality due to malignant cancers between 1980-1997, was 12.31% for lung cancer, 1.71% for larynx cancer, 1.55% for esophageal cancer, and 1.49% for oral cavity/pharyngeal cancer. Previous tobacco smoking was correlated with the mortality rate trends for lung cancer (beta: 0.910, IC 95%: 1.097-1.797, R2 0.827). For the poorest social groups by federal entity, the correlation was inverted (beta: -0.510, IC 95% -0.170, -0.039, R2: 0.260). In Mexico, increased tobacco smoking, improved cancer diagnosis, and the demographic transition, are probably the main factors determining cancer mortality rates. However, other lifestyle associated variables, such as urbanization, physical activity, carotenoid intake, and other dietary and toxic substances like alcohol, may also influence the morbidity and mortality rates. Although tobacco-related cancer is a fast-growing public health problem having a poor prognosis, tobacco smoking, the main risk factor, could be eliminated by health education and promotion, together with publicity regulation and healthy taxation policies.


Infantilizing Autism.
When members of the public envision the disability of autism, they most likely envision a child, rather than an adult. In this empirically based essay, three authors, one of whom is an autistic self-advocate, analyzed the role played by parents, charitable organizations, the popular media, and the news industry in infantilizing autism. Parents portrayed the face of autism to be that of a child 95% of the time on the homepages of regional and local support organizations. Nine of the top 12 autism charitable organizations restricted descriptions of autism to child-referential discourse. Characters depicted as autistic were children in 90% of fictional books and 68% of narrative films and television programs. The news industry featured autistic children four times as often as they featured autistic adults in contemporary news articles. The cyclical interaction between parent-driven autism societies, autism fundraising charities, popular media, and contemporary news silences adult self-advocates by denying their very existence. Society's overwhelming proclivity for depicting autism as a disability of childhood poses a formidable barrier to the dignity and well-being of autistic people of all ages.


Ergonomics Analysis for Subjective and Objective Fatigue Between Laparoscopic and Robotic Surgical Skills Practice Among Surgeons.
Introduction. Our aim was to determine how self-reported and objectively measured fatigue of upper limb differ between laparoscopic and robotic surgical training environments. Methods. Surgeons at the 2016 SAGES Conference Learning Center and at our institution were enrolled. Two standardized surgical tasks (peg transfer [PT] and needle passing [NP]) were performed twice in each surgical skills practical environments: (1) laparoscopic training-box environment (Fundamentals of Laparoscopic Surgery [FLS]) and (2) Mimic dV-trainer (MIMIC). Muscle activation of upper trapezius (UT), anterior deltoid (AD), flexor carpi radialis, and extensor digitorum were recorded using surface electromyography (EMG; Trigno, Delsys, Inc, Natick, MA). Subjective fatigue was self-reported using Piper Fatigue Scale-12. Analysis was done using SPSS v25.0, α = .05. Results. Demographics were similar between FLS (N = 14) and MIMIC (N = 12). For PT, MIMIC had a significant increase in EMGRMS of UT (P < .001) and AD (P < .001). Conversely, FLS led to significant decreased muscle fatigue in UT (P = .015). For NP, MIMIC had a significant increase in EMGRMS for UT (P = .034) and AD (P = .031), but FLS induced more muscle fatigue for AD (P = .004). There was significant decrease in self-reported fatigue after performing FLS tasks (P = .030) but not after MIMIC (P = .663). Conclusion. Our results showed that practice with MIMIC resulted in greater activation of shoulder muscles, while FLS caused more significant muscle fatigue in the same muscles. This could be due to ergonomic disadvantages and nonoptimal ergonomic settings. Further studies are needed to understand the optimal ergonomics and its impact on fatigue and muscle activation during use of both the FLS and MIMIC training systems.


Modeling of reflectometric and ellipsometric spectra from the skin in the terahertz and submillimeter waves region.
The human skin is modeled as a stack of homogeneous layers in the terahertz and submillimeter waves regions with some anisotropy due to the helical sweat glands and other elongated entities. A dielectric model for the skin is presented, valid for a wider frequency range (up to the terahertz region) taking into account the dispersive nature of the effective conductivity. Polarized reflectivity and generalized ellipsometric parameters are calculated versus angle and wavelength. Recent studies have claimed that the helical sweat ducts act as an array of low-Q helical antennae and are dominant in shaping the spectral response in the sub-terahertz region. We found that water absorption, dispersion and multiple interference effects play the major role in shaping the spectrum without the need for the assumption of the sweat ducts acting as low-Q helical antennae. High sensitivities to the water content are found particularly in the ellipsometric parameters at large incidence angles. Hence a new methodology is proposed to detect skin cancer using variable angle ellipsometry or polarized reflectometry. The parameter found with the highest sensitivity to water content is cos Δ(pp) with Δ(pp) being the phase of the on-diagonal reflection matrix ratio between p-to-p polarization.


Simultaneous operative repair of multilevel lower extremity occlusive disease.
Sixty-two patients (39 men (63%), 23 women (27%), mean age 68 years) with multilevel lower extremity arterial occlusive disease underwent simultaneous inflow and outflow operative arterial repair consisting of aortofemoral bypass in 22 (35%), axillofemoral bypass in 17 (28%), femorofemoral bypass in 15 (24%), iliac endarterectomy in 7 (11%), and unilateral aortoiliac bypass in 1 (2%), combined with 69 outflow procedures (unilateral in 55 patients, 89%), including above-knee femoropopliteal in 12 (17%), below-knee femoropopliteal in 35 (51%), femoroinfrapopliteal in 20 (29%), popliteal tibial in 1 (1%), and femoropedal bypass in 1 (1%). Multiple criteria were used to identify patients with multilevel disease likely to benefit from multilevel procedures. The operations were performed by two operating teams in a median time of 240 minutes. Prosthetic grafts were used for eight (13%) distal bypasses, the remainder were autogenous vein. There was one operative death (1.8%). The mortality rate, morbidity rate, and operative time were not significantly different from a group of patients who underwent concurrent, isolated inflow operations (aortofemoral, axillobifemoral, femorofemoral bypass or iliac endarterectomy). Mean follow-up was 14.9 months (range, 0 to 120). The life-table primary patency for the inflow procedures was 92.6% at 24 months, the outflow was 94.9% at 24 months. Cumulative limb salvage was 90.9% at 48-month follow-up. All patients with claudication were relieved of their symptoms. We conclude that complete correction of multilevel disease can be accomplished with operative time, morbidity rate, and patency equal to that of single level repair. Multilevel procedures provide complete relief of symptoms in a higher percentage of patients than has been reported after single level repair.


Local staging of pancreatic cancer: criteria for unresectability of major vessels as revealed by pancreatic-phase, thin-section helical CT.
This study was conducted to determine the criteria for unresectability of major peripancreatic vessels in patients with pancreatic carcinoma as revealed by optimally enhanced, pancreatic-phase thin-section helical CT. Twenty-five patients with pancreatic adenocarcinoma who underwent local dissection during curative or palliative surgery also underwent preoperative pancreatic-phase thin-section helical CT (40- to 70-sec delay, 2.5- to 3-mm collimation). Tumor involvement of the portal and superior mesenteric veins and the celiac, hepatic, and superior mesenteric arteries was prospectively graded on a 0-4 scale based on circumferential contiguity of tumor to vessel. Subsequent surgical results were then correlated with the CT grades. At surgery, definitive evaluation was possible for 80 vessels. Forty-eight of 48 vessels graded 0 and three of three vessels graded 1 were resectable. Four of seven vessels graded 2, seven of eight vessels graded 3, and 14 of 14 vessels graded 4 were unresectable. A threshold of between grades 2 and 3, which corresponded to tumor involvement of one-half circumference of the vessel, yielded the lowest number of false-negatives and an acceptable number of false-positives for unresectability. Such a threshold would have yielded a sensitivity of 84%, a specificity of 98%, a positive predictive value of 95%, and a negative predictive value of 93% for unresectability of the vessels studied. A grading system for tumor involvement of the major vessels in patients with pancreatic adenocarcinoma can be based on the degree of circumferential contiguity of tumor to vessel. Involvement of vessel to tumor that exceeds one-half circumference of the vessel is highly specific for unresectable tumor.


Tetrodotoxin-resistant sodium channels contribute to directional responses in starburst amacrine cells.
The biophysical mechanisms that give rise to direction selectivity in the retina remain uncertain. Current evidence suggests that the directional signal first arises within the dendrites of starburst amacrine cells (SBACs). Two models have been proposed to explain this phenomenon, one based on mutual inhibitory interactions between SBACs, and the other positing an intrinsic dendritic mechanism requiring a voltage-gradient depolarizing towards the dendritic tips. We tested these models by recording current and voltage responses to visual stimuli in SBACs. In agreement with previous work, we found that the excitatory currents in the SBACs were directional, and remained directional when GABA receptors were blocked. Contrary to the mutual-inhibitory model, stimuli that produce strong directional signals in ganglion cells failed to reveal a significant inhibitory input to SBACs. Suppression of the tonic excitatory conductance, proposed to generate the dendritic voltage-gradient required for the dendrite autonomous model, failed to eliminate the directional signal in SBACs. However, selective block of tetrodotoxin-resistant sodium channels did reduce the strength of the directional excitatory signal in the SBACs. These results indicate that current models of direction-selectivity in the SBACs are inadequate, and suggest that voltage-gated excitatory channels, specifically tetrodotoxin-resistant sodium channels, are important elements in directional signaling. This is the first physiological evidence that tetrodotoxin-resistant sodium channels play a role in retinal information processing.


DNA recognition by the brinker repressor--an extreme case of coupling between binding and folding.
The Brinker (Brk) nuclear repressor is a major element of the Drosophila Decapentaplegic morphogen signaling pathway. Its N-terminal part has weak homology to the Antennapedia homeodomain and binds to GC-rich DNA sequences. We have investigated the conformation and dynamics of the N-terminal 101 amino acid residues of Brk in the absence and in the presence of cognate DNA by solution NMR spectroscopy. In the absence of DNA, Brk is unfolded and highly flexible throughout the entire backbone. Addition of cognate DNA induces the formation of a well-folded structure for residues R46 to R95. This structure consists of four helices forming a helix-turn-helix motif that differs from homeodomains, but has similarities to the Tc3 transposase, the Pax-6 Paired domain, and the human centromere-binding protein. The GC-rich DNA recognition can be explained by specific major groove hydrogen bonds from the N-terminal end of helix alpha3. The transition from a highly flexible, completely unfolded conformation in the absence of DNA to a well-formed structure in the complex presents a very extreme case of the "coupling of binding and folding" phenomenon.


Predictors of functional recovery following surgery for foot drop due to degenerative lumbar disease.
Surgery for foot drop secondary to lumbar degenerative disease is not always associated with postoperative functional improvement. It is still unclear whether early decompression results in better functional recovery and how soon surgery should be performed. This study aimed to evaluate predicting factors that affect short- and long-term recovery outcomes and to explore the relationship between timing of lumbar decompression and recovery from foot drop in an attempt to identify a cutoff time from symptom onset until decompression for optimal functional improvement. The authors collected demographic, clinical, and radiographic data on patients who underwent surgery for foot drop due to lumbar degenerative disease. Clinical data included tibialis anterior muscle (TAM) strength before and after surgery, duration of preoperative motor weakness, and duration of radicular pain until surgery. TAM strength was recorded at the immediate postoperative period and 1 month after surgery while long-term follow-up on functional outcomes were obtained at ≥ 2 years postsurgery by telephone interview. Data including degree and duration of preoperative motor weakness as well as the occurrence of pain and its duration were collected to analyze their impact on short- and long-term outcomes. The majority of patients (70%) showed functional improvement within 1 month postsurgery and 40% recovered to normal or near-normal strength. Univariate analysis revealed a trend toward lower improvement rates in patients with preoperative weakness of more than 3 weeks (33%) compared with patients who were operated on earlier (76.5%, p = 0.034). In a multivariate analysis, the only significant predictor for maximal strength recovery was TAM strength before surgery (OR 6.80, 95% CI 1.38-33.42, p = 0.018). Maximal recovery by 1 month after surgery was significantly associated with sustained long-term functional improvement (p = 0.006). Early surgery may improve the recovery rate in patients with foot drop caused by lumbar degenerative disease, yet the strongest predictor for the extent of recovery is the severity of preoperative TAM weakness. Maximal recovery in the short-term postoperative period is associated with sustained long-term functional improvement and independence.


A double-blind, placebo-controlled trial assessing the efficacy of levetiracetam extended-release in very heavy drinking alcohol-dependent patients.
Despite advances in the development of medications to treat alcohol dependence, few medications have been approved by the U.S. Food and Drug Administration. The use of certain anticonvulsant medications has demonstrated potential efficacy in treating alcohol dependence. Previous research suggests that the anticonvulsant levetiracetam may be beneficial in an alcohol-dependent population of very heavy drinkers. In this double-blind, randomized, placebo-controlled clinical trial, 130 alcohol-dependent patients who reported very heavy drinking were recruited across 5 clinical sites. Patients received either levetiracetam extended-release (XR) or placebo and a Brief Behavioral Compliance Enhancement Treatment intervention. Levetiracetam XR was titrated during the first 4 weeks to 2,000 mg/d. This target dose was maintained during weeks 5 through 14 and was tapered during weeks 15 and 16. No significant differences were detected between the levetiracetam XR and placebo groups in either the primary outcomes (percent heavy drinking days and percent subjects with no heavy drinking days) or in other secondary drinking outcomes. Treatment groups did not differ on a number of nondrinking outcomes, including depression, anxiety, mood, and quality of life. The only difference observed was in alcohol-related consequences. The levetiracetam XR treatment group showed significantly fewer consequences than did the placebo group during the maintenance period (p = 0.02). Levetiracetam XR was well tolerated, with fatigue being the only significantly elevated adverse event, compared with placebo (53% vs. 24%, respectively; p = 0.001). This multisite clinical trial showed no efficacy for levetiracetam XR compared with placebo in reducing alcohol consumption in heavy drinking alcohol-dependent patients.


A Molecular Host Response Assay to Discriminate Between Sepsis and Infection-Negative Systemic Inflammation in Critically Ill Patients: Discovery and Validation in Independent Cohorts.
Systemic inflammation is a whole body reaction having an infection-positive (i.e., sepsis) or infection-negative origin. It is important to distinguish between these two etiologies early and accurately because this has significant therapeutic implications for critically ill patients. We hypothesized that a molecular classifier based on peripheral blood RNAs could be discovered that would (1) determine which patients with systemic inflammation had sepsis, (2) be robust across independent patient cohorts, (3) be insensitive to disease severity, and (4) provide diagnostic utility. The goal of this study was to identify and validate such a molecular classifier. We conducted an observational, non-interventional study of adult patients recruited from tertiary intensive care units (ICUs). Biomarker discovery utilized an Australian cohort (n = 105) consisting of 74 cases (sepsis patients) and 31 controls (post-surgical patients with infection-negative systemic inflammation) recruited at five tertiary care settings in Brisbane, Australia, from June 3, 2008, to December 22, 2011. A four-gene classifier combining CEACAM4, LAMP1, PLA2G7, and PLAC8 RNA biomarkers was identified. This classifier, designated SeptiCyte Lab, was validated using reverse transcription quantitative PCR and receiver operating characteristic (ROC) curve analysis in five cohorts (n = 345) from the Netherlands. Patients for validation were selected from the Molecular Diagnosis and Risk Stratification of Sepsis study (ClinicalTrials.gov, NCT01905033), which recruited ICU patients from the Academic Medical Center in Amsterdam and the University Medical Center Utrecht. Patients recruited from November 30, 2012, to August 5, 2013, were eligible for inclusion in the present study. Validation cohort 1 (n = 59) consisted entirely of unambiguous cases and controls; SeptiCyte Lab gave an area under curve (AUC) of 0.95 (95% CI 0.91-1.00) in this cohort. ROC curve analysis of an independent, more heterogeneous group of patients (validation cohorts 2-5; 249 patients after excluding 37 patients with an infection likelihood of "possible") gave an AUC of 0.89 (95% CI 0.85-0.93). Disease severity, as measured by Sequential Organ Failure Assessment (SOFA) score or Acute Physiology and Chronic Health Evaluation (APACHE) IV score, was not a significant confounding variable. The diagnostic utility of SeptiCyte Lab was evaluated by comparison to various clinical and laboratory parameters available to a clinician within 24 h of ICU admission. SeptiCyte Lab was significantly better at differentiating cases from controls than all tested parameters, both singly and in various logistic combinations, and more than halved the diagnostic error rate compared to procalcitonin in all tested cohorts and cohort combinations. Limitations of this study relate to (1) cohort compositions that do not perfectly reflect the composition of the intended use population, (2) potential biases that could be introduced as a result of the current lack of a gold standard for diagnosing sepsis, and (3) lack of a complete, unbiased comparison to C-reactive protein. SeptiCyte Lab is a rapid molecular assay that may be clinically useful in managing ICU patients with systemic inflammation. Further study in population-based cohorts is needed to validate this assay for clinical use.


Non-cell-autonomous regulation of petal initiation in Arabidopsis thaliana.
In many flowering plants, petals initiate in alternate positions from first whorl sepals, suggesting possible signaling between sepal boundaries and petal initiation sites. PETAL LOSS (PTL) and RABBIT EARS (RBE) regulate petal initiation in Arabidopsis thaliana and their transcripts are expressed in sepal boundary and petal initiation sites, respectively, suggesting that PTL acts in a non-cell-autonomous manner. Here, we determined that cells expressing PTL and RBE fusion proteins did not overlap but were adjacent, confirming the non-cell-autonomous function of PTL. Genetic ablation of intersepal cells by expressing the diphtheria toxin-A chain gene driven by the PTL promoter resulted in flowers lacking petals, suggesting these cells are required for petal initiation. Transcriptome analysis combined with a PTL induction system revealed 42 genes that were upregulated under PTL activation, including UNUSUAL FLORAL ORGANS (UFO), which likely plays an important role in petal initiation. These findings suggest a molecular mechanism in which PTL indirectly regulates petal initiation and UFO mediates positional signaling between the sepal boundary and petal initiation sites.


Individual placement and support for young adults at risk of early work disability (the SEED trial). A randomized controlled trial.
Objectives Individual placement and support (IPS) is an effective approach for helping people with severe mental illness gain employment. This study aimed to investigate if IPS can be effectively repurposed to support young adults at risk of early work disability due to various social and health related problems. Methods A randomized controlled trial including 96 young adults (18‒29 years; 68% men) was conducted in Norway. Participants were not in employment, education, or training, received temporary benefits due to social or health-related problems, and were eligible for traditional vocational rehabilitation (TVR). Participants were randomized to IPS (N=50) or TVR (N=46). Self-reported data were collected at baseline and at 6- and 12-months follow-up. The primary outcome was obtaining any paid employment in the competitive labor market during follow-up. Secondary outcomes were physical and mental health, well-being, coping, alcohol consumption, and drug use. Results Significantly more IPS participants obtained competitive employment compared to TVR participants during 12-months follow-up (48% versus 8%; odds ratio 10.39, 95% confidence interval 2.79‒38.68). The IPS group reported significantly better outcomes than the TVR group in subjective health complaints, helplessness, and hopelessness. In post hoc analyses adjusted for baseline and missing data, the IPS group reported significantly better outcomes on these measures in addition to level of disability, optimism about future well-being, and drug use. Conclusions IPS is effective for young adults at risk of early work disability. IPS was superior to TVR in increasing competitive employment and promoted improvements in some non-vocational outcomes. IPS services should be offered to improve employment rates in this vulnerable group.


Understanding influences on physical activity participation by older adults: A qualitative study of community-dwelling older adults from the Hertfordshire Cohort Study, UK.
The health benefits of physical activity (PA) participation in later life are widely recognised. Understanding factors that can influence the participation of community-dwelling older adults in PA is crucial in an ageing society. This will be paramount in aiding the design of future interventions to effectively promote PA in this population. The main aim of this qualitative study was to explore influences on PA among community-dwelling older people, and the secondary aim was to explore gender differences. Qualitative data were collected in 2014 by conducting focus group discussions using a semi-structured discussion guide with older people resident in Hertfordshire, UK. Discussions were audio-recorded, transcribed verbatim and transcripts analysed thematically. Ninety-two participants were recruited to the study (47% women; 74-83 years) and a total of 11 focus groups were conducted. Findings indicated six themes that appeared to affect older adults' participation in PA: past life experiences; significant life events; getting older; PA environment; psychological/personal factors; and social capital. Overall, the findings emphasised the role of modifiable factors, namely psychological factors (such as self-efficacy, motivation, outcome expectancy) and social factors (such as social support and social engagement). These factors exerted their own influence on physical activity participation, but also appeared to mediate the effect of other largely non-modifiable background and ageing-related factors on participants' engagement with PA in later life. In view of these findings, intervention designers could usefully work with behavioural scientists for insight as to how to enhance psychological and social factors in older adults. Our data suggest that interventions that aim to build self-efficacy, motivation and social networks have the potential to indirectly promote PA participation in older adults. This would be best achieved by developing physical activity interventions through working with participants in an empowering and engaging way.


Optimizing global health experiences in emergency medicine residency programs: a consensus statement from the Council of Emergency Medicine Residency Directors 2011 Academic Assembly global health specialty track.
An increasing number of emergency medicine (EM) residency training programs have residents interested in participating in clinical rotations in other countries. However, the policies that each individual training program applies to this process are different. To our knowledge, little has been done in the standardization of these experiences to help EM residency programs with the evaluation, administration and implementation of a successful global health clinical elective experience. The objective of this project was to assess the current status of EM global health electives at residency training programs and to establish recommendations from educators in EM on the best methodology to implement successful global health electives. During the 2011 Council of Emergency Medicine Residency Directors (CORD) Academic Assembly, participants met to address this issue in a mediated discussion session and working group. Session participants examined data previously obtained via the CORD online listserve, discussed best practices in global health applications, evaluations and partnerships, and explored possible solutions to some of the challenges. In addition a survey was sent to CORD members prior to the 2011 Academic Assembly to evaluate the resources and processes for EM residents' global experiences. Recommendations included creating a global health working group within the organization, optimizing a clearinghouse of elective opportunities for residents and standardizing elective application materials, site evaluations and resident assessment/feedback methods. The survey showed that 71.4% of respondents have global health partnerships and electives. However, only 36.7% of programs require pre-departure training, and only 20% have formal competency requirements for these global health electives. A large number of EM training programs have global health experiences available, but these electives and the trainees may benefit from additional institutional support and formalized structure.


Morphological description of Amblyomma brasiliense Aragão, 1908 (Acari: Ixodidae) larvae and nymphs.
The immature stages of Neotropical ticks are poorly known and in many cases have not been described. Tis work presents a morphological description of Amblyomma brasiliense larvae (F1) and a redescription of nymphs (F1). A. brasiliense is reported as one of the most aggressive ticks to humans in Brazil. Immature ticks obtained from a laboratory colony initiated from adult specimens collected in the Parque Estadual Intervales (24 degrees 18 degrees S and 48 degrees 24' W), São Paulo, Brazil, were analyzed under scanning electron microscope, and also under light and stereoscopic microscopes. A. brasiliense larvae present basis capituli rectangular; short palpi; rounded idiosoma; coxa I with two spurs, the external one being longer than the internal one; and coxae II and III each with one short spur. Nymphs present basis capituli rectangular with a sharp pointed cornua; oval idiosoma with scutum reaching coxa III; coxa I with two evident spurs, the external one being longer than the internal one; coxae II-III each with one short spur; and coxa IV with a very short spur and chitinous tubercles on internal surface of posterior border of idiosoma. These morphological features, in association with chaetotaxy and porotaxy, should make possible the identification of immature Amblyomma ticks.


Resistance caused by decreased penetration of beta-lactam antibiotics into Enterobacter cloacae.
Strains of Enterobacter cloacae were selected on the basis of resistance to aztreonam, ceftazidime, moxalactam, or imipenem. All strains produced the same E2 beta-lactamase, with an isoelectric point greater than 9.5 and with high hydrolytic activity in the presence of cephaloridine. Resistance to beta-lactams could not be correlated with the amount of beta-lactamase present in the various strains. beta-Lactamase activity was induced strongly by moxalactam and imipenem in the wild-type and moxalactam-resistant strains, with beta-lactamase representing as much as 4% of the total cellular protein after induction (2 X 10(5) molecules per cell). Ceftazidime and aztreonam were poor inducers. None of the antibiotics studied was readily hydrolyzed by the E2 beta-lactamase; aztreonam and moxalactam inhibited the enzyme with apparent Ki values of 1.2 and 100 nM, respectively. Aztreonam, which bound covalently to the E2 beta-lactamase with a half-life of 2.3 h at 25 degrees C, was used to measure penetrability of beta-lactam into the periplasmic space of the resistant E. cloacae strains. In all of the E2-producing organisms studied, a significant permeability barrier existed. A maximum concentration of 0.02 microgram of aztreonam per ml should have saturated the periplasmic beta-lactamase in the highest enzyme producers studied. However, fully active beta-lactamase was observed in the periplasm of cells exposed to aztreonam at concentrations at least 1,000-fold higher than that theoretically necessary to inhibit the total enzyme within the cell. Thus, the major cause for resistance to beta-lactam antibiotics in these E. cloacae strains was lack of penetration across the outer membrane.


Nucleotide sequence and expression of the catechol 2,3-dioxygenase-encoding gene of phenol-catabolizing Pseudomonas CF600.
Pseudomonas CF600 degrades phenol and some of its methylated derivatives via a plasmid-encoded catabolic pathway. The catechol 2,3-dioxygenase (C23O) enzyme of this pathway catalyses the conversion of catechol to 2-hydroxymuconic semialdehyde. We have determined the nucleotide (nt) sequence of the dmpB structural gene for this enzyme, and expressed and identified its polypeptide product in Escherichia coli. The xylE gene of TOL plasmid pWWO and the nahH gene of plasmid NAH7 encode analogous C23O enzymes. Comparison of these three genes shows homology of 78-81% on the nt level and 83-87% homology on the amino acid level.


Cardiovascular disease in women: challenge of the middle years.
Although heart disease remains the primary cause of death among women of all races and ethnic groups in the United States, many women are still unaware of these facts. Health care providers may also not be aware. Lack of awareness among providers may limit or delay the education and intervention to prevent future cardiovascular disease for women in their middle years. In addition, the metabolic syndrome may develop in these years of a woman's life because of today's lifestyle issues and the reported increases in obesity. The appearance of the metabolic syndrome already places a woman in a precarious position for the development of risk factors such as frank hypertension and diabetes. The metabolic syndrome presents an opportunity for providers to enhance surveillance and to educate their patients in the prevention of future events. Complications of pregnancy may also foreshadow the development of hypertension as well as type 2 diabetes mellitus later in life. Some investigators consider pregnancy as an excursion into the metabolic syndrome and as a failed "stress test," if complications such as gestational diabetes or preeclampsia develop. The emergence of those complications may give a more recognized prediction of future cardiovascular events. All primary care providers, including obstetricians and gynecologists, need to be alert as to the follow-up of these women, and incorporate continued vigilance within the standard office visit including waist measurement, annual lipid measurement, glucose monitoring, blood pressure assessment, and intense education and referrals when needed.


COVID-19: viral-host interactome analyzed by network based-approach model to study pathogenesis of SARS-CoV-2 infection.
Epidemiological, virological and pathogenetic characteristics of SARS-CoV-2 infection are under evaluation. A better understanding of the pathophysiology associated with COVID-19 is crucial to improve treatment modalities and to develop effective prevention strategies. Transcriptomic and proteomic data on the host response against SARS-CoV-2 still have anecdotic character; currently available data from other coronavirus infections are therefore a key source of information. We investigated selected molecular aspects of three human coronavirus (HCoV) infections, namely SARS-CoV, MERS-CoV and HCoV-229E, through a network based-approach. A functional analysis of HCoV-host interactome was carried out in order to provide a theoretic host-pathogen interaction model for HCoV infections and in order to translate the results in prediction for SARS-CoV-2 pathogenesis. The 3D model of S-glycoprotein of SARS-CoV-2 was compared to the structure of the corresponding SARS-CoV, HCoV-229E and MERS-CoV S-glycoprotein. SARS-CoV, MERS-CoV, HCoV-229E and the host interactome were inferred through published protein-protein interactions (PPI) as well as gene co-expression, triggered by HCoV S-glycoprotein in host cells. Although the amino acid sequences of the S-glycoprotein were found to be different between the various HCoV, the structures showed high similarity, but the best 3D structural overlap shared by SARS-CoV and SARS-CoV-2, consistent with the shared ACE2 predicted receptor. The host interactome, linked to the S-glycoprotein of SARS-CoV and MERS-CoV, mainly highlighted innate immunity pathway components, such as Toll Like receptors, cytokines and chemokines. In this paper, we developed a network-based model with the aim to define molecular aspects of pathogenic phenotypes in HCoV infections. The resulting pattern may facilitate the process of structure-guided pharmaceutical and diagnostic research with the prospect to identify potential new biological targets.


[Screening colorectal cancer. Perception and behavior of the population].
There is strong evidence favoring colorectal cancer screening. Preliminary data suggests that it is not included in routine practice with the adequate frequency. We intended to recognize in a Health Care System (HCS) that provides the needed resources, the facilitators and barriers related with the implementation of this preventive practice, how many individuals have carried out one procedure and what this preventive activity predicts. A telephone survey was administered to a sample of affiliates from an HCS that serves employees of the University of Buenos Aires; 132 completed the questionnaire (70% response rate). Facilitators obtained an affirmative response in a 64 to 97%, and barriers from 11 to 27%. In the latter category a special subgroup (39%) was afraid of adverse events, and there was a feeling of embarrassment in others (30%); 33% of respondents had carried out a screening procedure, mainly FOBT 27, sigmoidoscopy 11 and colonoscopy 20. A majority (95%) stated that they "would do the procedure if doctors recommend it", or "not do it unless my doctor advises to do it" (87%). Answering affirmatively that "physicians will do the best for their patients" was associated with having had a CRC screening test, OR 1.55 (95% CI: 1.02-2.37) p: 0.04. Studied individuals showed good predisposition for colorectal cancer screening, but to put it into practice, medical advice seems to be a prominent determinant.


Towards high capacity latex-coated porous polymer monoliths as ion-exchange stationary phases.
The preparation of high capacity agglomerated monolithic ion-exchangers for capillary ion chromatography is described. Post-modification of reactive monoliths was investigated as an alternative to co-polymerisation of a suitable functional monomer with an overarching goal of increasing ion-exchange capacity. Direct sulfonation of poly styrene-co-divinyl benzene monoliths using concentrated sulfuric acid or chlorosulfonic acid was unsuccessful even for monoliths containing as low as 8% crosslinker. In contrast, chemical transformation of reactive monoliths containing glycidyl methacrylate was used to increase the ion-exchange capacity by up to more than thirty-fold with ion exchange capacities of 14-29 microequiv g(-1) achieved. Three different reactions were considered, including reaction with 4-hydroxybenenesulfonic acid under basic conditions; reaction with thiobenzoic acid followed by transformation to a reactive thiol and the subsequent oxidation to the sulfonic acid; and direct sulfonation with sodium sulfite. Of these, the reaction with sodium sulfite resulted in the most significant increase in the capacity and the best separation performance. In the isocratic mode separation efficiencies of over 13,500 plates m(-1) were observed (for iodate). The separation of seven inorganic anions was also demonstrated using a hydroxide gradient.


Comparative role of disc degeneration and ligament failure on functional mechanics of the lumbar spine.
Understanding spinal kinematics is essential for distinguishing between pathological conditions of spine disorders, which ultimately lead to low back pain. It is of high importance to understand how changes in mechanical properties affect the response of the lumbar spine, specifically in an effort to differentiate those associated with disc degeneration from ligamentous changes, allowing for more precise treatment strategies. To do this, the goals of this study were twofold: (1) develop and validate a finite element (FE) model of the lumbar spine and (2) systematically alter the properties of the intervertebral disc and ligaments to define respective roles in functional mechanics. A three-dimensional non-linear FE model of the lumbar spine (L3-sacrum) was developed and validated for pure moment bending. Disc degeneration and sequential ligament failure were modelled. Intersegmental range of motion (ROM) and bending stiffness were measured. The prediction of the FE model to moment loading in all three planes of bending showed very good agreement, where global and intersegmental ROM and bending stiffness of the model fell within one standard deviation of the in vitro results. Degeneration decreased ROM for all directions. Stiffness increased for all directions except axial rotation, where it initially increased then decreased for moderate and severe degeneration, respectively. Incremental ligament failure produced increased ROM and decreased stiffness. This effect was much more pronounced for all directions except lateral bending, which is minimally impacted by ligaments. These results indicate that lateral bending may be more apt to detect the subtle changes associated with degeneration, without being masked by associated changes of surrounding stabilizing structures.


[Methylene blue to treat anaphylaxis during anesthesia: case report.].
The risk of perioperative anaphylaxis should always be considered. The incidence of anesthetic allergic reactions is controversial, varying from 1/3,000 to 1/20,000, with mortality range between 3 and 9%. This report describes the use of methylene blue as coadjuvant drug to treat anaphylaxis refractory to conventional therapy. A 53-year-old male patient was submitted to inguinal hernia correction under spinal anesthesia. After receiving 1.5 g intravenous dipirone at surgery completion, he immediately developed bronchospasm, cyanosis, decreased SpO2 and SBP, culminating with cardiac arrest. Resuscitation was started with external cardiac massage followed by tracheal intubation, as well as 1 mg epinephrine and 1 mg atropine injections. Heart rate returned (150 bpm) with no palpable pulse though. Additional 1 mg epinephrine and 1 g hydrocortisone were administered with central pulse recovery (8 minutes). Although receiving dopamine (20 microg.kg-1.min-1), patient remained hypotensive (60 mmHg) until 80 minutes. Intravenous 100 mg methylene blue was then administered with increased SBP to 85 and 105 mmHg after the second dose. Dopamine dose was tapered from 10 to 7, 5 and finally 2 microg.kg-1.min-1. Histamine is the major anaphylaxis mediator. Inducing nitric oxide (NO) production, it consequently increases guanylate cyclase, which promotes arteriolar vasodilation by increasing cyclic GMP. Methylene blue may be helpful in such situations because it inhibits guanylate cyclase and consequently vasodilation, resulting in hemodynamic improvement.


An atypical acute exacerbation of COPD due to Aspergillus fumigatus.
A 64-year-old male with a history of stabile chronic obstructive pulmonary disease (COPD) presented with increasing dyspnea and sputum for the last two months. Complete blood count showed WBC 14x103/ml, Hgb: 14.2 g/dL and eosinophilia. Blood biochemistry was normal. Chest x-ray showed hyperlucency while thorax computed tomography (CT) revealed obstructive lung disease and bronchiectasis. Pulmonary function tests demonstrated severe obstructive lung disease and a negative bronchoreversibility with a moderately reduced diffusing capacity/alveolar volume (DLCO/VA). ABG gases revealed significant hypoxemia. Sputum culture was negative. Total IgE was 1140 ng/ml. Aspergillus RAST, precipitins and aspergillusgalactomannan antigen were positive. CF genetic screening tests gave negative results. Allergic bronchopulmonary aspergillosis (ABPA) is a hypersensitivity reaction that occurs due to bronchial aspergillus colonization. It is most common in patients with asthma and cystic fibrosis. We present a COPD case with an acute exacerbation due to Aspergillus fumigatus that lead to an aberrant clinical profile unresponsive to conventional treatment. Clinicians should consider Aspergillus fumigatus as an etiologic agent in an atypical and severe COPD exacerbation.


Gamma c-signaling cytokines induce a regulatory T cell phenotype in malignant CD4+ T lymphocytes.
In this study, we demonstrate that malignant mature CD4(+) T lymphocytes derived from cutaneous T cell lymphomas (CTCL) variably display some aspects of the T regulatory phenotype. Whereas seven cell lines representing a spectrum of primary cutaneous T cell lymphoproliferative disorders expressed CD25 and TGF-beta, the expression of FOXP3 and, to a lesser degree, IL-10 was restricted to two CTCL cell lines that are dependent on exogenous IL-2. IL-2, IL-15, and IL-21, all of which signals through receptors containing the common gamma chain, induced expression of IL-10 in the IL-2-dependent cell lines as well as primary leukemic CTCL cells. However, only IL-2 and IL-15, but not IL-21, induced expression of FOXP3. The IL-2-triggered induction of IL-10 and FOXP3 expression occurred by signaling through STAT3 and STAT5, respectively. Immunohistochemical analysis of the CTCL tissues revealed that FOXP3-expressing cells were common among the CD7-negative enlarged atypical and small lymphocytes at the early skin patch and plaque stages. Their frequency was profoundly diminished at the tumor stage and in the CTCL lymph node lesions with or without large cell transformation. These results indicate that the T regulatory cell features are induced in CTCL T cells by common gamma chain signaling cytokines such as IL-2 and do not represent a fully predetermined, constitutive phenotype independent of the local environmental stimuli to which these malignant mature CD4(+) T cells become exposed.


A new genus of Stenetriidae Hansen, 1905 (Asellota: Isopoda: Crustacea) from the Great Barrier Reef, Australia and the southwestern Pacific.
Onychatrium gen. nov. is described, with five included species: Onychatrium forceps sp. nov., the type species and Onychatrium torosus sp. nov., both from the Great Barrier Reef; Onychatrium entale (Nordenstam, 1946) comb. nov., from Tapateuen (= Tabiteue Island), Gilbert Islands; Onychatrium thomasi (Bolstad & Kensley, 1999) comb. nov., from Madang, Papua New Guinea; and Onychatrium echiurum (Nobili, 1906) comb. nov., and species inquirenda from the Tumaotu Islands, Eastern French Polynesia. The primary distinguishing characters for Onychatrium gen. nov. are a trapezoid pseudosrostrum, the male pereopod 1 with elongate dactylus (4.7-7.3 as long as proximal width), propodus with strongly produced and acute lobe, carpus with a distally acute, flat, ventrally directed process (except O. torosus sp. nov., which has a short and truncate process) and the merus with a distally directed inferodistal lobe. The genus is known only from the southern Pacific, from the Tuamotus (eastern French Polynesia) to the Great Barrier Reef and northern Papua New Guinea.


[Analysis of malignancy of human brain glioma with silver colloid-staining technique].
The nucleolar organizer regions are loops of DNA that exist in cellular nucleolus and possess ribosomal ribonucleic acid genes. Ribonucleic acid polymerase I acts on these areas. This experience collects 52 primary human gliomas at TSGH from 1984 to 1989. These included 23 astrocytomas, 8 anaplastic astrocytomas and 21 glioblastoma multiforms. After thin cutting on slides and deparaffinizing, we stained each by the silver colloid technique. We counted the total quantity of Ag-NOR dots of over 400 tumor cells of each case and average of Ag-NOR dots per cell (+/- standard error of the mean) with X1000 oil lens. The results are 1.68 +/- 0.05, 2.00 +/- 0.10; 2.55 +/- 0.13 respectively, showing significant differences among these. The average Ag-NOR paralleled the degree of histopathological malignancy. With the average Ag-NOR of tumor cells to predict the patients prognosis, we found that the group containing patients with less than 2.00 Ag-NOR per cell had better prognosis than the group with 2.00 Ag-NOR or more. The silver colloid-staining technique is a simple, rapid and reproducible method to evaluate the malignancy of human brain gliomas and to estimate the long-term prognosis of patients.


Mechanical Properties and Chemical Reactivity of Li xSiO y Thin Films.
Silicon (Si) is a commonly studied candidate material for next-generation anodes in Li-ion batteries. A native oxide SiO2 on Si is often inevitable. However, it is not clear if this layer has a positive or negative effect on the battery performance. This understanding is complicated by the lack of knowledge about the physical properties of the SiO2 lithiation products and by the convolution of chemical and electrochemical effects during the anode lithiation process. In this study, Li xSiO y thin films as model materials for lithiated SiO2 were deposited by magnetron sputtering at ambient temperature, with the goal of (1) decoupling chemical reactivity from electrochemical reactivity and (2) evaluating the physical and electrochemical properties of Li xSiO y. X-ray photoemission spectroscopy analysis of the deposited thin films demonstrate that a composition close to previous experimental reports of lithiated native SiO2 can be achieved through sputtering. Our density functional theory calculations also confirm that the possible phases formed by lithiating SiO2 are very close to the measured film compositions. Scanning probe microscopy measurements show that the mechanical properties of the film are strongly dependent on lithium concentration, with a ductile behavior at a higher Li content and a brittle behavior at a lower Li content. The chemical reactivity of the thin films was investigated by measuring the AC impedance evolution, suggesting that Li xSiO y continuously reacts with the electrolyte, in part because of the high electronic conductivity of the film determined from solid-state impedance measurements. The electrochemical cycling data of the sputter-deposited Li xSiO y/Si films also suggest that Li xSiO y is not beneficial in stabilizing the Si anode surface during battery operation, despite its favorable mechanical properties.


Bisphenol AP is anti-estrogenic and may cause adverse effects at low doses relevant to human exposure.
A recent increase in the use of bisphenol A (BPA) alternatives to manufacture plastics has led to safety concerns. Here, we evaluated the estrogenic and anti-estrogenic activities of bisphenol AP (BPAP), a poorly studied BPA alternative, using in vitro, in vivo and in silico tools. BPAP exhibited weak estrogenicity but strong anti-estrogenicity (IC50 = 2.35 μM) in a GeneBLAzer™ β-lactamase reporter gene assay. BPAP, when administered alone or in combination with E2 (50 μg kg-1 bw d-1) for 3 d, significantly decreased the uterine weights of post-weaning CD-1 mice at doses of 10 mg kg-1 bw d-1 and higher. When administered alone to prepubertal CD-1 mice for 10 d, BPAP significantly decreased the uterine weights at doses of 80 μg kg-1 bw d-1 and higher. Toxicogenomic analysis showed that BPAP regulated an opposite patterns of gene expression than that of E2 in mouse uteri. In a glucose tolerance test using male mice, BPAP was found to disrupt the blood glucose homeostasis at low doses relevant to human exposure (1 and 100 μg kg-1 bw d-1). Our results suggest that BPAP should be of great concern which might affect the sexual development in immature feminine and disrupt the blood glucose homeostasis at very low doses.


Impaired mesial frontal and putamen activation in Parkinson's disease: a positron emission tomography study.
Selection of movement in normal subjects has been shown to involve the premotor, supplementary motor, anterior cingulate, posterior parietal, and dorsolateral prefrontal areas. In Parkinson's disease (PD), the primary pathological change is degeneration of the nigrostriatal dopaminergic projections, and this is associated with difficulty in initiating actions. We wished to investigate the effect of the nigral abnormality in PD on cortical activation during movement. Using C15O2 and positron emission tomography (PET), we studied regional cerebral blood flow in 6 patients with PD and 6 control subjects while they performed motor tasks. Subjects were scanned while at rest, while repeatedly moving a joystick forward, and while freely choosing which of four possible directions to move the joystick. Significant increases in regional cerebral blood flow were determined with covariance analysis. In normal subjects, compared to the rest condition, the free-choice task activated the left primary sensorimotor cortex, left premotor cortex, left putamen, right dorsolateral prefrontal cortex and supplementary motor area, anterior cingulate area, and parietal association areas bilaterally. In the patients with PD, for the free-choice task, compared with the rest condition, there was significant activation in the left sensorimotor and premotor cortices but there was impaired activation of the contralateral putamen, the anterior cingulate, supplementary motor area, and dorsolateral prefrontal cortex. Impaired activation of the medial frontal areas may account for the difficulties PD patients have in initiating movements.


Investigating neural correlates of behavior in freely behaving rodents using inertial sensors.
Simultaneous behavior and multielectrode neural recordings in freely behaving rodents holds great promise to study the neural bases of behavior and disease models in combination with genetic manipulations. Here, we introduce the use of three-axis accelerometers to characterize the behavior of rats and mice during chronic neural recordings. These sensors were small and light enough to be worn by rodents and were used to record three-axis acceleration during freely moving behavior. A two-layer neural network-based pattern recognition algorithm was developed to extract the natural behavior of mice from the acceleration data. Successful recognition of resting, eating, grooming, and rearing are shown using this approach. The inertial sensors were combined with continuous 24-h recordings of neural data from the striatum of mice to characterize variations in neural activity with circadian cycles and to study the neural correlates of spontaneous action initiation. Finally, accelerometers were used to study the performance of rodents in traditional operant conditioning, where they were used to extract the reaction time of rodents. Thus the addition of accelerometer recordings of rodents to chronic multielectrode neural recordings provides great value for a number of neuroscience applications.


Patients' health in contract and fee-for-service care. I. A descriptive comparison.
Fee-for-service care, paying afterwards for services provided, is the traditional adult patient financial system in dentistry in Sweden. The public dental health service (PDHS) in the county of Värmland has since 1999 also an alternative system, contract care. There, a fixed sum of money is paid annually for dental care, which then is received without additional costs. This study compares the demographics, general health and oral health-related quality of life (OHRQoL) in the patient financial systems fee-for-service and contract care in the PDHS in Värmland. A questionnaire was answered by 1,324 patients, response rate 57%. A non-response analysis was undertaken. The non-response analysis showed that the likelihood for answering the questionnaire was higher for women, for respondents in contract care and for increasing age. Further analyses revealed that the non-respondents were healthier than the respondents and that experience of pain in the mouth was the only variable increasing the likelihood of response. General health was studied with the SF-36 and OHRQoL with the OHIP-14. The demographics studied were gender, age, birth country, marital status, education and social network. The results showed that there were differences in patients' health between the patient financial systems. Respondents in contract care had better OHRQoL than those in fee-for-service care. They also had better general health in four of the dimensions of SF-36, were younger, better educated, born in Sweden and were married/living with somebody to a larger extent than fee-for-service care respondents. Fee-for-service care respondents experienced higher social affinity with their housing area. In conclusion, patients in contract care had better general health and OHRQoL than patients in fee-for-service care. There were social differences in choice of financial system and biased non-response.


Intradialytic protein supplementation reduces inflammation and improves physical function in maintenance hemodialysis patients.
Protein malnutrition is both a cause and consequence of inflammation and related comorbidities for maintenance hemodialysis (MHD) patients. This study sought to determine if oral supplementation with soy or whey protein during dialysis treatment reduces inflammation and improves physical function and body composition in MHD patients. The design used in the study was randomized controlled trial, and the setting used was hemodialysis clinics in Champaign and Chicago, Illinois. Patients who received treatment ≥3 days/week, were ages ≥30 years did not have congestive heart failure or chronic obstructive pulmonary disease, and were receiving dialysis treatment for ≥3 months were eligible for inclusion. Patients were randomized to oral supplementation with a whey protein, soy protein, or placebo beverage. Patients (WHEY, n = 11; SOY, n = 12; CON, n = 15) consumed their assigned beverage before every dialysis session for 6 months. Body composition was measured by dual-energy x-ray absorptiometry, physical function by gait speed and shuttle walk test, and markers of inflammation (C-reactive protein and interleukin 6) using commercially available enzyme-linked immunosorbent assay kits before and after the 6-month intervention. Dietary intake was assessed by 24-hour dietary recalls. Six months of whey or soy supplementation significantly reduced predialysis interleukin 6 levels (P < .05 for both), whereas there was a trend for a reduction in C-reactive protein when both protein groups were combined (P = .062). Gait speed and shuttle walk test performance also significantly improved in the protein groups (P < .05 for both). No changes in body composition were observed. However, alkaline phosphatase, a marker of bone turnover, was significantly reduced in the protein groups. Intradialytic protein supplementation during a 6-month intervention reduced inflammation and improved physical function and represents an affordable intervention to improve the health of MHD patients.


A Comparison of QM/MM Simulations with and without the Drude Oscillator Model Based on Hydration Free Energies of Simple Solutes.
Maintaining a proper balance between specific intermolecular interactions and non-specific solvent interactions is of critical importance in molecular simulations, especially when predicting binding affinities or reaction rates in the condensed phase. The most rigorous metric for characterizing solvent affinity are solvation free energies, which correspond to a transfer from the gas phase into solution. Due to the drastic change of the electrostatic environment during this process, it is also a stringent test of polarization response in the model. Here, we employ both the CHARMM fixed charge and polarizable force fields to predict hydration free energies of twelve simple solutes. The resulting classical ensembles are then reweighted to obtain QM/MM hydration free energies using a variety of QM methods, including MP2, Hartree⁻Fock, density functional methods (BLYP, B3LYP, M06-2X) and semi-empirical methods (OM2 and AM1 ). Our simulations test the compatibility of quantum-mechanical methods with molecular-mechanical water models and solute Lennard⁻Jones parameters. In all cases, the resulting QM/MM hydration free energies were inferior to purely classical results, with the QM/MM Drude force field predictions being only marginally better than the QM/MM fixed charge results. In addition, the QM/MM results for different quantum methods are highly divergent, with almost inverted trends for polarizable and fixed charge water models. While this does not necessarily imply deficiencies in the QM models themselves, it underscores the need to develop consistent and balanced QM/MM interactions. Both the QM and the MM component of a QM/MM simulation have to match, in order to avoid artifacts due to biased solute⁻solvent interactions. Finally, we discuss strategies to improve the convergence and efficiency of multi-scale free energy simulations by automatically adapting the molecular-mechanics force field to the target quantum method.


Dynamic interactions and mutual synchronization of sinoatrial node pacemaker cells. A mathematical model.
Dynamic interactions and mutual entrainment of coupled sinoatrial pacemaker cells with different intrinsic frequencies were investigated using a computerized mathematical model. Transmembrane potentials were simulated using equations of individual membrane currents based on voltage clamp data for the sinoatrial node. The intrinsic frequency of a given cell was altered by applying bias hyperpolarizing current, or by changing the amount of slow inward current. Cells were coupled through simple ohmic resistances to form linear arrays of two or more cells. Simulations closely reproduced previous experimental work showing that the mutual interactions between pacemakers are mediated electrotonically and show phase dependence. Results from the present simulations provide an explanation for the ionic basis of these phase-dependent interactions. In addition, it is demonstrated that the mutual entrainment of coupled pacemakers can lead to their coordinated behavior (synchronization). Two pacemaker cells can synchronize at simple harmonic (i.e., 1:1, 2:1, etc.) or more complex ratios (3:2, 5:3, etc.), depending on the differences in intrinsic frequencies and the degree of electrical coupling between cells. Simulations using larger numbers of linearly connected cells yielded various patterns of pacemaker activity including 2:1 sinoatrial block and complex dysrhythmic activity. The overall results may be used to predict higher order interactions of thousands of cells comprising the sinus node. Under such a scheme, synchronization occurs not by the conducted influence of a dominant pacemaker cell, but by the mutual "democratic" interaction of individual pacemaker cells.


Crossover Assessment of Intraoral and Cuffed Ventilation by Emergency Responders.
A cuffed bag valve mask (BVM) is the most common device used by emergency medical responders to ventilate patients. The BVM can be difficult for users to seal around the patient's mouth and nose. An intraoral mask (IOM) with snorkel-like design may facilitate quicker and better ventilation particularly under austere conditions. Both a BVM and IOM were utilized by 27 trained emergency medical technicians and paramedics to ventilate a lightly embalmed cadaver. Ventilation efficacy, workload, and usability were assessed for both devices across four study conditions. The IOM was superior to the BVM in delivered tidal volume ratio (measure of leak, p < 0.03) and minute ventilation (p < 0.0001). Workload, ergonomic and usability assessments indicated that the IOM facilitated gripping through the reduced hand interface size (p < 0.01), decreased user effort (p < 0.001), and reduced upper limb workload (p = 0.0088). In the assessed model, the IOM represented a better choice for airway management than the standard cuffed BVM. An emergency medical device that is intuitive, efficacious and less demanding has the potential to reduce responder stress and improve resuscitation efforts, especially during austere rescue and patient transport.


Calibration of the Computer Science and Applications, Inc. accelerometer.
We established accelerometer count ranges for the Computer Science and Applications, Inc. (CSA) activity monitor corresponding to commonly employed MET categories. Data were obtained from 50 adults (25 males, 25 females) during treadmill exercise at three different speeds (4.8, 6.4, and 9.7 km x h(-1)). Activity counts and steady-state oxygen consumption were highly correlated (r = 0.88), and count ranges corresponding to light, moderate, hard, and very hard intensity levels were < or = 1951, 1952-5724, 5725-9498, > or = 9499 cnts x min(-1), respectively. A model to predict energy expenditure from activity counts and body mass was developed using data from a random sample of 35 subjects (r2 = 0.82, SEE = 1.40 kcal x min(-1)). Cross validation with data from the remaining 15 subjects revealed no significant differences between actual and predicted energy expenditure at any treadmill speed (SEE = 0.50-1.40 kcal x min(-1)). These data provide a template on which patterns of activity can be classified into intensity levels using the CSA accelerometer.


Protein kinase Calpha negatively regulates cell spreading and motility in MDA-MB-231 human breast cancer cells downstream of epidermal growth factor receptor.
Previous work has shown that phorbol esters modulate chemotaxis. Here, we demonstrate that PKC activation via phorbol 12-myristate 13-acetate (PMA) treatment of MDA-MB-231 cells inhibits EGF-induced cell spreading, the initial event of motility and chemotaxis. Of five PKC isoforms (alpha,iota,lambda,delta,and epsilon) identified in this cell line, PMA treatment only induced PKCalpha translocation from the cytosol to the membrane, an event that correlated with the development of the rounded morphology. Cell recovery was linked to PKCalpha downregulation in part via the proteasome pathway since treatment with MG101 in the presence of PMA did not lead to PKCalpha degradation and cell recovery. Co-immunoprecipitation and immunolocalization demonstrated that EGF co-localized with PKCalpha and EGFR, however, PMA did not abrogate EGFR transactivation. This work suggests that PKCalpha is the primary target of PMA acting as a transient negative regulator of cell spreading and motility in MDA-MB-231 breast cancer cells.


Characteristics of Internet use in relation to game genre in Korean adolescents.
As the number of internet users increases, a new game genre using the internet as a networking tool is emerging. Some game genres are regarded as having greater addiction potentials than others. Games and the internet are closely related. We investigated games frequently used by adolescents and classified each of them with the help of game professionals. We also examined internet use patterns to identify relationships between game genre and internet use patterns. 627 middle school and high school students (male 488, female 139) completed questionnaires concerning computer and game use patterns and Korean internet addiction scales. Game genres were divided into eight criteria (simulation, role playing game, web board, community, action, adventure, shooting, and sports). Using Korean internet addiction scales, 627 participants were divided into a normal group (474), a potential risk group (128), and a high-risk group (25). Each group showed significant differences in total internet addiction scores. We classified players into specific game users based upon the game types they most prefer. Role playing game users showed significantly higher internet addiction scores than web board and sports game users. Game and internet addictions are also connected with interpersonal relationship patterns. We suggest that users of some game genre have unique psychological addiction potentials that are different from others and that this influences both game selection and internet use.


Overexpression of Sarcoendoplasmic Reticulum Calcium ATPase 2a Promotes Cardiac Sympathetic Neurotransmission via Abnormal Endoplasmic Reticulum and Mitochondria Ca2+ Regulation.
Reduced cardiomyocyte excitation-contraction coupling and downregulation of the SERCA2a (sarcoendoplasmic reticulum calcium ATPase 2a) is associated with heart failure. This has led to viral transgene upregulation of SERCA2a in cardiomyocytes as a treatment. We hypothesized that SERCA2a gene therapy expressed under a similar promiscuous cytomegalovirus promoter could also affect the cardiac sympathetic neural axis and promote sympathoexcitation. Stellate neurons were isolated from 90 to 120 g male, Sprague-Dawley, Wistar Kyoto, and spontaneously hypertensive rats. Neurons were infected with Ad-mCherry or Ad-mCherry-hATP2Aa (SERCA2a). Intracellular Ca2+ changes were measured using fura-2AM in response to KCl, caffeine, thapsigargin, and carbonylcyanide-p-trifluoromethoxyphenylhydrazine to mobilize intracellular Ca2+ stores. The effect of SERCA2a on neurotransmitter release was measured using [3H]-norepinephrine overflow from 340 to 360 g Sprague-Dawley rat atria in response to right stellate ganglia stimulation. Upregulation of SERCA2a resulted in greater neurotransmitter release in response to stellate stimulation compared with control (empty: 98.7±20.5 cpm, n=7; SERCA: 186.5±28.41 cpm, n=8; P<0.05). In isolated Sprague-Dawley rat stellate neurons, SERCA2a overexpression facilitated greater depolarization-induced Ca2+ transients (empty: 0.64±0.03 au, n=57; SERCA: 0.75±0.03 au, n=68; P<0.05), along with increased endoplasmic reticulum and mitochondria Ca2+ load. Similar results were observed in Wistar Kyoto and age-matched spontaneously hypertensive rats, despite no further increase in endoplasmic reticulum load being observed in the spontaneously hypertensive rat (spontaneously hypertensive rats: empty, 0.16±0.04 au, n=18; SERCA: 0.17±0.02 au, n=25). In conclusion, SERCA2a upregulation in cardiac sympathetic neurons resulted in increased neurotransmission and increased Ca2+ loading into intracellular stores. Whether the increased Ca2+ transient and neurotransmission after SERCA2A overexpression contributes to enhanced sympathoexcitation in heart failure patients remains to be determined.


Community healthcare worker response to childhood disorders: Inadequacies and needs.
Community healthcare workers (CHWs) play a vital role in linking health facilities and communities where there is a high prevalence of childhood disorders. However, there is limited literature on whether this cadre of workers is adequately prepared for this task. This study explored the training needs of CHWs working in the field of childhood disorders and disabilities to improve the future training of CHWs and service delivery. This study was conducted in an urban district in KwaZulu-Natal, South Africa. This qualitative study used purposive sampling to recruit 28 CHWs and 4 key informants working in health facilities in one district of the KwaZulu-Natal Province in South Africa. Data were collected via semi-structured interviews and focus groups. Interviews were conducted in the first language (isiZulu) of the CHWs. Data were analysed thematically. Ethical clearance was obtained from a Biomedical Science Research Ethics Committee. There was an evident lack of knowledge and skill in managing childhood disorders and disabilities by CHWs. Enablers and restrictors affecting service delivery were highlighted. Moreover, the training needs of CHWs have raised critical concerns because of the variable nature of training and perceived inadequate preparation for service delivery. The challenges raised were also generic to the holistic role of CHWs and not particularly specific to the CHW role in childhood disorders and disabilities. Training of CHWs in childhood disorders may assist in improving CHWs' competence and confidence in the field, which may enhance service delivery and thus may assist in contributing towards improving healthcare for children at this level of care.


Parameters affecting the chemical work output of a hybrid photoelectrochemical biofuel cell.
A hybrid photoelectrochemical biofuel cell employing the photoanode architecture of a dye-sensitized solar cell has been assembled. A porphyrin dye sensitizes a TiO(2) semiconductor over the visible range to beyond 650 nm. Photoinduced charge separation at the dye-TiO(2) interface results in electron migration to a cathode, and the holes generated on surface bound dyes oxidize soluble electron mediators. The increased [Ox] : [Red] ratio of the mediator drives the solution-based enzymatic oxidation of appropriate substrates. In this report we investigate how the accumulation of anodic and cathodic products limits cell performance. The NAD(+)/NADH and benzoquinone/hydroquinone redox couples were studied as sacrificial electron donors in the absence of appropriate enzymes or substrates. Comparatively poor cell performance was observed using the benzoquinone/hydroquinone couple. This effect is explained in terms of rapid charge recombination by electron donation from the electrode to benzoquinone in solution, as compared to much less recombination with NAD(+). With the NAD(+)/NADH couple the cell performance is relatively independent of the redox poise of the anode solution, but limited by accumulation of reduction products in the cathodic compartment. Using the NAD(+)/NADH couple, the photochemical reforming of ethanol to hydrogen was demonstrated under conditions where the process would be endergonic in the dark.


TORO: ninety-six-week virologic and immunologic response and safety evaluation of enfuvirtide with an optimized background of antiretrovirals.
The additional 48-week optional treatment extension of the T-20 versus Optimized Regimen Only (TORO) studies evaluated long-term safety and efficacy of enfuvirtide (ENF) through week 96 in patients receiving ENF plus optimized background (OB) and patients switching to ENF plus OB from OB alone. Patient randomization was 2:1 to ENF plus OB (n = 663) and OB (n = 334), of which 89.7% and 89.8% were male, 89.3% and 88.6% were Caucasian, and median age was 41 and 42 years, respectively. HIV risk factors were comparable between the ENF plus OB and OB groups with the major factors being 65.2% versus 66.2% homosexual contact, 17.8% versus 19.8% heterosexual contact, 4.1% versus 4.8% bisexual contact, respectively, and 6.9% injection drug use in both groups. OB patients were allowed to switch to ENF plus OB at virologic failure before week 48 and required to switch at week 48 to continue in the study (n = 230). Efficacy and safety assessments were conducted for each group. At week 96, 55% of ENF plus OB subjects completed the study and 26.5% achieved a viral load of less than 400 copies per milliliter (17.5% achieved less than 50 copies per milliliter). Viral load and CD4 mean change from baseline was -2.1 and -1.1 log(10) HIV-1-RNA copies per milliliter and +166 and +116 CD4 cells/mm(3) for ENF plus OB and switch patients, respectively. No new ENF-related safety issues emerged in weeks 48-96. Injection site reactions led to discontinuation in 7% and 10% of ENF plus OB and switch patients, respectively. In conclusion, these data demonstrate durable efficacy and safety of ENF over 96 weeks and that early use of ENF in combination with other agents for the treatment of antiretroviral-experienced HIV-infected subjects is beneficial.


Cancer cell death induced by nanomagnetolectin.
Magnetic nanoparticles represent a new paradigm for molecular targeting therapy in cancer. However, the transformative targeting potential of magnetic nanoparticles has been stymied by a key obstacle-safe delivery to specified target cells in vivo. As cancer cells grow under nutrient deprivation and hypoxic conditions and decorate cell surface with excessive sialoglycans, sialic acid binding lectins might be suitable for targeting cancer cells in vivo. Here we explore the potential of magnetic nanoparticles functionalized with wheat germ lectin (WGA) conjugate, so-called nanomagnetolectin, as apoptotic targetable agents for prostate cancer. In the presence of magnetic field (magnetofection) for 15min, 2.46nM nanomagnetolectin significantly promoted apoptosis (∼12-fold, p value <0.01) of prostate cancer cells (LNCaP, PC-3, DU-145) compared to normal prostate epithelial cells (PrEC, PNT2, PZ-HPV-7), when supplemented with 10mM sialic acid under nutrient deprived condition. Nanomagnetolectin targets cell-surface glycosylation, particularly sialic acid as nanomagnetolectin induced apoptosis of cancer cells largely diminished (only 2 to 2.5-fold) compared to normal cells. The efficacy of magnetofected nanomagnetolectin was demonstrated in orthotopically xenografted (DU-145) mice, where tumor was not only completely arrested, but also reduced significantly (p value <0.001). This was further corroborated in subcutaneous xenograft model, where nanomagnetolectin in the presence of magnetic field and photothermal heating at ∼42°C induced apoptosis of tumor by ∼4-fold compared to tumor section heated at ∼42°C, but without magnetic field. Taken all together, the study demonstrates, for the first time, the utility of nanomagnetolectin as a potential cancer therapeutic.


Cellular factors promoting resistance to effective treatment of glioma with oncolytic myxoma virus.
Oncolytic virus therapy is being evaluated in clinical trials for human glioma. While it is widely assumed that the immune response of the patient to the virus infection limits the utility of the therapy, investigations into the specific cell type(s) involved in this response have been performed using nonspecific pharmacologic inhibitors or allogeneic models with compromised immunity. To identify the immune cells that participate in clearing an oncolytic infection in glioma, we used flow cytometry and immunohistochemistry to immunophenotype an orthotopic glioma model in immunocompetent mice after Myxoma virus (MYXV) administration. These studies revealed a large resident microglia and macrophage population in untreated tumors, and robust monocyte, T-, and NK cell infiltration 3 days after MYXV infection. To determine the role on the clinical utility of MYXV therapy for glioma, we used a combination of knockout mouse strains and specific immunocyte ablation techniques. Collectively, our experiments identify an important role for tumor-resident myeloid cells and overlapping roles for recruited NK and T cells in the clearance and efficacy of oncolytic MYXV from gliomas. Using a cyclophosphamide regimen to achieve lymphoablation prior and during MYXV treatment, we prevented treatment-induced peripheral immunocyte recruitment and, surprisingly, largely ablated the tumor-resident macrophage population. Virotherapy of cyclophosphamide-treated animals resulted in sustained viral infection within the glioma as well as a substantial survival advantage. This study demonstrates that resistance to MYXV virotherapy in syngeneic glioma models involves a multifaceted cellular immune response that can be overcome with cyclophosphamide-mediated lymphoablation.