[Localization, kinetic parameters, and functions of cholinesterase of the starfish ampulla].
The starfish amplullae cholinesterase was shown to represent acetylcholinesterase and enhance its activity along with increasing motility of the starfish. Bundles of muscle fibres containing cholinesterase were found in the ampullae. Cholinesterase was shown to be localized in the muscle cells and in collagen layer in vicinity of the muscle cells. The data obtained suggest participation of the starfish ampullae cholinesterase in non-synaptic cholinergic transmission between the radial nerve axons and the muscle fibre extension. Besides, the enzyme could take part in functional relationship between the muscle cells and the outer epithelial cells of the starfish ampullae.


Three-dimensional architecture of murine rod cilium revealed by cryo-EM.
The connecting cilium of the rod photoreceptor is a tubular structure that bridges two adjacent cellular compartments, the inner segment, the major site of biosynthesis and energy metabolism, and the outer segment, a highly specialized ciliary structure responsible for phototransduction. The connecting cilium allows for active processes of protein sorting and transport to occur between them. Mutations affecting the cargo, their transporters, and the structural components of the primary cilium and basal body lead to aberrant trafficking and photoreceptor cell death. Understanding the overall design of the cilium, its architectural organization, and the function of varied protein complexes within the structural hierarchy of the cilium requires techniques for visualizing their native three-dimensional structures at high magnification. Here we describe methods for isolating retinas from mice, purifying fragments of rod cells that include much of the inner segment and the rod photoreceptor cilia, vitrifying the cell fragments, and determining their structures by cryo-electron tomography.


Susceptibility of Xenopus laevis tadpoles to infection by the ranavirus Frog-Virus 3 correlates with a reduced and delayed innate immune response in comparison with adult frogs.
Xenopus laevis adults mount effective immune responses to ranavirus Frog Virus 3 (FV3) infections and clear the pathogen within 2-3 weeks. In contrast, most tadpoles cannot clear FV3 and succumb to infections within a month. While larval susceptibility has been attributed to ineffective adaptive immunity, the contribution of innate immune components has not been addressed. Accordingly, we performed a comprehensive gene expression analysis on FV3-infected tadpoles and adults. In comparison to adults, leukocytes and tissues of infected tadpoles exhibited modest (10-100 time lower than adult) and delayed (3 day later than adult) increase in expression of inflammation-associated (TNF-α, IL-1β and IFN-γ) and antiviral (Mx1) genes. In contrast, these genes were readily and robustly upregulated in tadpoles upon bacterial stimulation. Furthermore, greater proportions of larval than adult PLs were infected by FV3. Our study suggests that tadpole susceptibility to FV3 infection is partially due to poor virus-elicited innate immune responses.


Systematic review and meta-analysis of renal replacement therapy modalities for acute kidney injury in the intensive care unit.
To compare clinical outcomes among critically ill adults with acute kidney injury (AKI) treated with continuous renal replacement therapy (CRRT), intermittent hemodialysis (IHD) or sustained low efficiency dialysis (SLED). We completed a systematic review and meta-analysis of studies published in 2015 or earlier using MEDLINE®, EMBASE®, Cochrane databases and grey literature. Eligible studies included randomized clinical trials (RCTs) or prospective cohort studies comparing outcomes of mortality, dialysis dependence or length of stay among critically ill adults receiving CRRT, IHD or SLED to treat AKI. Mortality and dialysis dependence from RCTs were pooled using meta-analytic techniques. Length of stay from RCTs and results from prospective cohort studies were described qualitatively. Twenty-one studies were eligible. RRT modality was not associated with in-hospital mortality (CRRT vs IHD: RR 1.00 [95% CI, 0.92-1.09], CRRT vs SLED: RR 1.23 [95% CI, 1.00-1.51]) or dialysis dependence (CRRT vs IHD: RR 0.90 [95% CI, 0.59-1.38], CRRT vs SLED: RR 1.15 [95% CI, 0.67-1.99]). We did not find a definitive advantage for any RRT modality on short-term patient or kidney survival. Well-designed, adequately-powered trials are needed to better define the role of RRT modalities for treatment of critically ill patients with AKI.


[Therapeutic options in advanced cancer of the prostate].
Drug therapy modalities in locally advanced and metastatic carcinoma of the prostate are primarily palliative. Therefore, the goal of systemic therapy is prevention or palliation of complications, such as pain, obstructive symptoms and bleeding. Thus far, there seems to be no survival benefit from hormonal treatment. The first step is always withdrawal of male sex hormones by means of orchiectomy or administration of LH-RH analogues, 70 to 80% of the patients respond favorably. Orchiectomy and LH-RH analogues are considered equal with regard to effectiveness and side effects, however, in the latter case an antiandrogen must be administered during the two weeks prior to start of treatment to prevent the flare-up phenomenon. Estrogens are rarely used anymore, because they can cause cardiovascular complications. In asymptomatic patients, the question remains to be answered if androgen withdrawal should be performed immediately at the time of diagnosis or delayed in case of possible symptoms. Antiandrogen agents block directly the androgen receptors in the prostatic cell. However, monotherapy with antiandrogens is not yet an established procedure. Instead, since 5% of circulating androgens are of adrenal origin, antiandrogens are combined with either orchiectomy or LH-RH analogues for total androgen suppression. The benefit of such a combined androgen suppression could not be proven conclusively and might be minimal at best. Novel modalities of hormonal therapy like intermittent androgen suppression are currently being investigated. In most cases, tumor progression after hormonal therapy is due to hormone-refractory cell lines. Cytotoxic chemotherapy is largely ineffective in treating prostatic cancer. Commonly used chemotherapeutic substances lead to temporary remission in 10 to 20% of the patients at most. External beam irradiation or Strontium-89 therapy are useful in palliation of painful bone metastases.


Risk factors for BK polyomavirus nephritis in renal allograft recipients.
Recurrent episodes of acute rejection (AR) and/or the intense immunosuppression used for their treatment have been proposed as risk factors for BK nephritis (BKN; BK refers to the initials of the first patient from whom this polyomavirus was isolated). To further examine the relationship between AR and BKN, we analyzed all kidney transplants performed at our center between January 1999 and August 2001 (n = 286). After a mean follow-up of 737 +/- 22 d, we identified nine cases of BKN (3.1%). The mean time to diagnosis of BKN was 326 +/- 56 d. No patient with BKN had a prior history of AR. During the same period, 62 patients were diagnosed with AR (22%). The mean time to diagnosis of AR was 197 +/- 40 d (p = 0.01 vs. time to diagnosis of BKN). Despite aggressive therapy with methylprednisolone and, in some cases, anti-lymphocyte antibody, none of these patients with AR developed BKN. We compared the baseline characteristics of patients in both groups and found that BKN patients were more likely to be white people (78 vs. 44%, p = 0.05) and male (89 vs. 53%, p = 0.04). Moreover, the mean tacrolimus (TAC) levels before diagnosis were higher in BKN than in AR patients (11.7 +/- 0.5 vs. 6.5 +/- 0.6 ng/mL, p < 0.001). In summary, our study shows that BKN often occurs in the absence of prior episodes of AR. In addition, our findings suggest that white males exposed to higher TAC levels are at greater risk of developing BKN.


Preliminary studies of fluorescent pseudomonads capable of growth at 41 C in swimming pool waters.
During the summer of 1973 cultures of Pseudomonas aeruginosa and other fluorescent pseudomonads capable of growth at 41 C were obtained from swimming pool waters at a training center for the mentally retarded. Isolates were subjected to selected physiological tests, pyocine typing, and immunotyping. High counts of P. aeruginosa or other fluorescent pseudomonads consisted mainly of single predominant types. Both P. aeruginosa strains and unidentified fluorescent Pseudomonas strains predominated in pool waters at different times.


Relationship of posttraumatic growth to symptoms of posttraumatic stress disorder and depression: A pilot study of Iraqi students.
Posttraumatic growth (PTG) and psychopathology are common outcomes following exposure to adversity and trauma. We examined the relationship of PTG to posttraumatic stress disorder (PTSD) and depression in a group of young Iraqi students with war trauma exposure. These young Iraqis had experienced an average of 13 different war-related adversities. The prevalence of probable PTSD was 17.2% and probable depression 23.1%. PTSD was associated with higher and depression with lower PTG. In addition, the relationship between PTG and PTSD was stronger among males than females. Although PTSD and depression were relatively common, they were related to PTG in opposite directions.


The Impact of Community-Based Palliative Care on Utilization and Cost of Acute Care Hospital Services in the Last Year of Life.
Community-based palliative care may potentially benefit patients by offering their preferred care at the end of life and benefit systems by reducing hospital use. To compare place of death and acute care hospital use in the last year of life between cancer decedents who did and did not access a community-based palliative care service (PCS). Retrospective observational cohort study using linked individual administrative records from cancer registry, hospital, emergency department (ED), mortality, and PCS databases. Propensity score-weighted regression methods were used. Whole of population study incorporating 28,561 West Australian cancer decedents from 2001 to 2011. Exposure was defined as ever/never accessed PCS. Outcomes were place of death (in/out of hospital) and the number, length of stay, and cumulative cost of hospital admissions at the end of life. Decedents who accessed the service (n = 16,530) had triple (adjusted odds ratio 3.19 [3.01-3.38]) the odds of dying out of hospital compared with those who did not. Unplanned hospitalizations were reduced in the last year (adjusted incidence rate ratio [IRR] 0.94 [0.91-0.97]) and last week of life (adjusted [IRR] 0.35 [0.33-0.38]), as were ED presentations (adjusted RR 0.92 [0.98-0.95], adjusted RR 0.26 [0.23-0.28]) in the last year and last week of life, respectively. There were significant reductions in average total bed days (-7.60 [-8.34 to -6.87]) and acute care costs (-A$5,491 [-A$6,155 to -A$4,827]) over the last year of life. In addition to supporting people to die out of hospital, PCS was associated with reduced acute care admissions, bed days, and costs over the last year of life. The provision of high-quality palliative care in the community alleviates the burden on acute care hospitals and, thus, may partially offset public funding of this model.


Fine-needle aspiration cytology in nonpalpable mammographic abnormalities in breast cancer screening: results from the breast cancer screening programme in Oslo 1996-2001.
Fine-needle aspiration cytology (FNAC) of nonpalpable mammographic lesions has been under attack from two sides for some years. There has been much discussion and controversy as to the ability to differentiate between in situ and invasive carcinomas in cytological material. A further issue is that of optimal sampling to obtain adequate cell material in sufficient quantity. We present the results of FNAC from 832 nonpalpable mammographic abnormalities detected in the course of the breast cancer screening programme in Oslo during 1996-2001. In 11.6% of cases the smears were inadequate, and there were 7% false negatives (FN) and 1.3% false positives. Of the FN, 64% represented microcalcifications and 86% were due to sampling errors. Absolute sensitivity was 74%, complete sensitivity 88% and specificity 88%. In 255 carcinomas a cytological diagnosis of them as in situ or invasive was made. In 93% of the invasive cases (190/205) these had been correctly identified as invasive on FNAC. In 78% of cases proper follow-up could be resolved by cytology/radiology alone. Suboptimal sampling and localization remains the main cause of FN FNAC results. Problems in differentiating between in situ and invasive breast carcinomas can be significantly reduced by applying strict criteria for in situ lesions.


Length of post-traumatic amnesia and its prediction of neuropsychological outcome following severe to extremely severe traumatic brain injury in a litigating sample.
Primary Objective: The aim of the current study was to examine the relationship between duration of post-traumatic amnesia (PTA) and neuropsychological outcome at one or more years following severe to extremely severe traumatic brain injury (TBI) in a litigating sample. Research Design: Retrospective study design, using data collected from 2010 to 2017. Methods and Procedures: Data from 41 cases obtained from a private medicolegal neuropsychological database was examined. The database comprised information pertaining to TBI etiology, TBI severity parameters, demographic variables, neuropsychological test results and scores on psychological questionnaires. PTA duration was examined as a continuous variable. All cases that demonstrated non-credible effort were excluded. Main Outcomes and Results: Continuous PTA duration was not found to be a significant predictor of cognitive impairment across domains of verbal intellect, non-verbal intellect, working memory, information processing speed, immediate memory, delayed memory, and executive functioning. Conclusions: The predictive relationship between duration of PTA and cognitive impairment that has been reported in non-litigating populations did not exist in a litigating TBI sample. Findings illustrate the importance of investigating the relationships between injury variables and cognitive outcome in a population undergoing litigation, to provide better understanding of outcome in this subgroup of patients with TBI.


Thromboelastography (TEG) or thromboelastometry (ROTEM) to monitor haemostatic treatment versus usual care in adults or children with bleeding.
Severe bleeding and coagulopathy are serious clinical conditions that are associated with high mortality. Thromboelastography (TEG) and thromboelastometry (ROTEM) are increasingly used to guide transfusion strategy but their roles remain disputed. This review was first published in 2011 and updated in January 2016. We assessed the benefits and harms of thromboelastography (TEG)-guided or thromboelastometry (ROTEM)-guided transfusion in adults and children with bleeding. We looked at various outcomes, such as overall mortality and bleeding events, conducted subgroup and sensitivity analyses, examined the role of bias, and applied trial sequential analyses (TSAs) to examine the amount of evidence gathered so far. In this updated review we identified randomized controlled trials (RCTs) from the following electronic databases: Cochrane Central Register of Controlled Trials (CENTRAL; 2016, Issue 1); MEDLINE; Embase; Science Citation Index Expanded; International Web of Science; CINAHL; LILACS; and the Chinese Biomedical Literature Database (up to 5 January 2016). We contacted trial authors, authors of previous reviews, and manufacturers in the field. The original search was run in October 2010. We included all RCTs, irrespective of blinding or language, that compared transfusion guided by TEG or ROTEM to transfusion guided by clinical judgement, guided by standard laboratory tests, or a combination. We also included interventional algorithms including both TEG or ROTEM in combination with standard laboratory tests or other devices. The primary analysis included trials on TEG or ROTEM versus any comparator. Two review authors independently abstracted data; we resolved any disagreements by discussion. We presented pooled estimates of the intervention effects on dichotomous outcomes as risk ratio (RR) with 95% confidence intervals (CIs). Due to skewed data, meta-analysis was not provided for continuous outcome data. Our primary outcome measure was all-cause mortality. We performed subgroup and sensitivity analyses to assess the effect based on the presence of coagulopathy of a TEG- or ROTEM-guided algorithm, and in adults and children on various clinical and physiological outcomes. We assessed the risk of bias through assessment of trial methodological components and the risk of random error through TSA. We included eight new studies (617 participants) in this updated review. In total we included 17 studies (1493 participants). A total of 15 trials provided data for the meta-analyses. We judged only two trials as low risk of bias. The majority of studies included participants undergoing cardiac surgery.We found six ongoing trials but were unable to retrieve any data from them. Compared with transfusion guided by any method, TEG or ROTEM seemed to reduce overall mortality (7.4% versus 3.9%; risk ratio (RR) 0.52, 95% CI 0.28 to 0.95; I(2) = 0%, 8 studies, 717 participants, low quality of evidence) but only eight trials provided data on mortality, and two were zero event trials. Our analyses demonstrated a statistically significant effect of TEG or ROTEM compared to any comparison on the proportion of participants transfused with pooled red blood cells (PRBCs) (RR 0.86, 95% CI 0.79 to 0.94; I(2) = 0%, 10 studies, 832 participants, low quality of evidence), fresh frozen plasma (FFP) (RR 0.57, 95% CI 0.33 to 0.96; I(2) = 86%, 8 studies, 761 participants, low quality of evidence), platelets (RR 0.73, 95% CI 0.60 to 0.88; I(2) = 0%, 10 studies, 832 participants, low quality of evidence), and overall haemostatic transfusion with FFP or platelets (low quality of evidence). Meta-analyses also showed fewer participants with dialysis-dependent renal failure.We found no difference in the proportion needing surgical reinterventions (RR 0.75, 95% CI 0.50 to 1.10; I(2) = 0%, 9 studies, 887 participants, low quality of evidence) and excessive bleeding events or massive transfusion (RR 0.38, 95% CI 0.38 to 1.77; I(2) = 34%, 2 studies, 280 participants, low quality of evidence). The planned subgroup analyses failed to show any significant differences.We graded the quality of evidence as low based on the high risk of bias in the studies, large heterogeneity, low number of events, imprecision, and indirectness. TSA indicates that only 54% of required information size has been reached so far in regards to mortality, while there may be evidence of benefit for transfusion outcomes. Overall, evaluated outcomes were consistent with a benefit in favour of a TEG- or ROTEM-guided transfusion in bleeding patients. There is growing evidence that application of TEG- or ROTEM-guided transfusion strategies may reduce the need for blood products, and improve morbidity in patients with bleeding. However, these results are primarily based on trials of elective cardiac surgery involving cardiopulmonary bypass, and the level of evidence remains low. Further evaluation of TEG- or ROTEM-guided transfusion in acute settings and other patient categories in low risk of bias studies is needed.


Perspectives on perinatal brain damage.
The author presents a perspective on perinatal brain injury from the point of brain development, and discusses subsequent adverse outcomes. It is stressed that injury is frequently attributed to perinatal causes and avoidable circumstances when in fact the obstetrician could not possibly foresee or circumvent it. Evidence of the dramatic recuperative powers of many infants, even when dying and being resuscitated, are compared with the minimal provocations frequently associated with handicap, underscoring the concept of prior injury or unique fragility. The review is presented to provoke new insights and perspectives.


Utilization of soy protein isolate mixed with rice protein in Japanese women.
Utilization and requirement of soy protein isolate (SPI) and SPI-rice combination were examined in twenty-five female students. After 1 day on protein-free diet, each subject received a low-protein diet for 10 days. The protein sources were SPI for ten subjects and SPI-rice combination, in which the ratio of two proteins was 6:4, for fifteen subjects. The nitrogen intakes were about 45, 65 and 85 mg/kg in both the two series of experiments. Energy intake was at an approximate maintenance level of 36.1 +/- 3.0 kcal/kg. Apparent nitrogen balance improved with increase in nitrogen intake. The regression equations between nitrogen intake (X, mg/kg) and balance (Y, mg/kg) are shown in the following: SPI: Y = 0.411 X - 40.8 (n = 10, r = 0.812) SPI and rice protein: Y = 0.392 X -32.7 (n = 15, r = 0.739) From the above equations, the maintenance intakes of SPI and SPI-rice combination for an apparent nitrogen equilibrium were calculated to be 99 and 83 mg N/kg, respectively. Digestibilities were 98.2 +/- 5.0% for SPI and 93.1 +/- 6.1% for SPI-rice combination. The NPUs of SPI at intake levels of 40, 60 and 80 mg N/kg were 47 +/- 24 (n = 4), 49 (n = 2) and 44 +/- 3 (n = 4), respectively. The NPUs of SPI and rice mixed protein at intake levels of 45, 70 and 90 mg N/kg were 67 +/- 13 (n = 5), 51 +/- 7 (n = 5) and 54 +/- 12 (n = 5), respectively. It was concluded from the present study that both SPI and the SPI-rice combination had a high nutritive efficiency comparable with that of egg protein.


Induction of ICAM-1 on human neural cells and mechanisms of neutrophil-mediated injury.
To define mechanisms by which inflammatory cells damage neural tissue, the author investigated stimuli that promote leukocyte adherence and injury to cultured human cortical neuron (HCN-1) and neuroblastoma cells (LAN-1 and SK-N-SH). Neutrophils do not adhere to unstimulated neural cells but will bind to neural cells that have been exposed to tumor necrosis factor alpha (TNF alpha) and in some cases other cytokines such as gamma interferon (gamma IFN) or interleukin-1 (IL-1). Tumor necrosis factor alpha induces synthesis of intercellular adhesion molecule-1 (ICAM-1) mRNA and cell surface expression of ICAM-1 on cultured neural cells. Adherence of neutrophils to cytokine-stimulated neural cells is mediated primarily by ICAM-1:LFA-1 interactions, because 70% to 90% of the binding can be blocked by monoclonal antibodies to either ligand. Prior introduction of an oxidizable dye, 5-(and 6-)carboxy-2',7' dichlorofluorescin diacetate into the LAN-1 cells demonstrates that adherent neutrophils can release oxidizing radicals into the neural cell cytoplasm. These results suggest that cytokines released in the course of inflammation may induce expression of ICAM-1 on neurons, allowing them to be targeted by leukocytes expressing the appropriate receptors. The resulting adhesive interactions may facilitate introduction of various toxic agents into the neural cytoplasm.


Patient Experiences and Perceptions Associated with the Use of Desiccated Thyroid Extract.
Background and objectives: It is unclear why many patients with hypothyroidism prefer the use of desiccated thyroid extract (DTE) as a thyroid hormone replacement formulation over levothyroxine (LT4) treatment, as recommended by clinical practice guidelines. We analyzed patient-reported information from patient online forums to better understand patient preferences for and attitudes toward the use of DTE to treat hypothyroidism. Materials and Methods: We conducted a mixed-methods study by evaluating the content of online posts from three popular hypothyroidism forums from patients currently taking DTE (n = 673). From these posts, we extracted descriptive information on patient demographics and clinical characteristics and qualitatively analyzed posts' content to explore patient perceptions on DTE and other therapies further. Results: Nearly half (46%) of the patients reported that a clinician initially drove their interest in trying DTE. Patients described many reasons for switching from a previous therapeutic approach to DTE, including lack of improvement in hypothyroidism-related symptoms (58%) and the development of side effects (22%). The majority of patients described DTE as moderately to majorly effective overall (81%) and more effective than the previous therapy (77%). The most frequently described benefits associated with DTE use were an improvement in symptoms (56%) and a change in overall well-being (34%). One-fifth of patients described side effects related to the use of DTE. Qualitative analysis of posts' content supported these findings and raised additional issues around the need for individualizing therapy approaches for hypothyroidism (e.g., a sense of each patient has different needs), as well as difficulties obtaining DTE (e.g., issues with pharmacy availability). Conclusions: Lack of individualized treatment and a feeling of not been listened to were recurrent themes among DTE users. A subset of patients may prefer DTE to LT4 for many reasons, including perceived better effectiveness and improved overall well-being, despite the risks associated with DTE.


DAPD Set of Pd-Containing Diatomic Molecules: Accurate Molecular Properties and the Great Lengths to Obtain Them.
In the present study, we obtained reliable bond energy, bond length, and zero-point vibrational frequency for a set of diatomic Pd species (the DAPD set). It includes PdH, Pd2, and PdX (X = B, C, N, O, F, Al, Si, P, S, and Cl). Our highest-level protocol (W4X-L) represents scalar and spin-orbit relativistic, valence- and inner-valence correlated, extrapolated CCSDTQ(5) energy. The DAPD set of molecules is challenging for computational chemistry methods in different manners; for Pd2, the spin-orbit contribution to the bond energy is fairly large, whereas for PdC and PdSi, the post-CCSD(T) correlation components are considerable. The diverse range of requirements represents a significant challenge for lower-level methods. While density functional theory (DFT) methods generally yield good agreements for bond lengths and vibrational frequencies, large deviations are found for bond energies. In general, hybrid DFT methods are more accurate than nonhybrid functionals, but the agreement in individual cases varies. This illustrates the critical role that new high-quality reference data would play in the continual development of lower-cost methods.


Optimal illumination pattern for transport-of-intensity quantitative phase microscopy.
The transport-of-intensity equation (TIE) is a well-established non-interferometric phase retrieval approach, which enables quantitative phase imaging (QPI) of transparent sample simply by measuring the intensities at multiple axially displaced planes. Nevertheless, it still suffers from two fundamentally limitations. First, it is quite susceptible to low-frequency errors (such as "cloudy" artifacts), which results from the poor contrast of the phase transfer function (PTF) near the zero frequency. Second, the reconstructed phase tends to blur under spatially low-coherent illumination, especially when the defocus distance is beyond the near Fresnel region. Recent studies have shown that the shape of the illumination aperture has a significant impact on the resolution and phase reconstruction quality, and by simply replacing the conventional circular illumination aperture with an annular one, these two limitations can be addressed, or at least significantly alleviated. However, the annular aperture was previously empirically designed based on intuitive criteria related to the shape of PTF, which does not guarantee optimality. In this work, we optimize the illumination pattern to maximize TIE's performance based on a combined quantitative criterion for evaluating the "goodness" of an aperture. In order to make the size of the solution search space tractable, we restrict our attention to binary-coded axis-symmetric illumination patterns only, which are easier to implement and can generate isotropic TIE PTFs. We test the obtained optimal illumination by imaging both a phase resolution target and HeLa cells based on a small-pitch LED array, suggesting superior performance over other suboptimal patterns in terms of both signal-to-noise ratio (SNR) and spatial resolution.


Expression of herpes simplex virus 1 microRNAs in cell culture models of quiescent and latent infection.
To facilitate studies of herpes simplex virus 1 latency, cell culture models of quiescent or latent infection have been developed. Using deep sequencing, we analyzed the expression of viral microRNAs (miRNAs) in two models employing human fibroblasts and one using rat neurons. In all cases, the expression patterns differed from that in productively infected cells, with the rat neuron pattern most closely resembling that found in latently infected human or mouse ganglia in vivo.


Enhanced mucosal immunoglobulin A response of intranasal adenoviral vector human immunodeficiency virus vaccine and localization in the central nervous system.
Replication-defective adenovirus (ADV) vectors represent a promising potential platform for the development of a vaccine for AIDS. Although this vector is typically administered intramuscularly, it would be desirable to induce mucosal immunity by delivery through alternative routes. In this study, the immune response and biodistribution of ADV vectors delivered by different routes were evaluated. ADV vectors expressing human immunodeficiency virus type 1 (HIV-1) Gag, Pol, and Env were delivered intramuscularly or intranasally into mice. Intranasal immunization induced greater HIV-specific immunoglobulin A (IgA) responses in mucosal secretions and sera than in animals with intramuscular injection, which showed stronger systemic cellular and IgG responses. Administration of the vaccine through an intranasal route failed to overcome prior ADV immunity. Animals exposed to ADV prior to vaccination displayed substantially reduced cellular and humoral immune responses to HIV antigens in both groups, though the reduction was greater in animals immunized intranasally. This inhibition was partially overcome by priming with a DNA expression vector expressing HIV-1 Gag, Pol, and Env before boosting with the viral vector. Biodistribution of recombinant adenovirus (rADV) vectors administered intranasally revealed infection of the central nervous system, specifically in the olfactory bulb, possibly via retrograde transport by olfactory neurons in the nasal epithelium, which may limit the utility of this route of delivery of ADV vector-based vaccines.


A review of select recent patents on novel nanocarriers.
Nanocarriers offer several advantages in drug delivery including increased retention time in the body, the ability to solubilize hydrophobic cargo, and the ability to target specific tissues. These attributes have led to a large volume of research being dedicated to the development of novel nanocarriers and their use in treating disease. Many advances have been made in the synthesis and formulation of nanocarriers. For instance, flash precipitation and the use of rotary evaporator provide new methods for nanocarrier synthesis. Biomolecules like heparin and sarcosine have been successfully utilized for the synthesis of unique copolymers for use in nanocarrier systems. Also, the efficacy of nanocarrier targeting has been increased by taking advantage of unique microenvironments present in specific pathologies, utilization of phage properties, and scaffold systems for multiple targeting moieties. Finally, nanocarriers have been shown to provide immunomodulatory effects. This article provides a focused review of several recent patents covering the synthesis, composition and the use of novel nanocarriers.


[Problems of multiple tests and evaluations in drug research].
The well known inferential statistical procedures only take into account the test of a single hypothesis or the estimation of a single parameter. In practical drug research the scientist in most cases is interested in multiple tests of several hypotheses or a simultaneous estimation of several parameters. After introducing the principal problems of simultaneous statistical inference several statistical methods are discussed. These methods can be applied to a variety of problems in drug research.


Bipedicle Flaps for Posterior Elbow Reconstruction.
Chronic posterior elbow soft tissue defects often require soft tissue reconstruction because of exposure of the underlying triceps tendon and proximal ulna. Current options for soft tissue coverage require sacrifice of a local muscle or microsurgery. The purpose of this study is to evaluate patient and surgical outcomes after reconstruction of small-sized to medium-sized (<50 cm) posterior elbow defects with bipedicle advancement flaps. A retrospective chart review was performed for 3 patients who underwent posterior elbow reconstruction with bipedicle flaps. The etiology of the soft tissue elbow defect was chronic infected olecranon bursitis (n=2) and exposed olecranon plate after open fracture (n=1). Patient comorbidities included: diabetes, CREST (Calcinosis, Raynaud phenomenon, Esophageal dysmotility, Sclerodactyly, Telangiectasia) syndrome, and rheumatoid arthritis. The mean patient age at time of reconstruction was 44 years (39 to 51 y), and the mean area of soft tissue defect was 39.3 cm (24 to 54 cm) after debridement of involved tissue. All patients had positive intraoperative cultures and were treated with culture-directed long-term intravenous antibiotics. There were no surgical complications or flap loss. All patients had reepithelialization of the donor site by postoperative week 8 and stable soft tissue coverage of the elbow after surgery at final follow-up with full preoperative elbow range of motion. This technique offers a simple, reliable solution for soft tissue coverage of the posterior elbow using excess local tissue for primary closure of posterior elbow wounds with minimal donor-site morbidity.


Inhalation technique of 166 adult asthmatics prior to and following a self-management program.
Self-management of asthma and self-treatment of exacerbations are considered important in the treatment of asthma. For successful self-treatment, medication has to be inhaled correctly, but the percentage of patients inhaling effectively varies widely. As part of a self-management program we checked and corrected inhalation technique. This paper addresses differences among inhalers in relation to patient characteristics and the effect of instruction, 1 year after enrollment. Maneuvers that are essential for adequate inhalation were identified. When errors in inhalation technique were observed, patients were instructed in the correct use of their devices. One year later, inhalation technique was checked again. Only patients who used the same inhaler throughout the entire study period were analyzed. Of the 245 adult asthmatic patients who were enrolled in the self-management program, 166 used the same inhaler throughout the study period. One hundred twenty patients (72%) performed all key items correctly at baseline and this increased to 80% after 1 year. At follow-up, older patients were less likely to demonstrate a perfect inhalation. Patients with a Diskhaler made fewest errors. Adjustment for differences in patient characteristics did not significantly change the results. Because many patients with asthma use their inhaler ineffectively, there is a need to know which inhaler leads to fewest errors. Diskhaler was nominated by this study. When patients are not able to demonstrate adequate inhalation technique in a "tranquil" setting, it is doubtful that they can do so when they experience an exacerbation. Therefore, inhalation instruction should be considered an essential ingredient, not only of self-management programs, but also of asthma patient care in general.


Activation-induced cell death in T cell hybridomas is due to apoptosis. Morphologic aspects and DNA fragmentation.
Some T cell hybridomas, upon activation via the TCR, rapidly undergo cell death. In this paper, we demonstrate that this activation-induced cell death (AICD) is accompanied by morphologic changes seen at the electron and light microscopy levels. The most striking changes are an extensive condensation of the chromatin and formation of membrane blebs. In addition to the morphologic changes, a significant portion of genomic DNA is broken at an interval of approximately 200 bp, producing a ladder of oligonucleosome-sized fragments after gel electrophoresis. Taken together, these observations indicate that AICD proceeds via apoptosis, or programmed cell death. This is additionally supported by the observation that AICD-associated phenomena are at least partially inhibited by cycloheximide or actinomycin D. Curiously, AICD and its associated DNA fragmentation are completely inhibited by aurintricarboxylic acid, a known nuclease inhibitor. The possible relationship between AICD in vitro, and the negative selection process (wherein selection may proceed via AICD of developing, autoreactive thymocytes) is discussed.


Division pattern of a round mutant of Escherichia coli.
A round mutant of Escherichia coli, when grown in Methocel medium, forms chains of cells and does not form tetrads. This implies that successive division planes of the round mutant are parallel rather than perpendicular. These results differ from a previous proposal that division planes in this round mutant are perpendicular to the prior division plane (W. D. Donachie, S. Addinall, and K. Begg, Bioessays 17:569-576, 1995).


The use of an in-house modified double antibody sandwich ELISA to detect Aspergillus antigens in sera of immunosuppressed patients.
The purpose of this study was to retrospectively detect Aspergillus antigens in sera obtained from immunocompromised host using an in-house modified double antibody sandwich ELISA. The ELISA employed the use of polyclonal antibodies raised against a water-soluble (WS) mycelial component of a clinical isolate of Aspergillus fumigatus. Rabbit and guinea-pig anti-WS antibodies were used as the capture and detector antibodies respectively. This was followed by the addition of anti-guinea pig antibodies conjugated to horse radish peroxidase and orthophenylenediamine. Of 143 specimens obtained from 140 patients, only 5 (3.5%) specimens demonstrated the presence of antigen. The 5 antigen-positive sera were from three patients. One was a confirmed case of paranasal aspergillosis and the other two were probable cases of invasive aspergillosis. The clinical validity of the ELISA should be determined by prospective detection of Aspergillus antigens in multiple serum samples collected from proven cases of invasive aspergillosis.


Measuring the outcome of health resort programs.
To evaluate the metric properties and practicability of valid, internationally available outcome instruments in the special setting of health resort programs. A cohort study in a convenience sample of patients with low back pain, upper back pain, conditions of the lower extremities, and conditions of the upper extremities was conducted. Their functioning and health were assessed before and after a health resort program by the disease-specific North American Spine Society (NASS) instruments Lumbar NASS and Cervical NASS; WOMAC Osteoarthritis Index; Disabilities of Arm, Shoulder and Hand Questionnaire; and the general instrument, Medical Outcome Study Short Form-36 (SF-36). Completeness on the scale level ranged between 1% and 10%. Criterion validity of condition-specific instruments was confirmed by stronger associations of the pain and function scales to the Physical Health component of the SF-36 (r = -0.59 to -0.79, p < 0.001 for all scales) than to the Mental Health component (r = -0.11, NS, to r = -0.42, p < 0.001). Reliability (Cronbach's alpha coefficient) was higher than 0.8 for all scales of condition-specific instruments and for 6 of 8 SF-36 scales. Floor and ceiling effects ranged between 0% and 7%. The condition-specific instruments demonstrated a good responsiveness with an effect size ranging between 0.28 and 0.55 and with a standardized response mean between 0.32 and 0.94. The responsiveness of most SF-36 scales was similar, but the Physical Function scale showed a lower responsiveness than the condition-specific scales. The evaluated instruments can be recommended for use in clinical trials that assess the outcome of health resort programs.


Long-term follow-up of men with male pattern baldness treated with topical minoxidil.
Forty-one men with male pattern baldness completed 132 study weeks (2 years 9 months) with topical minoxidil and had follow-up 1-inch target-area vertex scalp hair counts. Initially these men were treated with either twice-daily 2% topical minoxidil for 12 months or 3% topical minoxidil for 8 to 12 months (one third of the subjects received placebo for the first 4 months). After 12 months all subjects continued to apply 3% topical minoxidil twice daily for 1 more year, after which they were randomized to once- versus twice-daily topical minoxidil for an additional 9 months. Those subjects who changed to once-daily application of topical minoxidil at 2 years had a mean change from baseline nonvellus hair count at 1 year of 291.2 (range of hairs four to 553) and at 2 years 9 months of 235 (two to 592 hairs). Those subjects who continued with twice-daily application of topical minoxidil throughout the study had a mean change from baseline nonvellus hair count at 1 year of 323 (15 to 589 hairs) and 335 (13 to 808 hairs) at 2 years 9 months with maintenance topical minoxidil. There were subjects on both maintenance schedules of topical minoxidil who lost some of the nonvellus hair they had initially gained with topical minoxidil; however, there was a greater mean loss in those patients following the once-daily versus twice-daily topical minoxidil regimen (p = 0.05). No subject lost nonvellus target hair as compared with baseline.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of TRPV1 gene mutation on bronchial asthma in children before and after treatment.
Bronchial asthma is a worldwide disease with high incidence. It not only harms children's physical and mental health, but it also brings a heavy burden to their families as well as the society. However, the trigger and pathogenesis of the disease remain unclear. This study aimed to analyze TRPV1 gene mutation and expression of cytokines in children with acute bronchial asthma before and after treatment, thus providing theoretical guidance for the diagnosis and treatment of bronchial asthma in children. Real-time quantitative polymerase chain reaction was adopted to detect TRPV1 mRNA expression level and enzyme-linked immuno sorbent assay was used to detect the serum total IgE level, eosinophil (EOS) number, IL-4, IL-5, and interferon (IFN) gamma levels in peripheral venous blood of children in the healthy control group and asthma group before and after treatment. Logistic regression analysis was applied to analyze the most essential factor inducing bronchial asthma in children. TRPV1 mRNA level of peripheral blood in the asthma group was higher than that in the control group before treatment (p < 0.01). The IL-4, IL-5, and EOS levels in serum were markedly higher than those in the control group (p < 0.01), whereas the IFN-gamma level was lower than that in the control group (p < 0.01). After conventional treatment, TRPV1 mRNA level increased significantly (p < 0.01). The levels of serum IL-4, IL-5, and EOS were significantly lower than those before treatment (p < 0.01), whereas, IFN-gamma level was higher than that before treatment (p < 0.01). Compared with that before treatment, the expression level of IgE showed a significant decrease after treatment (p < 0.01). The results of logistic regression analysis indicated that TRPV1 expression level, IL-4 level, and rs4790522 site mutation were the main risk factors inducing bronchial asthma in children. TRPV1 gene mutation was closely related to bronchial asthma in children, which provided a theoretical basis for the treatment and prognosis of children with bronchial asthma.


Combatting infectious diseases; nanotechnology as a platform for rational vaccine design.
Currently, several successful vaccines are available. However, for pathogens with a highly variable genetic composition, and for which serum IgG antibodies are not a useful correlate of protection, effective vaccines are yet to be developed. This is due to a lack of both the understanding of the immunological pathways leading to long-term protection and the ability to translate the available knowledge into a suitable vaccine formulation. Regarding the latter, nanoparticles can be an attractive platform for vaccine development, as they offer multiple options for improving safety and efficacy. For example, side effects might be decreased upon encapsulation of the adjuvant and the concomitant delivery of antigen and adjuvant is a very promising tool for increasing efficacy. In addition to the many promises, the use of nanoparticles as vaccine carriers should be implemented with caution: the more sophisticated a particle, the more parameters need to be controlled during production and storage.


Efficacy of nasal Staphylococcus aureus eradication by topical nasal mupirocin in patients with perennial allergic rhinitis.
Staphylococcus aureus (SA) colonization is frequent in patients with perennial allergic rhinitis (PAR). Mupirocin has well-recognized antistaphylococcal activity, and its nasal formulation is approved for the eradication of SA nasal colonization. To investigate the frequency of SA nasal carriage, its possible influence on AR severity, and nasal mupirocin's role on AR clinical severity. Sixty patients, aged 5 to 60 years, with AR were included, and 55 healthy individuals served as a control group. Nasal smear specimens were drawn from both nares. A skin prick test to inhalational allergens and a score-graded clinical evaluation of AR were performed. Carriers of SA were treated with topical nasal mupirocin. The SA carrier (SAC) state was found in 23 (38%) of the patients with AR and in 8 (15%) of the healthy controls (P = .004). Comparing SACs with AR SA noncarriers, nasal symptom scores (SSs) tended to be higher in the SAC group (mean [SD], 11.09 [2.16] vs 8.86 [1.43]; P < .005). Treatment with topical nasal mupirocin diminished the SAC state to 10% (P = .009) but did not change AR clinical severity, as assessed by the SS. In patients with PAR, the SAC state is more prevalent compared with the healthy population. Topical nasal mupirocin reduces the SAC state but fails to clinically improve PAR, as assessed by the SS.


Automated 3D coronary sinus catheter detection using a scanning-beam digital x-ray system.
Scanning-beam digital x-ray (SBDX) is an inverse geometry x-ray fluoroscopy system capable of tomosynthesis-based 3D tracking of catheter electrodes concurrent with fluoroscopic display. To facilitate respiratory motion-compensated 3D catheter tracking, an automated coronary sinus (CS) catheter detection algorithm for SBDX was developed. The technique uses the 3D localization capability of SBDX and prior knowledge of the catheter shape. Candidate groups of points representing the CS catheter are obtained from a 3D shape-constrained search. A cost function is then minimized over the groups to select the most probable CS catheter candidate. The algorithm was implemented in MATLAB and tested offline using recorded image sequences of a chest phantom containing a CS catheter, ablation catheter, and fiducial clutter. Fiducial placement was varied to create challenging detection scenarios. Table panning and elevation was used to simulate motion. The CS catheter detection method had 98.1% true positive rate and 100% true negative rate in 2755 frames of imaging. Average processing time was 12.7 ms/frame on a PC with a 3.4 GHz CPU and 8 GB memory. Motion compensation based on 3D CS catheter tracking was demonstrated in a moving chest phantom with a fixed CS catheter and an ablation catheter pulled along a fixed trajectory. The RMS error in the tracked ablation catheter trajectory was 1.41 mm, versus 10.35 mm without motion compensation. A computationally efficient method of automated 3D CS catheter detection has been developed to assist with motion-compensated 3D catheter tracking and registration of 3D cardiac models to tracked catheters.


Management of large-angle esotropia.
If large-angle esotropia is defined as 80 D or more, and if the surgery induces no limitation of movement or incomitance, there is an excellent likelihood that four-muscle surgery in one operation will result in a residual deviation between 15 D of convergent strabismus and 15 D of divergent strabismus.


Cobalt phosphonates: an unusual polymeric cobalt phosphonate containing a clathrated phosphonate anion and a layered bisphosphonate.
Novel cobalt phosphonates [Co(H(2)O)(4)(H(4)L)][H(2)L].2H(2)O, 1, and Co(2)(H(2)O)(2)(L), 2, have been synthesized from 1,8-octylenediphosphonic acid (H(4)L). 1 has been fully characterized by X-ray single-crystal data, TGA, IR spectroscopy, and chemical analysis. The compound crystallizes in the triclinic space group P1 with a = 5.5415(8) A, b = 8.6382(8) A, c = 16.794 (2) A, alpha = 87.694(2) degrees, beta = 80.859(2) degrees, gamma = 76.005(2) degrees, V = 770.11(19) A(3), and Z = 1. A cobalt atom lies in the center of symmetry and is octahedrally coordinated by two oxygen atoms from two undissociated diphosphonic ligands H(4)L and four molecules of water. The cobalt atom and undissociated ligand H(4)L are combined to form polymeric chains along the c-axis, resulting in the formation of a one-dimensional framework. The positive charge on the cobalt atom remains upon coordination and is balanced by a negatively charged uncoordinated ligand (H(2)L) found as a clathrate in the lattice. Two lattice water molecules, hydrogen-bonded with the coordinated and uncoordinated ligands, complete the structure. The metal phosphonate chains are held together and bridge the uncoordinated anionic ligands by a number of strong hydrogen bonds, which make the structure possible. Cobalt phosphonate 2 has been characterized by TGA measurements, IR spectroscopy, and chemical analysis. The compound has a layered structure with an interlayer spacing of 14.26 A. Metal phosphonate layers are cross-linked by hydrocarbon chains. The water molecules are coordinated to the metal atom. According to IR data, compound 2 contains two equivalent PO bonds and one different PO bond, which may be a result of the different types of Co-O-P connectivity within one phosphonic group.


Concomitant Pulmonary Tuberculosis and Invasive Aspergillosis Infection in an Immunocompetent Host.
Pulmonary tuberculosis (TB) is a common infection in developing countries and is associated with low socioeconomic status. It is considered a serious infection due it its long-term sequelae. Post-TB lung disease decreases life expectancy and increases the risk of recurrent TB infection. It also significantly increases the risk of other bacterial and fungal infections. Aspergillosis develops in the cavitary lesions of TB, worsening them and resulting in a deteriorating clinical situation. Concomitant pulmonary TB (PTB) and invasive aspergillosis is uncommon, particularly in the absence of factors that suppress the immune system. In this report, we describe the case of a healthy young adult without previous structural lung damage who presented with primary PTB and acute invasive aspergillosis infection, discuss the treatment dilemma and provide a literature review. Aspergillosis can mimic tuberculosis (TB) infection.Concomitant pulmonary TB and invasive aspergillosis is uncommon.The co-administration of antifungal and anti-TB medications presents significant therapeutic challenges.


GDSL esterase/lipases OsGELP34 and OsGELP110/OsGELP115 are essential for rice pollen development.
Pollen exine contains complex biopolymers of aliphatic lipids and phenolics. Abnormal development of pollen exine often leads to plant sterility. Molecular mechanisms regulating exine formation have been studied extensively but remain ambiguous. Here we report the analyses of three GDSL esterase/lipase protein genes, OsGELP34, OsGELP110, and OsGELP115, for rice exine formation. OsGELP34 was identified by cloning of a male sterile mutant gene. OsGELP34 encodes an endoplasmic reticulum protein and was mainly expressed in anthers during pollen exine formation. osgelp34 mutant displayed abnormal exine and altered expression of a number of key genes required for pollen development. OsGELP110 was previously identified as a gene differentially expressed in meiotic anthers. OsGELP110 was most homologous to OsGELP115, and the two genes showed similar gene expression patterns. Both OsGELP110 and OsGELP115 proteins were localized in peroxisomes. Individual knockout of OsGELP110 and OsGELP115 did not affect the plant fertility, but double knockout of both genes altered the exine structure and rendered the plant male sterile. OsGELP34 is distant from OsGELP110 and OsGELP115 in sequence, and osgelp34 and osgelp110/osgelp115 mutants were different in anther morphology despite both were male sterile. These results suggested that OsGELP34 and OsGELP110/OsGELP115 catalyze different compounds for pollen exine development.


Genes involved in pericyte-driven tumor maturation predict treatment benefit of first-line FOLFIRI plus bevacizumab in patients with metastatic colorectal cancer.
Pericytes are crucial for angiogenesis. The impact of pericyte function to bevacizumab efficacy in mCRC treatment has not been comprehensively examined. This retrospective study investigated germline polymorphisms in genes related to early pericyte maturation to predict bevacizumab efficacy in 424 patients of two clinical trials treated first line with FOLFIRI+bevacizumab. Eight single-nucleotide polymorphisms (SNPs) were tested for potential biomarker value: RGS5 (regulator of G-protein signaling 5; rs1056515, rs2661280), PDGFR-β (platelet-derived growth factor receptor-β; rs2229562, rs2302273), CSPG4 (chondroitin sulfate proteoglycan NG2; rs8023621, rs1127648) and RALBP1 (RalA binding protein 1; rs10989, rs329007). For progression-free survival (PFS), PDGFR-β (rs2302273) was able to define significantly different patient cohorts in uni- and multivariate testing. RALPB1 (rs329007) showed predictive value for tumor response. The C allele in RGS5 (rs2661280) predicted longer overall survival and CSPG4 rs1127648 was associated with differences in PFS, but for both value was lost when multivariate analysis was applied. A comprehensive statistical analysis revealed that the biomarker value of the SNPs was dependent on primary tumor location. This is the first study to identify pericyte germline polymorphisms associated with clinical outcome in mCRC patients treated first line with FOLFIRI+bevacizumab. The differences seen with regard to primary tumor location may lead to further research to understand the clinical outcome differences seen in right- and left-sided colon cancer.


Point-of-care lung ultrasound in young children with respiratory tract infections and wheeze.
Characterise lung ultrasound (LUS) findings, diagnostic accuracy and agreement between novice and expert interpretations in young children with respiratory tract infections and wheeze. Prospective cross-sectional study in a paediatric ED. Patients ≤2 years with a respiratory tract infection and wheeze at triage were recruited unless in severe respiratory distress. Prior to clinical management, a novice sonologist performed the LUS using a six-zone scanning protocol. The treating physician remained blinded to ultrasound findings; final diagnoses were extracted from the medical record. An expert sonologist, blinded to all clinical information, assessed the ultrasound video clips at study completion. Positive LUS was defined as the presence of ≥1 of the following findings: ≥3 B-lines per intercostal space, consolidation and/or pleural abnormalities. Ninety-four patients were enrolled (median age 11.1 months). LUS was positive in 42% (39/94) of patients (multiple B-lines in 80%, consolidation in 64%, pleural abnormalities in 23%). The proportion of positive LUS, along with their diagnostic accuracy (sensitivity (95% CI), specificity (95% CI)), were as follows for children with bronchiolitis, asthma, pneumonia and asthma/pneumonia: 46% (45.8% (34.0% to 58.0%), 72.7% (49.8% to 89.3%)), 0% (0% (0.0% to 23.3%), 51.3% (39.8% to 62.6%)), 100% (100% (39.8% to 100.0%), 61.1% (50.3% to 71.2%)), 50% (50% (6.8% to 93.2%), 58.9% (48.0% to 69.2%)), respectively. There was good agreement between the novice and expert sonographers for a positive LUS (kappa 0.68 (95% CI 0.54 to 0.82)). Among children with respiratory tract infections and wheeze, a positive LUS seems to distinguish between clinical syndromes by ruling in pneumonia and ruling out asthma. If confirmed in future studies, LUS may emerge as a point-of-care tool to guide diagnosis and disposition in young children with wheeze.


Evaluation of the stability of pin configuration in K-wire fixation of displaced supracondylar fractures in children.
Precutaneous Kirschner wire fixation of displaced Gartlands types II and III supracondylar fractures with image guidance remains a preferred procedure for most orthopaedic units. Various K-wire configurations have been in use, but recent objective evaluation of pin stability has favoured two techniques; the 2 crossed pins placed from the medial and lateral epicondyles, and 2 parallel lateral pins when the former technique is impracticable because of swelling. We reviewed patients who had K-wire fixation precutaneously over a 2-year period (Feb 1996 to Feb 1998). There were 44 children, 30 males, 14 females, ages between 1-15 years. 19 patients had (medial and lateral pins) 15, (2 parallel lateral) and 10 patients (2 lateral crossed) precutaneous pin placement respectively. Their postoperative course indicated that maximum stability was obtained with two opposite crossed pins, followed by the 2 paralleled pin method. These observation determined the choice of technique by the surgeons who operated on these patients.


The testosterone-dependent and independent transcriptional networks in the hypothalamus of Gpr54 and Kiss1 knockout male mice are not fully equivalent.
Humans and mice with loss of function mutations in GPR54 (KISS1R) or kisspeptin do not progress through puberty, caused by a failure to release GnRH. The transcriptional networks regulated by these proteins in the hypothalamus have yet to be explored by genome-wide methods. We show here, using 1 million exon mouse arrays (Exon 1.0 Affymetrix) and quantitative polymerase chain reaction (QPCR) validation to analyse microdissected hypothalamic tissue from Gpr54 and Kiss1 knockout mice, the extent of transcriptional regulation in the hypothalamus. The sensitivity to detect important transcript differences in microdissected RNA was confirmed by the observation of counter-regulation of Kiss1 expression in Gpr54 knockouts and confirmed by immunohistochemistry (IHC). Since Gpr54 and Kiss1 knockout animals are effectively pre-pubertal with low testosterone (T) levels, we also determined which of the validated transcripts were T-responsive and which varied according to genotype alone. We observed four types of transcriptional regulation (i) genotype only dependent regulation, (ii) T only dependent regulation, (iii) genotype and T-dependent regulation with interaction between these variables, (iv) genotype and T-dependent regulation with no interaction between these variables. The results implicate for the first time several transcription factors (e.g. Npas4, Esr2), proteases (Klk1b22), and the orphan 10-transmembrane transporter TMEM144 in the biology of GPR54/kisspeptin function in the hypothalamus. We show for the neuronal activity regulated transcription factor NPAS4, that distinct protein over-expression is seen in the hypothalamus and hippocampus in Gpr54 knockout mice. This links for the first time the hypothalamic-gonadal axis with this important regulator of inhibitory synapse formation. Similarly we confirm TMEM144 up-regulation in the hypothalamus by RNA in situ hybridization and western blot. Taken together, global transcriptional profiling shows that loss of GPR54 and kisspeptin are not fully equivalent in the mouse hypothalamus.


The transformation of utilities for health states worse than death: consequences for the estimation of EQ-5D value sets.
Utilities for health are measured on an interval scale, where 1 refers to full health and 0 refers to death. No theoretical lower boundary on the utilities for states worse than death exists. As a consequence, negative values receive greater weight in the calculation of mean utilities. To avoid this, negative values often are bound at -1. The objective of this study was to compare the effect of 3 methods to bound negative values at -1 on the estimation of EQ-5D value sets: truncation, monotonic, and linear transformation. Data of the Dutch EQ-5D valuation study were used. A total of 298 respondents directly valued 17 EQ-5D health states using the time trade-off (TTO) method. Random effects regression analysis was used to interpolate TTO values for all possible EQ-5D states. In the regression analysis the dependent variable is 1 minus the TTO value and the independent variables describe the health state. Two widely used models to estimate EQ-5D value were applied after truncation of negative values and monotonic and linear transformation of negative values. Both models also were estimated on medians. Truncation of negative values gave the largest mean absolute error (MAE); the linear transformation resulted in the smallest MAE. When medians were used for estimation, the MAEs were comparable with the estimation on means. The choice of a method to bound negative values is arbitrary and affects the resulting value set. For the estimation of EQ-5D value sets from a societal perspective the use of medians should be considered.


Immunorecognition of the active form of the oestrogen receptor by using a monoclonal antibody.
In previous studies, two forms (alpha and beta) of the oestrogen receptor, with different immunological characteristics, were observed in the cytosol fraction of fetal guinea-pig uterus, by using a monoclonal antibody to the human oestrogen receptor (D547Sp gamma). Only the alpha form was recognized by the antibody, shifting its sedimentation coefficient in high-salt sucrose gradients. The present work investigated the effect of several factors (time, temperature, high salt concentrations and Na2MoO4) on the interconversion of these two forms. Only the beta form was observed when cytosol was incubated with oestradiol for only 2-3 h, but 20 h later, 40-60% of the total oestradiol-receptor complexes were found as the alpha form. The transformation from the beta to the alpha form was accelerated by temperature (25 degrees C, 15 min) and exposure to high salt concentrations (0.4 M-KCl). On the other hand, Na2MoO4 completely blocked the transformation induced by time and temperature, but had little effect on that induced by KCl. The appearance of the alpha form always correlated with an increase in receptor binding to nuclei and DNA-cellulose. Finally, it was found that the isolated beta form, recovered from the gradient, was transformed into the alpha form after overnight dialysis under reduced pressure. The present data suggest that the alpha form, which is recognized by the monoclonal antibody, is the activated form of the oestrogen receptor.


[State of lipid peroxidation in patients with ischemic heart disease during aortocoronary bypass surgery using epidural anesthesia].
Effects of epidural anesthesia (EA) on free-radical lipid oxidation and the antioxidant system were studied in coronary patients during aortocoronary bypass operations and in the immediate postoperative period in comparison with patients operated on under common anesthesia (without EA). Thirty-six patients (men) aged 42.4 +/- 1.4 years were examined. EA as a component of total anesthesia in aortocoronary bypass surgery was conducive to a lesser activation of free-radical lipid oxidation. After surgery EA sooner normalized free-radical oxidation and the antioxidant system, which was observed at the end of the operation and during the first 24 h after it. During artificial circulation EA did not appreciably affect the degree of activation of free-radical oxidation.


Effect of recovery period of mixture pasture on cattle behaviour, pasture biomass production and pasture nutritional value.
Pasture management that considers pasture growth dynamics remains an open question. Conceptually, such management must allow for grazing only after the recuperation of the pasture between two separate timely grazing periods when pasture reaches optimum recovery, as per the first law of Voisin's rational grazing system. The optimum recovery period not only implies a pasture with better nutritional value and higher biomass yield but one that also reduces the production of enteric methane (CH4) to improve the grazing efficiency of cattle. Therefore, this study aimed to evaluate three different recovery periods (RP) of mixed grasses on the grazing behaviour of heifers, as well as herbage selectivity, herbage yield and nutritional value, in vitro degradability and CH4 production. Based on these criteria, three pasture RPs of 24 (RP24), 35 (RP3) and 46 (RP46) days were evaluated in six blocks using a randomized block design. At each predetermined RP, samples of the pasture were taken before the animals were allowed to graze. Right after collecting the pasture samples, heifers accessed the pasture during 4 h consecutively for grazing simulation and behavioural observations. We also measured the bite rate of each animal. The pasture growing for 24 days had the highest biomass production, best nutritional value, best efficiency of in vitro CH4 relative emission (ml) per DM degraded (g) and bite rate of the three RPs. Heifers all selected their herbage, irrespective of RP, but with different nutritional value and higher in vitro degradability. However, this did not change the production of in vitro CH4. Considering the growth conditions of the area where the study was performed, we recommend the shorter RP24 as the most suitable during the summer season. The study's findings support the idea of management intervention to increase the quality of grazing systems.


Predication of reflex recovery after stroke using quantitative assessments of motor impairment at 1 month.
The objective of this study was to characterize the time-course of changes reflex stiffness after stroke, and to use the Fugl-Meyer Assessment (FMA) at 1 month to predict the ensuing recovery patterns over 1 year. We quantified the modulation of reflex stiffness as a function of elbow joint angles at 1, 2, 3, 6, and 12 months after stroke, using a parallel cascade system identification technique. We then used the "growth mixture" and logistic regression models to characterize recovery patterns over 1 year and to predict these patterns, based on the FMA score at 1 month. We observed two major distinct recovery classes for the relationship between reflex stiffness and elbow angle. The FMA at 1 month was a significant predictor of reflex stiffness as a function of elbow angle at different time points in the first year. The logistical regression class membership may enable us to accurately predict reflex behavior during the first year, information of great potential value for planning targeted therapeutic interventions. Finally, the findings suggest that abnormal reflex function could contribute to functional motor impairment.


The role of small or single-institution prospective studies in thoracic research.
Interventional studies are designed specifically to test the impact of a treatment or management strategy on an outcome of interest. In this article, we will highlight two viewpoints regarding the role of small or single-institution prospective interventional studies within the field of thoracic surgery: first, that these studies are beneficial and can positively impact the field, and second, that these studies can have such significant limitations that they may have no meaningful impact. We will also review elements that can make for successful design and execution of small prospective interventional studies.


Calcified cartilage in patients with osteoarthritis of the hip compared to that of healthy subjects. A design-based histological study.
Calcified cartilage is suggested to be involved in the pathogenesis of osteoarthritis (OA) by facilitating endochondral ossification at the bone-cartilage unit. Therefore, the objective was to quantify the volume and surface area of the calcified cartilage in the femoral head in OA patients and healthy subjects. We used design-based stereological principles, i.e., systematic uniform random sampling and vertical uniform random sections of the entire femoral head. We investigated the articular and calcified cartilage and femoral head surface area and volume, excluding fovea capitis and marginal osteophytes, in 20 patients with OA and 15 healthy subjects. The volume of the calcified cartilage was significantly larger for the patients with OA compared with the healthy subjects (mean difference [95% CI]) (284 [110,457] mm3, p = 0.002). The upper and lower surface area of the calcified cartilage, i.e. the tidemark and cement line, were both significantly larger for OA patients compared with the healthy subjects (17.8 [8.4,27.3] cm2, p < 0.001) and (38.7 [20.8,56.7] cm2, p = 0.002), respectively. The volume of the calcified cartilage and the volume of the femoral head were significantly correlated for the patients with OA (Spearman's ρ = 0.51, p = 0.021), but not for the healthy subjects (ρ = 0.41, p = 0.123). Patients with OA had a larger femoral head surface area and more calcified cartilage compared to healthy subjects. The volume of the calcified cartilage correlated positively with the volume of the femoral head for patients with OA, but not for healthy subjects. This strongly supports the existing view that bone growth in OA is associated with endochondral ossification.


"Extracorporeal loop" (blind loop): an alternative microsurgical technique for prelaminated flap transfer.
Free tissue transfer is frequently used for reconstruction in the head and neck region. In patients who suffer tissue loss due to a high-energy trauma, the reconstructive surgeon is often faced with a large gap between the defect and the site of anastomosis outside the zone of injury. Utilization of long vein grafts for reestablishment of blood flow to the transferred tissue vessels is a common procedure. Arteriovenous fistula (AVF) formation on the side table is an alternative solution. In this paper, an "extracorporeal loop" (blind loop) technique is presented as an alternative microsurgical method for prelaminated flap transfer to two different traumatic recipient areas in the face (nose and ear).


Nail-extraction device mismatch: an issue in developing countries intramedullary nail removal practice.
Intramedullary nail is the gold standard in the management of long bone diaphyseal fractures of tibia and femur. The jig of these nails has corresponding extraction device whose pitch for nail coupling come in various sizes. This unlike plate and screws may be difficult to predict preoperatively and may pose a problem during removal. Difficulties in removal may arise due to the proliferation of nail brands especially in developing countries. The study aims to identify the incidence of extraction device mismatch among orthopaedic surgeons in Nigeria as well as the indications and complications associated with intramedullary nail removal. A two-page questionnaire was administered to 87 orthopaedic surgeons attending the Annual General Meeting of the Nigerian Medical Association. The attitudes of the participants towards intramedullary nail were assessed. All participants agree to asymptomatic removal. Patients wish was the leading indication for asymptomatic removal among the participants. Sixty-one percent of the surgeons have had the need to remove a nail different from the brand in the hospital their practice. The commonest indication for symptomatic removal was infections. Forty-seven percent of the participant encountered nail extraction-device mismatch. The incidence of extraction device mismatch may portend a public health issue. There may be need for patient who had intra medullary nail insertion to be told their brand. We advocate for standardization of extraction device pitch for intramedullary nail.


Processing scarce biological samples for light and transmission electron microscopy.
Light microscopy (LM) and transmission electron microscopy (TEM) aim at understanding the relationship structure-function. With advances in biology, isolation and purification of scarce populations of cells or subcellular structures may not lead to enough biological material, for processing for LM and TEM. A protocol for preparation of scarce biological samples is presented. It is based on pre-embedding the biological samples, suspensions or pellets, in bovine serum albumin (BSA) and bis-acrylamide (BA), cross-linked and polymerized. This preparation provides a simple and reproducible technique to process biological materials, present in limited quantities that can not be amplified, for light and transmission electron microscopy.


Plasticity of neonatal neuronal networks in very premature infants: Source localization of temporal theta activity, the first endogenous neural biomarker, in temporoparietal areas.
Temporal theta slow-wave activity (TTA-SW) in premature infants is a specific signature of the early development of temporal networks, as it is observed at the turning point between non-sensory driven spontaneous local processing and cortical network functioning. The role in development and the precise location of TTA-SW remain unknown. Previous studies have demonstrated that preterms from 28 weeks of gestational age (wGA) are able to discriminate phonemes and voice, supporting the idea of a prior genetic structural or activity-dependent fingerprint that would prepare the auditory network to compute auditory information at the onset of thalamocortical connectivity. They recorded TTA-SW in 26-32 wGA preterms. The rate of TTA-SW in response to click stimuli was evaluated using low-density EEG in 30 preterms. The sources of TTA-SW were localized by high-density EEG using different tissues conductivities, head models and mathematical models. They observed that TTA-SW is not sensory driven. Regardless of age, conductivities, head models and mathematical models, sources of TTA-SW were located adjacent to auditory and temporal junction areas. These sources become situated closer to the surface during development. TTA-SW corresponds to spontaneous transient endogenous activities independent of sensory information at this period which might participate in the implementation of auditory, language, memory, attention and or social cognition convergent and does not simply represent a general interaction between the subplate and the cortical plate. Hum Brain Mapp 38:2345-2358, 2017. © 2017 Wiley Periodicals, Inc.


A 4-year follow-up of two complete mandibular implant-supported removable prostheses in a patient with severe rheumatoid polyarthritis: case report.
Rheumatoid polyarthritis is a systemic autoimmune disease characterized by chronic synovitis and bone damage associated with significant functional disability and morbidity. This case report describes a 4-year follow-up of a 56-year-old female receiving polymedication for severe rheumatoid polyarthritis and osteoporosis with a fully edentulous maxilla treated with two osseointegrated implants supporting a removable mandibular prosthesis. No practitioner wanted to use implants to stabilize her mandibular prosthesis because of the health risks involved. This report encourages the dental practitioner to be familiar with the symptoms and oral manifestations of rheumatoid polyarthritis in order to help manage this disease when patients lose all of their teeth.


Medical management of asthma and folk medicine in a Hispanic community.
To describe beliefs about asthma and asthma treatment in a Hispanic (Dominican-American) community to determine how alternative belief systems affect compliance with medical regimens. Twenty-five mothers of children with asthma were interviewed in their homes, in their primary language, Spanish. Mothers were questioned about their beliefs regarding asthma etiology, treatment, prevention of acute episodes, and use of prescribed medications. Most mothers (72%) said that they did not use prescribed medicines for the prevention of asthma; instead, they substituted folk remedies called "zumos." The home remedies were derived from their folk beliefs about health and illness. Most mothers (60%) thought that their child did not have asthma in the absence of an acute episode. Eighty-eight percent said that medications are overused in this country and that physicians hide therapeutic information from them. Mothers' reliance on home remedies for asthma prevention leads to a high rate of noncompliance with prescribed regimens. Yet they perceive themselves as compliant with an effective regimen that differs from standard medical practice. Further studies should explore ways of promoting physician/patient communication in order to find ways of coordinating medical and folk beliefs to enhance compliance with medically prescribed regimens.


Treatment of esophageal and gastric tumors.
Endoscopic mucosal resection (EMR) has gained increasing popularity in the treatment of esophageal and gastric mucosal cancers in Japan, for complete local resection of the cancer-bearing mucosa. In Western countries, the concept of mucosal cancer and the necessity of treating it are gradually but steadily coming to be accepted. As a treatment for inoperable cases of advanced cancer, on the other hand, intubation with self-expanding metal stents has been widely accepted as a less invasive form of palliation for malignant obstruction of the upper gastrointestinal tract. Tumor ingrowth into the stent can be mechanically avoided by adding a membrane coating to it. In the last two years, applications for therapeutic endoscopy in the treatment of esophageal and gastric tumors have continued to widen.


Perspectives on the Early Quality of Evidence Guiding the Therapeutic Management of SARS-CoV-2: A Systematic Literature Review.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak is a serious health concern. Repurposing of existing drugs indicated for other conditions seems to be the first choice for immediate therapeutic management. The quality of early evidence favoring the different treatment options needs to be apprised for informed decision-making. In this systematic literature review, we apprised the quality of available evidence for different therapeutic options and also the basis for different treatment guidelines. To include all studies that are in different stages of publication, we also included studies from the preprint servers BioRxiv and MedRxiv and published studies from PubMed. We retrieved 5621 articles and included 22 studies for the systematic review. Based on our study, chloroquine/hydroxychloroquine, either alone or in combination with azithromycin, remdesivir, corticosteroids, convalescent sera, ritonavir/lopinavir, tocilizumab and arbidol were evaluated as therapeutic options. The data from different study designs reveal contradictory findings except for convalescent sera for which the evidence available is only from case series. Based on this early evidence, various national guidelines recommend remdesivir, convalescent sera, corticosteroids and hydroxychloroquine in different subsets of patients. Establishing consensus with respect to the end points to be assessed for respiratory viruses may enhance the quality of evidence in case of future pandemics. The systematic review highlighted the lacuna and methodologic deficiency in early clinical evidence and included an update on different therapeutic management guidelines. Further clinical evidence from the ongoing trials may lead to evolution of treatment guidelines with the addition of more therapeutic options.


Injectable tissue-engineered cartilage with different chondrocyte sources.
Injectable engineered cartilage that maintains a predictable shape and volume would allow recontouring of craniomaxillofacial irregularities with minimally invasive techniques. This study investigated how chondrocytes from different cartilage sources, encapsulated in fibrin polymer, affected construct mass and volume with time. Swine auricular, costal, and articular chondrocytes were isolated and mixed with fibrin polymer (cell concentration of 40 x 10 cells/ml for all groups). Eight samples (1 cm x 1 cm x 0.3 cm) per group were implanted into nude mice for each time period (4, 8, and 12 weeks). The dimensions and mass of each specimen were recorded before implantation and after explantation. Ratios comparing final measurements and original measurements were calculated. Histological, biochemical, and biomechanical analyses were performed. Histological evaluations (n = 3) indicated that new cartilaginous matrix was synthesized by the transplanted chondrocytes in all experimental groups. At 12 weeks, the ratios of dimension and mass (n = 8) for auricular chondrocyte constructs increased by 20 to 30 percent, the ratios for costal chondrocyte constructs were equal to the initial values, and the ratios for articular chondrocyte constructs decreased by 40 to 50 percent. Constructs made with auricular chondrocytes had the highest modulus (n = 3 to 5) and glycosaminoglycan content (n = 4 or 5) and the lowest permeability value (n = 3 to 5) and water content (n = 4 or 5). Constructs made with articular chondrocytes had the lowest modulus and glycosaminoglycan content and the highest permeability value and water content (p < 0.05). The amounts of hydroxyproline (n = 5) and DNA (n = 5) were not significantly different among the experimental groups (p > 0.05). It was possible to engineer injectable cartilage with chondrocytes from different sources, resulting in neocartilage with different properties. Although cartilage made with articular chondrocytes shrank and cartilage made with auricular chondrocytes overgrew, the injectable tissue-engineered cartilage made with costal chondrocytes was stable during the time periods studied. Furthermore, the biomechanical properties of the engineered cartilage made with auricular or costal chondrocytes were superior to those of cartilage made with articular chondrocytes, in this model.


Severe Lactic Acidosis Caused by Thiamine Deficiency in a Child with Relapsing Acute Lymphoblastic Leukemia: A Case Report.
Lactic acidosis is characterized by an excessive production of lactic acid or by its impaired clearance. Thiamine deficiency is an uncommon cause of lactic acidosis, especially in countries where malnutrition is rare. We describe the case of a 5-year-old boy who presented with a central nervous system relapse of acute lymphoblastic leukemia. During the chemotherapy regimen, the patient developed drug-induced pancreatitis with paralytic ileus requiring prolonged glucosaline solution infusion. In the following days, severe lactic acidosis (pH 7.0, lactates 253 mg/dL, HCO3- 8 mmol/L) was detected, associated with hypoglycemia (42 mg/dL) and laboratory signs of acute liver injury. Due to the persistent hypoglycemia, the dextrose infusion was gradually increased. Lactates, however, continued to raise, so continuous venovenous hemodiafiltration was started. While lactates initially decreased, 12 h after CVVHDF suspension, they started to raise again. Assuming that it could have been caused by mitochondrial dysfunction due to vitamin deficiency after prolonged fasting and feeding difficulties, parenteral nutrition and thiamine were administered, resulting in a progressive reduction in lactates, with the normalization of pH during the next few hours. In the presence of acute and progressive lactic acidosis in a long-term hospitalized patient, thiamine deficiency should be carefully considered and managed as early as possible.


CIDR: Ultrafast and accurate clustering through imputation for single-cell RNA-seq data.
Most existing dimensionality reduction and clustering packages for single-cell RNA-seq (scRNA-seq) data deal with dropouts by heavy modeling and computational machinery. Here, we introduce CIDR (Clustering through Imputation and Dimensionality Reduction), an ultrafast algorithm that uses a novel yet very simple implicit imputation approach to alleviate the impact of dropouts in scRNA-seq data in a principled manner. Using a range of simulated and real data, we show that CIDR improves the standard principal component analysis and outperforms the state-of-the-art methods, namely t-SNE, ZIFA, and RaceID, in terms of clustering accuracy. CIDR typically completes within seconds when processing a data set of hundreds of cells and minutes for a data set of thousands of cells. CIDR can be downloaded at https://github.com/VCCRI/CIDR .


Factors for self-assessment score of interprofessional team collaboration in community hospitals in Japan.
Interprofessional collaboration is considered an important strategy in overcoming the complex issues associated with healthcare outcomes. A nationwide, community-based integrated care system developed for the care of older people in individual communities in Japan requires community hospitals to deliver integrated care to coordinate efforts for creating effective environments for health. This study aimed to explore the factors associated with the self-assessment score of interprofessional collaboration in community hospitals. Cross-sectional study using the Assessment of Interprofessional Team Collaboration Scale (AITCS). This study was conducted in three small community hospitals in Japan. All healthcare staff in the hospitals via research collaborators were asked to complete the anonymous self-administered questionnaire of the AITCS comprising questions related to individual factors (age, gender, profession), hospital to which they belonged, relationships with neighbouring facilities, job burden and job satisfaction from July to October 2018. The association between the questionnaire items as explanatory variables and AITCS score as an objective variable was determined using univariate followed by multiple regression analyses. The data from 325 of 630 participants were analysed, of whom 252 were female (77.5%) and 240 were nurses (73.8%). The mean of the total AITCS score was 117.6 (range 37-185), and the Visual Analogue Scale for relationships with neighbouring facilities, job satisfaction and job burden was 53.0 mm (0-100), 46.1 mm (0-100) and 64.3 mm (0-100), respectively. In univariate analyses of the association with AITCS score, explanatory variables with a significance level of <0.05 were relationships with neighbouring facilities, job satisfaction and job burden. On multiple regression analysis, the total AITCS score was independently associated with age, profession (nurse/non-nurse), relationships with neighbouring facilities and job satisfaction. Better self-assessment score of interprofessional collaboration is more strongly associated with younger age, a nursing profession, better relationships with neighbouring facilities and greater job satisfaction than with the hospital to which the participant belonged. These findings may help community hospitals enhance the integration of service delivery and benefit to the community through interprofessional collaboration.


The biphasic expression pattern of miR-200a and E-cadherin in epithelial ovarian cancer and its correlation with clinicopathological features.
Epithelial ovarian cancer (EOC) is the leading cause of death among gynecologic malignancies. Despite great efforts to improve early detection and optimize chemotherapeutic regimens, the 5-year survival rate is only 30% for patients presenting with late-stage ovarian cancer. The high mortality of this disease is due to late diagnosis in over 70% of ovarian cancer cases. A class of small noncoding RNAs, or microRNAs, was found to regulate gene expression at the post-transcriptional level. Some, but not all, of the data indicated that the miR-200 family was dysregulated in a variety of malignancies. In this study, we demonstrated that miR-200a and E-cadherin were significantly upregulated in EOC compared to benign epithelial ovarian cysts and normal ovarian tissues. However, further stratification of the subject indicated that the expression levels of miR-200a were significantly downregulated in late-stage (FIGO III+V) and grade 3 groups compared with early stage (FIGO I+II) and grade 1 to 2 groups. Similarly, relatively low levels of miR-200a were observed in the lymph compared to the node-negative group. E-cadherin expression was found to be absent in normal ovarian tissue and was frequently expressed in benign epithelial ovarian cysts, with absence or low levels observed in late-stage ovarian cancers. There was a significantly positive correlation between miR-200a and E-cadherin in EOC. The biphasic expression pattern suggested that miR-200a levels may serve as novel biomarkers for the early detection of EOC, and miR-200a and E-cadherin are candidate targets for the development of new treatment modalities against ovarian cancer.


The Effects of Vaccinium myrtillus Extract on Hamster Pial Microcirculation during Hypoperfusion-Reperfusion Injury.
The present study was aimed to assess the in vivo hamster pial microvessel alterations due to 30 min transient bilateral common carotid artery occlusion (BCCAO) and reperfusion (60 min); moreover, the neuroprotective effects of Vaccinium myrtillus extract, containing 34.7% of anthocyanins, were investigated. Two groups of male hamsters were used: the first fed with control diet and the other with Vaccinium myrtillus supplemented diet. Hamster pial microcirculation was visualized by fluorescence microscopy through an open cranial window. Pial arterioles were classified according to Strahler's method. In age-matched control diet-fed hamsters, BCCAO caused a decrease in diameter of all arterioles. At the end of reperfusion, the reduction of diameter in order 3 arterioles was by 8.4 ± 3.1%, 10.8 ± 2.3% and 12.1 ± 1.1% of baseline in the 2, 4 and 6 month control diet-fed hamsters, respectively. Microvascular permeability and leukocyte adhesion were markedly enhanced, while perfused capillary length (PCL) decreased. The response to acetylcholine and papaverine topical application was impaired; 2'-7'-dichlorofluoresceine-diacetate assay demonstrated a significant ROS production. At the end of BCCAO, in age-matched Vaccinium myrtillussupplemented diet-fed hamsters, the arteriolar diameter did not significantly change compared to baseline. After 60 min reperfusion, order 3 arterioles dilated by 9.3 ± 2.4%, 10.6 ± 3.1% and 11.8 ± 2.7% of baseline in the 2, 4 and 6 month Vaccinium myrtillus supplemented diet-fed hamsters, respectively. Microvascular leakage and leukocyte adhesion were significantly reduced in all groups according to the time-dependent treatment, when compared with the age-matched control diet-fed hamsters. Similarly, the reduction in PCL was progressively prevented. Finally, the response to acetylcholine and papaverine topical application was preserved and there was no significant increase in ROS production in all groups. In conclusion, Vaccinium myrtillusextract protected pial microcirculation during hypoperfusion-reperfusion, preventing vasoconstriction, microvascular permeability, leukocyte adhesion, reduction in PCL and preserving the endothelium function.


Distribution and fate of HBCD and TBBPA brominated flame retardants in North Sea estuaries and aquatic food webs.
Tetrabromobisphenol A (TBBPA) and hexabromocyclododecane diastereoisomers (alpha-, beta/-, and gamma-HBCD) were investigated in effluents from sewage treatment works, landfill leachates, sediments, and food web organisms of the North Sea basin. Residues were quantified by liquid chromatography-mass spectrometry. Both flame retardants were enriched in sewage sludges, where a maximum total (sigma) HBCD concentration of 9.1 mg/kg (dry weight; d.w.) was found; TBBPA was at levels of 102 microg/kg. Landfill leachates from The Netherlands showed up to 36 mg (sigmaHBCD)/ kg (d.w.). gamma-HBCD dominated isomeric profiles in sediments, and concentrations were elevated near to a site of HBCD manufacture. alpha-HBCD was the primary congener detected in marine mammals; however, very few samples exhibited TBBPA. sigmaHBCD ranged from 2.1 to 6.8 mg/kg (lipid weight; l.w.) in liver and blubber of harbor porpoises (Phocoena phocoena) and seals (Phoca vitulina). TBBPA levels in cormorant (Phalacrocorax carbo) livers were up to 1 order of magnitude lower compared to sigmaHBCD. HBCD in eels (Anguilla anguilla) from the Scheldt basin (Belgium) reflected the spatial distribution of concentrations in local sediments. This study shows evidence of HBCD bioaccumulation at the trophic level and biomagnification in the ascending aquatic food chain, and these findings justify risk assessment studies at the ecosystem level.


9 cis,11 trans conjugated linoleic acid (CLA) is synthesised and desaturated into conjugated 18:3 in bovine adipose tissues.
Although endogenous synthesis of conjugated linoleic acid (CLA) in the mammary gland of lactating cows has been already well documented, no study has determined so far as to which tissue and/or organ is involved in CLA synthesis in the growing ruminant except one study showing that CLA synthesis does not occur in ruminant liver. In this context, adipose tissue appears to be a good candidate for endogenous synthesis of CLA in the growing ruminant. The aim of this study was to compare the respective metabolisms of 11trans 18:1 (vaccenic acid, VA) and 9cis,11trans 18:2 (rumenic acid) to that of stearic acid (the preferential substrate of Δ9 desaturase) in adipose tissues (subcutaneous, SC and intermuscular, IM) of six Charolais steers by using the in vitromethod of incubated tissue slices. Samples of SC and IM adipose tissues were incubated at 37°C for 16 h under an atmosphere of 95% O2/5% CO2 in a medium supplemented with 0.75 mM of fatty acid (FA) mixture (representative of circulating non-esterified FA) and 186 μM [1-14C]-18:0 or 58.6 μM [1-14C]-VA or 56 μM [1-14C]-9cis,11trans CLA. Viability of explants was verified by measuring metabolic functions (glucose uptake and glucose-6-phosphate dehydrogenase activity). After 16 h of incubation, FA uptake was similar for all FA (18:0, VA and 9cis,11trans 18:2) in both SC and IM adipose tissues (around 40%). Once in adipose tissue, all FA were preferentially esterified (>80% of cell FA) favouring neutral lipid synthesis (around 90% of esterified FA). Stearic acid was highly (27%) desaturated into oleic acid in SC adipose tissue whereas this desaturation was much lower (6.8%) in IM adipose tissue (P < 0.0001). VA was desaturated into 9cis,11trans CLA at a low extent of about 2.5% to 4.4% in both adipose tissues probably because of a limited affinity of Δ9 desaturase for VA. 9cis,11trans CLA was itself converted by desaturation into 6cis, 9cis,11trans 18:3 at the intensity of 10.8% and 14.5% of cell 9cis,11trans CLA in SC and IM adipose tissues, respectively. In conclusion, bovine adipose tissues of the growing ruminant were especially involved in the endogenous synthesis of CLA from VA and in its desaturation into conjugated derivative, mainly 6cis, 9cis,11trans 18:3, of which biological properties need to be elucidated.


Neurophysiology study of early visual processing of face and non-face recognition under simulated prosthetic vision.
Behavioral researches have shown that the visual function can be partly restored by phosphene-based prosthetic vision for the non-congenital blinds. However, the early visual processing mechanisms of phosphene object recognition is still unclear. This paper aimed to investigate the electro-neurophysiology underlying the phosphene face and non-face recognition. The modulations of latency and amplitude of N170 component in the event-related potential (ERP) were analyzed. Our preliminary results showed that (1) both normal and phosphene face stimuli could elicit prominent N170; nevertheless, phosphene stimuli caused notable latency delay and amplitude suppression on N170 compared with normal stimuli and (2) under phosphene non-face stimuli, a slight but significant latency delay occurred compared with normal stimuli, while amplitude suppression was not observed. Therefore, it was suggested that (1) phosphene perception caused a disruption of the early visual processing for non-canonical images of objects, which was more profound in phosphene face processing; (2) the face-specific processing was reserved under prosthetic vision and (3) holistic processing was the major stage in early visual processing of phosphene face recognition, while part-based processing was attenuated due to the loss of the details.


Perfusion therapy for extremity melanoma.
Because of their initial appearance on extremities, malignant melanomas lend themselves to isolated chemotherapeutic perfusions. Perfusion is attractive because one can deliver effective cytotoxic drugs without systemic toxicity. We are reviewing 20 patients treated between 1960 and 1973 with isolated perfusion. Melphalan (L-phenylalanine mustard) was the drug of choice. Eleven of the 20 patients had previous surgical treatment. Three of the 11 patients are still alive from 27 to 72 months postperfusion. Eight died after an average survival time of 33 months. Of the seven patients who underwent perfusion as primary therapy, four patients are alive from 25 to 76 months postperfusion, and three died after an average survival time of 34 months. There is direct correlation between stages and levels of melanoma, and perfusion and prolonged survival time.