Prevalence and determinants of genital shedding of herpes simplex virus among women attending Italian colposcopy clinics.
To determine the prevalence of herpes simplex virus (HSV) shedding among women attending Italian colposcopy clinics and describe their lifestyle, demographic characteristics, genital symptoms and signs. A cross-sectional study was performed to assess shedding of HSV among 4565 women requiring a gynecological consultation. An amplified enzyme immunoassay that detects an HSV type-common glycoprotein D was used to reveal HSV shedding in cervical specimens. Statistical analysis was performed using Chi-square test and Student's t test. A prevalence of 7.8% was found among colposcopy clinic patients. No significant differences regarding patients' average age, age at first sexual intercourse, contraceptive method used, and number of sexual partners in the previous year were found between subjects with and without viral shedding (P > 0.05). The detection of a concomitant genital infection with Trichomonas vaginalis as well as the report of previous episodes of genital herpes (GH) were significantly higher in the positive group (P < 0.01). Only 2.8% of the patients shedding HSV presented with vesicles and ulcers, with the majority of them being asymptomatic. This is the first Italian survey on genital herpes conducted among colposcopy clinic patients. Our data show that the prevalence of HSV shedding in this study population is high and confirms that the disease is often asymptomatic. The demographics and behavioural variables of women shedding HSV seem to differ from the ones assessed in high risk patients.


Chronic illness adjustment scale (CIAS): Development and validation.
The objectives of this study were to (i) develop a self-report instrument to measure adjustment to chronic illness, (ii) evaluate its core structure and (iii) study various psychometric properties in the development of this instrument. The entire process of developing and validating the instrument is conducted in different phases: item writing and content validation; exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) to extract dimensions of the instrument; reliability and validity testing. A total of 1095 participants were included in the study. The EFA was run using IBM SPSS 23 and CFA was done by AMOS 21. The Cronbach alpha of the chronic illness adjustment scale was found to be .70. The instrument correlates positively with illness perception and well-being which supports the notion that the chronic illness adjustment scale has satisfactory convergent validity. The multifaceted nature of the chronic illness adjustment scale can serve as a global indicator of adjustment to chronic illness, allowing various interventions to flow toward the healthcare arena.


Cooperative Mn(i)-complex catalyzed transfer hydrogenation of ketones and imines.
The synthesis and reactivity of Mn(i) complexes bearing bifunctional ligands comprising both the amine N-H and benzimidazole fragments are reported. Among the various ligands, the N-((1H-benzimidazol-2-yl)methyl)aniline ligand containing Mn(i) complex presented higher reactivity in the transfer hydrogenation (TH) of ketones in 2-propanol. Experimentally, it was established that both the benzimidazole and amine N-H proton played a vital role in the enhancement of the catalytic activity. Utilizing this system a wide range of aldehydes and ketones were reduced efficiently. Notably, the TH of several imines, as well as chemoselective reduction of unsaturated ketones, was achieved in the presence of this catalyst. DFT calculations were carried out to understand the plausible reaction mechanism which disclosed that the transfer hydrogenation reaction followed a concerted outer-sphere mechanism.


Establishment of a murine leukaemia cell line resistant to the growth-inhibitory effect of bryostatin 1.
Bryostatin 1 is a novel macrocyclic lactone activator of protein kinase C (PKC) which has clinical potential as an anti-cancer agent. The mechanism of action of this agent is unknown, but protein kinase C has been implicated. In order to investigate this possibility, we have developed P388 sublines resistant to bryostatin 1, by continuous challenge of the parent cell line with increasing incremental concentrations of the drug over 4 months. Cell lines were established at monthly intervals yielding four sublines: P388/BR/A, which were removed at 1 month; P388/BR/B, obtained after 2 months; P388/BR/C, obtained after 3 months; and P388/BR/D, which were established after 4 months. All four P388/BR sublines show an equal degree of resistance to the growth inhibitory effects of bryostatin 1, with a relative resistance ratio (RR) IC50 of approximately 4,000. The ability of the cytosol of cells to phosphorylate PKC-specific substrate is decreased by 41% for BR/A, 57% for BR/B 80% for BR/C and 94% for BR/D compared with the parental cell line, even when grown in the absence of bryostatin 1 for up to 4 weeks. Similar decreases are seen for cytosolic phorbol ester binding and whole-cell PKC isoenzyme expression. All four P388/BR sublines show high and equal levels of cross-resistance to the PKC activatory phorbol ester, phorbol 12-myristate 13-acetate (PMA). There is no loss of resistance to either bryostatin 1 or PMA up to 3 months after termination of exposure of the sublines to bryostatin 1. There was no significant degree of cross-resistance to daunorubicin in the bryosatin 1-resistant cell lines, P388/BR/A, B, C or D, when compared with the parent cell line, P388.


Future trends which will influence waste disposal.
The disposal and management of solid wastes are ancient problems. The evolution of practices naturally changed as populations grew and sites for disposal became less acceptable. The central search was for easy disposal at minimum costs. The methods changed from indiscriminate dumping to sanitary landfill, feeding to swine, reduction, incineration, and various forms of re-use and recycling. Virtually all procedures have disabilities and rising costs. Many methods once abandoned are being rediscovered. Promises for so-called innovations outstrip accomplishments. Markets for salvage vary widely or disappear completely. The search for conserving materials and energy at minimum cost must go on forever.


Cardiac dysfunction in a rat model of chronic bacteremia.
Cardiac function was examined in vivo and in vitro in rats to determine if cardiac dysfunction could be demonstrated in a nonlethal model of infection. Bacteremic rats (n = 6) had a subcutaneous polymicrobial abscess produced via repeated inoculations of an encapsulated foreign body with Escherichia coli, Bacteroides fragilis, and Staphylococcus aureus while control rats (n = 6) had the same subcutaneous, encapsulated foreign body (an inflammatory focus) but were not inoculated with bacteria. Cardiovascular function was assessed indirectly in vivo by measuring the maximal O2 uptake during a progressive exercise test in both groups before and 14 days after the initiation of inoculations. Cardiac function was also assessed in vitro in the same rats by measuring stroke volumes generated at six different preloads with constant heart rate and afterload. Bacteremic rats had a significantly different fever curve and leukocytotic response than control rats over the 14 day period. The majority of rats that received inoculations demonstrated bacteremias, while none of the control animals had positive cultures for the inoculated organisms. Although in vivo assessment of cardiovascular function showed no evidence of dysfunction, in vitro assessment demonstrated a significant rightward shift of the Starling curve in bacteremic rats. These data suggest that LV dysfunction occurs even during nonlethal infections but may be masked in vivo by compensatory mechanisms.


An investigation into UV light exposure as an experimental model for artificial aging on tensile strength and force delivery of elastomeric chain.
This study investigated the effect of ultraviolet type A light (UVA) exposure on the tensile properties of elastomeric chain. UVA light exposure was used as model for artificial aging, simulating prolonged storage of elastomeric chain. Tensile strength (n = 60) was measured after exposing Ormco, Forestadent and 3M chains to UVA light for 0, 2, 3, and 4 weeks. Force decay was measured (n = 60) using chain exposed for 5, 10, and 14 days. The chains were subsequently stretched at a constant distance and the resulting forces measured at 0, 1, 24 hours and 7, 14, 21, and 28 days. This test simulated a clinical scenario of pre-stretching and subsequent shortening of elastomeric chain. Tensile strength had statistically significant difference and was directly related to the duration of ultraviolet (UV) light exposure. Forestadent chain, which had the second highest value for the 'as received' product, showed the most consistent values over time with the lowest degradation. Ormco showed the lowest values for 'as received' as well as after UV exposure; 3M chain had the highest loss of tensile strength. Force decay was also significantly different. UV light exposure of 10 days or more appears to mark a 'watershed' between products: 3M had most survivors, Forestadent chain had some survivors, depending on the time the chain was stretched for. None of the Ormco product survived UV light exposure for more than 5 days. UVA light exposure may be used as a model for artificial aging as it reduces force delivery and tensile strength of exposed chains.


A PAX5-OCT4-PRDM1 developmental switch specifies human primordial germ cells.
Dysregulation of genetic pathways during human germ cell development leads to infertility. Here, we analysed bona fide human primordial germ cells (hPGCs) to probe the developmental genetics of human germ cell specification and differentiation. We examined the distribution of OCT4 occupancy in hPGCs relative to human embryonic stem cells (hESCs). We demonstrated that development, from pluripotent stem cells to germ cells, is driven by switching partners with OCT4 from SOX2 to PAX5 and PRDM1. Gain- and loss-of-function studies revealed that PAX5 encodes a critical regulator of hPGC development. Moreover, an epistasis analysis indicated that PAX5 acts upstream of OCT4 and PRDM1. The PAX5-OCT4-PRDM1 proteins form a core transcriptional network that activates germline and represses somatic programmes during human germ cell differentiation. These findings illustrate the power of combined genome editing, cell differentiation and engraftment for probing human developmental genetics that have historically been difficult to study.


Long-term efficacy and safety of canakinumab in patients with colchicine-resistant familial Mediterranean fever: results from the randomised phase III CLUSTER trial.
To evaluate the long-term efficacy and safety of canakinumab to treat patients with colchicine-resistant familial Mediterranean fever (crFMF) during Epoch 4 (weeks 41 to 113) of the CLUSTER study. Patients received open-label canakinumab 150 or 300 mg, every 4 or 8 weeks during a 72-week period. We evaluated disease activity every 8 weeks using the physician global assessment (PGA) of disease activity, counting the number of flares, and measuring concentrations of C reactive protein (CRP) and serum amyloid A (SAA). Safety was studied by determination and classification of observed adverse events (AEs). We analysed safety and efficacy separately in two subgroups of patients receiving a cumulative dose of less than 2700 mg, or equal or more than 2700 mg. Of the 61 patients that started the CLUSTER study, 60 entered Epoch 4 and 57 completed it. During the 72-week period, 35/60 (58.3%) patients experienced no flares, and 23/60 (38.3%) had one flare, as compared with a median of 17.5 flares per year reported at baseline. PGA scores indicated no disease activity for the majority of patients throughout the study. Median CRP concentrations were always lower than 10 mg/L, while median SAA concentrations remained over the limit of normal (10 mg/L) but under the 30 mg/L threshold. No new or unexpected AEs were reported. crFMF patients treated with canakinumab during 72 weeks experienced a minimal incidence of flares and good control of clinical disease activity, with no new safety concerns reported.


Attributes and Organizational Factors that Enabled Innovation in Health Care Service Delivery during the COVID-19 Pandemic - Case Studies from Brazil, Canada and Japan.
Innovation by health service organizations can enable adaptation to and transformation of challenges caused by health shocks. Drawing on results from case studies in Brazil, Canada, and Japan, this study looked at innovations the study hospitals introduced in response to challenges caused by COVID-19 to identify: 1) attributes of the innovations that make them conducive to adoption; and 2) organizational factors that facilitate the creation and implementation of innovative health care approaches during health system shocks. Qualitative information was gathered using key informant interviews, participatory observations at the study hospitals and a review of relevant documentation. A thematic approach was used for analysis, and a cross-country comparison framework was prepared to synthesize findings from the case studies in the three countries. In response to the disruptions caused by COVID-19, the study hospitals undertook innovative changes in services, processes, organizational structures, and operational policy. The driving force behind the innovations was the need and urgency generated by the unprecedented nature of the pandemic. With COVID-19, if an innovation met the perceived needs of hospitals and provided an operational advantage, some level of complexity in the implementation appeared to be acceptable. The study findings suggest that for hospitals to create and implement innovations in response to health shocks, they need to: have adaptive and flexible organizational structures; build and maintain functioning communication systems; have committed leadership; ensure all staff share an understanding of hospital organizational and professional missions; and establish social networks that facilitate the creation and implementation of new ideas.


Receptor-mediated endocytosis of alpha 2-macroglobulin: solubilization and partial purification of the fibroblast alpha 2-macroglobulin receptor.
Recent studies in our laboratory have been aimed at biochemically characterizing the alpha 2M receptor present on fibroblast membranes. The approach we have taken is to develop a means of assessing binding to solubilized alpha 2M binding sites. The binding activity was not removed by treatment with high salt concentration or treatment with chaotropic agents. Removal of the binding activity from membranes did occur using a variety of detergents which suggests that the receptor molecules may be "intrinsic" membrane proteins. The most useful detergent for solubilizing the alpha 2M receptor was octyl-beta-D-glucoside. The alpha 2M binding activity could be removed from NRK fibroblasts and Vero cells using this detergent and was found to remain in solution at 100,000 X g. Removal of the octyl-beta-D-glucoside by extensive dialysis resulted in formation of protein-lipid aggregates that bind to 125I-alpha 2M specifically and with high affinity. Such binding sites were not generated when KB cells (which lack receptors) were extracted with the detergent. Significantly, the observed affinities detected for both high- and low-affinity binding sites were similar to those reported with intact cells or membranes. In addition, binding to the solubilized sites could be inhibited using compounds known to block the receptor-mediated endocytosis of alpha 2M (bacitracin, IBHNA). Other compounds (monensin, dansylcadaverine), which did not directly inhibit the high-affinity binding sites, may exert their effects at different stages in receptor-mediated endocytosis (i.e., receptor recycling). alpha 2M binding sites from NIH-3T3 (spontaneous) tumors have been purified approximately 95-fold by a combination of ion exchange and gel permeation chromatography. The receptor appears to be an acidic protein that approximately coelutes with aldolase (45 A, 158,000 daltons) on gel filtration. Ion exchange chromatography appears to remove an endogenous inhibitor of alpha 2M binding and may also remove binding sites having lower affinity for 125I-alpha 2M. Recent studies using immobilized alpha 2M as an affinity resin suggest that the high-affinity alpha 2M receptor may have a subunit molecular weight of approximately 85,000. Studies are now in progress aimed at further characterizing these high-affinity binding sites. Once bound to the alpha 2M receptor, alpha 2M enters cells via coated pits and is rapidly transferred to receptosomes. These organelles carry the ligand into the Golgi region, from which it is transferred to lysosomes where it is slowly degraded.(ABSTRACT TRUNCATED AT 400 WORDS)


Minimum structural requirements for peptide presentation by major histocompatibility complex class II molecules: implications in induction of autoimmunity.
The precise mechanisms of failure of immunological tolerance to self proteins are not known. Major histocompatibility complex (MHC) susceptibility alleles, the target peptides, and T cells with anti-self reactivity must be present to cause autoimmune diseases. Experimental autoimmune encephalomyelitis (EAE) is a murine model of a human autoimmune disease, multiple sclerosis. In EAE, residues 1-11 of myelin basic protein (MBP) are the dominant disease-inducing determinants in PL/J and (PL/J x SJL/J)F1 mice. Here we report that a six-residue peptide (five of them native) of MBP can induce EAE. Using peptide analogues of the MBP-(1-11) peptide, we demonstrate that only four native MBP residues are required to stimulate MBP-specific T cells. Therefore, this study demonstrates lower minimum structural requirements for effective antigen presentation by MHC class II molecules. Many viral and bacterial proteins share short runs of amino acid similarity with host self proteins, a phenomenon known as molecular mimicry. Since a six-residue peptide can sensitize MBP-specific T cells to cause EAE, these results define a minimum sequence identity for molecular mimicry in autoimmunity.


[Occurrence of tardive dyskinesia during neuroleptic treatment].
The incidence of tardive dyskinesia under neuroleptic treatment is diversely estimated. The effectiveness of anticholinergics in its prevention is under discussion. Among 52 patients treated with neuroleptics for more than 5 years (mean: 9 years) only 2 developed tardive dyskinesia, and the disorder was transient in each case. The low doses of neuroleptic drugs administered and their systematic association with anticholinergics seem to be partly responsible for this low incidence of tardive dyskinesia.


Tumor biology, biomarkers, and liquid biopsy in pediatric renal tumors.
The expansion of knowledge regarding driver mutations for Wilms tumor (WT) and malignant rhabdoid tumor of the kidney (MRT) and various translocations for other pediatric renal tumors opens up new possibilities for diagnosis and treatment. In addition, there are growing data surrounding prognostic factors that can be used to stratify WT treatment to improve outcomes. Here, we review the molecular landscape of WT and other pediatric renal tumors as well as WT prognostic factors. We also review incorporation of circulating tumor DNA/liquid biopsies to leverage this molecular landscape, with potential use in the future for distinguishing renal tumors at the time of diagnosis and elucidating intratumor heterogeneity, which is not well evaluated with standard biopsies. Incorporation of liquid biopsies will require longitudinal collection of multiple biospecimens. Further preclinical research, identification and validation of biomarkers, molecular studies, and data sharing among investigators are crucial to inform therapeutic strategies that improve patient outcomes.


Molecular mechanism and function of CD40/CD40L engagement in the immune system.
During the generation of a successful adaptive immune response, multiple molecular signals are required. A primary signal is the binding of cognate antigen to an antigen receptor expressed by T and B lymphocytes. Multiple secondary signals involve the engagement of costimulatory molecules expressed by T and B lymphocytes with their respective ligands. Because of its essential role in immunity, one of the best characterized of the costimulatory molecules is the receptor CD40. This receptor, a member of the tumor necrosis factor receptor family, is expressed by B cells, professional antigen-presenting cells, as well as non-immune cells and tumors. CD40 binds its ligand CD40L, which is transiently expressed on T cells and other non-immune cells under inflammatory conditions. A wide spectrum of molecular and cellular processes is regulated by CD40 engagement including the initiation and progression of cellular and humoral adaptive immunity. In this review, we describe the downstream signaling pathways initiated by CD40 and overview how CD40 engagement or antagonism modulates humoral and cellular immunity. Lastly, we discuss the role of CD40 as a target in harnessing anti-tumor immunity. This review underscores the essential role CD40 plays in adaptive immunity.


Increased muscle expression of interleukin-17 in Duchenne muscular dystrophy.
Duchenne muscular dystrophy (DMD) is a degenerative muscle wasting disease caused by mutations in the dystrophin gene. Dystrophic muscle is characterized by chronic inflammation, and inflammatory mediators could be promising targets for innovative therapeutic interventions. We analyzed muscle biopsy samples of DMD-affected children to characterize interleukin (IL)-17 and Forkhead box P3 (Foxp3) expression levels and to identify possible correlations with clinical status. Expression levels of IL-17, Foxp3, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), IL-6, and transforming growth factor-β (TGF-β) were analyzed by real-time PCR in muscle biopsy samples from patients with DMD (n = 27) and juvenile dermatomyositis (JDM) (n = 8). Motor outcome of patients with DMD was evaluated by North Star Ambulatory Assessment score. In DMD, we found higher levels of IL-17 and lower levels of Foxp3 mRNA compared with those for a typical inflammatory myopathy, JDM. Moreover, the IL-17/Foxp3 ratio was higher in DMD than in JDM biopsy samples. IL-17 mRNA levels appeared to be related to the expression levels of other proinflammatory cytokines (TNF-α and MCP-1) and significantly associated with clinical outcome of patients. The association of IL-17 expression with levels of other inflammatory cytokines and with the clinical course of DMD suggests a possible pathogenic role of IL-17.


Endoscopic resection of benign breast tumors: retromammary space approach.
Endoscopic surgery is characterized by the creation of a working space. At our department, we have obtained good results with a retromammary space approach in which the tumor is resected after creation of a working space in the retromammary space. The special instruments used for this purpose comprise an endoscopic vein harvesting system to dissect the retromammary space, a dissecting balloon to compress the space to achieve hemostasis, and laparosonic coagulating shears to incise the tumor. This surgical technique provides a superior cosmetic result, and the level of patient satisfaction has been high.


Collapse dynamics of single proteins extended by force.
Single-molecule force spectroscopy has opened up new approaches to the study of protein dynamics. For example, an extended protein folding after an abrupt quench in the pulling force was shown to follow variable collapse trajectories marked by well-defined stages that departed from the expected two-state folding behavior that is commonly observed in bulk. Here, we explain these observations by developing a simple approach that models the free energy of a mechanically extended protein as a combination of an entropic elasticity term and a short-range potential representing enthalpic hydrophobic interactions. The resulting free energy of the molecule shows a force-dependent energy barrier of magnitude, DeltaE =epsilon(F - F(c))(3/2), separating the enthalpic and entropic minima that vanishes at a critical force F(c). By solving the Langevin equation under conditions of a force quench, we generate folding trajectories corresponding to the diffusional collapse of an extended polypeptide. The predicted trajectories reproduce the different stages of collapse, as well as the magnitude and time course of the collapse trajectories observed experimentally in ubiquitin and I27 protein monomers. Our observations validate the force-clamp technique as a powerful approach to determining the free-energy landscape of proteins collapsing and folding from extended states.


Providing potential solutions by using FT-IR spectroscopy for biofluid analysis: Clinical impact of optical screening and diagnostic tests.
Currently, the potential of FT-IR spectroscopy for rapid diagnosis of many pathologies has been demonstrated by numerous research studies including those targeting COVID-19 detection. However, the number of clinicians aware of this potential and who are willing to use spectroscopy in their clinics and hospitals is still negligible. In addition, lack of awareness creates a huge gap between clinicians and researchers involved in clinical translation of current FT-IR technology hence hindering initiatives to bring basic and applied research together for the direct benefit of patients. Knowledge and medical training on FT-IR on the side of clinicians should be one of the first steps to be able to integrate it into the list of complementary exams which may be requested by health professionals. Countless FT-IR applications could have a life-changing impact on patients' lives, especially screening and diagnostic tests involving biofluids such as blood, saliva and urine which are routinely non-invasively or minimally-invasively. Blood may be the most difficult to obtain by the invasive method of collection, but much can be evaluated in its components, and areas such as hematology, infectiology, oncology and endocrinology can be directly benefited. Urine with a relatively simple collection method can provide pertinent information from the entire urinary system, including the actual condition of the kidneys. Saliva collection can be simpler for the patient and can provide information on diseases affecting the mouth and digestive system and can be used to diagnose diseases such as oral cancer in its early-stages. An unavoidable second step is the active involvement of industries to design robust and portable instruments for specific purposes, as the medical community requires user-friendly instruments of advanced computational algorithms. A third step resides in the legal situation involving the global use of the technique as a new diagnostic modality. It is important to note that decentralized funds for variety of technologies hinders the training of clinical and medical professionals for the use of newly arising technologies and affect the engagement of these professionals with technology developers. As a result of decentralized funding, research efforts are spread out over a range of technologies which take a long time to get validated and translated to the clinic. Partnership over similar groups of technologies and efforts to test the same technologies while overcoming barriers posed to technology validation in different areas around the globe may benefit the clinical/medical, research and industry community globally.


Kassina senegalensis skin tachykinins: molecular cloning of kassinin and (Thr2, Ile9)-kassinin biosynthetic precursor cDNAs and comparative bioactivity of mature tachykinins on the smooth muscle of rat urinary bladder.
Tachykinins are among the most widely-studied families of regulatory peptides characterized by a highly-conserved C-terminal -Phe-X-Gly-Leu-Met.amide motif, which also constitutes the essential bioactive core. The amphibian skin has proved to be a rich source of these peptides with physalaemin from the skin of Physalaemus fuscomaculatus representing the archetypal aromatic tachykinin (X=Tyr or Phe) and kassinin from the skin of Kassina senegalensis representing the archetypal aliphatic tachykinin in which X=Val or Ile. Despite the primary structures of both mature peptides having been known for at least 30 years, neither the structures nor organizations of their biosynthetic precursors have been reported. Here we report the structure and organization of the biosynthetic precursor of kassinin deduced from cDNA cloned from a skin secretion library. In addition, a second precursor cDNA encoding the novel kassinin analog (Thr(2), Ile(9))-kassinin was identified as was the predicted mature peptide in skin secretion. Both transcripts exhibited a high degree of nucleotide sequence similarity and of open-reading frame translated amino acid sequences of putative precursor proteins. The translated preprotachykinins each consisted of 80 amino acid residues encoding single copies of either kassinin or its site-substituted analog. Synthetic replicates of each kassinin were found to be active on rat urinary bladder smooth muscle at nanomolar concentrations. The structural organization of both preprotachykinins differs from that previously reported for those of Odorrana grahami skin indicating a spectrum of diversity akin to that established for amphibian skin preprobradykinins.


The Use of Poly-L-Lysine as a Capture Agent to Enhance the Detection of Antinuclear Antibodies by ELISA.
Antibodies to nuclear antigens (antinuclear antibodies or ANAs) are the serological hallmark of systemic lupus erythematosus (SLE). These antibodies bind diverse nuclear antigens that include DNA, histones and non-histone proteins as well as complexes of proteins with DNA and RNA. Because of the frequency of ANA expression in SLE, testing is an important component of clinical evaluation as well as determination of eligibility for clinical trials or utilization of certain therapies. Immunofluorescence assays have been commonly used for this purpose although this approach can be limited by issues of throughput, variability and difficulty in determining positivity. ELISA and multiplex assays are also useful approaches although these assays may give an incomplete picture of antibodies present. To develop a sensitive and quantitative ANA assay, we have explored an ELISA platform in which plates are pre-coated with a positively charged nucleic acid binding polymer (NABP) to increase adherence of antigens containing DNA or RNA. As a source of antigens, we have used supernatants of Jurkat cells undergoing apoptosis in vitro. As results presented show, a poly-L-lysine (PLL) pre-coat significantly enhances detection of antibodies to DNA as well as antigens such as histones, SSA, SSB and RNP. Comparison of the ELISA assay with the PLL pre-coat with a multiplex assay using the BioPlex® 2200 system indicated good agreement in results for a panel of lupus sera. Together, these studies indicate that a pre-coat with a positively charged polymer can increase the sensitivity of an ANA ELISA using as antigens molecules released from dead and dying cells. This assay platform may facilitate ANA testing by providing an ensemble of antigens more similar in composition and structure with antigens present in vivo, with a NABP promoting adherence via charge-charge interactions.


The role of alpha(5)beta(1)-integrin in the IGF-I-induced migration of extravillous trophoblast cells during the process of implantation.
The role of integrins in the processes of adhesion and migration makes them attractive potential participants in the complex events of embryo implantation and placentation. Recently, the role of the alpha(v)beta(3)-integrin pathway was shown in the insulin-like growth factor-I (IGF-I)-stimulated migration of extravillous trophoblast (EVT) cells. This study was designed to investigate the role of alpha(5)beta(1)-integrin in this respect. Using cultured EVT cells, migration assays were carried out for IGF-I-treated or untreated cells in the presence or absence of the GRGDSP and GRGESP hexapeptides, alphaIR3, and a blocking antibody against alpha(5)beta(1)-integrin. Immuno-electron microscopy and immunofluorescent staining were performed to localize the distribution of alpha(5)beta(1)- and alpha(v)beta(3)-integrins, Rab5a, paxillin, phospho-FAK (pFAK), and vinculin. The results showed that IGF-I-induced migration of EVT cells was abolished following treatment with GRGDSP hexapeptide, alphaIR3, and a blocking antibody against alpha(5)beta(1)-integrin. Further, statistical analysis showed that the area-related numerical density of the alpha(5)beta(1)-integrin in the perinuclear regions was significantly higher than in the cell extensions. Immunocytochemical experiments demonstrated an up-regulation in internalization rate of alpha(5)beta(1)-integrin in IGF-I-stimulated EVT cells. Furthermore, alpha(5)beta(1)-integrin exhibited co-localization with Rab5a, but not with alpha(v)beta(3)-integrin, pFAK, paxillin, and vinculin at the focal adhesions of the EVT cells. Taken together, these findings suggest an essential role for alpha(5)beta(1)-integrin in IGF-I-promoted migration of EVT cells. It is possible therefore that IGF-I-induced internalization of alpha(5)beta(1)-integrin may be an important event during the migration of EVT cells in the complex processes of implantation and placentation.


Noninvasive detection of hypoxic myocardium using fluorine-18-fluoromisonidazole and positron emission tomography.
Fluoromisonidazole (FMISO) is metabolically trapped in viable cells as a function of reduced cellular pO2. Therefore [18F]-FMISO is potentially useful for evaluating patients with hypoxic but viable myocardium. The goal of this study was to investigate [18F]FMISO uptake in ischemic myocardium non-invasively using positron emission tomography (PET). Studies were performed in 10 open-chest dogs subjected to either complete (Group 1, n = 5) or partial (Group 2, n = 5) occlusion of the left anterior descending coronary artery. The tracer was administered by intravenous bolus following the onset of ischemia and serial PET images were acquired for the next 4 hr. In Group 1, viability was assessed using histochemical staining (nitroblue tetrazolium, NBT) and 99mTc-pyrophosphate (Tc-PYP). In Group 2, viability was assessed using measurements of regional wall motion, histochemical staining and histology (two animals). In each study, PET images obtained at times between 2 and 4 hr postinjection showed specific enhancement of tracer activity in the distal anterior wall and apex of the left ventricle. At 4 hr, the tissue-to-blood pool count ratio was significantly higher in ischemic regions; 1.8 +/- 0.4 for Group 1 and 1.6 +/- 0.2 for Group 2 versus 1.0 +/- 0.1 in nonischemic regions. Postmortem tissue sampling of Group 1 hearts showed significant FMISO retention in samples without evidence for infarction, either by NBT or Tc-PYP deposition, as well as in more severely ischemic regions. In Group 2 animals, FMISO was retained in myocardial regions with reduced blood flow (microspheres), which exhibited improved contraction following reperfusion. We conclude that PET imaging of [18F]FMISO is a promising technique for the noninvasive identification of viable hypoxic myocardium.


The genus Clubiona Latreille, 1904 (Araneae: Clubionidae) in the Maghreb, with notes on the genevensis group and new records from the Mediterranean Region.
A survey of the members of the genus Clubiona Latreille, 1904 in the Maghreb is presented. The presence of Clubiona comta C. L. Koch, 1839, C. dinienis Simon, 1878, C. leucaspis Simon, 1932, C. phragmitis C. L. Koch 1843 and C. vegeta Simon, 1918 is confirmed. Clubiona pseudosimilis Mikhailov, 1990, from the eastern Mediterranean is new to Africa and Portugal. A specimen of C. neglecta O. Pickard-Cambridge, 1862, cited from Morocco in the past, was misidentified and appears to be C. pseudoneglecta Wunderlich, 1994. The species is new to Algeria and Spain. Two new synonyms are revealed: Clubiona baborensis Denis, 1937 from Algeria = C. diniensis Simon, 1878 N. Syn. and Clubiona venusta Pavesi, 1880 from Tunisia = Selamia reticulata (Simon, 1870) N. Syn. Clubiona mandibularis Lucas, 1846 is considered a Nomen dubium. The comta group is redefined and the "genevensis subgroup" is elevated to species group, including two subgroups. A key and illustrations to the species of the genevensis group are presented and all the species occurring in the Maghreb are illustrated.


The impact of frailty on failure-to-rescue in geriatric trauma patients: A prospective study.
Failure-to-rescue (FTR) (defined as death from a major complication) is considered as an index of hospital quality in trauma patients. However, the role of frailty in FTR events remains unclear. We hypothesized that FTR rate is higher in elderly frail trauma patients. We performed a prospective cohort study of all elderly (age ≥ 65 years) trauma patients presenting at our level one trauma center. Patient's frailty status was calculated utilizing the Trauma Specific Frailty Index (TSFI) within 24 hours of admission. Patients were stratified into non-frail, pre-frail, and frail. FTR was defined as death from a major complication (respiratory, infectious, cardiac, and renal). Binary logistic regression analysis was performed after adjusting for age, gender, injury severity (ISS), and vital parameters to assess the relationship between frailty status and FTR. A total of 368 elderly trauma patients were evaluated of which 25% (n = 93) were non-frail, 38% (n = 139) pre-frail, and 37% (n = 136) frail. Overall, 30% of the patients developed in-hospital complications; of them, mortality occurred in 26% of the patients (FTR group). In the FTR group, 69% of the patients were frail compared to 17% pre-frail and 14% non-frail (p = 0.002). On multivariate regression analysis for predictors of FTR, frail status was an independent predictor of FTR (OR [95% CI] = 2.67 [1.37-5.20]; p = 0.004). On sensitivity analysis, positive predictive value of TSFI for FTR was 69% and negative predictive value for FTR was 67%. In elderly trauma patients, the presence of frailty increased the odds of FTR almost threefold as compared to non-frail. Although FTR has been considered as an indicator of health care quality, the findings of this study suggest that frailty status independently contributes to FTR. This needs to be considered in the future development of quality metrics, particularly in the case of geriatric trauma patients. Prognostic study, level II.


Temozolomide-induced myelodysplasia.
A patient who had received temozolomide (TMZ) as a single agent in treatment of malignant glioma developed therapy-induced myelodysplasia (T-MDS). TMZ is an orally active imidazotetrazine which methylates guanine residues in DNA, ultimately causing single and double-strand DNA breaks leading to apoptotic cell death. TMZ does not chemically cross-link DNA and is considered a nonclassical alkylating agent, similar in structure and activity to dacarbazine. Observations on this patient, and on similarly treated others, suggest that the cumulative dose threshold (CDT) for TMZ that predisposes to T-MDS and which may potentially lead to acute myeloid leukemia (T-AML) is around 18000 to 20000 mg/sq m. Although the incidence of T-MDS and the predisposing CDT of TMZ may differ from that of other potentially leukemogenic compounds currently and formerly used as chemotherapeutic agents, all alkylating agents, including TMZ, should be considered potentially leukemogenic when administered long term.


Pulsed NMR study of network formation in the course of bulk polymerization of methyl acrylate.
The proton spin-spin relaxation time (T2) during the bulk polymerization of methyl acrylate was measured as a function of the reaction time at various temperatures. Three kinds of T2 (T2L (long), T2S (short) and T2M (intermediate)) were obtained as the polymerization proceeded. The fraction of T2S (FS) increased sigmoidally at a certain reaction time, while that of T2L (FL) decreased reciprocally. The former corresponded to the amount of a polymer whose molecular weight was sufficiently high enough to cause a tight entanglement that produced a transient network structure; the latter reflected a decrease in the mixture of the monomer and the low molecular weight of the polymer. T2M is considered to arise from a relatively mobile region of the entanglement. The relationship between the fractions of T2S + T2M and the polymer yield was found to be linear, which led us to monitor the polymer yield in real time during the polymerization in a non-distractive manner. 13C DD (dipolar decoupling)/MAS (magic angle spinning) NMR spectra were also measured to monitor the polymerization process in terms of the molecular motions between the main chain and the side chain in the formation of a network structure. The 13C DD/MAS NMR spectra show that the side chain motion became restricted as well as the main chain when the "Trommosdorff effect" (gel effect) was observed, and a part of the monomers were trapped in the network structure.


Diffraction mechanism of a light-diffusing film with an alternate-polymer-layer structure.
A diffraction mechanism, and an optical model to reflect that mechanism, for a light-diffusing film with an alternate-polymer-layer structure was proposed and validated. According to this model, the film forms an angular distribution of light intensity that is almost constant in a certain scattering-angle (cutoff angle) range and drastically decreases outside that range; that is, the profile is similar to a trapezoid. Although the trapezoid intensity distribution (TID) is a fundamental distribution of the film, the mechanism to form the TID has not previously been clarified. A key mechanism is that the refractive-index distribution of the layer structure is expressed as stacked phase gratings, some of which should diffract zeroth-order light to higher orders strongly, and the repetition of this multiple diffraction spreads light over the whole cutoff angle range, thereby generating the TID. To verify the proposed mechanism, intensity distributions were calculated by using the proposed model. The calculation results indicate that the proposed model reproduces the TID.


Efficacy and safety of a switch from twice-daily tacrolimus to once-daily generic tacrolimus in stable liver transplant patients.
Once-daily tacrolimus reduces non-compliance relative to twice-daily tacrolimus. However, little is known about the safety and efficacy of conversion from twice-daily tacrolimus to generic once-daily tacrolimus in liver transplantation (LT). Herein, we investigated the efficacy and safety of a switch from twice-daily tacrolimus to generic once-daily tacrolimus in patients with stable liver graft function. This prospective, multicenter, open-label, single-arm study was conducted in 17 medical centers for 1 year from July 2019 to July 2020 (NCT04069065). Primary endpoint was the incidence of biopsy-proven acute rejection (BPAR) for 24 weeks after conversion. Secondary endpoints were graft failure, patient death, and adverse events (AEs). Of 151 screened LT patients, 144 patients were enrolled. BPAR, graft failure, and patient death did not occur in this patient population. There were no statistical differences in blood tests, liver function tests, or biochemical tests between visits in any of the patients. Median tacrolimus trough level decreased abruptly from 4.7 ng/mL to 3.2 ng/mL after generic once-daily tacrolimus conversion, but median tacrolimus dose increased due to low tacrolimus trough level. Ninety-two adverse events occurred in 54 patients. Liver enzyme levels increased in seven patients (4.9%) after the switch to generic once-daily tacrolimus, but the liver function tests of these patients normalized thereafter. There were three cases of severe AEs not related to investigational drug. Present study suggests that conversion from twice-daily tacrolimus to generic once-daily tacrolimus is effective and safe in stable LT patients.


The Standardization of Hospital-Acquired Infection Rates Using Prediction Models in Iran: Observational Study of National Nosocomial Infection Registry Data.
Many factors contribute to the spreading of hospital-acquired infections (HAIs). This study aimed to standardize the HAI rate using prediction models in Iran based on the National Healthcare Safety Network (NHSN) method. In this study, the Iranian nosocomial infections surveillance system (INIS) was used to gather data on patients with HAIs (126,314 infections). In addition, the hospital statistics and information system (AVAB) was used to collect data on hospital characteristics. First, well-performing hospitals, including 357 hospitals from all over the country, were selected. Data were randomly split into training (70%) and testing (30%) sets. Finally, the standardized infection ratio (SIR) and the corrected SIR were calculated for the HAIs. The mean age of the 100,110 patients with an HAI was 40.02 (SD 23.56) years. The corrected SIRs based on the observed and predicted infections for respiratory tract infections (RTIs), urinary tract infections (UTIs), surgical site infections (SSIs), and bloodstream infections (BSIs) were 0.03 (95% CI 0-0.09), 1.02 (95% CI 0.95-1.09), 0.93 (95% CI 0.85-1.007), and 0.91 (95% CI 0.54-1.28), respectively. Moreover, the corrected SIRs for RTIs in the infectious disease, burn, obstetrics and gynecology, and internal medicine wards; UTIs in the burn, infectious disease, internal medicine, and intensive care unit wards; SSIs in the burn and infectious disease wards; and BSIs in most wards were >1, indicating that more HAIs were observed than expected. The results of this study can help to promote preventive measures based on scientific evidence. They can also lead to the continuous improvement of the monitoring system by collecting and systematically analyzing data on HAIs and encourage the hospitals to better control their infection rates by establishing a benchmarking system.


Early markers of autism spectrum disorders in infants and toddlers prospectively identified in the Social Attention and Communication Study.
The Social Attention and Communication Study involved the successful implementation of developmental surveillance of the early markers of autism spectrum disorders in a community-based setting. The objective in the current study was to determine the most discriminating and predictive markers of autism spectrum disorders used in the Social Attention and Communication Study at 12, 18 and 24 months of age, so that these could be used to identify children with autism spectrum disorders with greater accuracy. The percentage of 'yes/no' responses for each behavioural marker was compared between children with autistic disorder (n = 39), autism spectrum disorder (n = 50) and developmental and/or language delay (n = 20) from 12 to 24 months, with a logistic regression also conducted at 24 months. Across all ages, the recurring key markers of both autistic disorder and autism spectrum disorder were deficits in eye contact and pointing, and from 18 months, deficits in showing became an important marker. In combination, these behaviours, along with pretend play, were found to be the best group of predictors for a best estimate diagnostic classification of autistic disorder/autism spectrum disorder at 24 months. It is argued that the identified markers should be monitored repeatedly during the second year of life by community health-care professionals.


Dynamics of polymer chains confined to a periodic cylinder: molecular dynamics simulation vs. Lifson-Jackson formula.
The dynamics of polymer chains confined to a periodic cylinder is explored using molecular dynamics simulation and theoretical analysis. The cylinder is divided into two cavities in one periodicity: one cavity consists of a channel of length L1 and diameter D1 and another cavity is a channel of length L2 and diameter D2. For L1 = L2 = L/2, the diffusion coefficient D of a single confined polymer chain decreases rapidly with increase in periodicities L. For a fixed periodicity with L = L1 + L2 = constant, the diffusion coefficient D of a single confined polymer chain shows strong dependence on L1 (or L2). Moreover, for a multi-chain system with L1 = L2, the diffusion coefficient D shows strong non-monotonic dependence on the chain monomer density ρ, and the confined polymer chains diffuse fastest for ρ = 0.068, in which there are three polymer chains in two periodicities as well as two opposing effects: one is that the excluded volume effect between polymer chains can reduce the free energy barrier, and another is that when the chain monomer density ρ increases further, the entanglement effect increases, which leads to that the diffusion coefficient D decreases as ρ increases. Finally, we found that the diffusion coefficient D has a similar oscillation relationship with the ratio of R/L for different chain lengths N and different periodicity L, and the oscillation amplitude decreases gradually as R/L increases; here R is the mean end-to-end distance of a single confined polymer chain, i.e., . From the view of free energy potential, both the width of the free energy potential well and the height of the free energy potential barrier govern simultaneously the diffusion behavior of confined polymer chains. According to the mean force potential (PMF) based on the weighted histogram analysis method (WHAM), we found that our results agree very well with the theoretical analysis using the Lifson-Jackson formula. Our investigation may help us understand the dynamics of particles in a periodic medium, which is one of the interesting problems in many different fields of science, such as physics, chemistry and biology.


Delayed recovery from post-thyroidectomy hypoparathyroidism: a case report.
Thirteen hours after a subtotal thyroidectomy was performed for hyperthyroidism, a patient developed carpopedal spasms, parathesias and hypocalcemia to 6.9 mg/dL. After initial stabilization with intravenous calcium administration, oral calcium carbonate and calcitriol were required. Ten months postoperatively serum calcium levels rose and supplementation was gradually discontinued. The serum parathyroid hormone (PTH) level was 1.0 pg/mL on the second postoperative day and levels were undetectable despite sensitive testing 3 months later (normal 10-65 pg/mL). Two years after surgery, the PTH level has increased to 36 pg/mL, but remains relatively low considering the patient's continued mild hypocalcemia. To our knowledge, there has been no previously reported case of long-term post-thyroidectomy hypocalcemia documenting undetectable parathyroid function and subsequent spontaneous improvement. This case suggests that delayed recovery of parathyroid function and discontinuation of vitamin D and calcium supplementation may be possible in some post-thyroidectomy patients with hypocalcemia due to severe hypoparathyroidism.


Fertility and union formation during crisis and societal consolidation in the Western Balkans.
Fertility decline in central and eastern Europe (CEE) since the fall of the communist regimes has been driven by both stopping and postponement of childbearing: two processes that have been related to crisis and economic development, respectively. In the Western Balkans these economic and political contexts followed each other in the form of a biphasic transition. I examine whether this sequence triggered fertility responses like those observed elsewhere. Relying on three independent data sources, I cross-validate the levels of, and describe the trends in, union formation and fertility (by birth order) between 1980 and 2010. Results do not reveal widespread declines in fertility to lowest-low levels during the most acute period of crisis. The subsequent postponement of marriage and first birth was also limited, and the two-child family remains the norm. This relative resilience of childbearing patterns compared with other CEE countries is discussed with reference to the institutional context.


Experimental approach to the kinetic study of unstable site-directed irreversible inhibitors: kinetic origin of the apparent positive co-operativity arising from inactivation of trypsin by p-amidinophenylmethanesulphonyl fluoride.
Experimental characterization of enzyme inactivation by unstable irreversible inhibitors has only previously been carried out by using discontinuous methods involving preincubation, removal of samples and further residual activity assays. A continuous method for the kinetic study of these inhibitors in the presence of an auxiliary substrate was recently proposed in a theoretical study. This method was based on approximate expressions for the evolution of the product concentration, which contained series expansions with five or more exponential terms, seriously complicating their use in practice. In the present paper, a new experimental method has been developed for the kinetic study of unstable and site-directed irreversible inhibitors, considering two different ranges of inhibitor concentration. Thus at low inhibitor concentrations, the system evolves from an initial to a final steady state, the rates of which are described by exact analytical equations. At high inhibitor concentrations, however, the product accumulation can be described by an exact uniexponential equation. This simple and efficient method has been applied to the kinetic study of trypsin inactivation by p-amidinophenylmethanesulphonyl fluoride, near the optimum pH of the enzyme. The dependence of the final steady-state rate on the substrate concentration shows apparent positive co-operativity which has not previously been reported. The kinetic origin of this type of co-operativity is predicted by one of the exact analytical equations derived here. The instability of new protein and non-protein irreversible inhibitors has to be carefully characterized to prevent true unstable irreversible inhibitors being wrongly described as allosteric reversible inhibitors.


Transcriptome evidence reveals enhanced autophagy-lysosomal function in centenarians.
Centenarians (CENs) are excellent subjects to study the mechanisms of human longevity and healthy aging. Here, we analyzed the transcriptomes of 76 centenarians, 54 centenarian-children, and 41 spouses of centenarian-children by RNA sequencing and found that, among the significantly differentially expressed genes (SDEGs) exhibited by CENs, the autophagy-lysosomal pathway is significantly up-regulated. Overexpression of several genes from this pathway, CTSB, ATP6V0C, ATG4D, and WIPI1, could promote autophagy and delay senescence in cultured IMR-90 cells, while overexpression of the Drosophila homolog of WIPI1, Atg18a, extended the life span in transgenic flies. Interestingly, the enhanced autophagy-lysosomal activity could be partially passed on to their offspring, as manifested by their higher levels of both autophagy-encoding genes and serum beclin 1 (BECN1). In light of the normal age-related decline of autophagy-lysosomal functions, these findings provide a compelling explanation for achieving longevity in, at least, female CENs, given the gender bias in our collected samples, and suggest that the enhanced waste-cleaning activity via autophagy may serve as a conserved mechanism to prolong the life span from Drosophila to humans.


Is it possible to "achieve a balance" and meet the "safety net"? Paediatric Advisory Subcommittee to the North West Thames Regional Health Authority.
The government's document Hospital Medical Staffing--Achieving a Balance--Plan for Action introduced the concept of a "safety net"--that is, a minimum safe level of staffing--of junior doctors in the acute specialties. The North West Thames Paediatric Advisory Group has therefore reviewed the implications and consequences of implementing the safety net in respect of children's services. The group found that if a reasonable safety net was to be provided that enabled the services to stay within the projected junior staffing levels, maintain a balance, meet training needs, and reduce junior doctors' hours of work, then changes in the organisation of the services would be required. Examining the options available showed that to achieve a safety net within the projected numbers of junior staff at least six paediatric units in the region would have to close. It is doubtful if there is the political will to support the very radical changes that would be needed in the distribution of services if the government's recommendations in Achieving a Balance were to be implemented. The profession, the Department of Health, and the public must be made aware that the proposed changes in medical staffing will cause a fundamental change in the traditional pattern of delivery of health care.


The impact of HIV/AIDS on children's educational outcome: a critical review of global literature.
The number of children losing one or both parents to HIV/AIDS has continued to rise in the past decade, with most of them being school-aged children. This study reviews global literature on the effects of HIV/AIDS (e.g., parental HIV-related illness or death) on children's schooling. Systematic review procedures generated 23 studies for examination. Existing studies show educational disadvantages among children affected by AIDS in various educational outcomes, including school enrollment and attendance, school behavior and performance, school completion, and educational attainment. A number of individual and contextual factors potentially moderate or mediate the effect of HIV/AIDS on children's education. These factors include gender of child, pattern of parental loss (maternal vs. paternal vs. dual), living arrangement (relationship with caregivers, gender of the household head), and household poverty. Current literature indicates limitations in number and scope of existing studies and in educational outcome measurements. There is a lack of studies with longitudinal design and data collection from multiple sources (e.g., students, teachers, caregivers), and a lack of studies on the relationship between psychosocial well-being of children affected by AIDS and their educational outcomes. Future studies need to employ more rigorous methodology and incorporate both individual and contextual factors for children affected by AIDS in various regions. More efforts are needed to design and implement culturally appropriate and context-specific approaches to improve the educational outcomes of children affected by AIDS.


Decoupling geometrical and chemical cues directing epidermal stem cell fate on polymer brush-based cell micro-patterns.
The intricacy of the different parameters involved in cell adhesion to biomaterials and fate decision (e.g. proliferation, differentiation, apoptosis) makes the decoupling of the respective effects of surface properties, extra-cellular matrix protein adsorption and ultimately cell behaviour difficult. This work presents a micro-patterned polymer brush platform to control the adsorption of extra-cellular matrix (ECM) proteins to well defined micron-size areas and consequently control cell adhesion, spreading and shape independently of other chemical and physical surface properties. Protein patterns can be readily generated with brushes presenting a range of hydrophilicity and surface charge density. The surface properties of the selected brushes are fully characterised using a combination of FTIR, XPS, ellipsometry, atomic force microscopy, water contact goniometry, dynamic light scattering and ζ-potential measurements. Interactions of proteins relevant to cell patterning and culture with these brushes are studied by surface plasmon resonance, dynamic light scattering, ellipsometry and immuno-fluorescence microscopy. Finally this platform is used in an assay investigating the relative contributions of matrix geometry and surface chemistry on epidermal stem cell differentiation. It is found that moderate hydrophobicity does not impact stem cell commitment, whereas strongly negative surface potential increases the incidence of differentiation. This correlates with a marked decrease in the formation of focal adhesions (but not cell spreading).


An NHC-Stabilized Silicon Analogue of Acylium Ion: Synthesis, Structure, Reactivity, and Theoretical Studies.
The silicon analogues of an acylium ion, namely, sila-acylium ions 2a and 2b [RSi(O)(NHC)2]Cl stabilized by two N-heterocyclic carbenes (NHC = 1,3,4,5-tetramethylimidazol-2-ylidene), and having chloride as a countercation were successfully synthesized by the reduction of CO2 using the donor stabilized silyliumylidene cations 1a and 1b [RSi(NHC)2]Cl (1a, 2a; R = m-Ter = 2,6-Mes2C6H3, Mes = 2,4,6-Me3C6H2 and 1b, 2b; R = Tipp = 2,4,6-iPr3C6H2). Structurally, compound 2a features a four coordinate silicon center together with a double bond between silicon and oxygen atoms. The reaction of sila-acylium ions 2a and 2b with water afforded different products which depend on the bulkiness of aryl substituents. Although the exposure of 2a to H2O afforded a stable silicon analogue of carboxylate anion as a dimer form, [m-TerSi(O)O]2(2-)·2[NHC-H](+) (3), the same reaction with the less bulkier triisopropylphenyl substituted sila-acylium ion 2b afforded cyclotetrasiloxanediol dianion [{TippSi(O)}4{(O)OH}2](2-)·2[NHC-H](+) (4). Metric and DFT (Density Functional Theory) evidence support that 2a and 2b possess strong Si═O double bond character, while 3 and 4 contain more ionic terminal Si-O bonds. Mechanistic details of the formation of different (SiO)n (n = 2, 3, 4) core rings were explored using DFT to explain the experimentally characterized products and a proposed stable intermediate was identified with mass spectrometry.


Vitamin D insufficiency in COVID-19 and influenza A, and critical illness survivors: a cross-sectional study.
The steroid hormone vitamin D has roles in immunomodulation and bone health. Insufficiency is associated with susceptibility to respiratory infections. We report 25-hydroxy vitamin D (25(OH)D) measurements in hospitalised people with COVID-19 and influenza A and in survivors of critical illness to test the hypotheses that vitamin D insufficiency scales with illness severity and persists in survivors. Cross-sectional study. Plasma was obtained from 295 hospitalised people with COVID-19 (International Severe Acute Respiratory and emerging Infections Consortium (ISARIC)/WHO Clinical Characterization Protocol for Severe Emerging Infections UK study), 93 with influenza A (Mechanisms of Severe Acute Influenza Consortium (MOSAIC) study, during the 2009-2010 H1N1 pandemic) and 139 survivors of non-selected critical illness (prior to the COVID-19 pandemic). Total 25(OH)D was measured by liquid chromatography-tandem mass spectrometry. Free 25(OH)D was measured by ELISA in COVID-19 samples. Receipt of invasive mechanical ventilation (IMV) and in-hospital mortality. Vitamin D insufficiency (total 25(OH)D 25-50 nmol/L) and deficiency (<25 nmol/L) were prevalent in COVID-19 (29.3% and 44.4%, respectively), influenza A (47.3% and 37.6%) and critical illness survivors (30.2% and 56.8%). In COVID-19 and influenza A, total 25(OH)D measured early in illness was lower in patients who received IMV (19.6 vs 31.9 nmol/L (p<0.0001) and 22.9 vs 31.1 nmol/L (p=0.0009), respectively). In COVID-19, biologically active free 25(OH)D correlated with total 25(OH)D and was lower in patients who received IMV, but was not associated with selected circulating inflammatory mediators. Vitamin D deficiency/insufficiency was present in majority of hospitalised patients with COVID-19 or influenza A and correlated with severity and persisted in critical illness survivors at concentrations expected to disrupt bone metabolism. These findings support early supplementation trials to determine if insufficiency is causal in progression to severe disease, and investigation of longer-term bone health outcomes.


Counselling adolescents and parents about cannabis: A primer for health professionals.
While cannabis use among adolescents is frequent in Canada, youth do recognize the potential harms, and increasingly expect knowledgeable health care providers to discuss substance use in everyday practice. This practice point provides sound, evidence-based tools to help health professionals address nonmedical (recreational) cannabis use and its related risks. After highlighting how to make the clinical setting a safe space for youth to talk about psychoactive substances, specific strategies for approaching cannabis use in effective, developmentally appropriate ways are described. Consistent with current literature, screening questionnaires to help structure discussion and identify adolescents who may benefit from more specialized interventions are recommended. Because one in six adolescents who experiments with cannabis goes on to misuse it, appraising their willingness to change risky behaviours is a key aspect of care, along with supportive goal-setting and helping families. Recommended resources for practitioners and parents are included.


MR imaging assessment of brain and cervical cord damage in patients with neuroborreliosis.
Neuroborreliosis is frequently indistinguishable from multiple sclerosis (MS) on both clinical and radiologic grounds. By using MR imaging, we assessed "occult" brain white matter (WM), brain gray matter (GM), and cervical cord damage in patients with neuroborreliosis in an attempt to achieve a more accurate picture of tissue damage in these patients, which might contribute to the diagnostic work-up. We studied 20 patients with neuroborreliosis and 11 sex- and age-matched control subjects. In all subjects, we acquired dual echo, T1-weighted, diffusion tensor (DT) and magnetization transfer (MT) MR imaging scans of the brain and fast short-tau inversion recovery and MT MR imaging scans of the cervical cord. T2-visible lesion load was measured by using a local thresholding segmentation technique. Mean diffusivity and fractional anisotropy histograms of the brain and cervical cord MT ratio histograms were produced. Normalized brain volumes (NBV) were measured by using SIENAx. Brain T2-visible lesions were detected in 12 patients, whereas no occult damage in the normal-appearing WM and GM was disclosed by using MT and DT MR imaging. No macroscopic lesions were found in the cervical cord, which was also spared by occult pathology. NBV did not differ between patients with neuroborreliosis and control subjects. This study shows that, contrary to what happens in MS, occult brain tissue damage and cervical cord pathology are not frequent findings in patients with neuroborreliosis. These observations might be useful in the diagnostic work-up of patients with neuroborreliosis and T2 brain lesions undistinguishable from those of MS.


Assessing spatial variability of soil petroleum contamination using visible near-infrared diffuse reflectance spectroscopy.
Visible near-infrared (VisNIR) diffuse reflectance spectroscopy (DRS) is a rapid, non-destructive method for sensing the presence and amount of total petroleum hydrocarbon (TPH) contamination in soil. This study demonstrates the feasibility of VisNIR DRS to be used in the field to proximally sense and then map the areal extent of TPH contamination in soil. More specifically, we evaluated whether a combination of two methods, penalized spline regression and geostatistics could provide an efficient approach to assess spatial variability of soil TPH using VisNIR DRS data from soils collected from an 80 ha crude oil spill in central Louisiana, USA. Initially, a penalized spline model was calibrated to predict TPH contamination in soil by combining lab TPH values of 46 contaminated and uncontaminated soil samples and the first-derivative of VisNIR reflectance spectra of these samples. The r(2), RMSE, and bias of the calibrated penalized spline model were 0.81, 0.289 log(10) mg kg(-1), and 0.010 log(10) mg kg(-1), respectively. Subsequently, the penalized spline model was used to predict soil TPH content for 128 soil samples collected over the 80 ha study site. When assessed with a randomly chosen validation subset (n = 10) from the 128 samples, the penalized spline model performed satisfactorily (r(2) = 0.70; residual prediction deviation = 2.0). The same validation subset was used to assess point kriging interpolation after the remaining 118 predictions were used to produce an experimental semivariogram and map. The experimental semivariogram was fitted with an exponential model which revealed strong spatial dependence among soil TPH [r(2) = 0.76, nugget = 0.001 (log(10) mg kg(-1))(2), and sill 1.044 (log(10) mg kg(-1))(2)]. Kriging interpolation adequately interpolated TPH with r(2) and RMSE values of 0.88 and 0.312 log(10) mg kg(-1), respectively. Furthermore, in the kriged map, TPH distribution matched with the expected TPH variability of the study site. Since the combined use of VisNIR prediction and geostatistics was promising to identify the spatial patterns of TPH contamination in soils, future research is warranted to evaluate the approach for mapping spatial variability of petroleum contaminated soils.


CYP3A4 and CYP2C19 genetic polymorphisms and zolpidem metabolism in the Chinese Han population: a pilot study.
Zolpidem (ZPD) is an imidazopyridine hypnotic and little is known about the pharmacogenetics of ZPD. Our objective was to evaluate inter-individual genetic variation in conjunction with metabolic ratios of ZPD found in a toxicological analysis. Healthy individuals (n=300) were genotyped for CYP2D6, CYP2C19, CYP2C9, CYP3A4 and CYP1A2 by allele-specific primer extension followed by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). Twenty-four Chinese volunteers were chosen and divided into the following four groups (n=6/group): group 1: CYP3A4*18 (wild-type, W), CYP2C19*2 (W); group 2: CYP3A4*18 (mutant, M), CYP2C19*2 (W); group 3: CYP3A4*18 (W), CYP2C19*2 (M); and group 4: CYP3A4*18 (M), CYP2C19*2 (M). ZPD and its major metabolites zolpidem 6-carboxylic acid (ZCA) and zolpidem phenyl-4-carboxylic acid (ZPCA) were determined after oral administration of ZPD (10mg), using an UPLC-MS/MS method. Positive correlations between CYP3A4 and CYP2C19 alleles and ZPD metabolism were found. The results of this study show that CYP3A4*18 increases CYP3A4 activity while CYP2C19*2 reduces CYP2C19 activity; the latter mutation is associated with the poor metabolism of ZPD in the Chinese Han population. The results also suggest that genetic factors play a major role in the metabolism of individual drugs with implications for both forensic science and clinical pharmacogenetics.


Are Active Video Games Effective at Eliciting Moderate-Intensity Physical Activity in Children, and Do They Enjoy Playing Them?
Despite current physical activity (PA) guidelines, children spend an average of 1-3 hours/day playing video games. Some video games offer physically active components as part of gameplay. We sought to determine if these active video games (AVGs) can elicit at least moderate PA in children, identify game elements important for PA, and determine if they are fun to play. Twenty children aged 8 to 16 years underwent cardiopulmonary exercise testing to determine their heart rate (HR) at ventilatory threshold. Participants played 2 different AVGs, and the gaming time that each participant's HR was above the HR thresholds for moderate and vigorous PA was determined. Gameplay elements that supported or inhibited active gameplay were also identified. Participants also completed questionnaires on physical activity, game engagement, and game experience. The Dance Central Spotlight and Kung-Fu for Kinect AVGs produced at least moderate PA, for a mean of 54.3% ± 29.5% and 87.8% ± 21.8% of gameplay time, respectively. Full-body movements, player autonomy, and self-efficacy were observed to be important elements of good AVG design. Although participants enjoyed these AVGs, they still preferred their favorite games (game engagement score of 1.82 ± 0.67 vs 0.95 ± 0.70 [Dance Central Spotlight] and 1.39 ± 0.37 [Kung Fu for Kinect]). AVGs can provide at least moderate PA and are enjoyable to play, but most popular video games do not incorporate active components. The implementation of government policies and a rating system concerning PA in video games may help address the widespread sedentary lifestyle of children.


Ultra-compact TE-pass polarizer with graphene multilayer embedded in a silicon slot waveguide.
A novel silicon slot waveguide TE-pass polarizer with graphene multilayer embedded in the slot is proposed and demonstrated by utilization of the fact that the variation of the modal characteristics for the TM mode is more than that for the TE mode. The designed polarizer is shown to have the ability to significantly suppress the transmission of the TM mode, while well guiding the TE mode. We numerically demonstrate a 7-μm-long polarizer has an ultra-high insertion loss of 31.5 dB for the TM mode and as little insertion loss as 0.2 dB for the TE mode at 1.55 μm. The presented polarizer offers the performance merits including high extinction ratio, ultra-low insertion loss, ultra-compactness, and easy integration with silicon slot waveguides without using any taper.


Fecal Calprotectin Test Performed at Home: A Prospective Study of Pediatric Patients With Inflammatory Bowel Disease.
Measuring fecal calprotectin (FC) in a laboratory is time-consuming and that is why home tests have been developed. We studied the use of an FC home test in pediatric patients with inflammatory bowel disease (PIBD) in real-life settings. The patients were asked to perform the IBDoc FC home test monthly for 6 months and to report their clinical disease activity at testing. Clinical decision-making, however, was guided by routine FC enzyme-linked immunosorbent assay (ELISA) for patients with raised IBDoc values. Spare frozen samples were analyzed using ELISA and IBDoc in the laboratory. The participants completed a questionnaire about FC testing at the start and end of the study. Of the 52 patients, 35 (67%) ages 5 to 18 years completed the study, and 197 home tests were performed. Of these, 15% failed, mainly because of technical reasons. Just under half of the patients (47%) considered home testing comparable or superior to routine testing. In contrast, the parents were unsatisfied (61%), mostly because the IBDoc results were significantly different from ELISA and they found the phone application difficult to handle but whenever the IBDoc was performed by a laboratory professional it was comparable with ELISA, suggesting that practical issues hampered home testing. Despite their reservations, more than 80% of parents felt that home testing would improve disease management. PIBD patients and their families were interested in FC home monitoring and willing to adopt testing as a part of their disease management, but this approach requires thorough guidance.


Discovery, adaptation and transcriptional activity of two tick promoters: Construction of a dual luciferase reporter system for optimization of RNA interference in rhipicephalus (boophilus) microplus cell lines.
Dual luciferase reporter systems are valuable tools for functional genomic studies, but have not previously been developed for use in tick cell culture. We evaluated expression of available luciferase constructs in tick cell cultures derived from Rhipicephalus (Boophilus) microplus, an important vector of bovine babesiosis and anaplasmosis. Commercial promoters were evaluated for transcriptional activity driving luciferase expression in the tick cell lines. The human phosphoglycerate kinase (PGK) promoter resulted in detectable firefly luciferase activity within 2 days post-transfection of the R. microplus cell line BME26, with maximal activity at 5 days post-transfection. Several other promoters were weaker or inactive in the tick cells, prompting identification and assessment of transcriptional activity of the homologous ribosomal protein L4 (rpL4, GenBank accession no.: KM516205) and elongation factor 1α (EF-1α, GenBank accession no.: KM516204) promoters cloned from R. microplus. Evaluation of luciferase expression driven by various promoters in tick cell culture resulted in selection of the R. microplus rpL4 promoter and the human PGK promoter driving transcription of sequences encoding modified firefly and NanoLuc® luciferases for construction of a dual luciferase reporter system for use in tick cell culture.


Usefulness of real-time three-dimensional echocardiography to quantify global left ventricular function and mechanical dyssynchrony after heart transplantation.
The purpose of this study was to examine left ventricular global function and left ventricular mechanical dyssynchrony (LVMD) after heart transplantation using three-dimensional echocardiography (RT-3DE) and to evaluate the value for RT-3DE in predicting cardiac allograft rejection (CAR). A total of 95 consecutive patients undergoing orthotropic heart transplantation, of whom 20 had CAR and 75 had no CAR, were enrolled in this study. Forty healthy volunteers were included as the control group. All patients underwent RT-3DE examination. Time to minimum systolic volume (Tmsv) of each left ventricular segment was measured. The parameters of LVMD including the standard deviation (SD) of Tmsv of 16 segments (Tmsv 16-SD), 12 segments (Tmsv 12-SD), and 6 basal segments (Tmsv 6-SD) were automatically calculated.The parameters of LVMD were adjusted by cardiac cycle and presented in terms of percentage as Tmsv 16-SD%, Tmsv 12-SD%, and Tmsv 6-SD%. The excursion average (Avg), excursion max (Max) and left ventricular ejection fraction (LVEF) were lower in patients with CAR than in those without CAR (all P < 0.05), while not different between patients without CAR and control subjects (all P > 0.05).The LVMD parameters, including Tmsv 16-SD, Tmsv 12-SD, Tmsv 6-SD, Tmsv 16-SD%, Tmsv 12-SD%, and Tmsv 6-SD%, were greater in patients with CAR than in those without CAR, while not different between patients without CAR and control subjects. All the RT-3DE parameters (LVEF and LVMD parameters) can predict CAR. However, Tmsv 16-SD (AUC 0.89 +/- 0.039, P < 0.001; sensitivity 85% and specificity 68%) and Tmsv 16-SD% (AUC 0.89 +/- 0.037, P < 0.001; sensitivity 95% and specificity 73%) offered the strongest power for detecting CAR. CAR can induce LVMD. LVMD parameters obtained by 3D-RTE, especially Tmsv 16-SD and Tmsv 16-SD%, provides a good sensitivity and specificity for predicting CAR after heart transplantation.


Specific binding of the human monocytic cell line U937 to the alternatively spliced connecting segment (IIICS) of fibronectin.
U937 cells attach to the RGDS-containing 80-kD fragment of fibronectin (Fn). The present report examined whether these cells recognize other domains of Fn. U937 cells attach to a 38-kD fragment derived from the A chain of Fn, which includes the Hep II domain and most of the alternatively spliced IIICS region. U937 did not bind to a 58-kD fragment derived from the B chain (which lacks IIICS) and has the Hep II site. They also did not bind to a 31-kD COOH-terminal fibrin-binding fragment or to a 29-kD fragment containing the Hep I domain. Cell adhesion to the 38-kD fragment was not inhibited by the 80-kD fragment, by GRGDSPC synthetic peptides, or by a mAb directed to the RGDS-containing domain of Fn. Attachment was completely inhibited by the 38-kD fragment and by the synthetic peptide CS-1, comprising the first 25 amino acid residues of IIICS. These results indicate that U937 cells interact with two sites of Fn, the RGDS-containing region, and the IIICS region.


Phase space determination from measured dose data for intraoperative electron radiation therapy.
A procedure to characterize beams of a medical linear accelerator for their use in Monte Carlo (MC) dose calculations for intraoperative electron radiation therapy (IOERT) is presented. The procedure relies on dose measurements in homogeneous media as input, avoiding the need for detailed simulations of the accelerator head. An iterative algorithm (EM-ML) has been employed to extract the relevant details of the phase space (PHSP) of the particles coming from the accelerator, such as energy spectra, spatial distribution and angle of emission of particles. The algorithm can use pre-computed dose volumes in water and/or air, so that the machine-specific tuning with actual data can be performed in a few minutes. To test the procedure, MC simulations of a linear accelerator with typical IOERT applicators and energies, have been performed and taken as reference. A solution PHSP derived from the dose produced by the simulated accelerator has been compared to the reference PHSP. Further, dose delivered by the simulated accelerator for setups not included in the fit of the PHSP were compared to the ones derived from the solution PHSP. The results show that it is possible to derive from dose measurements PHSP accurate for IOERT MC dose estimations.


A critical review on the occurrence and distribution of the uranium- and thorium-decay nuclides and their effect on the quality of groundwater.
This critical review presents the key factors that control the occurrence of natural elements from the uranium- and thorium-decay series, also known as naturally occurring radioactive materials (NORM), including uranium, radium, radon, lead, polonium, and their isotopes in groundwater resources. Given their toxicity and radiation, elevated levels of these nuclides in drinking water pose human health risks, and therefore understanding the occurrence, sources, and factors that control the mobilization of these nuclides from aquifer rocks is critical for better groundwater management and human health protection. The concentrations of these nuclides in groundwater are a function of the groundwater residence time relative to the decay rates of the nuclides, as well as the net balance between nuclides mobilization (dissolution, desorption, recoil) and retention (adsorption, precipitation). This paper explores the factors that control this balance, including the relationships between the elemental chemistry (e.g., solubility and speciation), lithological and hydrogeological factors, groundwater geochemistry (e.g., redox state, pH, ionic strength, ion-pairs availability), and their combined effects and interactions. The various chemical properties of each of the nuclides results in different likelihoods for co-occurrence. For example, the primordial 238U, 222Rn, and, in cases of high colloid concentrations also 210Po, are all more likely to be found in oxic groundwater. In contrast, in reducing aquifers, Ra nuclides, 210Pb, and in absence of high colloid concentrations, 210Po, are more mobile and frequently occur in groundwater. In highly permeable sandstone aquifers that lack sufficient adsorption sites, Ra is often enriched, even in low salinity and oxic groundwater. This paper also highlights the isotope distributions, including those of relatively long-lived nuclides (238U/235U) with abundances that depend on geochemical conditions (e.g., fractionation induced from redox processes), as well as shorter-lived nuclides (234U/238U, 228Ra/226Ra, 224Ra/228Ra, 210Pb/222Rn, 210Po/210Pb) that are strongly influenced by physical (recoil), lithological, and geochemical factors. Special attention is paid in evaluating the ability to use these isotope variations to elucidate the sources of these nuclides in groundwater, mechanisms of their mobilization from the rock matrix (e.g., recoil, ion-exchange), and retention into secondary mineral phases and ion-exchange sites.


Predominant affection of the blue cone pathway in Parkinson's disease.
Luminance contrast sensitivity and colour contrast thresholds were determined in 26 Parkinson patients and 17 normal controls of comparable age. They were psychophysically tested with a colour monitor system. Stimuli consisted of Gaussian enveloped luminance modulated or colour modulated (protan and tritan axis) vertical sine wave gratings with a spatial frequency of 1 cycle/degree. The stimuli subtended 4 degrees in diameter. Thresholds were determined using a two alternative forced choice method. Three different experimental conditions were explored: the detectability of stationary gratings, of moving gratings at velocities of 0, 2.5 and 5.0 cycles/s, and the detectability of horizontal square wave displacement at a frequency of 5 Hz for gratings of specified contrast levels. Intergroup differences were evaluated using two-tailed t tests with Satterthwaite corrections. Consistent and significant differences between normals and patients were found for tritan stimuli in the static and both dynamic conditions, and for luminance contrast stimuli in the displacement condition. Protan stimuli were much less apt to detect differences between the groups. We conclude that the retinal deficit of dopamine in Parkinson's disease is reflected in diminished centre/surround inhibition and that these changes are primarily apparent when vision is tested along the tritan axis, because blue cones are sparsely distributed.


Profiles of Great Lakes critical pollutants: a sentinel analysis of human blood and urine. The Great Lakes Consortium.
To determine the contaminants that should be studied further in the subsequent population-based study, a profile of Great Lakes (GL) sport fish contaminant residues were studied in human blood and urine specimens from 32 sport fish consumers from three Great Lakes: Lake Michigan (n = 10), Lake Huron (n = 11), and Lake Erie (n = 11). Serum was analyzed for 8 polychlorinated dioxin congeners, 10 polychlorinated furan congeners, 4 coplanar and 32 other polychlorinated biphenyl (PCB) congeners, and 11 persistent chlorinated pesticides. Whole blood was analyzed for mercury and lead. Urine samples were analyzed for 10 nonpersistent pesticides (or their metabolites) and 5 metals. One individual was excluded from statistical analysis because of an unusual exposure to selected analytes. Overall, the sample (n = 31) consumed, on average, 49 GL sport fish meals per year for a mean of 33 years. On average, the general population in the GL basin consume 6 meals of GL sport fish per year. The mean tissue levels of most persistent, bioaccumulative compounds also found in GL sport fish ranged from less than a twofold increase to that of PCB 126, which was eight times the selected background levels found in the general population. The overall mean total toxic equivalent for dioxins, furans, and coplanar PCBs were greater than selected background levels in the general population (dioxins, 1.8 times; furans, 2.4 times; and coplanar PCBs, 9.6 times). The nonpersistent pesticides and most metals were not identified in unusual concentrations. A contaminant pattern among lake subgroups was evident. Lake Erie sport fish consumers had consistently lower contaminant concentrations than consumers of sport fish from Lake Michigan and Huron. These interlake differences are consistent with contaminant patterns seen in sport fish tissue from the respective lakes; GL sport fish consumption was the most likely explanation for observed contaminant levels among this sample. Frequent consumers of sport fish proved to be effective sentinels for identifying sport fish contaminants of concern. In the larger study to follow, serum samples will be tested for PCBs (congener specific and coplanar), DDE, dioxin, and furans.


Electrocardiographic changes during treatment of leishmaniasis with pentavalent antimony (sodium stibogluconate).
Serial electrocardiograms (ECGs) were obtained during 65 courses of sodium stibogluconate treatment in 59 Kenyan patients with leishmaniasis (56 visceral and 3 cutaneous). ECG abnormalities developed during 54% of the treatment courses. The frequency with which abnormalities occurred was related to the total daily dose of antimony (Sb), increasing from 2/9 patients treated with 10 mg Sb/kg/d to 25/48 treated with 20-30 mg Sb/kg/d and 8/8 treated with 40-60 mg Sb/kg/d. The frequency with which ECG abnormalities developed was also related to the duration of treatment, increasing from 11/65 patients after 7 days to 18/44 after 15 days, 26/39 after 30 days and 11/12 after 60 days. ECG abnormalities were similar to those previously described during treatment with trivalent antimonial drugs, the most common being flattening and/or inversion of T waves. Prolongation of the corrected QT interval occurred in 13 patients, all of whom were treated for more than 30 days or with more than 20 mg Sb/kg/d. One patient died suddenly during the fourth week of treatment with 60 mg Sb/kg/d, and 2 patients died of measles after 9 or 10 days of treatment with 30 mg Sb/kg/d. QT prolongation and a concave ST segment developed in all 3 patients who died. We conclude that minor ECG abnormalities are common when sodium stibogluconate is used at doses above 20 mg Sb/kg/d for more than 15 days, and that life-threatening arrhythmias may occur if very high doses are used.


Scalp Tissue Expansion Above a Custom-Made Hydroxyapatite Cranial Implant to Correct Sequelar Alopecia on a Transposition Flap.
Resection of cranial tumors involving both bone and scalp tissue may require the recruitment of soft tissue using a flap above the bone reconstruction. When a transposition flap has been chosen, the alopecia zone on the donor site may be difficult to treat afterward. Scalp expansion is the gold standard in these situations, but this has never been described above cranial implants. We report the first case of a patient who underwent a scalp tissue expansion above a custom-made hydroxyapatite cranial implant to correct sequelar alopecia. A 30-year-old man presented with a dermatofibrosarcoma of the scalp with bone invasion. A cranioplasty with a custom-made hydroxyapatite implant and a transposition flap were performed. Although healing was achieved, the donor site of the transposition flap left a 9 × 13 cm sequelar alopecia area on the vertex. To correct it, a rectangular 340-cm3 expander was partially placed above the cranial implant and under the transposition flap. A second 120-cm3 expander was put on the contralateral temporal region. The expansion was successful. No expander infection, cranial implant displacement, or fracture on imaging performed during the follow-up period was detected. The alopecia zone was entirely excised. The patient was very satisfied with the cosmetic result. Tissue expansion above hydroxyapatite implants may be of concern to the physician because of the risk of infection and rupture of the cranial implant. With this clinical case, we emphasize some precautions to prevent these issues.


Is frontotemporal lobar degeneration a rare disorder? Evidence from a preliminary study in Brescia county, Italy.
Frontotemporal Lobar Degeneration (FTLD) has always been considered a rare disorder, but only a few epidemiologic studies are available. The aim of the present work was to ascertain all FTLD patients in a Northern Italy area from January 2001 to December 2008, and to estimate the disease prevalence. On the census day, 213 FTD patients were still alive, resulting in an overall prevalence of 17.6 per 100,000 inhabitants. The prevalence of FTLD in patients aged 45-65 years was 22 per 100,000 inhabitants (95% CI=17-27). The prevalence of FTLD was the highest in patients aged 66-75 (78 per 100,000 inhabitants, 95% CI=56-100), and it was still high over 75 years (54 per 100,000 inhabitants, 95% CI=36-69). FTLD is a more common form of dementia than previously recognized. Our results claimed that FTLD is not only an early-onset disorder, but it is frequent in advanced age as well.


Clinical presentations in combat veterans diagnosed with posttraumatic stress disorder.
This article investigated subtypes of symptom patterns among male combat veterans diagnosed with posttraumatic stress disorder (PTSD) through a cluster analysis of their Minnesota Multiphasic Personality Inventory-2 (MMPI-2; Butcher, Graham, Ben-Porath, Tellegen, Dahlstrom, & Kaemmer, 2001) clinical and validity scales. Participants were 126 veterans seeking outpatient treatment for combat-related PTSD at a Veterans Affairs Medical Center. Two well-fitting MMPI-2 cluster solutions (a four-cluster solution and a three-cluster solution) were evaluated with several statistical methods. A four-cluster solution was determined to best fit the data. Follow-up analyses demonstrated between-cluster differences on MMPI-2 "fake bad" scales and content scales, the Beck Depression Inventory (BDI; Beck, Ward, Mendelson, Mock, & Erbaugh, 1961), Dissociative Experiences Scale (DES; Bernstein & Putnam, 1986), Mississippi Combat PTSD scale (M-PTSD; Keane, Caddall, & Taylor, 1988), and Clinician-Administered PTSD Scale (CAPS-1; Blake et al., 1990). Clusters also were different in disability-seeking status, employment status, and income. Implications for research and clinical practice using the MMPI-2 with combat veterans presenting with PTSD are briefly addressed.


Analytical performance, reference values and decision limits. A need to differentiate between reference intervals and decision limits and to define analytical quality specifications.
With the increasing use of decision limits (action limits, cut-off points) specified for a number of analytical components in diagnosis and for action in critical situations, formulated in national or international recommendations, the traditional interpretation of reference intervals has been uncertain, and sometimes the two concepts are being mixed up by incorporating risk calculations in the reference intervals. There is, therefore, a need to clarify the two concepts and to keep them definitely separated. Reference intervals are the 95% limits for the descriptions of the distributions of the values of analytical components measured on reference samples from reference individuals. Decision limits are based on guidelines from national and international expert groups defining specific concentrations of certain components as limits for decision about diagnosis or well-defined specific actions. Analytical quality specifications for reference intervals have been defined for bias since the 1990s, but in the recommendations specified in the clinical guidelines analytical quality specifications are only scarcely defined. The demands for negligible biases are, however, even more essential for decision limits, as the choice is no longer left to the clinician, but emerge directly from the concentration. Even a small bias will change the number of diseased individuals, so the demands for negligible biases are obvious. A view over the analytical quality as published gives a variable picture of bias for many components, but with many examples of considerable bias which must be critical--yet no specifications have been stipulated until now.


Diagnosis of Parkinson's disease based on disease-specific autoantibody profiles in human sera.
Parkinson's disease (PD), hallmarked by a variety of motor disorders and neurological decline, is the second most common neurodegenerative disease worldwide. Currently, no diagnostic test exists to identify sufferers, and physicians must rely on a combination of subjective physical and neurological assessments to make a diagnosis. The discovery of definitive blood-borne biomarkers would be a major step towards early and reliable diagnosis. Despite attention devoted to this search, such biomarkers have remained elusive. In the present study, we used human protein microarrays to reveal serum autoantibodies that are differentially expressed among PD and control subjects. The diagnostic significance of each of these autoantibodies was evaluated, resulting in the selection of 10 autoantibody biomarkers that can effectively differentiate PD sera from control sera with a sensitivity of 93.1% and specificity of 100%. PD sera were also distinguishable from sera obtained from Alzheimer's disease, breast cancer, and multiple sclerosis patients with accuracies of 86.0%, 96.6%, and 100%, respectively. Results demonstrate that serum autoantibodies can be used as highly specific and accurate biomarkers for PD diagnosis throughout the course of the disease.


[Agenesis of the gallbladder (diagnostic considerations. Apropos of a case].
The authors show a case of gall-bladder agenesis, in the preoperative diagnostics of which both echotomography and T.A.C. were used. From the data of the literature, as well as from the personal experience, one can understand the preoperative diagnosis presuming gall-bladder agenesis can be placed in presence od manifestations of biliary nature added to gall-bladder nonopacity at oral cholecystography and to absence of gall-bladder at echotomography and T.A.C. The certainty diagnosis, however, can be only obtained at laparotomy with the hand exploration, with the preoperative cholangiography and eventually the selective arteriography of hepatic artery.


Neonatal metrizamide gastrointestinal series in suspected necrotizing enterocolitis.
The diagnosis of necrotizing enterocolitis (NEC) in neonates may be made by clinical presentation, roentgenographic findings, or a combination of both. Diagnosis leads to immediate treatment including nasogastric suction, parenteral antibiotics, plasma, and close monitoring of clinical, roentgenographic, and laboratory findings. Occasionally, neither the clinical nor plain roentgenographic appearance of an infant allows the diagnosis of NEC to be made or excluded with confidence. In such infants portable isotonic metrizamide gastrointestinal (GI) series were used to help make the decision of whether to begin treatment for NEC or to continue feeding the patient. Of 15 patients examined, two exhibited signs of NEC and were successfully treated medically without GI (tract) sequelae. Twelve neonates had normal results of metrizamide GI series and ten were immediately fed with no GI complication. One of these 12 infants had feedings withheld for several days as a result of a positive blood culture. One infant with severe cardiac and pulmonary disease had profound adynamic ileus and could not be fed. We have found the metrizamide GI series to be a useful study in neonates suspected of having NEC.