[Persistent neonatal hyperinsulinism. Analysis of the differential diagnosis. Report of two cases].
Persistent neonatal hyperinsulinism is the most common cause of refractory hypoglycemia during the first year of life. Inadequate insulin secretion is associated to mutations of four different genes, that can be diagnosed to orient patient management. We report two patients: a female newborn that presented a hypoglycemia of 16 mg/dl two hours after birth, was subjected to a subtotal pancreatectomy that did not correct hypoglycemia, requiring a total pancreatectomy. Pathological study of the pancreas showed a focal adenomatous hyperplasia. At the present time, she is three years of age and maintains euglycemia with fractionated feeding. A male newborn that had seizures at 28 hours of life and a hypoglycemia of 15 mg/dl was detected. He was also subjected to a subtotal pancreatectomy, that did not correct hypoglycemia and bad to be extended to a total pancreatectomy. At the present time, he is 3 years and 11 months of age and has a normal psychomotor development.


BIOKID: randomized controlled trial comparing bicarbonate and lactate buffer in biocompatible peritoneal dialysis solutions in children [ISRCTN81137991].
Peritoneal dialysis (PD) is the preferred dialysis modality in children. Its major drawback is the limited technique survival due to infections and progressive ultrafiltration failure. Conventional PD solutions exert marked acute and chronic toxicity to local tissues. Prolonged exposure is associated with severe histopathological alterations including vasculopathy, neoangiogenesis, submesothelial fibrosis and a gradual loss of the mesothelial cell layer. Recently, more biocompatible PD solutions containing reduced amounts of toxic glucose degradation products (GDPs) and buffered at neutral pH have been introduced into clinical practice. These solutions contain lactate, bicarbonate or a combination of both as buffer substance. Increasing evidence from clinical trials in adults and children suggests that the new PD fluids may allow for better long-term preservation of peritoneal morphology and function. However, the relative importance of the buffer in neutral-pH, low-GDP fluids is still unclear. In vitro, lactate is cytotoxic and vasoactive at the concentrations used in PD fluids. The BIOKID trial is designed to clarify the clinical significance of the buffer choice in biocompatible PD fluids. The objective of the study is to test the hypothesis that bicarbonate based PD solutions may allow for a better preservation of peritoneal transport characteristics in children than solutions containing lactate buffer. Secondary objectives are to assess any impact of the buffer system on acid-base status, peritoneal tissue integrity and the incidence and severity of peritonitis. After a run-in period of 2 months during which a targeted cohort of 60 patients is treated with a conventional, lactate buffered, acidic, GDP containing PD fluid, patients will be stratified according to residual renal function and type of phosphate binding medication and randomized to receive either the lactate-containing Balance solution or the bicarbonate-buffered Bicavera solution for a period of 10 months. Patients will be monitored by monthly physical and laboratory examinations. Peritoneal equilibration tests, 24-h dialysate and urine collections will be performed 4 times. Peritoneal biopsies will be obtained on occasion of intraabdominal surgery. Changes in small solute transport rates, markers of peritoneal tissue turnover in the effluent, acid-base status and peritonitis rates and severity will be analyzed.


Increased glutathione-S-transferase activity in a cell line with acquired resistance to nitrogen mustards.
A Walker 256 rat mammary carcinoma cell line (WR) resistant to bifunctional nitrogen mustards has been shown to have an approximate twofold increase in bulk glutathione-S-transferase activity compared to the parent cell line. Substrate specificity studies suggest that higher levels of Yb subunit contribute to the increased activity. By exposing WR cells to additional chlorambucil, either as a single concentration (50 micrograms/ml) or at 5 micrograms/ml for 10 days, transferase activity was further increased by up to three times the normal WR level. By using colony-forming assays, mitotic index depression, or trypan blue exclusion, the increased transferase activity could be correlated with an increase in resistance of these cells to either subsequent chlorambucil or a different bifunctional nitrogen mustard, phosphoramide mustard.


Total Hip Arthroplasty and the Spinopelvic Relationship: What's the Latest!
Although total hip arthroplasty (THA) has proved to be a successful surgical procedure, both prosthetic and bone impingement resulting in dislocation continue to occur. Studies have shown that spine pathology resulting in lumbar stiffness and hip arthritis often coexist. Spinopelvic mobility patterns during postural changes affect three-dimensional acetabular component position, which affects the incidence of prosthetic impingement and THA instability. Several spinopelvic risk factors that may affect THA stability have been identified. Numerous reports recommend performing a preoperative spinopelvic mobility analysis to identify risk factors and adjust acetabular component position accordingly to lessen the risk of impingement. In doing so, acetabular component position is individualized based on spinopelvic mobility patterns. Additionally, functional femoral anteversion, affected by individual femoral rotation patterns during dynamic activities, may contribute to the incidence of impingement. It is important to review the interrelationship between spine and pelvic mobility and how it relates to THA and may reduce the incidence of instability.


Neutrophilic Asthma Is Associated with Increased Airway Bacterial Burden and Disordered Community Composition.
Neutrophilic asthma (NA) is an important asthma inflammatory phenotype associated with disease severity, airflow limitation, and steroid resistance, and its mechanism is still uncertain. Evidences suggest a potential role for bacteria in its pathogenesis, but, so far, this remains poorly understood. We sought to investigate airway bacterial burden, community composition, and inflammatory response in NA. Fifty-four stable asthmatics without infection were enrolled and separated into either NA group (n = 20) or non-NA group (n = 34). Subject demographics, Asthma Control Test (ACT) scores, medications, and pulmonary functions were documented. Sputum cytology, airway bacterial burden, microbial community composition, and inflammatory cytokines were assessed. The total airway bacterial burden was significantly increased in subjects with NA versus non-NA and was positively correlated with the sputum neutrophil percentage. Airway neutrophilia was associated with less airway bacterial community richness and diversity, along with a distinct community composition. In patients with NA, bacteria in phylum Proteobacteria, especially Haemophilus spp. and Moraxella spp., showed significant increases in both actual loads and relative abundances, while bacteria in phyla Firmicutes, Actinobacteria, and Saccharibacteria showed decreased relative abundances compared with non-NA. Patients with NA demonstrated higher levels of interleukin-1β (IL-1β), IL-6, IL-8, IL-12, IL-17A, and tumor necrosis factor-α (TNF-α) in sputum samples compared with non-NA. Increased bacterial burden and distinct microbiota composition were the key characters of neutrophilic phenotype in asthma, accompanied by excessive airway inflammation. Understanding the relationship between airway microbiota and neutrophilic inflammation may help in treatment and management of asthma, such as targeting airway microbiota.


Use of the chaplaincy in the neonatal intensive care unit.
The hospital chaplain is a relatively new member of the health care team, and there is little objective information on the chaplaincy's role in the hospital. To evaluate the chaplain's role, I collected information from the chaplain's daily logs and from a medical staff/parent questionnaire on the use of the chaplaincy in the neonatal intensive care unit (NICU). Both the medical/nursing staff and the chaplains viewed the chaplain's role as encompassing both traditional (religious functionary) and nontraditional functions. However, the medical/nursing staff consulted chaplains only for traditional roles. The discrepancy in chaplain use suggests a need for objective study about the chaplain's role. I offer hypotheses for testing the role of the chaplaincy in the NICU.


Blood pressure maintenance in awake dehydrated rats: renin, vasopressin, and sympathetic activity.
The role of vasopressin, the renin system, and sympathetic activity in sustaining blood pressure in the dehydrated state was investigated in normotensive nonanesthetized male Wistar rats. After 48-h dehydration, plasma arginine vasopressin was 14.0 +/- 1.7 pg/ml and plasma norepinephrine 0.46 +/- 0.05 ng/ml. In another group of rats in which the angiotensin converting enzyme inhibitor (MK 421, 5 mg po twice daily) was administered throughout the dehydration period, blood pressure was reduced by more than 20% (P less than 0.001), and both plasma arginine vasopressin and norepinephrine were higher at 23.4 +/- 3.9 pg/ml (P less than 0.01) and 0.83 +/- 0.07 ng/ml (P less than 0.01), respectively. Taken together, in rats with or without converting enzyme blockade, there was an inverse correlation between mean blood pressure and plasma arginine vasopressin (r = 0.67, P less than 0.01) as well as plasma norepinephrine (r = 0.82, P less than 0.01) levels. The acute administration of a specific vasopressin pressor inhibitor (dPVDAVP) reduced mean blood pressure in the rats with a blocked renin system by 16.9 mmHg (P less than 0.001). In rats without converting enzyme inhibition, the induced fall was only 6.4 mmHg. These results indicate that following 48-h dehydration the renin angiotensin system interacts with the vasopressin secretory mechanism to sustain blood pressure, with renin playing a predominant role. They further suggest that, following blockade of the renin system, activation of the sympathetic nervous system probably also contributes to blood pressure maintenance.


System for automatic activation of skinned muscle fibers.
A system is described for automatically changing the solutions surrounding skinned muscle fibers at a controlled temperature. The system consists of a complex muscle chamber and solenoid valve-controlled vacuum-driven syringe pumps. The chamber has a muscle trough, a conduit for circulating coolant, and channels for precooling solutions. After traveling through the cooling channels, the solutions are injected into one end of the muscle trough and removed by suction at the other. The volume of the trough (approximately 40 microliter) is kept as low as possible to minimize the amount of solution required for complete solution exchange (approximately 400 microliter). Solution changes are complete within 400 ms. The timing of the sequence of solution changes is precisely controlled by digital timers that activate the solenoid valves of the pumps. By exposing the fibers briefly to a high concentration of free calcium and then changing to a buffered calcium solution, it is possible to achieve steady tension levels in 1-2 s, even when partially activating the fibers. Such rapid partial activation is possible because the exposure to free calcium can be precisely timed. The timers also allow the solution changes to be synchronized with other perturbations.


Two-speed genomes of Epichloe fungal pathogens show contrasting signatures of selection between species and across populations.
Antagonistic selection between pathogens and their hosts can drive rapid evolutionary change and leave distinct molecular footprints of past and ongoing selection in the genomes of the interacting species. Despite an increasing availability of tools able to identify signatures of selection, the genetic mechanisms underlying coevolutionary interactions and the specific genes involved are still poorly understood, especially in heterogeneous natural environments. We searched the genomes of two species of Epichloe plant pathogen for evidence of recent selection. The Epichloe genus includes highly host-specific species that can sterilize their grass hosts. We performed selection scans using genome-wide SNP data from seven natural populations of two co-occurring Epichloe sibling species specialized on different hosts. We found evidence of recent (and ongoing) selective sweeps across the genome in both species. However, selective sweeps were more abundant in the species with a larger effective population size. Sweep regions often overlapped with highly polymorphic AT-rich regions supporting the role of these genome compartments in adaptive evolution. Although most loci under selection were specific to individual populations, we could also identify several candidate genes targeted by selection in sweep regions shared among populations. The genes encoded small secreted proteins typical of fungal effectors and cell wall-degrading enzymes. By investigating the genomic signatures of selection across multiple populations and species, this study contributes to our understanding of complex adaptive processes in natural plant pathogen systems.


Planarization of B20 clusters by Si and C atom substitution.
An optimization strategy combining a global semi-empirical quantum mechanical search and all-electron density functional theory was adopted to determine the lowest energy structures of B19Si and B19C clusters. The planarization of a B20 cluster by Si and C atom substitution was observed. The structural transition was from the double-ring tubular B20 to an almost perfect planar B19Si and a quasi-planar bowl B19C. B19Si possessed a geometry with a central B atom surrounded by a six-membered ring and a 13-atom outer ring. B19C adopted a geometry with a B5C six-membered hole. Both Si and C atoms occupied peripheral positions. The observed planarization may be attributed to sp2 hybridization, changes in the peripheral bonding, and structural mechanics. Some properties, including the HOMO-LUMO gaps, on-site charge on Si and C atoms, and deformed charge distribution, were discussed.


[Detection of antibodies against enterovirus 71 in the sera of Moscow residents].
Twenty three out of 55 serum samples obtained from subjects aged 20-55 years have proved to be seropositive, 15 of the positive serum samples (i.e. more than 50%) possessing the titer greater than or equal to 1:16. The presence of specific antibodies in the sera of the inhabitants of Moscow suggests that either enterovirus 71 or some other virus whose antigens are partly identical to those of enterovirus 71 apparently circulates among the population of Moscow.


Identification of the D2-dopamine receptor binding subunit in several mammalian tissues and species by photoaffinity labeling.
Photoaffinity labeling of the D2-dopamine receptor in plasma membrane preparations of various tissues from several mammalian species was performed using the recently developed D2-dopaminergic antagonist probe [125I]N-(p-azidophenethyl)spiperone ([125I]N3-NAPS). In tissues containing D2-receptors such as the corpus striatum from rat, dog, calf, hamster, guinea pig, and rabbit as well as the anterior pituitary of rat, bovine, and hamster, the probe covalently labels a peptide of Mr = 94,000. Specificity of the labeling is typically D2-dopaminergic in character. The covalent labeling is blocked by (+)-butaclamol but not by the inactive (-)isomer. Agonists block incorporation with the order of potency: N-n-propylnorapomorphine greater than apomorphine greater than dopamine. The D2-selective antagonist spiperone blocks labeling of the Mr = 94,000 peptide whereas the D1-selective antagonist SCH-23390 is ineffective. Thus, these results indicate that the ligand binding subunit of the D2-dopamine receptor resides on a Mr = 94,000 peptide in these various tissues from several species. Under conditions where proteolysis is not stringently controlled, peptides of lower Mr (32-38,000) are labeled at the expense of the Mr = 94,000 peptide. The most efficient protease inhibitor tested in these systems was EDTA, suggesting that the generation of these lower Mr receptor fragments might be the result of a metal-dependent proteolysis in the membrane preparations. In the rat neurointermediate lobe, a tissue containing D2-receptors, [125I]N3-NAPS specifically labels a major peptide of Mr approximately equal to 120,000 in addition to the Mr = 94,000 peptide.(ABSTRACT TRUNCATED AT 250 WORDS)


Anti-parasitic benzoxaboroles are ineffective against Theileria parva in vitro.
East Coast Fever (ECF) is a disease affecting cattle in sub-Saharan Africa, caused by the tick-borne Apicomplexan pathogen Theileria parva. The disease is a major problem for cattle farmers in affected regions and there are few methods of control, including a complex infection and treatment vaccine, expensive chemotherapy, and the more widespread tick control through acaricides. New intervention strategies are, therefore, sorely needed. Benzoxaboroles are a versatile class of boron-heterocyclic compounds with demonstrable pharmacological activity against a diverse group of pathogens, including those related to T. parva. In this study, the in vitro efficacy of three benzoxaboroles against the intracellular schizont stage of T. parva was investigated using a flow cytometry approach. Of the benzoxaboroles tested, only one showed any potency, albeit only at high concentrations, even though there is high protein sequence similarity in the CPSF3 protein target compared to other protozoan pathogen species. This finding suggests that benzoxaboroles currently of interest for the treatment of African animal trypanosomiasis, toxoplasmosis, cryptosporidiosis and malaria may not be suitable for the treatment of ECF. We conclude that testing of further benzoxaborole compounds is needed to fully determine whether any lead compounds can be identified to target T. parva.


Cationic type I amphiphiles as modulators of membrane curvature elastic stress in vivo.
Recently we proposed that the antineoplastic properties observed in vivo for alkyl-lysophospholipid and alkylphosphocholine analogues are a direct consequence of the reduction of membrane stored elastic stress induced by these amphiphiles. Here we report similar behavior for a wide range of cationic surfactant analogues. Our systematic structure-activity studies show that the cytotoxic properties of cationic surfactants follow the same pattern of activity we observed previously for alkyl-lysophospholipid analogues, indicating a common mechanism of action that is consistent with the theory that these amphiphiles reduce membrane stored elastic stress. We note that several of the cationic surfactant compounds we have evaluated are also potent antibacterial and antifungal agents. The similarity of structure-activity relationships for cationic surfactants against microorganisms and those we have observed in eukaryotic cell lines leads us to suggest the possibility that the antibacterial and antifungal properties of cationic surfactants may also be due to modulation of membrane stored elastic stress.


Age and Ageing journal 50th anniversary commentary seriesWhy illness is more important than disease in old age.
Clinical reasoning and research in modern geriatrics often prioritises the disease concept. This is understandable as it has brought impressive advances in medicine (e.g. antibiotics, vaccines, successful cancer treatment and many effective surgeries). However, so far the disease framework has not succeeded in getting us to root causes of many age-related chronic diseases (e.g. Alzheimer's disease, diabetes, osteoarthritis). Moreover, in aging and disease constructs alone fail to explain the variability in illness presentations. Therefore, we propose to apply the underused illness concept in a new way by reconsidering the importance of common symptoms in the form of a dynamic network of symptoms as a complementary framework. We show that concepts and methods of complex system thinking now enable to fruitfully monitor and analyse the multiple interactions between symptoms in such in networks, offering new routes for prognosis and treatment. Moreover, close attention to the symptoms that bother older persons may also improve weighing the therapeutic objectives of well-being and survival and aligning treatment targets with the patients' priorities.


Distinct functions for Aldh1 and Raldh2 in the control of ligand production for embryonic retinoid signaling pathways.
During vertebrate embryogenesis retinoic acid (RA) synthesis must be spatiotemporally regulated in order to appropriately stimulate various retinoid signaling pathways. Various forms of mammalian aldehyde dehydrogenase (ALDH) have been shown to oxidize the vitamin A precursor retinal to RA in vitro. Here we show that injection of Xenopus embryos with mRNAs for either mouse Aldh1 or mouse Raldh2 stimulates RA synthesis at low and high levels, respectively, while injection of human ALDH3 mRNA is unable to stimulate any detectable level of RA synthesis. This provides evidence that some members of the ALDH gene family can indeed perform RA synthesis in vivo. Whole-mount immunohistochemical analyses of mouse embryos indicate that ALDH1 and RALDH2 proteins are localized in distinct tissues. RALDH2 is detected at E7.5-E10.5 primarily in trunk tissue (paraxial mesoderm, somites, pericardium, midgut, mesonephros) plus transiently from E8.5-E9.5 in the ventral optic vesicle and surrounding frontonasal region. ALDH1 is first detected at E9.0-E10. 5 primarily in cranial tissues (ventral mesencephalon, dorsal retina, thymic primordia, otic vesicles) and in the mesonephros. As previous findings indicate that embryonic RA is more abundant in trunk rather than cranial tissues, our findings suggest that Raldh2 and Aldh1 control distinct retinoid signaling pathways by stimulating high and low RA biosynthetic activities, respectively, in various trunk and cranial tissues.


Efficacy and safety of blue-light scleral cross-linking.
To evaluate the efficacy of blue-light scleral cross-linking as well as its safety in preventing retinal damage beneath the treated sclera. Six rabbits were unilaterally treated with topical riboflavin (0.5%) and blue light (465 nm) on the equatorial sclera using a light emitting diode source with an exposure area of 9 mm in diameter. Four weeks after the treatment, the animals were euthanized and the exposed sclera and contralateral eye sclera excised for comparative testing of biomechanical rigidity and histologic retinal cellular damage. Extensiometry was performed to evaluate the stress-strain curve of treated versus untreated sclera, and light microscopy of the treated sclera and underlying retina were also comparatively evaluated. Blue-light scleral cross-linking showed a three-fold increased stiffening in all tested animals in the stress-strain curve. Histological investigation revealed no retinal damage in any of the treated eyes. Scleral cross-linking with riboflavin and blue light (465 nm) has a stiffening effect on the sclera without histological tissue damage to the retina.


How accurate are hypertension treatment decisions? Absolute risk assessment and prescribing for moderate hypertension--a study of Cambridge general practitioners.
To determine the factors influencing general practitioners' prescribing for patients with moderate hypertension, and to investigate their ability to assess patients' absolute risk of cardiovascular disease. Decision-making was assessed by a postal questionnaire involving five standardised example patients, send to all general practitioners in the CB-postcode area (Cambridge, UK). Patients varied in age, gender, diabetic and smoking status, blood pressure and total cholesterol, giving 5-year absolute risks ranging from less than 5% to greater than 20%. GP treatment decisions and risk factor influence, age bias, absolute risk calculations and cost-awareness were analysed with SPSS. Absolute risk was calculated using the New Zealand Core Services Committee (NZCSC) Guidelines, 1995. GP response rate was 66%. Sample demographics (mean age 42 years, 39% female) did not differ statistically from local health authority data. High blood pressure alone was the most common reason for instituting therapy (67%), independent of other risk factors. Drug choices did not differ statistically from Prescription Pricing Authority data. This supports the validity of the questionnaire. GP estimations of absolute risk were inconsistent, and there was a significant association between underestimation of elderly patient risk and overestimation of younger patient risk (p = 0.05). The Cambridge GPs more frequently based treatment decisions on patients' blood pressure level alone, rather than considering absolute risk as recommended by NZCSC and WHO guidelines. The relationship between patient age and risk estimations may indicate a "young age premium" influencing treatment. The data support the need for risk-based guidelines and education in treatment of hypertension.


Status epilepticus: an independent outcome predictor after cerebral anoxia.
Prognosis of status epilepticus (SE) depends on its cause, but there is uncertainty as to whether SE represents an independent outcome predictor for a given etiology. Cerebral anoxia is a relatively homogenous severe encephalopathy. Postanoxic SE is associated to a nearly 100% mortality in this setting; however, it is still unclear whether this is a severity marker of the underlying encephalopathy, or an independent factor influencing outcome. The goal of this study was to assess if postanoxic SE is independently associated with mortality after cerebral anoxia. This was a retrospective observation of consecutive comatose survivors of cardiac arrest, including subjects treated with hypothermia. On the subgroup with EEG recordings in the first hospitalization days, univariate and multivariate analyses were applied to potential determinants of in-hospital mortality, and included the following variables: age, gender, type and length of cardiac arrest, occurrence of circulatory shock, presence of therapeutic hypothermia, and electrographic SE. Out of 166 postanoxic patients, 107 (64%) had an EEG (median latency from admission, 2 days); in this group, therapeutic hypothermia was administered in 59%. Death occurred in 71 (67%) patients. Postanoxic SE was associated with mortality regardless of type of acute cardiac rhythm and administration of hypothermic treatment. In this hospital-based cohort, postanoxic status epilepticus (SE) seems to be independently related to death in cardiac arrest survivors, suggesting that SE might determine a bad prognosis for a given etiology. Confirmation of these results in a prospective assessment is needed.


Postsurgical Physical Therapy Management of Congenital Muscular Torticollis.
To present 2 cases with severe congenital muscular torticollis and describe postsurgical physical therapy management. Two children diagnosed with severe congenital muscular torticollis underwent surgical release of the involved sternocleidomastoid muscle with subsequent physical therapy. Presurgical examination findings, surgical technique, and postsurgical rehabilitation guidelines are described. Both cases achieved full range of motion and symmetrical muscle strength. Functionally, they were able to maintain consistent midline posture, perform transitions with symmetrical head righting, and demonstrate age-appropriate motor skills. Children with surgical management of congenital muscular torticollis tend to have more severe involvement of the sternocleidomastoid. Prolonged experience in atypical postures may result in potential asymmetrical development of visual, vestibular, and proprioceptive systems. Perception of midline was reestablished through integration of visual, vestibular, and proprioceptive stimuli. For more insights from the authors, access Supplemental Digital Content 1, at http://links.lww.com/PPT/A141.


Differential accessibility of bilirubin to erythrocyte membrane vesicles bearing different structural features.
Interaction of bilirubin with different types of erythrocyte membrane vesicles such as unsealed, heterogeneous, sealed and inside-out membrane vesicles prepared from human and goat erythrocytes was studied. Out of various types of membrane vesicles, in both species, unsealed membrane vesicles bound quantitatively higher amounts of bilirubin followed by heterogeneous and sealed membrane vesicles whereas inside-out membrane vesicles bound the lowest amount of bilirubin. These differences in the amount of bound bilirubin to different membrane vesicles were correlated well with the percentage accessibility of sialic acid to neuraminidase in these membranes suggesting that bilirubin bound preferentially to the outer layer of erythrocyte membranes than the inner layer. Further, membrane vesicles prepared from human erythrocytes bound higher amounts of bilirubin than those prepared from goat erythrocytes. This can be ascribed to different phospholipid composition of these membranes.


Sustainability Assessment of a District-Wide Quality Improvement on Newborn Care Program in Rural Rwanda: A Mixed-Method Study.
Neonatal mortality continues to be a global challenge, particularly in low- and middle-income countries. There is growing work to reduce mortality through improving quality of systems and care, but less is known about sustainability of improvements in the setting post initial implementation. We conducted a 12-month sustainability assessment of All Babies Count (ABC), a district-wide quality improvement project including mentoring and improvement collaborative designed to improve quality and reduce neonatal mortality in two districts in rural Rwanda. We measured changes in key neonatal process, coverage, and outcome indicators between the completion of ABC implementation and 12 months after the completion. In addition, we conducted 4 focus group discussions and 15 individual in-depth interviews with health providers and facility and district leaders to understand factors that influenced sustainability of improvements. We used an inductive, content analytic approach to derive six themes related to the ABC sustainability to explain quantitative results. Twelve months after the completion of ABC implementation, we found continued improvements in core quality, coverage, and neonatal outcomes. During ABC, the percentage of women with 4 antenatal visits increased from 12% to 30% and remained stable 12 months post-ABC (30%, p = 0.7) with an increase in facility-based delivery from 92.6% at the end of ABC to 95.8% (p = 0.01) at 12-month post-ABC. During ABC intervention, the 2 districts decreased neonatal mortality from 30.1 to 19.4 deaths per 1,000 live births with maintenance of the lower mortality 12 months post-ABC (19.4 deaths per 1,000 live births, p = 0.7). Leadership buy-in and development of self-reliance encouraging internally generated solutions emerged as key factors to sustain improvements while staff turnover, famine, influx of refugees, and unintended consequences of new national newborn care policies threatened sustainability. Despite discontinuity of key ABC support, health facilities kept the momentum of good practices and were able to maintain or increase the level of prenatal, neonatal quality of care and outcomes over a period of 12 months following the end of initial ABC implementation. Additional studies are needed to determine the longer-term sustainability beyond one year.


Incidence and pattern of Thrombocytopenia in cardiac surgery patients.
To observe the incidence and pattern of thrombocytopenia in cardiac surgery patients. This prospective, cohort study was conducted at the Aga Khan University Hospital, Karachi, from November 2014 to April 2015, and comprised adult cardiac patients. Patients with platelet count less than 150,000 x 109/L, history of malignancy, immune thrombocytopenic purpura and on chemo or radiotherapy were excluded. All information including demographics, platelet count, heparin doses, total cardiopulmonary bypass time, cross-clamp time, blood products transfused, any thromboembolic complication and the presence of infection were recorded on a pre-designed proforma. SPSS 19 was used for data analysis.. Of the 177 patients, 130(73.4%) were males and 47(26.6%) were females. The overall mean age was 59.21±10.99 years. Thrombocytopenia was observed in 167(94.4%) patients. Of them, platelet count dropped below 50% in 71(42.5%) patients, 30-50% in 68(40.7%) patients and 20-30% in 28(16.8%) from the baseline value. Regarding pattern of thrombocytopenia, maximum drop in platelet count was noticed on 2nd and 3rd day of surgery. Furthermore, 9(5.3%) patients developed severe thrombocytopenia (<50,000 x 109/L). The incidence of thrombocytopenia and its severity after cardiac surgery was very high in our study population when compared with western population.


An Alternative Splicing Variant of the Mixed-Lineage Leukemia 5 Protein Is a Cellular Adhesion Receptor for ScaA of Orientia tsutsugamushi.
Scrub typhus is a mite-borne disease caused by the obligately intracellular bacterium Orientia tsutsugamushi. We previously demonstrated that ScaA, an autotransporter membrane protein of O. tsutsugamushi, is commonly shared in various genotypes and involved in adherence to host cells. Here, we identified a mixed-lineage leukemia 5 (MLL5) mammalian trithorax group protein as a host receptor that interacts with ScaA. MLL5, identified by yeast two-hybrid screening, is an alternative splicing variant of MLL5 (vMLL5) which contains 13 exons with additional intron sequences encoding a tentative transmembrane domain. Indeed, vMLL5 is expressed on the plasma membrane as well as in intracellular compartments in eukaryotic cells and colocalized with adherent O. tsutsugamushi. In addition, ScaA-expressing Escherichia coli showed significantly increased adherence to vMLL5-overexpressing cells compared with vector control cells. We mapped the C-terminal region of the passenger domain of ScaA as a ligand for vMLL5 and determined that the Su(var)3-9, Enhancer of zeste, Trithorax (SET) domain of MLL5 is an essential and sufficient motif for ScaA binding. We observed significant and specific inhibition of bacterial adhesion to host cells in competitive inhibition assays using the C-terminal fragment of ScaA or the SET domain of vMLL5. Moreover, immunization with the C-terminal fragment of ScaA provided neutralizing activity and protective immunity against lethal challenge with O. tsutsugamushi as efficiently as vaccination with the whole passenger domain of ScaA. These results indicate that vMLL5 is a novel cellular receptor for ScaA-mediated adhesion of O. tsutsugamushi and facilitates bacterial adhesion to host cells, thereby enhancing bacterial infection. IMPORTANCE O. tsutsugamushi is a mite-borne pathogen that causes scrub typhus. As an obligately intracellular pathogen, its adhesion to and invasion of host cells are critical steps for bacterial growth. However, the molecular basis of the bacterial ligand and host receptor interaction is poorly defined. Here, we identified a splicing variant of MLL5 (vMLL5) as a cellular adhesion receptor of ScaA, an outer membrane autotransporter protein of O. tsutsugamushi. We mapped the interacting domains in the bacterial ligand and host receptor and confirmed their functional interaction. In addition, immunization with the C-terminal region of ScaA, which involves an interaction with the SET domain of vMLL5, not only induces enhanced neutralizing antibodies but also provides protective immunity against lethal challenge with O. tsutsugamushi.


Dexamethasone is associated with early deaths in light chain amyloidosis patients with severe cardiac involvement.
Cardiac light chain amyloidosis (AL-CA) patients often die within three months of starting chemotherapy. Chemotherapy for non-immunoglobulin M gammopathy with AL-CA frequently includes bortezomib (Bor), cyclophosphamide (Cy), and dexamethasone (D). We previously reported that NT-ProBNP levels can double within 24h of dexamethasone administration, suggesting a deleterious impact on cardiac function. In this study, we evaluate the role of dexamethasone in early cardiovascular mortality during treatment. We retrospectively assessed 100 de novo cardiac AL patients (62% male, mean age 68 years) treated at our institute between 2009 and 2018 following three chemotherapy regimens: CyBorDComb (all initiated on day 1; 34 patients), DCyBorSeq (D, day 1; Cy, day 8; Bor, day 15; 17 patients), and CyBorDSeq (Cy, day 1; Bor, day 8; D, day 15; 49 patients). The primary endpoint was cardiovascular mortality and cardiac transplantation at days 22 and 455. At day 22, mortality was 20.6% with CyBorDComb, 23.5% with DCyBorSeq, and 0% with CyBorDSeq (p = 0.003). At day 455, mortality was not significantly different between regimens (p = 0.195). Acute toxicity of dexamethasone was evaluated on myocardial function using a rat model of isolated perfused heart. Administration of dexamethasone induced a decrease in left ventricular myocardium contractility and relaxation (p<0.05), supporting a potential negative inotropic effect of dexamethasone in AL-CA patients with severe cardiac involvement. Delaying dexamethasone during the first chemotherapy cycle reduces the number of early deaths without extending survival. It is clear that dexamethasone is beneficial in the long-term treatment of patients with AL-CA. However, the initial introduction of dexamethasone during treatment is critical, but may be associated with early cardiac deaths in severe CA. Thus, it is important to consider the dosage and timing of dexamethasone introduction on a patient-severity basis. The impact of dexamethasone in the treatment of AL-CA needs further investigation.


The added value of bevacizumab concomitantly administered with carboplatin versus carboplatin alone in patients with recurrent glioblastomas.
Carboplatin (CBDCA) and bevacizumab (BEV) are active in glioblastoma (GBM) with different profiles of toxicity. To date, no study has compared the value of the addition of BEV to historical or traditional cytotoxic chemotherapy. We sought to determine the relative value of BEV in combination with CBDCA versus CBDCA alone in patients with recurrent GBM. Eligible patients with progressive GBM following surgery, radiotherapy and temozolomide received CBDCA either alone (group 1, n = 25) or in combination with BEV (group 2, n = 23) at 5 mg/kg once every 3 weeks between June 2010 and December 2013. Baseline characteristics and outcomes after treatment were recorded. The primary end points of this retrospective analysis were progression-free survival (PFS) and objective response rate. Secondary end points included safety and overall survival (OS). Forty-eight patients were enrolled. The median number of cycles was 4 in group 1 and 6 in group 2. No toxicities or intracerebral bleeding were observed. The objective response rate was higher in group 2 than group 1, 66% vs 24% (p = 0.003). The estimated median PFS and OS were 3.1 vs 6.7 months (p<0.0001) and 6.1 vs 8.6 months (p = 0.09) in group 1 vs group 2, respectively. The combination of BEV and CBDCA is associated with improved response rates and survival compared with CBDCA alone. These results highlight the value of BEV in recurrent GBM. However, the clinical benefit of this interesting approach needs validation in a larger patient cohort.


Mating disruption of Lobesia botrana (Lepidoptera: Tortricidae): effect of pheromone formulations and concentrations.
The reluctance of Israeli vine growers to adopt the mating disruption technique to control the moth Lobesia botrana Den. & Schiff. has been attributed to the high cost of this method compared with that of traditional insecticide control. In this study, we tested the possibility of reducing the cost, first by testing different pheromone formulations (and thus open the market for competition) and second by reducing the pheromone concentration used in vineyards. Comparisons were made between two pheromone formulations--Shin-Etsu (Tokyo, Japan) at 165 g/ha and Concep (Sutera, Bend, OR) at 150 g/ha--and between two concentrations of Shin-Etsu, 165 and 110 g/ha. Pheromone dispensers were placed at the onset of the second moth generation. Comparison of the numbers of clusters infested with eggs and larvae of L. botrana showed no significant differences in the performance, either between the two formulations, or between the two tested concentrations. The results suggest that 1) the two formulations are equally effective, and 2) a low pheromone concentration is sufficient to maintain good control of small populations of L. botrana. However, when the population is high, pest control efficacy is not improved by increasing the pheromone concentration. Therefore, in the interest of reducing the relatively high cost of mating disruption, we emphasize that increasing the pheromone concentration does not provide improved control of high populations of L. botrana. The cost of mating disruption can be diminished by reducing the applied pheromone concentration and by using the least expensive pheromone formulations


Host factor Rab11a is critical for efficient assembly of influenza A virus genomic segments.
It is well documented that influenza A viruses selectively package 8 distinct viral ribonucleoprotein complexes (vRNPs) into each virion; however, the role of host factors in genome assembly is not completely understood. To evaluate the significance of cellular factors in genome assembly, we generated a reporter virus carrying a tetracysteine tag in the NP gene (NP-Tc virus) and assessed the dynamics of vRNP localization with cellular components by fluorescence microscopy. At early time points, vRNP complexes were preferentially exported to the MTOC; subsequently, vRNPs associated on vesicles positive for cellular factor Rab11a and formed distinct vRNP bundles that trafficked to the plasma membrane on microtubule networks. In Rab11a deficient cells, however, vRNP bundles were smaller in the cytoplasm with less co-localization between different vRNP segments. Furthermore, Rab11a deficiency increased the production of non-infectious particles with higher RNA copy number to PFU ratios, indicative of defects in specific genome assembly. These results indicate that Rab11a+ vesicles serve as hubs for the congregation of vRNP complexes and enable specific genome assembly through vRNP:vRNP interactions, revealing the importance of Rab11a as a critical host factor for influenza A virus genome assembly.


Microglial glucocorticoid receptors play a pivotal role in regulating dopaminergic neurodegeneration in parkinsonism.
Among the pathogenic processes contributing to dopaminergic neuron (DN) death in Parkinson disease (PD), evidence points to non-cell-autonomous mechanisms, particularly chronic inflammation mounted by activated microglia. Yet little is known about endogenous regulatory processes that determine microglial actions in pathological states. We examined the role of glucocorticoid receptors (GRs), activated by glucocorticoids released in response to stress and known to regulate inflammation, in DN survival. Overall GR level was decreased in substantia nigra of PD patients and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-intoxicated mice. GR changes, specifically in the microglia after MPTP treatment, revealed a rapid augmentation in the number of microglia displaying nuclear localization of GR. Mice with selective inactivation of the GR gene in macrophages/microglia (GR(LysMCre)) but not in DNs (GR(DATCre)) showed increased loss of DNs after MPTP intoxication. This DN loss in GR(LysMCre) mice was not prevented by corticosterone treatment, in contrast to the protection observed in control littermates. Moreover, absence of microglial GRs augmented microglial reactivity and led to their persistent activation. Analysis of inflammatory genes revealed an up-regulation of Toll-like receptors (TLRs) by MPTP treatment, particularly TLR9, the level of which was high in postmortem parkinsonian brains. The regulatory control of GR was reflected by higher expression of proinflammatory genes (e.g., TNF-α) with a concomitant decrease in anti-inflammatory genes (e.g., IL-1R2) in GR(LysMCre) mice. Indeed, in GR(LysMCre) mice, alterations in phosphorylated NF-κB levels indicated its protracted activation. Together, our data indicate that GR is important in curtailing microglial reactivity, and its deregulation in PD could lead to sustained inflammation-mediated DN injury.


Translational therapeutic strategies in amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a clinically severe and fatal neurodegenerative disease characterized by a loss of both upper and lower motor neurons, resulting in progressive muscle loss and paralysis. While the exact cause of neuronal death in ALS remains unknown, it is proposed that multiple molecular defects trigger motor neuron cell death. These pathophysiological mechanisms include oxidative stress, mitochondrial impairment, protein aggregation, glutamate cytotoxicity, transcription dysfunction, inflammation, and apoptotic cell death. An understanding of how these potential therapeutic targets interrelate will provide direction both in the development of a pharmacotherapy and in the design of clinical trials in ALS. Important issues related to therapeutic development are the principals that should be followed in designing and conducting experiments using genetic animal models and what body of evidence is desirable to fully inform clinical decision making. In the context of ALS, we review some of the salient issues related to the use of genetic models in providing a guide to assessing studies in translating therapeutic strategies to patients with ALS and discuss therapeutic targets and pharmacological approaches to slowing disease progression. As in other neurodegenerative diseases, the most effective neuroprotection may result from combined treatment strategies.


Applying a risk-adjustment framework to primary care: can we improve on existing measures?
Outcome-based performance measurement and prospective payment are common features of the current managed care environment. Increasingly, primary care clinicians and health care organizations are being asked to assume financial risk for enrolled patients based on negotiated capitation rates. Therefore, the need for methods to account for differences in risk among patients enrolled in primary care organizations has become critical. Although current risk-adjustment measures represent significant advances in the measurement of morbidity in primary care populations, they may not adequately capture all the dimensions of patient risk relevant to primary care. We propose a risk-adjustment framework for primary care that incorporates clinical features related to patients' health status and nonclinical factors related to patients' health behaviors, psychosocial factors, and social environment. Without this broad perspective, clinicians with more unhealthy and more challenging populations are at risk of being inadequately compensated and inequitably compared with peers. The risk-adjustment framework should also be of use to health care organizations that have been mandated to deliver high-quality primary care but are lacking the necessary tools.


Epigenetic Analysis of Circulating Tumor DNA in Localized and Metastatic Prostate Cancer: Evaluation of Clinical Biomarker Potential.
Novel and minimally-invasive prostate cancer (PCa)-specific biomarkers are needed to improve diagnosis and risk stratification. Here, we investigated the biomarker potential in localized and de novo metastatic PCa (mPCa) of methylated circulating tumor DNA (ctDNA) in plasma. Using the Marmal-aid database and in-house datasets, we identified three top candidates specifically hypermethylated in PCa tissue: DOCK2,HAPLN3, and FBXO30 (specificity/sensitivity: 80%-100%/75-94%). These candidates were further analyzed in plasma samples from 36 healthy controls, 61 benign prostatic hyperplasia (BPH), 102 localized PCa, and 65 de novo mPCa patients using methylation-specific droplet digital PCR. Methylated ctDNA for DOCK2/HAPLN3/FBXO30 was generally not detected in healthy controls, BPH patients, nor in patients with localized PCa despite a positive signal in 98%-100% of matched radical prostatectomy tissue samples. However, ctDNA methylation of DOCK2,HAPLN3, and/or FBXO30 was detected in 61.5% (40/65) of de novo mPCa patients and markedly increased in high- compared to low-volume mPCa (89.3% (25/28) vs. 32.1% (10/31), p < 0.001). Moreover, detection of methylated ctDNA was associated with significantly shorter time to progression to metastatic castration resistant PCa, independent of tumor-volume. These results indicate that methylated ctDNA (DOCK2/HAPLN3/FBXO30) may be potentially useful for identification of hormone-naïve mPCa patients who could benefit from intensified treatment.


Enhanced thermoelectric properties of In-filled Co4Sb12 by dispersion of reduced graphene oxide.
Uniform dispersion of nanosized secondary phases in bulk thermoelectric materials has proven to be an effective strategy to reduce the lattice part of thermal conductivity and improve the thermoelectric efficiency. In this work, reduced graphene oxide (rGO) was uniformly dispersed in the In0.5Co4Sb12 bulk material by ultrasonication. The formation of impurity phases of InSb and CoSb2 in the In-filled Co4Sb12 is inevitable, as observed from XRD and EPMA analyses. The Raman spectra of the nanocomposites showed broad peaks suggesting phonon softening and additional peaks corresponding to rGO. Electron transport was not affected by rGO addition, resulting in little change in the electrical resistivity and Seebeck coefficient. The lattice thermal conductivity of the bulk material was significantly reduced by the addition of a small amount of rGO, primarily attributed to the interface scattering of phonons. Hence, the highest zT of ∼1.53 at 773 K was achieved for the In0.5Co4Sb12/0.25 vol% rGO composite in the temperature range from 723 K to 773 K.


Practical Considerations in High-Precision Compound-Specific Radiocarbon Analyses: Eliminating the Effects of Solvent and Sample Cross-Contamination on Accuracy and Precision.
Preparative capillary gas chromatography (pcGC) is widely used for the isolation of single compounds for radiocarbon determinations. While being effective at isolating compounds, there are still genuine concerns relating to contamination associated with the isolation procedure, such as incomplete removal of solvent used to recover isolated compounds from the traps and cross-contamination, which can lead to erroneous 14C determinations. Herein we describe new approaches to identifying and removing these two sources of contamination. First, we replaced the common "U" trap design, which requires recovery of compounds using organic solvent, with a novel solventless trapping system (STS), consisting of a simple glass tube containing a glass wool plug, allowing condensation of a target compound in the wool and its solventless recovery by pushing the glass wool directly into a foil capsule for graphitization. With the STS trap, an average of 95.7% of the target compound was recovered, and contamination from column bleed was reduced. In addition, comparison of 14C determinations of fatty acid methyl ester (FAME) standards determined offline to those isolated by pcGC in STS traps showed excellent reproducibility and accuracy compared to those isolated using the commercial "U" traps. Second, "coldspots" were identified in the instrument, i.e., the termini of capillaries in the preparative unit, which can be cleaned of compounds condensed from earlier runs using a heat gun. Our new procedure, incorporating these two modifications, was tested on archeological fat hoards, producing 14C dates on isolated C16:0 and C18:0 fatty acids statistically consistent with the bulk dates of the archeological material.


Pathophysiological features in the brains of female Spontaneously Diabetic Torii (SDT) fatty rats.
Diabetes mellitus (DM) and obesity are associated with neurodegenerative diseases such as Alzheimer's disease and psychiatric disorders such as major depression. In this study, we investigated pathophysiological changes in the brains of female Spontaneously Diabetic Torii (SDT) fatty rats with diabetes and obesity. Brains of Sprague-Dawley (SD), SDT and SDT fatty rats were collected at 58 weeks of age. The parietal cortical thickness was measured and the number of pyramidal cells in the hippocampal cornu ammonis 1 and 3 (CA1 and CA3) and the number of granule cells in the dentate gyrus (DG) regions were counted. The area of glial fibrillary acidic protein (GFAP) positivity in CA1, CA3 and DG regions were measured. The parietal cortical thickness and the number of cells in CA3 and DG regions of SDT and SDT fatty rats did not show obvious changes. On the other hand, in the CA1 region, the number of cells in SDT rats and SDT fatty rats was significantly lower than that in SD rats, and that in SDT fatty rats was significantly lower than that in SDT rats. The GFAP-positive area in SDT fatty rats was significantly reduced compared to that in SD rats only in the DG region. Preliminarily result showed that the expression of S100a9, an inflammation-related gene, was increased in the brains of SDT fatty rats. These results suggest that female SDT fatty rat may exhibit central nervous system diseases due to obesity and DM.


Nuclear membrane ruptures underlie the vascular pathology in a mouse model of Hutchinson-Gilford progeria syndrome.
The mutant nuclear lamin protein (progerin) produced in Hutchinson-Gilford progeria syndrome (HGPS) results in loss of arterial smooth muscle cells (SMCs), but the mechanism has been unclear. We found that progerin induces repetitive nuclear membrane (NM) ruptures, DNA damage, and cell death in cultured SMCs. Reducing lamin B1 expression and exposing cells to mechanical stress - to mirror conditions in the aorta - triggered more frequent NM ruptures. Increasing lamin B1 protein levels had the opposite effect, reducing NM ruptures and improving cell survival. Remarkably, raising lamin B1 levels increased nuclear compliance in cells and was able to offset the increased nuclear stiffness caused by progerin. In mice, lamin B1 expression in aortic SMCs is normally very low, and in mice with a targeted HGPS mutation (LmnaG609G), levels of lamin B1 decrease further with age while progerin levels increase. Those observations suggest that NM ruptures might occur in aortic SMCs in vivo. Indeed, studies in LmnaG609G mice identified NM ruptures in aortic SMCs, along with ultrastructural abnormalities in the cell nucleus that preceded SMC loss. Our studies identify NM ruptures in SMCs as likely causes of vascular pathology in HGPS.


Contributing Components of Metabolic Energy Models to Metabolic Cost Estimations in Gait.
As metabolic cost is a primary factor influencing humans' gait, we want to deepen our understanding of metabolic energy expenditure models. Therefore, this paper identifies the parameters and input variables, such as muscle or joint states, that contribute to accurate metabolic cost estimations. We explored the parameters of four metabolic energy expenditure models in a Monte Carlo sensitivity analysis. Then, we analysed the model parameters by their calculated sensitivity indices, physiological context, and the resulting metabolic rates during the gait cycle. The parameter combination with the highest accuracy in the Monte Carlo simulations represented a quasi-optimized model. In the second step, we investigated the importance of input parameters and variables by analysing the accuracy of neural networks trained with different input features. Power-related parameters were most influential in the sensitivity analysis and the neural network-based feature selection. We observed that the quasi-optimized models produced negative metabolic rates, contradicting muscle physiology. Neural network-based models showed promising abilities but have been unable to match the accuracy of traditional metabolic energy expenditure models. We showed that power-related metabolic energy expenditure model parameters and inputs are most influential during gait. Furthermore, our results suggest that neural network-based metabolic energy expenditure models are viable. However, bigger datasets are required to achieve better accuracy. As there is a need for more accurate metabolic energy expenditure models, we explored which musculoskeletal parameters are essential when developing a model to estimate metabolic energy.


Photopharmacologic Vision Restoration Reduces Pathological Rhythmic Field Potentials in Blind Mouse Retina.
Photopharmacology has yielded compounds that have potential to restore impaired visual responses resulting from outer retinal degeneration diseases such as retinitis pigmentosa. Here we evaluate two photoswitchable azobenzene ion channel blockers, DAQ and DAA for vision restoration. DAQ exerts its effect primarily on RGCs, whereas DAA induces light-dependent spiking primarily through amacrine cell activation. Degeneration-induced local field potentials remain a major challenge common to all vision restoration approaches. These 5-10 Hz rhythmic potentials increase the background firing rate of retinal ganglion cells (RGCs) and overlay the stimulated response, thereby reducing signal-to-noise ratio. Along with the bipolar cell-selective photoswitch DAD and second-generation RGC-targeting photoswitch PhENAQ, we investigated the effects of DAA and DAQ on rhythmic local field potentials (LFPs) occurring in the degenerating retina. We found that photoswitches targeting neurons upstream of RGCs, DAA (amacrine cells) and DAD (bipolar cells) suppress the frequency of LFPs, while DAQ and PhENAQ (RGCs) had negligible effects on frequency or spectral power of LFPs. Taken together, these results demonstrate remarkable diversity of cell-type specificity of photoswitchable channel blockers in the retina and suggest that specific compounds may counter rhythmic LFPs to produce superior signal-to-noise characteristics in vision restoration.


Benzenesulfonamides: a unique class of chemokine receptor type 4 inhibitors.
The interaction of CXCR4 with CXCL12 (SDF-1) plays a critical role in cancer metastasis by facilitating the homing of tumor cells to metastatic sites. Based on our previously published work on CXCR4 antagonists, we have synthesized a series of aryl sulfonamides that inhibit the CXCR4/CXCL12 interaction. Analogue bioactivities were assessed with binding affinity and Matrigel invasion assays. Computer modeling was employed to evaluate a selection of the new analogues docked into the CXCR4 X-ray structure and to rationalize discrepancies between the affinity and Matrigel in vitro assays. A lead compound displays nanomolar potency in the binding affinity assay (IC(50)=8.0 nM) and the Matrigel invasion assay (100 % blockade of invasion at 10 nM). These data demonstrate that benzenesulfonamides are a unique class of CXCR4 inhibitors with high potency.


Prenatal exposure to per- and polyfluoroalkyl substances and maternal and neonatal thyroid function in the Project Viva Cohort: A mixtures approach.
Maternal and neonatal thyroid function is critical for growth and neurodevelopment. Exposure to individual per- and polyfluoroalkyl substances (PFAS) can alter circulating thyroid hormone levels, but few studies have investigated effects of combined exposure to multiple PFAS. Estimate associations of exposure to multiple PFAS during early pregnancy with maternal and neonatal thyroid function. The study population consisted of 726 mothers and 465 neonates from Project Viva, a Boston, Massachusetts area longitudinal pre-birth cohort. We measured six PFAS [perfluorooctanoate (PFOA), perfluorooctane sulfonate (PFOS), perfluorononanoate (PFNA), perfluorohexane sulfonate (PFHxS), 2-(N-ethyl-perfluorooctane sulfonamido)acetate (EtFOSAA), and 2-(N-methyl-perfluorooctane sulfonamido)acetate (MeFOSAA)] and thyroxine (T4), Free T4 Index (FT4I), and thyroid stimulating hormone (TSH) in maternal plasma samples collected during early pregnancy, and neonatal T4 in postpartum heel sticks. We estimated individual and joint effects of PFAS exposure with thyroid hormone levels using weighted quantile sum (WQS) regression and Bayesian kernel machine regression (BKMR), and evaluated potential non-linearity and interactions among PFAS using BKMR. Higher concentrations of the PFAS mixture were associated with significantly lower maternal FT4I, with MeFOSAA, EtFOSAA, PFOA, and PFHxS contributing most to the overall mixture effect in BKMR and WQS regression. In infants, higher concentrations of the PFAS mixture were associated with lower T4 levels, primarily in males, with PFHxS and MeFOSAA contributing most in WQS, and PFHxS contributing most in BKMR. The PFAS mixture was not associated with maternal T4 or TSH levels. However, in maternal BKMR analyses, ln-PFOS was positively associated with T4 levels (Δ25th to 75th percentile: 0.21 µg/dL; 95% credible interval: -0.03, 0.47) and ln-PFHxS was associated with a non-linear effect on TSH levels. These findings support the hypothesis that there may be combined effects of prenatal exposure to multiple PFAS on maternal and neonatal thyroid function, but the direction and magnitude of these effects may vary across individual PFAS.


Copper deficiency-induced hypercholesterolemia: effects on HDL subfractions and hepatic lipoprotein receptor activity in the rat.
Male Sprague-Dawley rats (10 per group) were fed diets adequate (control) or deficient (CuDef) in copper for 6 wk. In the CuDef group, plasma total cholesterol, high density lipoprotein (HDL) cholesterol and apoA-I levels were significantly higher than in controls. Apolipoprotein analysis of the HDL fractions revealed a relative enrichment of apoE in the CuDef group. Size analysis of 1.21 density lipoproteins by nondenaturing gradient gel electrophoresis demonstrated the presence of more material migrating in the size range of HDL1 in the CuDef group. Separation of HDL into apoE-rich and apoA-I-rich fractions by heparin-affinity chromatography confirmed the presence of increased apoE-rich HDL in the CuDef group. To determine the mechanism responsible for higher apoE-rich HDL in the CuDef group, the lipoprotein receptor binding activity in hepatic membranes from the control and CuDef group was assayed. Kinetic analysis of the binding data revealed that the lipoprotein binding assay was primarily measuring the activity of the HDL receptor, which is not apoE mediated. When corrected for differential enrichment of plasma membrane, hepatic membranes from the CuDef group bound significantly fewer lipoproteins than did controls. Furthermore, the hepatic receptor binding activity was negatively correlated with the proportion of HDL enriched with apoE.


Immune responses in eight patients infected with Bacteroides fragilis.
Sera from eight patients with either septicemia or pyothorax caused by Bacteroides fragilis were assayed for specific antibody by the agglutination, immunofluorescence, and agar gel diffusion techniques. Antibodies were detected in the sera of all eight patients by all three serological tests employed; titers as high as 1:160 and 1:320 were detected by the agglutination test and the immunofluorescence technique, respectively, and all patients had precipitating antibody. Failure of B. fragilis sub-species thetaiotaomicron to react with any heterologous antisera from the other seven patients infected with B. fragilis subspecies fragilis thetaiotaomicron was not serologically related to any of the other strains studied. Demonstration of an immune response to anaerobic infection supported the pathogenesis of anaerobes in infectious disease processes and indicated the possible diagnostic role that serological tests may serve in the diagnosis of anaerobic infections in humans.


Modeling endface output patterns of optical micro/nanofibers.
Endface output patterns of micro/nanofibers (MNFs) are simulated using a Three-Dimension Finite-Difference Time-Domain (3D-FDTD) method. The intensity distribution and beam widths of near- or farfield output patterns of freestanding silica and tellurite MNFs with flat, angled, spherical and tapered endfaces in air and/or water are obtained. It shows that, for a subwavelength-diameter MNF, highly confined output beam can be obtained in the near field, and the beam width can be tuned by the ratio of fiber diameter and light wavelength with a minimum width smaller than the wavelength. Meanwhile, MNFs with shaped endfaces behave differently from standard fibers in reflection, redirection and focus of light beam at the endfaces. These results may offer valuable references for practical evaluation and application of terminated MNFs with wavelength- or subwavelength-scale endfaces.


Exploring the influence of workplace supports and relationships on safe medication practice: A pilot study of Australian graduate nurses.
Despite the growing awareness of the benefits of positive workplace climates, unsupportive and disruptive workplace behaviours are widespread in health care organisations. Recent graduate nurses, who are often new to a workplace, are particularly vulnerable in unsupportive climates, and are also recognised to be at higher risk for medication errors. Investigate the association between workplace supports and relationships and safe medication practice among graduate nurses. Exploratory study using quantitative survey with a convenience sample of 58 nursing graduates in two Australian States. Online survey focused on graduates' self-reported medication errors, safe medication practice and the nature of workplace supports and relationships. Spearman's correlations identified that unsupportive workplace relationships were inversely related to graduate nurse medication errors and erosion of safe medication practices, while supportive Nurse Unit Manager and supportive work team relationships positively influenced safe medication practice among graduates. Workplace supports and relationships are potentially both the cause and solution to graduate nurse medication errors and safe medication practices. The findings develop further understanding about the impact of unsupportive and disruptive behaviours on patient safety and draw attention to the importance of undergraduate and continuing education strategies that promote positive workplace behaviours and graduate resilience.


Combined effects of bisphenol A and cadmium on growth and nitrate assimilation of soybean seedling roots.
Bisphenol A (BPA) and cadmium (Cd) pollution exist simultaneously in many regions. However, little information is available regarding the combined effects of BPA and Cd pollution on plants. Plant roots are in direct contact with the soil, which is an important compartment of BPA and Cd. In the present study, the effects of combined BPA and Cd pollution on soybean seedling roots were evaluated in pot experiments. Combined treatment with BPA and Cd at low concentrations (1.5 mg/kg BPA and 0.2 mg/kg Cd) improved soybean seedling root growth. However, other combined BPA and Cd treatments, including combined treatment with BPA (Cd) at the low concentration and Cd (BPA) at the high concentration as well as combined treatment with BPA and Cd at the high concentration, inhibited soybean seedling root growth. The improvement or inhibition of soybean seedling root growth was greater in the combined BPA and Cd treatments than in single treatments. The effects of the combined BPA and Cd treatments on root growth resulted from changes in nitrate assimilation. In addition, the combined effects of BPA and Cd on the nitrate and ammonium contents in roots are discussed. The present research provides a basic understanding of the combined effects of BPA and Cd pollution on plant roots.


Anti-suicidal effects of IV ketamine in a real-world setting.
The current study evaluated the effectiveness of intravenous ketamine treatment for suicidality in a community-based clinical sample of 295 outpatients (mean age= 40.37; 58.6 % male). We conducted growth mixture modeling to estimate latent classes of changes in symptoms of suicidality measured by the Concise Health Risk Tracking - Self-Report (CHRT-SR) across five infusions in a two-week course of treatment. Best-fit indices indicated three trajectory groups demonstrating non-linear, quadratic changes in CHRT-SR scores during ketamine treatment. The largest group of patients (n= 170, 57.6 %) had moderate CHRT-SR scores at baseline and showed gradual improvement during treatment. The other two groups of patients had severe CHRT-SR scores at baseline and diverged into one group with no improvement throughout treatment (n = 63, 21 %) and one group with rapid improvement (n = 62, 21 %). Of the clinical and demographic variables available and tested, only higher scores pertaining to active thoughts of death and/or plan were found to predict which of the patients with severe CHRT-SR scores at baseline would not benefit from treatment. The present study provides an important contribution to the knowledge of ketamine's effects on symptoms related to suicide over time. providing support for the possible effectiveness of ketamine in a proportion of patients.


The expression of Argonaute2 and related microRNA biogenesis proteins in normal and hypoxic trophoblasts.
Endogenous microRNAs (miRNAs) post-transcriptionally regulate mRNA and protein expression during tissue development and function. Whereas adaptation to environmental insults are tightly regulated in human tissues, the role of miRNAs and miRNA biogenesis proteins in this context is inadequately explored. We sought to analyse the expression of the key RNAi enzyme Argonaute2 (Ago2) and other miRNA biogenesis proteins in human trophoblasts during differentiation and in hypoxic environment. Using an in vitro analysis of primary term human trophoblasts, we identified the expression of the core miRNA biogenesis proteins in human villous trophoblasts, with expression levels unaffected by cellular differentiation. We found that the miRNA biosynthetic pathway was functional and produced miRNAs, with miR-93 up-regulated and miR-424 down-regulated in hypoxic environment. In contrast, hypoxia did not alter the expression of key miRNA machinery proteins. The pivotal miRNA processing enzyme Ago2, along with its interacting protein DP103, were expressed in normal placentas as well as in placentas from pregnancies complicated by placental hypoperfusion that resulted in fetal growth restriction. Ago2 and DP103 co-immunoprecipitated, and did not limit trophoblast response to hypoxic stress. We concluded that the core miRNA machinery proteins are expressed and functional in human trophoblasts. The influence of hypoxia on the expression of a subset of placental miRNA species is unlikely to reflect altered expression of key miRNA biogenesis proteins.


Prevalence and correlates of past 30-day dual-vaping of nicotine and cannabis among adolescents in five New England states.
Adolescent vaping behavior includes vaping of multiple substances, including both nicotine and cannabis (dual-vaping). This study describes the prevalence and the sociodemographic correlates of past 30-day dual-vaping. We recruited adolescents ages 13-17 from five New England states (Massachusetts, Connecticut, Rhode Island, Vermont, New Hampshire) through the Prodege online survey panel from April 2021 to August 2022. Dual-vaping was defined as vaping both nicotine and cannabis (THC and/or CBD) in the past 30-days. We analyzed the prevalence of sole-nicotine, sole-cannabis, and dual-vaping of nicotine and cannabis and used multinomial logistic regression to examine associations between sociodemographic factors and sole- and dual-vaping of nicotine and cannabis. The analytic sample included 2013 observations from 1858 participants (mean age 15.1 years, 46.2% female, 74.1% White, 82.2% heterosexual). Among these observations, 5.6% reported past 30-day sole-nicotine vaping, 5.5% reported sole-cannabis vaping, and 7.3% had dual-vaped. Correlates for higher odds of past 30-day dual-vaping included total social media sites used and household tobacco use, in contrast with sole-cannabis vaping, which included older age and self-reported depression (all p's <0.05). Adolescent past 30-day dual-vaping of nicotine and cannabis was more prevalent than past 30-day sole-vaping of either nicotine or cannabis alone. Future studies should continue to collect detailed data on the type of substances, besides nicotine, that adolescents are vaping.


The mechanics of notochord elongation, straightening and stiffening in the embryo of Xenopus laevis.
We have examined the biomechanical development of the notochord of Xenopus early tail-bud embryos by: (1) quantifying morphological and mechanical changes in the embryo during stages 20-28, and (2) conducting manipulative experiments to elucidate mechanical roles of various components of the notochord. The notochord, which is composed of a stack of flat cells surrounded by a connective tissue sheath, elongates dramatically and begins straightening between stages 21 and 25. At this time the fiber density in the notochord sheath goes up, the osmotic activity of the notochord cells increases, vacuoles within these cells swell, the internal pressure of the notochord increases 2- to 3-fold, and the flexural stiffness of the notochord rises by an order of magnitude. We suggest that the tendency of the notochord cells to osmotically swell is resisted by the sheath, thereby permitting the internal pressure to rise. This pressure increase results in the greater stiffness that permits the notochord to elongate and straighten without being buckled by the surrounding tissues.


Level of heat shock proteins decreases in individuals carrying B-chromosomes in the grasshopper Eyprepocnemis plorans.
We analyzed the effect of B-chromosome presence on expression level of heat shock protein 70 (Hsp70) in cerebral ganglion and gonad in both males and females of the grasshopper Eyprepocnemis plorans. Two natural Spanish populations, Salobreña (Granada) and Torrox (Málaga) were assayed, the former harbouring a neutralized (non-driving) B-chromosome (B(2)) and the latter a parasitic (driving) B-chromosome (B(24)). The analysis was performed by Western blotting, immunostaining and densitometric measuring expression level of the Hsp70 family in adult individuals. The results showed that Hsp70 levels of testis were significantly higher in Salobreña than Torrox, and were significantly lower in testes of B-carrying males from both populations. A similar effect was observed in the ovary of females from Torrox. No effect was, however, observed in cerebral ganglia in any sex or population. B-chromosome effects in Torrox showed a dose-dependent pattern. The results point to an interesting interaction between B-chromosome and stress protein expression in reproductive tissue.


Hydrophilic modification of SLA 3D printed droplet generators by photochemical grafting.
Few droplet generators manufactured using desktop stereolithography 3D printers have been reported in the literature. Moreover, 3D printed microfluidic chips are typically hydrophobic, limiting their application to water in oil droplets. Herein, we present designs for concentric and planar 3D printed microfluidic devices suitable for making polymeric microparticles using an off-the-shelf commercial stereolithography printer and resin. The devices consist of a microscope slide, binder clips, and printed components. Channels were modified by an ultraviolet grafting of methacrylic acid to the surface of chips, yielding a hydrophilic coating without modification to the bulk polymer. The water contact angle decreased from 97.0° to 25.4° after grafting. The presence of the coating was confirmed by microscopy and spectroscopy techniques. Polystyrene microparticles in the <100 μm size range were generated with varying molecular weights using the described microfluidic chips. Our work provides a facile method to construct droplet generators from commercial stereolithography printers and resins, and a rapid surface modification technique that has been under-utilized in 3D printed microfluidics. A wide range of microfluidic devices for other applications can be engineered using the methods described.


Trends in freeze-drying equipment and materials.
The above discussion outlines some trends seen in the development of equipment for freeze-drying of various biological-like products. It has been pointed out that many similarities in equipment now exist. There are still areas in which a consensus on best design has not been reached and in these areas the user is offered some clear choices. Various changes have been introduced in recent years into freeze-drying equipment principally in an effort to increase reliability and improve the quality of the product produced. It is probable as more products are freeze-dried (and particularly as these products become more cost-sensitive) that manufacturers will also require that greater performance-capability be built into their equipment. This may perhaps translate into requirements for carrying high peak-vapor-rates and for automatic control in an effort to reduce drying cycles and increase throughput in a given machine.


Gene expression of islet cell antigen p69 in human, mouse, and rat.
Based on the detection of specific antibodies and T-cell sensitization in patients with IDDM, islet cell antigen p69 (ICAp69) has been suggested to be a target antigen of diabetic autoimmunity. The biological function, tissue expression, and developmental kinetics of ICAp69 are largely unknown. We analyzed ICAp69 expression at the gene transcription and protein level in human and rodent tissues. By using template-calibrated quantitative reverse transcriptase polymerase chain reaction (RT-PCR), high levels of ICAp69 mRNA were found in human pancreatic islets and brain. In mouse and rat, ICAp69 gene expression peaked in islet cell lines followed by testis, islets, and brain. ICAp69 mRNA was found at low levels in other organs by RT-PCR but not by Northern blot analysis. In mice, ICAp69 transcription becomes detectable in fetal life, and fetal and adult gene expression patterns are similar. Western blot analysis of human and mouse tissues showed high expression of ICAp69 in brain, testis, pancreatic tissue, and islet cell lines. In these organs, ICAp69 immunoreactivity is predominately localized at the blood brain barrier (capillary endothelium), at the blood testis barrier (Sertoli cells and spermatids), and in pancreatic islets (beta-cells). The subcellular localization of ICAp69 to endoplasmic reticulum, Golgi complex, and vesicles by immune electron microscopy suggests a role of this neuroendocrine molecule in cellular protein traffic and processing.off


(-)-Linalool inhibits in vitro NO formation: Probable involvement in the antinociceptive activity of this monoterpene compound.
Recent studies performed in our laboratory have shown that (-)-linalool, the natural occurring enantiomer in essential oils, possesses anti-inflammatory, antihyperalgesic and antinociceptive effects in different animal models. The antinociceptive and antihyperalgesic effect of (-)-linalool has been ascribed to the stimulation of the cholinergic, opioidergic and dopaminergic systems, to its local anaesthetic activity and to the blockade of N-Methyl-d-aspartate receptors (NMDA). Since nitric oxide (NO) and prostaglandin E(2) (PGE(2)) play an important role in oedema formation and hyperalgesia and nociception development, to investigate the mechanism of these actions of the (-)-linalool, we examined the effects of this compound on lipopolysaccharide (LPS)-induced responses in macrophage cell line J774.A1. Exposure of LPS-stimulated cells to (-)-linalool significantly inhibited nitrite accumulation in the culture medium without inhibiting the LPS-stimulated increase of inducible nitric oxide synthase (iNOS) expression, suggesting that the inhibitory activity of (-)-linalool is mainly due to the iNOS enzyme activity. In contrast, exposure of LPS-stimulated cells to (-)-linalool failed, if not at the highest concentration, both in inhibiting PGE(2) release and in inhibiting increase of inducible cyclooxygenase-2 (COX(2)) expression in the culture medium. Collectively, these results indicate that the reduction of NO production/release is responsible, at least partially, for the molecular mechanisms of (-)-linalool antinociceptive effect, probably through mechanisms where cholinergic and glutamatergic systems are involved.


[Detection of the products of several T4B phage genes among the proteins of infected Escherichia].
Proteins of wild-type phate T4 and its mutants in genes rII, su(30), stII, stIII, rVI-39 and 60 are studied by means of polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate of membrane fractions and lysates of infected bacteria. Physiological studies of mutants in these genes carried out by the authors and other investigators allow to suggest that when functions are realized, the products of these genes interact with the plasmatic membrane of the infected bacteria. The product of rIIB cystrone has been indentified as the protein of the membrane of an infected bacterium, its molecular weight being about 30 000. The products of genes su(30), stII, stIII and rVI could not be identified either in lysate or in membrane fraction. The function of gene stII is "superstoichiometric", which may be due to the involvement of a very small product amount when the function of gene stII is realized. Mutants in genes 39 and 60 lack one and the same protein with molecular weight of about 40 000 which is a protein of the membrane of an infected bacterium. This protein may be a product of gene 39, and the functional product of gene 60 is necessary to synthesize it.


Microarray analysis of gene expression in the kidneys of new- and post-onset diabetic NOD mice.
We profiled the expression of 5,760 clones from a kidney subtraction library in the kidneys of three groups of NOD mice: nondiabetic, new-onset, and long-term diabetic. A total of 27 genes had lower expression and 1 gene (Gpx3) had higher expression in the new-onset diabetic mice compared with nondiabetic control NOD mice (P < 0.001). Similarly, 19 of the above 27 genes and 7 additional genes had higher expression and the Gpx3 gene had lower expression in long-term diabetic mice compared with controls (P < 0.001). Interestingly, only three genes may be different between new-onset and long-term diabetic mice (P < 0.0004). These genes are from diverse functional groups, including oxidative phosphorylation, free radical neutralization, channels, pumps, lipid processing, transcription and translation machinery, protein trafficking, constitutive protein processing, and immune function. The majority of these genes fall into four signaling pathways: insulin, transforming growth factor-beta, tumor necrosis factor-alpha, and peroxisome proliferator-activated receptor. The most significant expression change was found for the stearoyl-coenzyme A desaturase 1 (SCD1) gene (P < 10(-7)). The lower expression levels of the SCD1 gene in both diabetic groups compared with controls were further confirmed by Northern blot analysis and immunohistochemistry.


Impact of occupational pesticide exposure on the human gut microbiome.
The rising use of pesticides in modern agriculture has led to a shift in disease burden in which exposure to these chemicals plays an increasingly important role. The human gut microbiome, which is partially responsible for the biotransformation of xenobiotics, is also known to promote biotransformation of environmental pollutants. Understanding the effects of occupational pesticide exposure on the gut microbiome can thus provide valuable insights into the mechanisms underlying the impact of pesticide exposure on health. Here we investigate the impact of occupational pesticide exposure on human gut microbiome composition in 7198 participants from the Dutch Microbiome Project of the Lifelines Study. We used job-exposure matrices in combination with occupational codes to retrieve categorical and cumulative estimates of occupational exposures to general pesticides, herbicides, insecticides and fungicides. Approximately 4% of our cohort was occupationally exposed to at least one class of pesticides, with predominant exposure to multiple pesticide classes. Most participants reported long-term employment, suggesting a cumulative profile of exposure. We demonstrate that contact with insecticides, fungicides and a general "all pesticides" class was consistently associated with changes in the gut microbiome, showing significant associations with decreased alpha diversity and a differing beta diversity. We also report changes in the abundance of 39 different bacterial taxa upon exposure to the different pesticide classes included in this study. Together, the extent of statistically relevant associations between gut microbial changes and pesticide exposure in our findings highlights the impact of these compounds on the human gut microbiome.


Personal hygiene and safety of governmental hospital staff in Shiraz, Islamic Republic of Iran.
Complying with infection control standards is essential to prevent nosocomial infections. We aimed to determine health workers' hygiene practices and compliance with recommended instructions for personal hygiene among staff in all 30 hospitals affiliated to Shiraz University of Medical Sciences. The results showed that physicians and nurses were less compliant with personal hygiene practices than cleaners. Availability of protective measures was better in teaching hospitals than nonteaching hospitals as were vaccination rates among staff (hepatitis B and tetanus/diphtheria) with physicians scoring highest. Measures are needed to improve health workers' compliance.


Clinical Review of Neuromusculoskeletal Complementary and Alternative Approaches for the Treatment of Chronic Pelvic Pain Syndrome.
Chronic pelvic pain syndrome (CPPS) is a functional pain disorder characterized by ongoing pain in the apparent absence of clinically identifiable causes. The prevalence of functional pain disorders demonstrates the importance of adequate management of ongoing symptomatology, but due to the uncertain etiology and myriad patient presentation phenotypes, reliable treatment options are difficult to implement. New interventions involving non-pharmacological approaches to pain management have been investigated across a spectrum of clinical and pre-clinical studies. Given that conservative care such as exercise, counseling, and musculoskeletal therapy is widely recommended as first-line treatment for CPPS, an updated review of these and related methodologies are needed. Familiarizing physicians and the public with the newest evidence for complementary and alternative medicine (CAM) and other conservative care treatments will assist with the promotion of evidence-based practices in a safe and reliable manner. This review aimed to summarize the current evidence and proposed mechanisms for non-pharmacological treatment specific to CAM and management of chronic pelvic pain centered on neuromusculoskeletal focused intervention such as acupuncture, auriculotherapy, manipulation, manual therapy, myofascial release, and phototherapy. The discussion suggests that reported improvements in pelvic pain or related symptomatology may be attributed to changes in the peripheral inflammasome and somatic origins of peripheral sensitization. Robustness of the included clinical studies is discussed throughout the review, and attention is paid to delineating inclusion criteria of formally diagnosed CPPS compared to general pelvic or abdominal pain. Overall, this review consolidates the current state of evidence regarding the utilization of non-traditional interventions using CAM techniques for the management of chronic pelvic pain and recommends a future direction for the field.


Imaging of congenital and developmental cystic lesions of the brain: a narrative review.
Congenital and developmental intracranial cysts represent a large heterogenous group with varied presentations and etiologies. They can range from normal variants to pathologic lesions often associated with known congenital syndromes or acquired insults. While some are incidentally found, others are symptomatic or may become symptomatic over time. The preferred type of neuroimaging for timely diagnosis helps determine appropriate management and treatment, if indicated. In this narrative review article, authors present a comprehensive description of developmental cystic lesions. Imaging descriptions are provided for each type of cystic lesion as well as several representative images. As advanced neuroimaging techniques become more ubiquitous in clinical use, more light may be shed on the natural history of certain intracranial cystic lesions throughout the lifespan. This includes prenatal imaging for early identification and prognostication to surveillance imaging into advanced age to ascertain associations of certain cystic lesions with age-related cognitive dysfunction.


Ergonomics and cytotechnologists: reported musculoskeletal discomfort.
Analysis of this survey of 244 responding cytotechnologists shows that a high percentage still suffer from musculoskeletal disorders commonly associated with poor ergonomic design in the workplace, despite the variety of ergonomically designed microscopes that have been introduced into the market. Although Kalavar and Hunting surveyed a small group of cytotechnologists in the Washington, DC, area in 1996, no broad-based study has focused specifically on these professionals. The landmark study of microscope ergonomics, published by Soderberg in 1980, documented widespread musculoskeletal complaints in the electronics industry. This article identifies and discusses the types of musculoskeletal discomforts experienced by cytotechnologists. We propose that both practicing and student cytotechnologists receive training in ergonomic principles and appropriate interventions, such as improved work practices, proper ergonomic aids, well-designed workstations, and ergonomically designed equipment. More than 85% of respondents reported some musculoskeletal discomfort. Among the symptoms presented are headache, neck pain and stiffness, pain of the lower and upper back, and upper-extremity discomfort. Neurological symptoms, such as numbness, tingling, and/or pain in the hand and fingers, are often associated with repetitive motion. More than one-third reported numbness, tingling, and/or pain in the left-hand fingers, usually used for the fine-focus control knob. Almost one-half of respondents reported such symptoms for the right hand and fingers, normally used to manipulate the mechanical stage controls. Only 9% of the respondents were left-handed. Most (91%) were right handed. Although just over one-third (34.4%) of respondents worked for an employer with an ergonomics program, ergonomic assessments had been performed for only about one-half (17.6%) of these. In addition, approximately 10% of respondents indicated that ergonomics assessments had been performed on their workstations, even though the employer did not have an ergonomics program. Because of the high prevalence of reported symptoms, we recommend that all cytologists be trained in basic ergonomic principles and the proper use of ergonomic aids. These interventions should reduce the prevalence of musculoskeletal discomfort in medical microscopists.


Stimulated bronchial epithelial cells release bioactive lysophosphatidylcholine 16:0, 18:0, and 18:1.
In human subjects and animal models with acute and chronic lung injury, the bioactive lysophosphatidylcholine (LPC) is elevated in lung lining fluids. The increased LPC can promote an inflammatory microenvironment resulting in lung injury. Furthermore, pathological lung conditions are associated with upregulated phospholipase A2 (PLA2), the predominant enzyme producing LPC in tissues by hydrolysis of phosphatidylcholine. However, the lung cell populations responsible for increases of LPC have yet to be systematically characterized. The goal was to investigate the LPC generation by bronchial epithelial cells in response to pathological mediators and determine the major LPC species produced. Primary human bronchial epithelial cells (NHBE) were challenged by vascular endothelial growth factor (VEGF) for 1 or 6 h, and condition medium and cells collected for quantification of predominant LPC species by high performance liquid chromatography-tandem mass spectrometry (LC-MS-MS). The cells were analyzed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for PLA2. The direct effects of LPC in inducing inflammatory activities on NHBE were assessed by transepithelial resistance as well as expression of interleukin-8 (IL-8) and matrix metalloproteinase-1 (MMP-1). VEGF stimulation of NHBE for 1 or 6 h, significantly increased concentrations of LPC16:0, LPC18:0, and LPC18:1 in condition medium compared to control. The sPLA2-selective inhibitor (oleyloxyethyl phosphorylcholine) inhibited the VEGF-induced release of LPC16:0 and LPC18:1 and PLA2 activity. In contrast, NHBE stimulated with TNF did not induce LPC release. VEGF did not increase mRNA of PLA2 subtypes sPLA2-X, sPLA2-XIIa, cPLA2-IVa, and iPLA2-VI. Exogenous LPC treatment increased expression of IL-8 and MMP-1, and reduced the transepithelial resistance in NHBE. Our findings indicate that VEGF-stimulated bronchial epithelial cells are a key source of extracellular LPCs, which can function as an autocrine mediator with potential to induce airway epithelial inflammatory injury.


The significance of intrauterine growth restriction is different from prematurity for the outcome of infants with gastroschisis.
Recent reviews of gastroschisis identify prematurity and low birth weight as predictors of morbidity and mortality. The authors compared the outcomes of intrauterine growth-restricted infants (IUGR) with gastroschisis to those without growth restriction because IUGR is different from prematurity. A retrospective analysis was performed for infants born with gastroschisis between 1990 and 2000 at 2 pediatric hospitals. Patients were segregated into 3 groups based on birth weight corrected for gestational age: group 1 (IUGR, <fifth percentile), group 2 (fifth to 25th percentile), and group 3 (>25th percentile). Patient demographics, method of closure, number of surgeries, presence of atresia, and time to full enteral feedings (FPO days) were assessed. Mortality rate, length of stay (LOS), and readmission rates were also compared. Analysis of variance (ANOVA)/Student's t test and Fisher's. Exact tests were used for statistical analysis (P <.05 significant). Regression analysis was also performed. One hundred thirteen patients were included (group 1 = 17; group 2 = 43; group 3 = 53). Overall, infants with IUGR had similar outcomes to non-IUGR infants, including FPO and total parenteral nutrition (TPN) days, LOS, readmission, and mortality rates. The method of closure did not affect outcome. Infants with atresia had significantly increased times to full feeding (95 v 34 days; P =.034), more surgeries (2.7 v 1.4; P =.002), and longer LOS (106 v 48 days; P =.011). Infants born at less than 37 weeks' gestation had significantly increased fasting (NPO) days (28 v 18 days; P =.005) and longer LOS (65 v 37 days; P =.006) when compared with infants born at greater than 37 weeks. Logistic regression analysis identified the presence of atresia as an independent risk factor for gastrointestinal dysfunction and the need for prolonged TPN. Prematurity also adversely affected these same parameters, although it did not reach statistical significance. Although infants with gastroschisis are generally small for gestational age, the outcomes of growth-restricted infants are similar to those of other infants. The type of closure does not affect outcome, regardless of birth weight. The presence of atresia or prematurity does lead to longer times for full feeding and LOS. Therefore, routine premature delivery of infants with gastroschisis should not be advocated, even in the context of IUGR.


Semiautomatic microscopy as a tool for cytologists.
Despite impressive advances in the application of computer image analysis to cytology, many of the identification tasks that cytologists are called on to perform remain refractory to automated image analysis. The major reason is that a large fraction of these images, though simple for a human to deal with, are too complex to yield to current image analysis methodologies. It may be years before automated computer image analysis is reduced to clinical practicality. Even then, it is not clear that all cytologic image analyses will prove amenable to automation. In the meantime, semiautomatic image analysis (computer-aided microscopy) can provide a viable alternative, especially to persistently difficult image analysis problems. In semiautomatic image analysis, the onerous tasks of data acquisition--e.g., stage movement, data entry and storage--are left to the computer, while the decision-making tasks-e.g., identifying a cell's morphologic class--are left to the observer. Such a system proves to be easy and flexible to use as well as economical to build. It can also provide a reliable data base for the later evaluation of fully automated systems as they are developed. One such semiautomatic system, the Image Combining Computer Microscope (ICCM), is described, and the range of its application is illustrated. Some of the examples of ICCM applications discussed are: neuronal cell plots, three-dimensional dendrite tracking, serial section reconstruction of axons and mapping of plaques and tangles in Alzheimer's disease. They illustrate how powerful a semiautomated system can be in handling complex image analysis problems. It is suggested that semiautomated image analysis provides a viable long-range alternative to many cytologic image analysis problems.