Systematic screen identifies miRNAs that target RAD51 and RAD51D to enhance chemosensitivity.
Homologous recombination mediates error-free repair of DNA double-strand breaks (DSB). RAD51 is an essential protein for catalyzing homologous recombination and its recruitment to DSBs is mediated by many factors including RAD51, its paralogs, and breast/ovarian cancer susceptibility gene products BRCA1/2. Deregulation of these factors leads to impaired DNA repair, genomic instability, and cellular sensitivity to chemotherapeutics such as cisplatin and PARP inhibitors. microRNAs (miRNA) are short, noncoding RNAs that posttranscriptionally regulate gene expression; however, the contribution of miRNAs in the regulation of homologous recombination is not well understood. To address this, a library of human miRNA mimics was systematically screened to pinpoint several miRNAs that significantly reduce RAD51 foci formation in response to ionizing radiation in human osteosarcoma cells. Subsequent study focused on two of the strongest candidates, miR-103 and miR-107, as they are frequently deregulated in cancer. Consistent with the inhibition of RAD51 foci formation, miR-103 and miR-107 reduced homology-directed repair and sensitized cells to various DNA-damaging agents, including cisplatin and a PARP inhibitor. Mechanistic analyses revealed that both miR-103 and miR-107 directly target and regulate RAD51 and RAD51D, which is critical for miR-103/107-mediated chemosensitization. Furthermore, endogenous regulation of RAD51D by miR-103/107 was observed in several tumor subtypes. Taken together, these data show that miR-103 and miR-107 overexpression promotes genomic instability and may be used therapeutically to chemosensitize tumors. These findings demonstrate a role for miR-103 and -107 in regulating DNA damage repair, thereby identifying new players in the progression of cancer and response to chemotherapy.


Central venous catheter placement in children with thrombocytopenia.
The use of central venous catheter (CVC) is essential in the management of chronically ill children. Thrombocytopenia is a common hematological finding in these patients, with an increased risk of bleeding during the insertion of CVC. No clear guidelines are reported regarding the CVC positioning in patients with disorders of hemostasis, and prophylactic platelet (PLT) transfusions are still controversial. Aim of this study was to report the bleeding risk in pediatric patients with thrombocytopenia who underwent positioning of CVC. A retrospective single-center study of all CVCs surgically inserted over a 2-year period (April 2011 - April 2013) at our institution was performed. Age, gender, diagnosis, type of CVC, hematological values (hemoglobin and PLT count, prothrombin international normalized ratio, active partial thromboplastin time) and post-operative bleeding complications were compared between patients with PLT count below (group A) and above 50×109/L (group B). Seventy-two CVC procedures were performed in 67 patients, with a median age of 45 months. Of these, 25 (35%) catheters were positioned in 25 patients included in group A and the remaining 47 (65%) in 42 patients in group B. All twenty-five cases in group A received a prophylactic PLT transfusion prior to the procedure. Bleeding complications were reported in only two cases in group A (8%). CVC placement in pediatric patients with thrombocytopenia can be safely performed. We believe a randomized multicenter study could be necessary to determine the benefit of PLT transfusions in children with a PLT count below the recommended level of 50×109/L.


Why do current strategies for optimal nutritional therapy neglect the microbiome?
Strategies for providing optimal nutritional therapy have evolved over time, with the emphasis on specific directives (such as route, use of immunonutrition, high protein, organ-specific formulas, etc.), achieving variable degrees of success for improving outcomes in the intensive care unit. As the largest immune organ in the body comprising the largest interface between the host and the external environment, the gut can have an amplifying effect on a pattern of dysbiosis, immune dysregulation, and multiple organ failure seen in the critically ill patient. Conversely, maintenance of gut integrity can serve to restore a pattern of homeostasis, appropriate immune responses, symbiosis, and clinical recovery. Simply providing refined polymeric formulas as enteral nutrition may not take full advantage of the potential for optimal outcome that could be derived by giving therapy designed to directly stimulate gut defenses and support the intestinal microbiota. This article describes a series of strategies (such as use of intact whole food formulas, soluble fiber, fecal microbial transplantation, serum bovine immunoglobulin, or agents to promote commensal behavior) that should modulate the gut microbiome and shift the critically ill patient toward a pattern of health and recovery.


Field evaluation of Cellognost indirect haemagglutination test in the diagnosis of human trypanosomiasis (sleeping sickness) in Kura local government area of Kano State, Nigeria.
The Cellognost indirect haemagglutination (IHA) test was compared with conventional techniques, namely, wet blood film examination, stained thin and thick films, animal inoculation and the microhaematocrit centrifugation, for detecting sleeping sickness cases. Sixty-four out of 245 serum samples collected from suspected and some diagnosed cases of the disease were positive by the Cellognost screening test, while only three cases were detected by stained thin film or microhaematocrit centrifugation. Of the 64 cases 45 strongly positive samples were subjected to quantitative assay. Twenty-five of these had anti-Trypanosoma gambiense antibody titres above 1:20 representing active infection in the population. This represented 10.2% of the total sample, and strongly suggested the persistence of the disease in the population.


Home-Based Intervention for the Prevention and Treatment of Malaria Among Children Younger Than 5 Years in the West Region of Cameroon: Protocol for a Randomized Controlled Trial.
Although malaria is preventable and curable, 1 child dies of this disease every 2 minutes in Africa. Home-based management of malaria reduces the progression of severe malaria by more than 50%. Scalable, efficacious, and cost-effective strategies are needed to empower the capacities of home caregivers of children younger than 5 years of age in health education, diagnosis, and treatment of malaria at home. The main objective of this trial is to assess the impact of the management provided by home caregivers on the prevention, diagnosis, and treatment of malaria in children younger than 5 years as compared to the home-based malaria management component of the integrated community-directed intervention (CDI) strategy of community health workers (CHWs). A randomized controlled trial will be conducted. CHWs have conducted a census of all households where there is at least one child younger than 5 years with their home caregivers. These children and their home caregivers have been randomly placed into the intervention or control groups among the households identified. The trial will allow malaria home-based prevention, diagnosis, and treatment of 350 children younger than 5 years old by home caregivers in the Fombap area (intervention group) where the integrated CDI strategy will not implemented. This group will be compared to the home-based malaria management component of the integrated CDI strategy in which 350 children in the same age group will be followed up by CHWs in the Baneghang area (control group). The primary outcomes will be the prevention, diagnosis, and treatment of malaria in children younger than 5 years of age by home caregivers at home. The secondary outcomes comprise the malaria follow-up indicators produced by home caregivers in the intervention group and those produced by CHWs in the control group. Both descriptive and one-way analysis of variance estimation techniques will be used to compare the mean difference in the 2 strategies. From September 2019 to October 2019, all home caregivers with children younger than 5 years of age were identified in the intervention and control group by CHWs. Following this, 203 home caregivers with their 350 children younger than 5 years were randomly selected and enrolled in the intervention group, while 225 home caregivers with their 350 children younger than 5 years were enrolled in the control group. In the intervention group, 203 home caregivers were trained in November 2019. This home treatment effectively started in December 2019 and will continue until May 2020. Findings from this randomized controlled trial will contribute to resolving the challenges of severe malaria and to limiting the death due to malaria of children younger than 5 years. This will bring benefits to home caregivers who will know how to promptly diagnose and properly treat malaria in their children at home. Pan African Clinical Trial Registry (PACTR) 202003487018009; https://pactr.samrc.ac.za/TrialDisplay.aspx?TrialID=9788. DERR1-10.2196/19633.


Fish Gelatin: Current Nutritional, Medicinal, Tissue Repair Applications, and as a Carrier of Drug Delivery.
Gelatin is obtained via partial denaturation of collagen and is extensively used in various industries. The majority of gelatin utilized globally is derived from a mammalian source. Several health and religious concerns associated with porcine/bovine gelatin have been reported. Therefore, gelatin from a marine source is widely being investigated for its efficiency and utilization in a variety of applications as a potential substitute for porcine/bovine gelatin. Although fish gelatin is less durable and possesses lower melting and gelling temperatures compared to mammal-derived gelatin, various modifications have been reported to promote its rheological and functional properties to be efficiently employed. The present review describes in detail the current innovative applications of fish gelatin involving the food industry, drug delivery, and possible therapeutic applications. Gelatin bioactive molecules may be utilized as carriers for drug delivery. Due to its versatility, gelatin can be used in different carrier systems, such as microparticles, nanoparticles, fibers, and hydrogels. The present review also provides a perspective on the other potential pharmaceutical applications of fish gelatin, such as tissue regeneration, antioxidant supplementation, and antihypertensive and anticancer treatments.


Short- and long-term treatment of stable effort angina with nicardipine, a new calcium channel blocker: a double-blind, placebo-controlled, randomised, repeated cross-over study.
This study evaluated 1 year the efficacy of therapy with nicardipine in patients with chronic stable angina pectoris. Twenty-five male patients were entered. After a placebo run-in phase, the patients received nicardipine 30 mg, nicardipine 40 mg, and placebo, three times daily given in random, double-blind manner for 8 weeks. A double-blind, cross-over study comparing nicardipine with placebo was then undertaken. After 5 months of open treatment with nicardipine 90 or 120 mg day-1, patients received either placebo or nicardipine for 3 weeks, each followed by the alternative treatment for an additional 3 weeks and further open-label treatment with nicardipine for another 3-5 months. There were no significant changes in the PR, QRS or QT intervals, or in the QRS pattern during the short-term and long-term studies. There were no significant differences in mean heart rate after nicardipine compared with baseline. During treatment with nicardipine 120 mg day-1, patients reported significantly fewer anginal attacks compared with placebo, and nitroglycerin consumption also decreased. Nicardipine increased treadmill time, time to onset of angina, and time to one mm ST segment depression. These effects were maintained after 6 months of continued nicardipine therapy. Adverse effects were minor and well tolerated and included headache, dizziness, gastrointestinal upset, flushing paraesthesia and pedal oedema. Abrupt withdrawal of nicardipine at the end of the study resulted in a rapid return of the original symptoms but without further deterioration from the baseline measurements. Nicardipine was effective in the treatment of stable effort angina pectoris; this benefit was maintained for the entire year of treatment.


Application of confocal single-molecule FRET to intrinsically disordered proteins.
Intrinsically disordered proteins (IDPs) are characterized by a large degree of conformational heterogeneity. In such cases, classical experimental methods often yield only mean values, averaged over the entire ensemble of molecules. The microscopic distributions of conformations, trajectories, or sequences of events often remain unknown, and with them the underlying molecular mechanisms. Signal averaging can be avoided by observing individual molecules. A particularly versatile method is highly sensitive fluorescence detection. In combination with Förster resonance energy transfer (FRET), distances and conformational dynamics can be investigated in single molecules. This chapter introduces the practical aspects of applying confocal single-molecule FRET experiments to the study of IDPs.


Exploration and modulation of brain network interactions with noninvasive brain stimulation in combination with neuroimaging.
Much recent work in systems neuroscience has focused on how dynamic interactions between different cortical regions underlie complex brain functions such as motor coordination, language and emotional regulation. Various studies using neuroimaging and neurophysiologic techniques have suggested that in many neuropsychiatric disorders, these dynamic brain networks are dysregulated. Here we review the utility of combined noninvasive brain stimulation and neuroimaging approaches towards greater understanding of dynamic brain networks in health and disease. Brain stimulation techniques, such as transcranial magnetic stimulation and transcranial direct current stimulation, use electromagnetic principles to alter brain activity noninvasively, and induce focal but also network effects beyond the stimulation site. When combined with brain imaging techniques such as functional magnetic resonance imaging, positron emission tomography and electroencephalography, these brain stimulation techniques enable a causal assessment of the interaction between different network components, and their respective functional roles. The same techniques can also be applied to explore hypotheses regarding the changes in functional connectivity that occur during task performance and in various disease states such as stroke, depression and schizophrenia. Finally, in diseases characterized by pathologic alterations in either the excitability within a single region or in the activity of distributed networks, such techniques provide a potential mechanism to alter cortical network function and architectures in a beneficial manner.


Arsenic removal from groundwater in Kütahya, Turkey, by novel calcined modified hydrotalcite.
Since arsenic is highly toxic and carcinogenic, it now causes serious health problems all over the world. Therefore, there is an urgent need to develop new techniques that are cost-effective and easily applicable to remove arsenic from contaminated waters. Layer double hydroxides have the potential to be a good adsorbent to remove arsenic from contaminated waters due to high surface area and high anion exchange capacity. In this paper, arsenic removal from water by calcined Fe-hydrotalcite (CFeHT) known as layered double hydroxide and prepared synthetically with coprecipitation method was researched. The study brings out that the effect of initial solution pH values was limited for the adsorption. The experimental study indicates that the adsorption of arsenic rapidly occured in comparison with other studies. It was determined that the pseudo-second-order kinetic model was more suitable than the first order. In isotherm studies, it was seen that the experimental data were compatible with Langmuir model. In this study was determined that CFeHT has a high arsenic removal potential. And also the concentration of the arsenic solution (600 µg/L) has been reduced below the allowable value by the World Health Organization (< 10 µg/L). The desorption test indicates that the desorption ratio of As(V) was obtained as 72.7.


Molecular cloning and transformation of cyclodiene resistance in Drosophila: an invertebrate gamma-aminobutyric acid subtype A receptor locus.
Cyclodiene resistance represents 60% of the reported cases of insecticide resistance and is also present in vertebrates. Resistance is due to insensitivity of the cyclodiene/picrotoxinin binding site on the gamma-aminobutyric acid subtype A (GABAA) receptor-chloride ionophore complex. Following isolation of cyclodiene-resistant Drosophila mutants, we report the cloning of the locus conferring resistance via a "chromosomal walk" and rescue of the susceptible phenotype by P-element-mediated germ-line transformation. Amino acid sequence analysis of a cDNA from the locus reveals homology with vertebrate GABAA subunits. To our knowledge, this represents the first cloning of an invertebrate GABA receptor and also allows us to manipulate the resistance status of an insect via germ-line transformation. This gene may be useful as a selectable marker in other insect systems.


[Role of innate macrophages in DNA and protein biosynthesis in cultured cells of murine (line A) ascitic hepatoma].
Joint effect of high-density lipoproteins (HDLP) and glucocorticoids presence is responsible for increased protein and DNA biosynthesis in the cells of the hepatic parenchyme. Hepatic innate macrophages devour HDLP and glucocorticoids and release the A-1 tetrahydrocorisol-apolipoprotein complex which in turn is channeled into interstitial spaces to be captured by hepatocytes, thus stimulating protein and DNA biosynthesis. This mechanism was observed in cultured cells of murine ascitic hepatoma; it was regulated by macrophages present in incubation medium. The rate of protein and DNA biosynthesis remained constant unless HDLP and glucocorticoids were present in the medium and in a macrophage-containing culture alone.


Testicular versus ejaculated spermatozoa in ICSI cycles of normozoospermic men with high sperm DNA fragmentation and previous ART failures.
As a part of male assessment, conventional sperm parameters including morphologic features have been dedicated as major factors influencing fertilisation and pregnancy rates in assisted reproductive technology (ART). Genomic integrity of spermatozoa has also been found to influence fertility prognosis, and hence, sperm DNA fragmentation index (DFI) has been adopted by many centres to document this entity. Despite several suggested approaches, there is lack of universal consensus on optimising fertility outcomes in males with high sperm DFI. In this context, the results from cycles using testicular spermatozoa (TESA) obtained by aspiration were compared with those of ejaculated spermatozoa (EJ) in normozoospermic subjects with high sperm DFI and previous ART failures. Clinical (41.9% versus 20%) and ongoing pregnancy rates (38.7% versus 15%) were significantly better and miscarriages were lower in TESA group when compared to EJ group. Sperm DFI should be a part of male partner's evaluation following unsuccessful ART attempts. When high DFI is detected (>30%), ICSI using testicular spermatozoa obtained by TESA seems an effective option particularly for those with repeated ART failures in terms of clinical, ongoing pregnancies and miscarriages even though conventional sperm parameters are within normal range.


A comparison of insight in body dysmorphic disorder and obsessive-compulsive disorder.
Insight/delusionality of beliefs is an important dimension of psychopathology across psychiatric disorders. This construct is of increasing interest in obsessive-compulsive and related disorders, including obsessive-compulsive disorder (OCD) and body dysmorphic disorder (BDD). Even though OCD and BDD are considered closely related, no prior study has compared these disorders across a range of categories of global insight (excellent, good, fair, poor, absent/delusional), and only one study has compared these disorders on individual components of insight. Using the reliable and valid Brown Assessment of Beliefs Scale (BABS), this study examined insight/delusionality of OCD- or BDD-related beliefs in 211 individuals with primary OCD versus 68 individuals with primary BDD. In both disorders, levels of insight spanned the full range, from excellent to absent (i.e., delusional beliefs). However, the distribution of BABS scores across insight categories differed significantly by disorder, with the majority of OCD subjects showing excellent or good insight, and the majority of BDD subjects showing poor or absent insight. Compared to OCD subjects, BDD subjects had significantly poorer insight both overall (total BABS score) and on all individual BABS items. BABS score was significantly correlated with BDD and OCD severity, but in regressions it accounted for only 21% of the variance in OCD and 28% in BDD. In summary, both global insight and its individual components are poorer in BDD than in OCD, which has implications for research and clinical care, as well as understanding of the relationship between these disorders. Disorder severity is associated with but not equivalent to insight/delusionality.


Congeneric variability in lifespan extension and onset of senescence suggest active regulation of aging in response to low temperature.
Lifespan extension under low temperature is well conserved across both endothermic and exothermic taxa, but the mechanism underlying this change in aging is poorly understood. Low temperature is thought to decrease metabolic rate, thus slowing the accumulation of cellular damage from reactive oxygen species, although recent evidence suggests involvement of specific cold-sensing biochemical pathways. We tested the effect of low temperature on aging in 11 strains of Brachionus rotifers, with the hypothesis that if the mechanism of lifespan extension is purely thermodynamic, all strains should have a similar increase in lifespan. We found differences in change in median lifespan ranging from a 6% decrease to a 100% increase, as well as differences in maximum and relative lifespan extension and in mortality rate. Low temperature delays reproductive senescence in most strains, suggesting an extension of healthspan, even in strains with little to no change in lifespan. The combination of low temperature and caloric restriction in one strain resulted in an additive lifespan increase, indicating these interventions may work via non- or partially-overlapping pathways. The known low temperature sensor TRPA1 is present in the rotifer genome, but chemical TRPA1 agonists did not affect lifespan, suggesting that this gene may be involved in low temperature sensation but not in chemoreception in rotifers. The congeneric variability in response to low temperature suggests that the mechanism of low temperature lifespan extension is an active genetic process rather than a passive thermodynamic one and is dependent upon genotype.


Pulsed laser deposition of metal films and nanoparticles in vacuum using subnanosecond laser pulses.
A study of silver, chromium, stainless-steel, and indium thin films prepared by subnanosecond laser deposition in vacuum is reported. We compare the laser ablation in vacuum at the weak- and tight-focusing conditions of a Ti:sapphire laser beam and analyze the nanoparticles synthesized in the latter case using absorption spectroscopy, x-ray fluorescence, atomic force microscopy, and scanning electron microscopy. Our results show that the nanoparticle formation can be accomplished using long laser pulses under tight-focusing conditions.


Comparison of Cytokine and Efflux Transporter Expression in Pediatric Versus Adult-onset Ulcerative Colitis.
Ulcerative colitis (UC), a chronic inflammation of the colon, is often more severe in children than adults. Identification of altered expression of efflux transporters, cytokines, and suppressor of cytokine signaling (SOCS) molecules in pediatric versus adult patients could provide insight into the differential molecular patterns related to the age and disease pathology. Mucosal samples from terminal ileum and colon in pediatric (9 UC-New, 4 UC-Remission) and adult (9 UC-New, 8 UC-Remission) patients were compared with healthy subjects (15 children and 10 adults) for mRNA expressions of several efflux transporters, cytokines, and SOCS molecules. The inflamed colon interleukin (IL)-6, IL-17A, and interferon-γ levels were elevated in UC-New subgroups but close to control values in UC-Remission. IL-1β expression was increased only in UC-New children. Interestingly, uninflamed ileum also showed increased IL-6 and IL-1β levels in UC-New subgroups. SOCS1/SOCS3 expression pattern followed a trend observed for inflammatory cytokines only in children. Both children and adults had decreased multidrug resistance protein 1 expression in colon, which inversely correlated with disease score, IL-6 and interferon-γ levels in UC-New children. IL-2 expression was upregulated in UC-Remission, compared with controls. Efflux transporter expression varies between UC children and adults except for decreased multidrug resistance protein 1. UC is characterized by a dysregulated TH1 and TH17 cytokine response irrespective of age at disease onset, with higher cytokine levels detected in children. Increased IL-2 levels in remission imply a protective role for regulatory T cells (Tregs).


Number Needed to Scan: Evidence-Based Point-of-Care Ultrasound (POCUS).
Interest and enthusiasm, regarding the use of point-of-care ultrasound (POCUS), continues to grow among clinicians in multiple medical specialties. Ultrasound machines technology has advanced to allow for smaller, even handheld machines. Integration of automated imaging technology has made these machines more user-friendly. However, one of the concerns with the widespread availability of POCUS is the overuse and misuse of this technology. In order to maximize the clinical impact of POCUS, this manuscript seeks to discuss a novel concept called the "Number needed to scan" (NNS). The NNS is an expression of the number of POCUS examinations needed to be performed to attain a benefit to the patient or to prevent an adverse outcome of a procedure. NNS serves a dual purpose: it can help clinicians understand the magnitude of clinical impact when they apply POCUS, and it can help clinicians explain this magnitude in layman terms to their patients. In this manuscript, we have focused our NNS calculations on landmark articles in three major categories: change in management; safety and accuracy; and catching a missed diagnosis. As clinicians seek to be good stewards of POCUS, NNS should be a concept used to consider which patients will be most likely to benefit from a clinician performed ultrasound.


Long-term outcomes of the Bankart and Latarjet repairs: a systematic review.
The most common surgical techniques for the treatment of recurrent anterior shoulder instability include the arthroscopic Bankart repair, the open Bankart repair and the open Latarjet procedure. The purpose of this study was to evaluate and compare the long-term outcomes following these procedures. A systematic review of modern procedures with a minimum follow-up of 5 years was completed. The objective outcome measures evaluated were post-operative dislocation and instability rate, the Rowe score, radiographic arthritis and complications. Twenty-eight studies with a total of 1652 repairs were analyzed. The estimated re-dislocation rate was 15.1% following arthroscopic Bankart repair, 7.7% following open Bankart repair and 2.7% following Latarjet repair, with the comparison between arthroscopic Bankart and open Latarjet reaching statistical significance (p<0.001). The rates of subjective instability and radiographic arthritis were consistently high across groups, with no statistical difference between groups. Estimated complication rates were statistically higher in the open Latarjet repair (9.4%) than in the arthroscopic Bankart (0%; p=0.002). The open Latarjet procedure yields the most reliable method of stabilization but the highest complication rate. There are uniformly high rates of post-operative subjective instability symptoms and radiographic arthritis at 5 years regardless of procedure choice.


Statewide survey of risk factor prevalence: the Ohio experience.
In 1982, a statewide survey was conducted to determine the prevalence of health risk factors among Ohio's population. The survey was mandated by a health education-risk reduction grant to the Ohio Department of Health. The background, development, and validation of the survey instrument are described. The four goals of "Health Ohio"--the collection of descriptive statistics on selected risk factors for adult Ohioans, the compilation of baseline data, the development of a standard methodology for a prevalence survey, and the reporting of these findings for potential users of the data--were achieved. The population sample consisted of 607 Ohioans aged 18 and older who were polled by telephone. Subjects were selected through a modified random digit dialing technique. As a result of this technique and the designation of a specific household respondent, demographic characteristics of the sample matched those of the State's population in the 1980 census. Among the implications of the survey findings were the needs to (a) remove economic barriers that apparently impede the installation of residential smoke detectors, (b) initiate health education at an earlier age to counter cigarette smoking trends, and (c) encourage adult self-determination in reducing health risk. The February 1982 "Health Ohio" data describing the need for intervention to reduce health risk factors have become the basis for health education-risk reduction efforts of the Ohio Department of Health. In addition, "Health Ohio" has spawned two local prevalence surveys in the State; these resulted in more precise local data on the prevalence of health risk factors. Other multiplier effects of health education-risk reduction projects should be documented for future reference.


Optic nerve regeneration in mammals: Regenerated or spared axons?
Intraorbital optic nerve crush in rodents is widely used as a model to study axon regeneration in the adult mammalian central nervous system. Recent studies using appropriate genetic manipulations have revealed remarkable abilities of mature retinal ganglion cell (RGC) axons to regenerate after optic nerve injury, with some studies demonstrating that axons can then go on to re-innervate a number of central visual targets with partial functional restoration. However, one confounding factor inherent to optic nerve crush injury is the potential incompleteness of the initial lesion, leaving spared axons that later on could erroneously be interpreted as regenerating distal to the injury site. Careful examination of axonal projection pattern and morphology may facilitate separating spared from regenerating RGC axons. Here we discuss morphological criteria and strategies that may be used to differentiate spared versus regenerated axons in the injured mammalian optic nerve.


The role of autophagy during development in higher eukaryotes.
Autophagy is a lysosome-mediated degradation pathway used by eukaryotes to recycle cytosolic components in both basal and stress conditions. Several genes have been described as regulators of autophagy, many of them being evolutionarily conserved from yeast to mammals. The study of autophagy-defective model systems has made it possible to highlight the importance of correctly functioning autophagic machinery in the development of invertebrates as, for example, during the complex events of fly and worm metamorphosis. In vertebrates, on the other hand, autophagy defects can be lethal for the animal if the mutated gene is involved in the early stages of development, or can lead to severe phenotypes if the mutation affects later stages. However, in both lower and higher eukaryotes, autophagy seems to be crucial during embryogenesis by acting in tissue remodeling in parallel with apoptosis. An increase of autophagic cells is, in fact, observed in the embryonic stages characterized by massive cell elimination. Moreover, autophagic processes probably protect cells during metabolic stress and nutrient paucity that occur during tissue remodeling. In light of such evidence, it can be concluded that there is a close interplay between autophagy and the processes of cell death, proliferation and differentiation that determine the development of higher eukaryotes.


Hierarchically organized honeycomb films through block copolymer directed self-assembly in "breath figure" templating and soft microwave-triggered annealing.
Hierarchically organized polymer films are produced with a high level of order from the combination of block copolymer nanophase segregation, "breath figure" methodology and microwave irradiation. A block copolymer based on poly(methyl methacrylate) and poly(n-butylacrylate) featuring cylindrical nanopatterns is involved in the "breath figure" process to create a microporous honeycomb structure. These films are submitted to microwave annealing to enhance the degree of ordering of the nano-segregation without the destruction of the honeycomb microstructure, which is not possible by classical thermal or solvent annealing. Ellipsometry, optical and atomic force microscopy are used to study three key parameters; the substrate nature, the film thickness and the microwave irradiation power. The silicon wafer is the substrate of choice to efficiently act as the heating transfer element and 60 seconds at 10 watts are enough to nicely order the 1 μm thick copolymer films. These conditions are eventually applied on hierarchically organized polymer films to obtain a hexagonal array of 100 nm deep holes within a matrix of perpendicularly aligned nano-cylinders.


Increased basigin in bleomycin-induced lung injury.
Basigin is expressed in many tissues during development, including lung. It is also found on tumor cells and in wounds where it is thought to stimulate adjacent fibroblasts to produce matrix metalloproteinases. To investigate whether basigin might be expressed in fibro-inflammatory lung processes, we generated bleomycin-induced lung injury in mice. At 14 d after intratracheal bleomycin, we found basigin prominently in areas of fibrosis, alveolar macrophages, and bronchiolar epithelium, whereas it was only weakly present in bronchiolar epithelium in untreated mice. Western blots of radioimmunoprecipitation assay RIPA-insoluble fractions of bleomycin-treated lungs showed increased basigin compared with RIPA-insoluble fractions of lung from untreated mice. By quantitative reverse transcriptase-polymerase chain reaction, lung basigin mRNA was significantly increased 14 d after bleomycin, and by in situ hybridization, basigin mRNA was prominent in bronchiolar epithelium. Western blots of bronchoalveolar lavage fluid (BALF) showed various forms of basigin after bleomycin that were not present in BALF from untreated lung. These results demonstrate that bleomycin-induced lung injury is associated with increased basigin expression in bronchiolar epithelium, deposition of basigin in fibrotic sites, and increased basigin in BALF. Accordingly, basigin may play a role in diffuse alveolar injury.


Size-and-Shape Space Gaussian Mixture Models for Structural Clustering of Molecular Dynamics Trajectories.
Determining the optimal number and identity of structural clusters from an ensemble of molecular configurations continues to be a challenge. Recent structural clustering methods have focused on the use of internal coordinates due to the innate rotational and translational invariance of these features. The vast number of possible internal coordinates necessitates a feature space supervision step to make clustering tractable but yields a protocol that can be system type-specific. Particle positions offer an appealing alternative to internal coordinates but suffer from a lack of rotational and translational invariance, as well as a perceived insensitivity to regions of structural dissimilarity. Here, we present a method, denoted shape-GMM, that overcomes the shortcomings of particle positions using a weighted maximum likelihood alignment procedure. This alignment strategy is then built into an expectation maximization Gaussian mixture model (GMM) procedure to capture metastable states in the free-energy landscape. The resulting algorithm distinguishes between a variety of different structures, including those indistinguishable by root-mean-square displacement and pairwise distances, as demonstrated on several model systems. Shape-GMM results on an extensive simulation of the fast-folding HP35 Nle/Nle mutant protein support a four-state folding/unfolding mechanism, which is consistent with previous experimental results and provides kinetic details comparable to previous state-of-the art clustering approaches, as measured by the VAMP-2 score. Currently, training of shape-GMMs is recommended for systems (or subsystems) that can be represented by ≲200 particles and ≲100k configurations to estimate high-dimensional covariance matrices and balance computational expense. Once a shape-GMM is trained, it can be used to predict the cluster identities of millions of configurations.


[Lung structure of Sphenodon punctatus. A comparative anatomic study].
After a critical review of the literature, the authors describe the structure of the lung of Sphenodon, dealing with form, arteries, veins, and septation. The external form, circulatory pattern, degree of complication, height of the septa, vascular structure, and decrease in complication (?) are discussed from the standpoint of comparative anatomy. In an attempt to explain the unusual findings, the capacity for development of the central and subpleural type of differentiation is compared and contrasted. Obvious relations between the unusual lung structure and the phylogenetic age of the Sphenodon were sought, with the conclusion that the Sphenodon cannot be included in the Reptilian oders since its physical structure forms a class by itself.


Studies on antagonism between porcine skin bacteria.
Strains of Staphylococcus warneri, Staph. epidermidis and Aerococcus viridans isolated from porcine skin were shown to produce substances that inhibited the growth of Staph. hyicus. Studies of interactions between the inhibitor-producing bacteria and Staph. hyicus on cellulose acetate membranes demonstrated a significant reduction (P less than 0.001) of the Staph. hyicus population at various stages of its growth cycle, depending on the inhibitor used. Inhibitor production profiles in batch culture showed a change in production with culture growth. Trypsin and an extracellular substance from Staph. hyicus had similar effects in reducing inhibitor production. Staphylococcus hyicus was shown to produce varying amounts of a proteolytic enzyme during growth in batch culture. Such in vitro models of bacterial interaction enable potential mechanisms of disease control using inhibitor-producing organisms to be investigated and provide a basis for studies in vivo.


YAP localization mediates mechanical adaptation of human cancer cells during extravasation in vivo.
Biophysical profiling of primary tumors has revealed that individual tumor cells fall along a highly heterogeneous continuum of mechanical phenotypes. One idea is that a subset of tumor cells is "softer" to facilitate detachment and escape from the primary site, a step required to initiate metastasis. However, it has also been postulated that cells must be able to deform and generate sufficient force to exit into distant sites. Here, we aimed to dissect the mechanical changes that occur during extravasation and organ colonization. Using multiplexed methods of intravital microscopy and optical tweezer based active microrheology, we obtained longitudinal images and mechanical profiles of cells during organ colonization in vivo . We determined that cells were softer, more liquid like upon exit of the vasculature but stiffened and became more solid like once in the new organ microenvironment. We also determined that a YAP mediated mechanogenotype influenced the global dissemination in our in vivo and in vitro models and that reducing mechanical heterogeneity could reduce extravasation. Moreover, our high throughput analysis of mechanical phenotypes of patient samples revealed that this mechanics was in part regulated by the external hydrodynamic forces that the cancer cells experienced within capillary mimetics. Our findings indicate that disseminated cancer cells can keep mutating with a continuum landscape of mechano-phenotypes, governed by the YAP-mediated mechanosensing of hydrodynamic flow.


Truncating CLCN1 mutations in myotonia congenita: variable patterns of inheritance.
Myotonia congenita due to protein truncating CLCN1 mutations is associated with variable patterns of inheritance. Three family kindreds are described, all of whom possess protein truncating mutations (Y33X, fs503X, R894X). One lineage also has coexistent R894X, A313T, and A320V mutations. The Y33X mutation kinship has autosomal recessive inheritance and a severe phenotype when homozygous. The fs503X family has autosomal dominant inheritance and a moderate-to-severe phenotype. The A313T mutation kindred also has autosomal dominant inheritance but expresses a mild phenotype, except for the more severely affected compound heterozygotes. Early truncating mutations precluding dimerization are expected to be autosomal recessive and express a severe phenotype, while later mutations may be variable. The pedigrees presented here demonstrate that intrafamilial phenotypic variability may result from a dosage effect of an additional mutation, not necessarily variable expressivity. Mutations that have unexpected patterns of inheritance may represent allelic variability.


Frequency domain analysis of renal autoregulation in the rat.
The frequency response of renal blood flow was estimated to test our suggestion that autoregulation had one fast and one slow component. Arterial pressure was varied periodically by a pump connected to the distal aorta or aperiodically by stimulating atrial fibrillation. There was only one regulatory mechanism, corresponding to the slow component seen earlier. Two apparent regulators were seen when pressure was measured far from the renal artery (right common carotid), rather than near (superior mesenteric). Simultaneous measurements of pressure in both arteries showed interference of the pressure wave from the heart with pressure waves from the pump in the distal aorta at frequencies near that of the heart rate. Measurements made in the superior mesenteric artery were likely to be correct. The earlier results were in error, and frequency response methods detect only one regulator. Animals made acutely hypertensive behaved similarly. The effect of the regulator appears as a phase shift at frequencies of 0.25 Hz and below and an attenuated flow magnitude at frequencies below 0.1 Hz. This frequency response is consistent with the flow propagation time to the macula densa. We conclude that macula densa feedback was the only flow-regulating mechanism detected.


A counter-swirl design concept for dry powder inhalers.
A swirling airflow is incorporated in several dry powder inhalers (DPIs) for effective powder de-agglomeration. This commonly requires the use of a flow-straightening grid in the DPI to reduce drug deposition loss caused by large lateral spreading of the emerging aerosol. Here, we propose a novel grid-free DPI design concept that improves the aerosol flow characteristics and reduces the aforementioned drug loss. The basis of this design is the implementation of a secondary airflow that swirls in the opposite direction (counter-swirl) to that of a primary swirling airflow. In-vitro deposition, computational fluid dynamics simulations and particle image velocimetry measurements are used to evaluate the counter-swirl DPI aerosol performance and flow characteristics. In comparison with a baseline-DPI that has only a primary swirling airflow, the counter-swirl DPI has 20% less deposition of the emitted drug dose in the induction port and pre-separator of a next generation impactor (NGI). This occurs as a result of the lower flow-swirl generated from the counter-swirl DPI which eliminates the axial reverse flow outside of the mouthpiece and substantially reduces lateral spreading in the exiting aerosol. Modifications to the counter-swirl DPI design were made to prevent drug loss from the secondary airflow tangential inlets, which involved the addition of wall perforations in the tangential inlets and the separation of the primary and secondary swirling airflows by an annular channel. These modified DPI devices were successful in that aspect but had higher flow-swirl than that in the counter-swirl DPI and thus had higher drug mass retained in the device and deposited in the induction port and pre-separator of the NGI. The fine particle fraction in the aerosols generated from all the counter-swirl-based DPIs and the baseline-DPI are found to be statistically similar to each other.


Femtosecond bond breaking and charge dynamics in ultracharged amino acids.
Historically, structure determination of nanocrystals, proteins, and macromolecules required the growth of high-quality crystals sufficiently large to diffract X-rays efficiently while withstanding radiation damage. The development of the X-ray free-electron laser has opened the path toward high resolution single particle imaging, and the extreme intensity of the X-rays ensures that enough diffraction statistics are collected before the sample is destroyed by radiation damage. Still, recovery of the structure is a challenge, in part due to the partial fragmentation of the sample during the diffraction event. In this study, we use first-principles based methods to study the impact of radiation induced ionization of six amino acids on the reconstruction process. In particular, we study the fragmentation and charge rearrangement to elucidate the time scales involved and the characteristic fragments occurring.


Chromosome 4 deletions are frequent in invasive cervical cancer and differ between histologic variants.
Patterns of discontinuous deletion of chromosome 4 have been described in histologic variants of lung carcinomas and may represent different "hotspot" targets for gene-environment interactions. Since similar environmental risks exist for cervical cancer, we investigated patterns of discontinuous deletion in two major histologic variants. Thirteen archival cases of squamous cell cancer (SCCA) and 11 cases of adenocarcinoma (AC) were precisely microdissected. Matched normal and tumor DNA were used for polymerase chain reaction (PCR) based loss of heterozygosity (LOH) analyses using 19 polymorphic markers spanning chromosome 4. Human papillomavirus (HPV) detection was determined by PCR using general and type-specific primers (HPV 16, 18). Differences in LOH between histologic tumor types and chromosomal regions were determined using Fisher's exact test. Loss at any chromosome 4 locus occurred in 92% of all tumors studied, with the majority of deletions occurring on the long arm of the chromosome. Four discrete minimal regions of discontinuous deletion (R) were identified. For these regions, LOH frequencies were 76% (R1, 4q34-q35), 48% (R2, 4q25-q26), 36% (R3, 4p15.1-p15.3), and 26% (R4, 4p16). Loss in SCCA predominated at 4q (4q34-q35; 83%) and in AC at 4p (4p15.3; 50%). Overall LOH on the p arm was significant in AC (82%) compared to SCCA (31%) (P = 0.02). HPV detection was similar in SCCA (85%) and AC (73%), and HPV 16/18 subtypes were similarly represented in both histologies. Chromosome 4 deletions are frequent in cervical carcinomas. Different patterns of deletion between SCCA and AC may represent gene regions targeted by different gene-environment interactions in these tumor subtypes.


Design of Metadata Services for Clinical Data Interoperability in Germany.
Secondary use of electronic health record (EHR) data requires a detailed description of metadata, especially when data collection and data re-use are organizationally and technically far apart. This paper describes the concept of the SMITH consortium that includes conventions, processes, and tools for describing and managing metadata using common standards for semantic interoperability. It deals in particular with the chain of processing steps of data from existing information systems and provides an overview of the planned use of metadata, medical terminologies, and semantic services in the consortium.


Intensive short-term dynamic residential treatment program for patients with treatment-resistant disorders.
The study investigated the effectiveness of an Intensive Short-Term Dynamic (ISTDP) residential treatment program for patients with treatment resistant anxiety- and/or depressive disorders, with and without comorbid personality disorders. A non-randomized controlled trial examined the effects of an eight week intensive residential treatment program based on principles from ISTDP. Patients (N=60), who had repeated prior treatment failure for current mental disorder, sufficient dysfunction to warrant hospitalization, and evidencing capacity to take an intrapsychic perspective on own problems, were included. Outcome variables included measures of target complaints (depression/anxiety, social role dysfunction, and interpersonal distress), general symptom distress, and interpersonal functioning. Measures were administered throughout and after treatment. Change was assessed by multilevel growth curve modeling. Changes during and after treatment were compared to those reported by a sub-sample of wait-list controls taking treatment as usual (N=30). The treatment group evidenced significant improvements on all measures. By contrast, receiving treatment as usual while on the wait-list did not yield significant changes. Effect sizes in the treatment group were consistently large at both termination and follow-up. Fourteen months after treatment 50.0% of patients had recovered in terms of target complaints. Approximately 53.3% and 48.3%, respectively, had recovered in terms of general symptom distress and interpersonal functioning. Limitations included a relatively small sample size, inability to discern the effectiveness of separate components of the treatment program, and lack of randomization of patients to wait-list and treatment. ISTDP-based residential treatment with an eight-week time-limit appears to be effective for alleviating common and severe, treatment resistant mental disorders. The treatment program was superior to receiving treatment as usual while on the wait-list. Participation in the program quickly reduced target complaints, symptoms and interpersonal problems for patients who, based on previous treatment experiences, were expected to fare poorly in treatment. Gains were consistently maintained or improved further at follow-up. Results are promising for patients with chronic debilitating problems who often do not profit from traditional psychiatric treatment.


Starch-binding domains as CBM families-history, occurrence, structure, function and evolution.
The term "starch-binding domain" (SBD) has been applied to a domain within an amylolytic enzyme that gave the enzyme the ability to bind onto raw, i.e. thermally untreated, granular starch. An SBD is a special case of a carbohydrate-binding domain, which in general, is a structurally and functionally independent protein module exhibiting no enzymatic activity but possessing potential to target the catalytic domain to the carbohydrate substrate to accommodate it and process it at the active site. As so-called families, SBDs together with other carbohydrate-binding modules (CBMs) have become an integral part of the CAZy database (http://www.cazy.org/). The first two well-described SBDs, i.e. the C-terminal Aspergillus-type and the N-terminal Rhizopus-type have been assigned the families CBM20 and CBM21, respectively. Currently, among the 85 established CBM families in CAZy, fifteen can be considered as families having SBD functional characteristics: CBM20, 21, 25, 26, 34, 41, 45, 48, 53, 58, 68, 69, 74, 82 and 83. All known SBDs, with the exception of the extra long CBM74, were recognized as a module consisting of approximately 100 residues, adopting a β-sandwich fold and possessing at least one carbohydrate-binding site. The present review aims to deliver and describe: (i) the SBD identification in different amylolytic and related enzymes (e.g., CAZy GH families) as well as in other relevant enzymes and proteins (e.g., laforin, the β-subunit of AMPK, and others); (ii) information on the position in the polypeptide chain and the number of SBD copies and their CBM family affiliation (if appropriate); (iii) structure/function studies of SBDs with a special focus on solved tertiary structures, in particular, as complexes with α-glucan ligands; and (iv) the evolutionary relationships of SBDs in a tree common to all SBD CBM families (except for the extra long CBM74). Finally, some special cases and novel potential SBDs are also introduced.


Lgr4 gene regulates corpus luteum maturation through modulation of the WNT-mediated EGFR-ERK signaling pathway.
Luteal-phase insufficiency is one of the major causes of female infertility, but the molecular mechanisms are still largely unknown. Here we found that disruption of Lgr4/Gpr48, the newly identified receptor for R-spondins, greatly reduced female fertility in mice. The expression of Lgr4 was induced specifically in granulosa-lutein cells during luteinization. In Lgr4-deficient female mice, the estrous cycle was prolonged and serum progesterone levels were dramatically downregulated. In Lgr4(-/-) corpora lutea, the expression of key enzymes for steroidogenesis as well as common luteal marker genes was significantly decreased. Additionally, the activity of epidermal growth factor receptor (EGFR)-ERK signaling was attenuated in Lgr4(-/-) granulosa-lutein cells. We found that the maturation of Lgr4(-/-) cells was impaired in cultured primary granulosa cells, but the defect was partially rescued by reactivation of EGFR signaling by heparin-binding EGF-like growth factor treatment. We found that the expression of wingless-type MMTV integration site family (WNT)/catenin (cadherin associated protein), beta 1 (CTNNB1) downstream targets, including matrix metalloproteinase 9, which is a critical matrix metalloproteinase for activation of EGF-like factors, was significantly downregulated in Lgr4(-/-) ovaries. Matrix metalloproteinase 9 inhibitor treatment attenuated human chorionic gonadotropin- but not heparin-binding EGF-like growth factor-induced ERK activation and luteinization in primary granulosa cells. Together, we report that Lgr4 modulates WNT-mediated EGFR-ERK signaling to facilitate corpus luteum maturation and ovarian steroidogenesis to maintain female reproduction.


Chemical shifts for the unusual DNA structure in Pf1 bacteriophage from dynamic-nuclear-polarization-enhanced solid-state NMR spectroscopy.
Solid-state NMR spectra, including dynamic nuclear polarization enhanced 400 MHz spectra acquired at 100 K, as well as non-DNP spectra at a variety of field strengths and at temperatures in the range 213-243 K, have allowed the assignment of the (13)C and (15)N resonances of the unusual DNA structure in the Pf1 virion. The (13)C chemical shifts of C3' and C5', considered to be key reporters of deoxyribose conformation, fall near or beyond the edges of their respective ranges in available databases. The (13)C and (15)N chemical shifts of the DNA bases have above-average values for AC4, AC5, CC5, TC2, and TC5, and below average values for AC8, GC8, and GN2, pointing to an absence of Watson-Crick hydrogen bonding, yet the presence of some type of aromatic ring interaction. Crosspeaks between Tyr40 of the coat protein and several DNA atoms suggest that Tyr40 is involved in this ring interaction. In addition, these crosspeak resonances and several deoxyribose resonances are multiply split, presumably through the effects of ordered but differing interactions between capsid protein subunits and each type of nucleotide in each of the two DNA strands. Overall, these observations characterize and support the DNA model proposed by Liu and Day and refined by Tsuboi et al., which calls for the most highly stretched and twisted naturally occurring DNA yet encountered.


Bioreconstructive poly-L/D-lactide implant compared with Swanson prosthesis in metacarpophalangeal joint arthroplasty in rheumatoid patients: a randomized clinical trial.
It was hypothesized that the bioresorbable interposition implant might offer a viable alternative to conventional silicone implant arthroplasty in rheumatoid metacarpophalangeal joint destruction. A randomized clinical study was performed to compare a stemless poly-L/D-lactide copolymer 96 : 4 (PLDLA) implant with the Swanson silicone implant. Results in 52 patients (53 hands and 175 joints) at a mean follow-up of 2 years (minimum 1 year) showed that the improvement in clinical assessments was comparable in both groups, except for better maintenance of palmar alignment in the Swanson group. The lack of implant fractures and intramedullary osteolysis were advantages of the PLDLA implant. The bioresorbable PLDLA interposition implant may offer an alternative tool for tailored reconstruction of rheumatoid metacarpophalangeal joints.


Chitosan magnetic nanoparticles for pH responsive Bortezomib release in cancer therapy.
The use of nanotechnology in cancer treatment offers exciting opportunities, including the possibility of destroying tumors with minimal damage to healthy tissue by novel targeted drug delivery systems. pH differences between healthy and tumor microenvironment provide pH responsive release of drugs at tumor site via smart nanoparticles. In this study, chitosan coated superparamagnetic iron oxide nanoparticles (CS MNPs) were in situ synthesized by ionic crosslinking method as nanocarrier systems and loaded with the drug Bortezomib (Velcade(®)). The drug loading capacity, drug release and stability of CS MNPs were analyzed. CS MNPs were visualized inside the cells by fluorescence microscopy. The cytotoxicity of Bortezomib, CS MNPs and Bortezomib loaded CS MNPs were tested by XTT analyses in vitro. Gene expression analyses revealed that pro-apoptotic PUMA and NOXA genes were upregulated while anti-apoptotic BCL-2, SURVIVIN and cIAP-2 genes were downregulated at Bortezomib loaded CS MNP treated cells. Immunocytochemical analyses demonstrated an increase in p53 tumor suppressor protein levels at treated cells, which supports the upregulation of PUMA and NOXA genes, while Survivin protein level did not significantly change. This study points out that the pH responsive magnetic targeting of Bortezomib is more efficacious than free drug treatment. Moreover, targeted delivery of Bortezomib would reduce the frequency of drug administration by lowering the required amount of drug dose.


Fingerprint vibrational spectra of protonated methyl esters of amino acids in the gas phase.
Infrared spectra of the protonated monomers of glycine, alanine, valine, and leucine methyl esters are presented. These protonated species are generated in the gas phase via matrix assisted laser desorption ionization (MALDI) within the cell of a Fourier transform ion cyclotron resonance spectrometer (FTICR) where they are subsequently mass selected as the only species trapped in the FTICR cell. Alternatively, they have also been generated by electrospray ionization and transferred to a Paul ion-trap mass spectrometer where they are similarly isolated. In both cases IR spectra are then derived from the frequency dependence of the infrared multiple photon dissociation (IRMPD) in the mid-infrared region (1000-2200 cm(-1)), using the free electron laser facility Centre de Laser Infrarouge d'Orsay (CLIO). IR bands are assigned by comparison with the calculated vibrational spectra of the lowest energy isomers using density functional theory (DFT) calculations. There is in general good agreement between experimental IRMPD spectra and calculated IR absorption spectra for the lowest energy conformer which provides evidence for conformational preferences. The two different approaches to ion generation and trapping yield IRMPD spectra that are in excellent agreement.


The odd couple of protistology: Edward Heron-Allen (1861-1943) and Arthur Earland (1866-1958).
Edward Heron-Allen and Arthur Earland were among the last great amateur foraminifera researchers. Their partnership began in 1907 and ended in 1932. While close in age to one another, they shared little more than a fascination for forams and a lack of any university training. In most other aspects, the two men were completely different. Heron-Allen was a famous upper class polymath, expert not only on forams, but also on the Persian language, violins, palm reading, history, asparagus, and barnacles. He was also an accomplished novelist and poet, who frequented literary circles. In contrast to the flamboyant Heron-Allen, Earland was a discrete civil servant who admitted to working on forams as a relief from the monotony of his job. Hence, the two were improbable partners. However, together they produced 39 substantial works on forams. Their studies concerned assemblages from Southern Ocean to the North Sea and they are today credited with the original description of 186 species. Here the distinct lives of the two men are presented, and their contributions to protistology, as partners as well as individuals, are reviewed. The case is made for considering Earland's work as neglected relative to that of Heron-Allen, except perhaps by foram taxonomists.


Post-establishment movement of western corn rootworm larvae (Coleoptera: Chrysomelidae) in Central Missouri corn.
If registered, transgenic corn, Zea mays L., with corn rootworm resistance will offer a viable alternative to insecticides for managing Diabrotica spp. corn rootworms. Resistance management to maintain susceptibility is in the interest of growers, the Environmental Protection Agency, and industry, but little is known about many aspects of corn rootworm biology required for an effective resistance management program. The extent of larval movement by the western corn rootworm, Diabrotica virgifera virgifera LeConte, that occurs from plant-to-plant or row-to-row after initial establishment was evaluated in 1998 and 1999 in a Central Missouri cornfield. Post-establishment movement by western corn rootworm larvae was clearly documented in two of four treatment combinations in 1999 where larvae moved up to three plants down the row and across a 0.46-m row. Larvae did not significantly cross a 0.91-m row after initial host establishment in 1998 or 1999, whether or not the soil had been compacted by a tractor and planter. In the current experiment, western corn rootworm larvae moved from highly damaged, infested plants to nearby plants with little to no previous root damage. Our data do not provide significant insight into how larvae might disperse after initial establishment when all plants in an area are heavily damaged or when only moderate damage occurs on an infested plant. A similar situation might also occur if a seed mixture of transgenic and isoline plants were used and if transgenic plants with rootworm resistance are not repellent to corn rootworm larvae.


Drosophila fizzy-related down-regulates mitotic cyclins and is required for cell proliferation arrest and entry into endocycles.
We demonstrate that fizzy-related (fzr), a conserved eukaryotic gene, negatively regulates the levels of cyclins A, B, and B3. These mitotic cyclins that bind and activate cdk1(cdc2) are rapidly degraded during exit from M and during G1. While Drosophila fizzy has previously been shown to be required for cyclin destruction during M phase, fzr is required for cyclin removal during G1 when the embryonic epidermal cell proliferation stops and during G2 preceding salivary gland endoreduplication. Loss of fzr causes progression through an extra division cycle in the epidermis and inhibition of endoreduplication in the salivary gland, in addition to failure of cyclin removal. Conversely, premature fzr overexpression down-regulates mitotic cyclins, inhibits mitosis, and transforms mitotic cycles into endoreduplication cycles.


Adoptive immunotherapy of cancer: accomplishments and prospects.
Adoptive immunotherapy is an approach to cancer treatment that has not received significant evaluation in cancer patients. Recent developments in modern cellular immunology have expanded the opportunities for utilizing adoptive immunotherapy in humans. A variety of animal models have been developed utilizing the adoptive transfer of immune cells that can mediate the regression of established murine tumors. Analyses of these models are defining the important criteria necessary for the application of this approach to humans. Studies of the immune response of human cells to autologous tumors are identifying cells with antitumor reactivity that may be of value in the treatment of human malignancy. Preliminary clinical studies have been performed demonstrating the feasibility of infusing large numbers of activated lymphoid cells into humans. Adoptive immunotherapy is an approach to the treatment of cancer that deserves further study.


Body image, weight management behavior, and women's interest in cosmetic surgery.
In the current study, we examined body image and weight management behavior as potential factors associated with young women's interest in obtaining cosmetic surgery. Research focusing on objectification theory, problem behavior syndrome, and positive body image provide support for considering these factors as predictors of interest in cosmetic surgery. Female undergraduates (N = 170; M = 20.27 years old) at a college in the northeastern United States completed assessments of interest in cosmetic surgery, body image, and weight management behavior. Regression analyses indicated that women who had higher BMIs, higher body surveillance, higher body shame, and who engaged in more unhealthy weight management behaviors were more interested in obtaining cosmetic surgery. Body appreciation was significantly negatively correlated with interest in cosmetic surgery but did not significantly predict it when other factors were considered in a regression model. Results suggest that self-objectification and engaging in a constellation of appearance-related risk behaviors may place women at risk for pursuing cosmetic surgery.


Emergence of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae in the Central African Republic: genetic characterization.
Cross-resistance to quinolones and beta-lactams is frequent in Enterobacteriaceae, due to the wide use of these antibiotics clinically and in the food industry. Prescription of one of these categories of antibiotic may consequently select for bacteria resistant to both categories. Genetic mechanisms of resistance may be secondary to a chromosomal mutation located in quinolone resistance determining region of DNA gyrase or topoisomerase IV or to a plasmid acquisition. The insertion sequence ISCR1 is often associated with qnr and may favour its dissemination in Gram-negative bacteria. The aim of this study was to determine the genetic mechanism of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in the Central African Republic. Among seventeen ESBL-producing Enterobacteriaceae isolated from urine, pus or stool between January 2003 and October 2005 in the Central African Republic, nine were resistant to ciprofloxacin (seven from community patients and two from hospitalized patients). The ESBL were previously characterized as CTX-M-15 and SHV-12. Susceptibility to nalidixic acid, norfloxacin and ciprofloxacin, and the minimal inhibitory concentrations of these drugs were determined by disc diffusion and agar dilution methods, respectively. The presence of plasmid-borne ISCR1-qnrA region was determined by PCR and amplicons, if any, were sent for sequencing. Quinolone resistance determining region of DNA gyrase gyrA gene was amplified by PCR and then sequenced for mutation characterization. We found that all CTX-M-producing strains were resistant to the tested quinolones. All the isolates had the same nucleotide mutation at codon 83 of gyrA. Two Escherichia coli strains with the highest MICs were shown to harbour an ISCR1-qnrA1 sequence. This genetic association might favour dissemination of resistance to quinolone and perhaps other antibiotics among Enterobacteriaceae. This study shows that at least two mechanisms might explain the emerging resistance of Enterobacteriaceae to quinolones in the CAR. Beside the classical topoisomerase mutation, the cause may be acquisition of a plasmid-borne qnrA1. Clinicians and bacteriologists should be made aware of possible dissemination of ISCR1-qnrA1 among Enterobacteriacae.


Anti-inflammatory therapy for cardiovascular disease.
Chronic subclinical inflammation is a central process in the pathogenesis of cardiovascular disease (CVD) and it has been linked with both the initiation and progression of atherosclerosis. Several pro-inflammatory cytokines, such as the C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) have been described as independent risk factors for coronary heart disease and promoters of atherogenesis. Thus, extensive research is being conducted to assess the role of anti-inflammatory therapy in the primary and secondary prevention of CVD. Our review aims to provide the clinical and scientific data pertaining to the effects of different anti-inflammatory agents administered in patients with CVD.


Two-dimensional spatial and spatial-frequency selectivity of motion-sensitive mechanisms in human vision.
Thresholds for detecting the direction of motion of drifting (8-Hz) vertical gratings [of spatial frequencies 0.1, 1.0, and 10.0 cycles per degree (c/deg)] were measured in the presence of masks that varied in both spatial frequency and orientation. Masks with different temporal properties were used. The specificity of masking was also measured for a stationary test grating of spatial frequency 3.0 c/deg. After suitable scaling and transformation, the masking data gave an estimate of the two-dimensional spatial-frequency tuning surface of cortical detector units in human vision. With the assumption of small-signal linearity and zero phase, the tuning surfaces were inverse Fourier transformed to give an indication of the size and structure of the psychophysical receptive fields of detector units. The results obtained with drifting test gratings and jittering (random phase) mask gratings indicate that motion-detector receptive fields increase in size (in cycles) with increasing spatial frequency but, at all spatial scales, have a length-width ratio of 1. These results are in close agreement with the summation results reported in J. Opt. Soc. Am. A 8, 1330 (1991). Using the same jittering mask stimuli and stationary test gratings, we confirm reports by Daugman [Vision Res. 24, 891 (1984)] and Harvey and Doan [J. Opt. Soc. Am. A 7, 116 (1990)] that motion-independent units have elongated receptive fields with a length-width ratio near 1.8. We conclude that the receptive fields of motion-dependent and -independent mechanisms in human vision are fundamentally different. The possibility that the orientation selectivity of a motion unit is sharpened by its selectivity for direction of motion is discussed.


Interleukin-1beta upregulates PTHrP-mRNA expression and protein production and decreases TGF-beta in normal human osteoblast-like cells.
Osteoblasts are regulated by complex interactions among systemic hormones, cytokines, and local growth factors. Bone resorption, at the level of the basic multicellular unit, is initiated by stimulation of osteoblast activity. The stimulatory effect of interleukin-1beta (IL-1beta) on bone resorption has not been fully clarified. We have therefore studied the influence of IL-1beta on the local production and secretion of parathyroid hormone-related protein (PTHrP) and transforming growth factor-beta (TGF-beta) from normal human osteoblast-like cells (hOB cells). Using a quantitative PCR-assay following reverse transcription of RNA, in situ hybridization, and a two-site immunofluorometric assay for PTHrP, we demonstrate that IL-1beta in a dose- and time-dependent manner increases PTHrP-mRNA expression and PTHrP-protein secretion. In addition, IL-1beta decreased the TGF-beta protein concentration in conditioned medium. Our results suggest that the actions of IL-1beta on bone may be mediated by novel mechanisms involving both local increase of PTHrP, a potent stimulator of bone resorption, and a decrease of TGF-beta, an important anabolic and coupling factor for bone turnover.


Anti-angiogenic therapeutic strategies in hereditary hemorrhagic telangiectasia.
Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant vascular dysplastic disorder, characterized by recurrent nosebleeds (epistaxis), multiple telangiectases and arteriovenous malformations (AVMs) in major organs. Mutations in Endoglin (ENG or CD105) and Activin receptor-like kinase 1 (ACVRL1 or ALK1) genes of the TGF-β superfamily receptors are responsible for HHT1 and HHT2 respectively and account for the majority of HHT cases. Haploinsufficiency in ENG and ALK1 is recognized at the underlying cause of HHT. However, the mechanisms responsible for the predisposition to and generation of AVMs, the hallmark of this disease, are poorly understood. Recent data suggest that dysregulated angiogenesis contributes to the pathogenesis of HHT and that the vascular endothelial growth factor, VEGF, may be implicated in this disease, by modulating the angiogenic-angiostatic balance in the affected tissues. Hence, anti-angiogenic therapies that target the abnormal vessels and restore the angiogenic-angiostatic balance are candidates for treatment of HHT. Here we review the experimental evidence for dysregulated angiogenesis in HHT, the anti-angiogenic therapeutic strategies used in animal models and some patients with HHT and the potential benefit of the anti-angiogenic treatment for ameliorating this severe, progressive vascular disease.


Efficacy and safety of omitting chest drains after video-assisted thoracoscopic surgery: a systematic review and meta-analysis.
The aim of this systematic review and meta-analysis was to determine the efficacy and safety of omitting chest drains compared to routine chest drain placement after video-assisted thoracoscopic surgery (VATS). Five bibliographic databases, ClinicalTrials.gov and PROSPERO were comprehensively searched from inception to July 29, 2020 (no language restrictions). Postoperative outcomes were extracted and synthesized complying with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Risk of bias (RoB) including non-reporting bias, heterogeneity, and sensitivity were assessed. Subgroup analyses were performed based on study design. Of 7,166 identified studies, 10 studies [four randomized controlled trials (RCTs) and six non-RCTs] with 1,079 patients were included. There were 561 patients in the no chest drain group (NCD) and 518 patients in the standard chest drain group (CD). In pairwise analysis the NCD group had significant shorter length of stay (LOS) [weighted mean difference (WMD) -1.53 days, P<0.001], less postoperative pain scores (WMD -1.09, P=0.002), but higher risk of drain insertion or thoracocentesis [risk radio (RR) 3.02, P=0.040]. There were no significant differences on the incidence of minor pneumothorax (RR 1.77, P=0.128), minor pleural effusion (RR 1.88, P=0.219), minor subcutaneous emphysema (RR 1.37, P=0.427) or pneumonia (RR 0.53, P=0.549). No mortality was observed in either group during the observation period (in-hospital or 30-day mortality). Omitting chest drains in selected patients after VATS seems effective leading to enhanced recovery with shorter length of postoperative stay and less pain but with a higher risk of drain insertion or thoracocentesis. However, a major part of the evidence comes from observational studies with high RoB. Further RCTs are needed to improve the current evidence.


Diagnostic and Prognostic Significance of Ki-67 Immunohistochemical Expression in Surface Epithelial Ovarian Carcinoma.
The Surface Epithelial Ovarian Carcinoma (SEOC) at the moment of diagnosis, the disease is extended beyond the structures of the pelvis. Ki-67 is one of the prognostic marker which determines the growth fraction of a tumour and its over expression is associated with malignancy, tumour aggression, reserved prognosis and metastasis. To evaluate the proliferative activity using Ki-67 immuno-staining in SEOC and to correlate with histological subtype, grade, Federation of Gynecology and Obstetrics (FIGO) stage, CA125 levels for diagnostic and prognostic purpose. The study was conducted in JSS Medical College and Hospital, JSS University, Mysuru. It was a descriptive cross-sectional study involving 40 cases of SEOC over a period of two years. The proliferation expression related to Ki-67 antigen was evaluated by immunohistochemical monoclonal MIB-1 antibody. In each case, the Ki-67 labeling index (Ki-67 LI) was articulated as percentage of positively stained cells using high power objective of the microscope (x400). Among the 40 carcinomas, 26 were serous, five mucinous, four each of clear cell and undifferentiated and one transitional cell carcinoma. A total of 75% were high grade tumours. High Ki-67 LI was associated with high grade tumours (69.9%), high grade serous tumours (65.34%) and advanced FIGO staging (70.6%) with the p-value of <0.001. CA 125 levels did not have a significant correlation with Ki-67 LI. Ki-67 is an exceptionally a cost effective marker to determine the growth fraction of a tumour cell population. In SEOC histological grade and FIGO stage when combined with Ki-67 LI in histopathology report would help in diagnostic differentiation of subtypes, prognostication, deciding the need for adjuvant chemotherapy and in predicting survival analysis.


Phenotypic correction of ornithine transcarbamylase deficiency using low dose helper-dependent adenoviral vectors.
Helper-dependent adenoviral vectors (HDAd) can mediate long-term phenotypic correction in the ornithine transacarbamylase (OTC)-deficient mice model with negligible chronic toxicity. However, the high doses required for metabolic correction will result in systemic inflammatory response syndrome in humans. This acute toxicity represents the major obstacle for clinical applications of HDAd vectors for the treatment of OTC deficiency. Strategies for reducing the dose necessary for disease correction are highly desirable because HDAd acute toxicity is clearly dose-dependent. We analysed a potent expression cassette and the hydrodynamic injection for the ability to reduce the HDAd dose necessary for phenotypic correction in OTC-deficient spf-ash mice. We have developed a vector containing a potent expression cassette expressing the OTC transgene, which allowed phenotypic correction at lower doses. Our results suggest that vector expressing greater OTC levels allows correction of orotic acid overproduction at lower doses that make clinical translation more relevant. We were able to further reduce the minimal effective dose by delivering the vector through the hydrodynamic injection technique. Vectors containing the expression cassette used in the present study, combined with other strategies for improving HDAd therapeutic index, will likely permit application of these vectors for the treatment of OTC deficiency as well as other urea cycle disorders.


Ethnic and socioeconomic differences in variability in nutritional biomarkers.
Several studies have reported ethnic, education, and income differentials in concentrations of selected nutritional biomarkers in the US population. Although biomarker measurements are not subject to biased self-reports, biologic variability due to individual characteristics and behaviors related to dietary exposures contributes to within-subject variability and measurement error. We aimed to establish whether the magnitude of components of variance for nutritional biomarkers also differs in these high-risk groups. We used data from 2 replicate measurements of serum concentrations of vitamins A, C, D, and E; folate; carotenoids; ferritin; and selenium in the third National Health and Nutrition Examination Survey second examination subsample (n = 948) to examine the within-subject and between-subject components of variance. We used multivariate regression methods with log-transformed analyte concentrations as outcomes to estimate the ratios of the within-subject to between-subject components of variance by categories of ethnicity, income, and education. In non-Hispanic blacks, the within-subject to between-subject variance ratio for beta-cryptoxanthin concentration was higher (0.23; 95% CI: 0.17, 0.29) relative to non-Hispanic whites (0.13; 0.11, 0.16) and Mexican Americans (0.11; 0.07, 0.14), and the lutein + zeaxanthin ratio was higher (0.29; 0.21, 0.38) relative to Mexican Americans (0.15; 0.10, 0.19). Higher income was associated with larger within-subject to between-subject variance ratios for serum vitamin C and red blood cell folate concentrations but smaller ratios for serum vitamin A. Overall, there were few consistent up- or down-trends in the direction of covariate-adjusted variability by ethnicity, income, or education. Population groups at high risk of adverse nutritional profiles did not have larger variance ratios for most of the examined biomarkers.


Analysis of Edg-Like LPA Receptor-Ligand Interactions.
The phospholipid derivative lysophosphatidic acid (LPA) serves as a signalling molecule through the activation of LPA receptors, which belong to the G-protein-coupled receptors. From a pharmacological point of view, the ('EDG-like') LPA1-3 receptors have attracted much attention, therefore we have also been focusing in our study on these subtypes. The LPA1receptors are widely expressed in the human body; interestingly, LPA1 might have a role in the pathomechanism of obesity. In order to recognize key structural features of the molecular interactions of human LPA1with its agonists, we built up the 3D structure of the LPA1 through homology modeling. Next, LPA1 agonists and antagonists were docked into the model. The mode of binding and the interactions between ligands and key amino acids (R3.28 and Q3.29) were consistent with mutagenesis assays and previously published models, indicating that this model is able to discriminate high-affinity compounds and may be useful for the development of novel agonists of LPA1. Homology models were also constructed for LPA2 and LPA3. All available agonists with published EC50 values, antagonists with IC50 values and compounds with Ki values for either of LPA1, LPA2 or LPA3 were collected from the ChEMBL database and were docked into the corresponding model.Ourmodels for the LPA1-3 receptors can discriminate high-affinity compounds identified in silico HTS studies and may be useful for the development of novel agonistsof LPA receptors. With a better understanding of the differences between LPA1-3 receptors new, selective agonists and antagonist could be designed, which could be used in the therapy of various diseases with a better side-effect profile.


Do Psychological Interventions Work for Psychosis in Adolescents?
There is emerging evidence to show that psychological interventions such as cognitive remediation therapy (CRT), psychoeducation, family therapy, and group psychotherapies may be useful for adolescents with psychosis. The current review is on the effects of various psychological interventions for adolescents with psychosis compared with treatment as usual (TAU) or other psychological interventions. We undertook a comprehensive search for all randomized controlled trials on the topic as per predefined criteria. For binary data, a standard estimation of risk ratio, and, for continuous data, the mean difference between groups were estimated. GRADE approach was used to assess studies. "Risk of Bias" was calculated, and finally random-effects model was used for analyses. The review included 7 studies (n = 317). Two studies compared CRT and TAU with TAU alone. CRT showed improvement in short-term memory compared with those in the TAU group (relative risk 0.58, 95% CI 0.37 to 0.89, participants = 31, very low-certainty evidence). When comparing group psychosocial therapy with TAU, global state scores measured using Children's Global Assessment Scale (CGAS) were clearly higher in the intervention arm (mean difference 5.10, 95% CI 1.35 to 8.85, participants = 56, very low-certainty evidence) as compared with the TAU group. None of the other interventions were found to be significantly effective for the treatment of psychosis in adolescents. Evidence suggests that psychological interventions may have beneficial effects in the treatment of adolescents with psychosis, but the evidence is of low or very low certainty.


[A case report of cerebral amyloid angiopathy-related inflammation treated with cyclophosphamide].
A 78-year-old woman was admitted to another hospital with progressive cognitive dysfunction and right hemiparesis. Magnetic resonance imaging (MRI) of the brain revealed a high-intensity area in bilateral frontal lobes on T2-weighted imaging (T2WI) and fluid-attenuated inversion recovery (FLAIR). Brain biopsy revealed no significant findings. She was transferred to our hospital for further examination. Although the etiology of the white matter lesion was not specified, the lesion and symptoms improved with steroid pulse therapy. Three months later, disturbance of consciousness and right hemiparesis worsened again, and the white matter lesion was expanded on MRI. Steroid pulse therapy was again performed, but her condition gradually deteriorated and akinetic mutism developed. T2-based imaging showed microbleeds and analysis of the apolipoprotein E (ApoE) genotype showed ε4/ε4; therefore, cerebral amyloid angiopathy-related inflammation was diagnosed. Two cycles of cyclophosphamide pulse therapy were performed, resulting in improvement of the white matter lesion, disorder of consciousness, and hemiparesis after the first cycle. The patient was discharged from our hospital in an ambulatory state. Steroid therapy for cerebral amyloid angiopathy-related inflammation is relatively well-known in Japan, but cyclophosphamide therapy appears useful for steroid-resistant amyloid angiopathy-related inflammation in the early stage of the disease.


Crisis centers and their role in treatment: suicide prevention versus health promotion.
Suicide is a major health problem. In the industrialized nations, it remains among the 10 leading causes of death. There has been a massive effort to reduce suicide mortality through establishment of prevention centers. These centers have proliferated despite a lack of evidence concerning their efficacy. The development and evaluation of suicide prevention centers, primarily community-based, telephone interventions, are discussed. The evidence regarding the effectiveness of suicide prevention centers and the presence of a causal relationship between exposure to crisis counseling and a reduction in suicide rates is reviewed. Some cautions and recommendations are made. Finally, a model of health promotion program planning and evaluation is examined in the context of suicide prevention.


Medical informatics and problem-based learning in conjunction.
At Dalhousie, we integrated medical informatics as a horizontal theme in the problem-based learning undergraduate medical education curriculum. The rationales for these initiatives can be addressed along the following dimensions: political, philosophical, psychological, educational, and professional practice. Student attitudes towards computers are changing, and incoming students are increasingly computer-literate. Of those Med I students surveyed at the end of the 1993-94 academic year, 94% felt that computers were moderately important or essential to medical education; 86% of Med II students surveyed were of the same opinion. The education of future physicians can be enhanced when medical informatics is introduced into a curriculum in conjunction with the problem-based learning approach.


Guided-mode resonant polarization-controlled tunable color filters.
We demonstrate efficient guided-mode resonant polarization-controlled tunable color filters. The devices consist of subwavelength gratings that are partially etched into a thin silicon-nitride film deposited on a glass substrate. Two color filters with grating periods of 300 nm and 370 nm are designed and fabricated. The 300-nm device exhibits green and blue colors and the 370-nm device generates red and yellow colors for TE and TM polarization, respectively. The pixels have a spectral bandwidth of ~12 nm with efficiencies exceeding 90% for TE polarization and 80% for TM polarization. The devices may find application in displays, image sensors, and biomedical imaging technologies.


Exploring valence bias as a metric for frontoamygdalar connectivity and depressive symptoms in childhood.
Negativity bias is a core feature of depression that is associated with dysfunctional frontoamygdalar connectivity; this pathway is associated with emotion regulation and sensitive to neurobiological change during puberty. We used a valence bias task (ratings of emotional ambiguity) as a potential early indicator of depression risk and differences in frontoamygdalar connectivity. Previous work using this task demonstrated that children normatively have a negative bias that attenuates with maturation. Here, we test the hypothesis that persistence of this negativity bias as maturation ensues may reveal differences in emotion regulation development, and may be associated with increased risk for depression. In children aged 6-13 years, we tested the moderating role of puberty on relationships between valence bias, depressive symptoms, and frontoamygdalar connectivity. A negative bias was associated with increased depressive symptoms for those at more advanced pubertal stages (within this sample) and less regulatory frontoamygdalar connectivity, whereas a more positive bias was associated with more regulatory connectivity patterns. These data suggest that with maturation, individual differences in positivity biases and associated emotion regulation circuitry confer a differential risk for depression. Longitudinal work is necessary to determine the directionality of these effects and explore the influence of early life events.


Alterations in auditory brain stem response distinguish occasional and constant tinnitus.
BACKGROUNDThe heterogeneity of tinnitus is thought to underlie the lack of objective diagnostic measures.METHODSLongitudinal data from 20,349 participants of the Swedish Longitudinal Occupational Survey of Health (SLOSH) cohort from 2008 to 2018 were used to understand the dynamics of transition between occasional and constant tinnitus. The second part of the study included electrophysiological data from 405 participants of the Swedish Tinnitus Outreach Project (STOP) cohort.RESULTSWe determined that with increasing frequency of the occasional perception of self-reported tinnitus, the odds of reporting constant tinnitus after 2 years increases from 5.62 (95% CI, 4.83-6.55) for previous tinnitus (sometimes) to 29.74 (4.82-6.55) for previous tinnitus (often). When previous tinnitus was reported to be constant, the odds of reporting it as constant after 2 years rose to 603.02 (524.74-692.98), suggesting that once transitioned to constant tinnitus, the likelihood of tinnitus to persist was much greater. Auditory brain stem responses (ABRs) from subjects reporting nontinnitus (controls), occasional tinnitus, and constant tinnitus show that wave V latency increased in constant tinnitus when compared with occasional tinnitus or nontinnitus. The ABR from occasional tinnitus was indistinguishable from that of the nontinnitus controls.CONCLUSIONSOur results support the hypothesis that the transition from occasional to constant tinnitus is accompanied by neuronal changes in the midbrain leading to a persisting tinnitus, which is then less likely to remit.FUNDINGThis study was supported by the GENDER-Net Co-Plus Fund (GNP-182), the European Union's Horizon 2020 grants no. 848261 (Unification of Treatments and Interventions for Tinnitus [UNITI]) and no. 722046 (European School for Interdisciplinary Tinnitus Research [ESIT]).


Visual recognition memory. Neurophysiological evidence for the role of temporal white matter in man.
A novel event-related potential (ERP) elicited by a visuospatial recognition memory task was recorded in 20 patients with temporal lobe epilepsy using depth electrodes sited in the temporal lobes. The ERPs comprised two components, an N400 and a P600, and were similar in morphology to the previously reported ERP to verbal recognition memory tasks. The two ERP components in both verbal and visuospatial tasks were dependent on stimulus type and our data suggest that they do not simply represent delayed P300 ERP responses. In 17/20 patients robust, reliable bilaterally present ERPs were elicited by both verbal and visuospatial memory tasks. N400 amplitude was larger in response to novel stimuli, whereas P600 amplitude was larger to repeated stimuli. P600 amplitude was larger in the right temporal lobe to both visuospatial and verbal stimulus material. N400 and P600 latencies did not vary with task, stimulus type or side of recording. In 3/20 patients, no ERPs were elicited by either memory task. In all 3 cases, unilateral temporal white matter abnormalities were demonstrated by magnetic resonance imaging. Behavioural measures, expressed in the form of standardized accuracy scores, did not differ from those of a normal control group, and hence are unlikely to account for the abnormalities in ERPs. These results are discussed with reference to the primate visual recognition memory pathway and suggest that ERPs to recognition memory tasks are generated by an interaction between the two homologous inferotemporal recognition memory pathways.


Neurological principles and rehabilitation of action disorders: rehabilitation interventions.
This third chapter discusses the evidence for the rehabilitation of the most common movement disorders of the upper extremity. The authors also present a framework, building on the computation, anatomy, and physiology (CAP) model, for incorporating some of the principles discussed in the 2 previous chapters by Frey et al and Sathian et al in the practice of rehabilitation and for discussing potentially helpful interventions based on emergent neuroscience principles.


Midazolam inhibits hippocampal long-term potentiation and learning through dual central and peripheral benzodiazepine receptor activation and neurosteroidogenesis.
Benzodiazepines (BDZs) enhance GABA(A) receptor inhibition by direct actions on central BDZ receptors (CBRs). Although some BDZs also bind mitochondrial receptors [translocator protein (18 kDa) (TSPO)] and promote the synthesis of GABA-enhancing neurosteroids, the role of neurosteroids in the clinical effects of BDZs is unknown. In rat hippocampal slices, we compared midazolam, an anesthetic BDZ, with clonazepam, an anticonvulsant/anxiolytic BDZ that activates CBRs selectively. Midazolam, but not clonazepam, increased neurosteroid levels in CA1 pyramidal neurons without changing TSPO immunostaining. Midazolam, but not clonazepam, also augmented a form of spike inhibition after stimulation adjacent to the pyramidal cell layer and inhibited induction of long-term potentiation. These effects were prevented by finasteride, an inhibitor of neurosteroid synthesis, or 17PA [17-phenyl-(3α,5α)-androst-16-en-3-ol], a blocker of neurosteroid effects on GABA(A) receptors. Moreover, the synaptic effects were mimicked by a combination of clonazepam with FGIN (2-[2-(4-fluorophenyl)-1H-indol-3-yl]-N,N-dihexylacetamide), a selective TSPO agonist, or a combination of clonazepam with exogenous allopregnanolone. Consistent with these in vitro results, finasteride abolished the effects of midazolam on contextual fear learning when administrated 1 d before midazolam injection. Thus, dual activation of CBRs and TSPO appears to result in unique actions of clinically important BDZs. Furthermore, endogenous neurosteroids are shown to be important regulators of pyramidal neuron function and synaptic plasticity.


Distribution of calcium-activated protease activity and endogenous substrates in the squid nervous system.
Calcium-activated protease (CAP) activity was studied in various neural tissues of the squid using endogenous (neurofilament protein) and exogenous ([14C]casein) substrate assays. Both assays demonstrated a significant CAP activity in perikarya from stellate ganglia, in axoplasm extruded from the giant axon, and in squid retinal fibers. The endogenous protein substrates in the perikarya and axons were 60,000 and 200,000 daltons, respectively. The Km for the CAP degradation of [14C] casein in axoplasm was about 2 x10-6 m. In contrast, both assays detected no CAP activity nor endogenous substrate in nerve terminals (synaptosomes from squid optic lobe). The absence of both CAP activity and endogenous substrate in nerve endings suggests that the axonal neurofilaments are degraded by CAP at the axon-nerve ending junction, followed by an autoinactivation of the CAP. Consistent with this hypothesis is that exposure of axoplasmic CAP to calcium leads to a rapid degradation of axonal neurofilament protein (t 1/2 less than 2 min) and a slower inactivation of the CAP (t 1/2 = 90 min). Axonal CAP requires a relatively high concentration of CA2+ sensitivity form of CAP found in other tissues.