Effects of aging and target location on reaction time and accuracy of lateral precision stepping during walking.
Older adults have poorer lateral balance and deficits in precision stepping accuracy, but the way these deficits manifest with lateral step distance is unclear. The purpose of this study was to investigate aging effects on lateral precision stepping performance in reaction to near and distant foot placement targets during treadmill walking. We hypothesized that older adults would step to targets later and less accurately than young adults, and that these difference would be more pronounced for distant targets. During the study, young and older adults stepped on lateral targets projected onto the surface of a treadmill one stride prior to their targeting step. We measured stepping accuracy to the target, the time when the swing foot diverged from its normal swing trajectory, and swing phase gluteus medius activity. Both groups had similar performance stepping to near targets, suggesting that giving older subjects a full stride to react to target location mitigates visuomotor processing delays that have contributed to deficits in stepping performance in prior studies. However, when stepping to distant targets, older adults had larger errors and later divergence times than young adults. This suggests that age-related deficits other than those in visuomotor processing contribute to poorer performance for more difficult stepping tasks. Furthermore, while young adults increased early swing gluteus medius activity with lateral target distance, older adults did not. This is the first study to show a potential neuromuscular basis for precision stepping deficits in older adults.


Stimulation of the thiol-dependent ADP-ribosyltransferase and NAD glycohydrolase activities of Bordetella pertussis toxin by adenine nucleotides, phospholipids, and detergents.
Pertussis toxin catalyzed ADP-ribosylation of the guanyl nucleotide binding protein transducin was stimulated by adenine nucleotide and either phospholipids or detergents. To determine the sites of action of these agents, their effects were examined on the transducin-independent NAD glycohydrolase activity. Toxin-catalyzed NAD hydrolysis was increased synergistically by ATP and detergents or phospholipids; the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was more effective than the nonionic detergent Triton X-100 greater than lysophosphatidylcholine greater than phosphatidylcholine. The A0.5 for ATP in the presence of CHAPS was 2.6 microM; significantly higher concentrations of ATP were required for maximal activation in the presence of cholate or lysophosphatidylcholine. In CHAPS, NAD hydrolysis was enhanced by ATP greater than ADP greater than AMP greater than adenosine; ATP was more effective than MgATP or the nonhydrolyzable analogue adenyl-5'-yl imidodiphosphate. GTP and guanyl-5'-yl imidodiphosphate were less active than the corresponding adenine nucleotides. Activity in the presence of CHAPS and ATP was almost completely dependent on dithiothreitol; the A0.5 for dithiothreitol was significantly decreased by CHAPS alone and, to a greater extent, by CHAPS and ATP. To determine the site of action of ATP, CHAPS, and dithiothreitol, the enzymatic (S1) and binding components (B oligomer) were resolved by chromatography. The purified S1 subunit catalyzed the dithiothreitol-dependent hydrolysis of NAD; activity was enhanced by CHAPS but not ATP. The studies are consistent with the conclusion that adenine nucleotides, dithiothreitol, and CHAPS act on the toxin itself rather than on the substrate; adenine nucleotides appear to be involved in the activation of toxin but not the isolated catalytic unit.


[Toxicity of dioxins in humans].
Dioxins are produced as pollutants in industrial production processes and during incineration (e.g. of waste). Thanks to their highly lipophilic nature and long half-life in fat tissue, they accumulate in the food chain. One of the most important initial steps in the toxicity of dioxins is the activation of a cytosol receptor. This induces the production of a number of proteins, including cytochrome P450 CYP1A1. Our knowledge about the effects of dioxins in humans is based primarily on long-term epidemiological studies of people who were exposed to high concentrations of dioxins, either occupationally or as the result of an accident. The best known and most definitely established effect of exposure to dioxins is chloracne. Whether there is a relation between exposure to dioxins and other diseases, such as cancer, is uncertain.


β sheets not required: combined experimental and computational studies of self-assembly and gelation of the ester-containing analogue of an Fmoc-dipeptide hydrogelator.
In our work toward developing ester-containing self-assembling peptides as soft biomaterials, we have found that a fluorenylmethoxycarbonyl (Fmoc)-conjugated alanine-lactic acid (Ala-Lac) sequence self-assembles into nanostructures that gel in water. This process occurs despite Fmoc-Ala-Lac's inability to interact with other Fmoc-Ala-Lac molecules via β-sheet-like amide-amide hydrogen bonding, a condition previously thought to be crucial to the self-assembly of Fmoc-conjugated peptides. Experimental comparisons of Fmoc-Ala-Lac to its self-assembling peptide sequence analogue Fmoc-Ala-Ala using a variety of microscopic, spectroscopic, and bulk characterization techniques demonstrate distinct features of the two systems and show that while angstrom-scale self-assembled structures are similar, their nanometer-scale size and morphological properties diverge and give rise to different bulk mechanical properties. Molecular dynamics simulations were performed to gain more insight into the differences between the two systems. An analysis of the hydrogen-bonding and solvent-surface interface properties of the simulated fibrils revealed that Fmoc-Ala-Lac fibrils are stronger and less hydrophilic than Fmoc-Ala-Ala fibrils. We propose that this difference in fibril amphiphilicity gives rise to differences in the higher-order assembly of fibrils into nanostructures seen in TEM. Importantly, we confirm experimentally that β-sheet-type hydrogen bonding is not crucial to the self-assembly of short, conjugated peptides, and we demonstrate computationally that the amide bond in such systems may act mainly to mediate the solvation of the self-assembled single fibrils and therefore regulate a more extensive higher-order aggregation of fibrils. This work provides a basic understanding for future research in designing highly degradable self-assembling materials with peptide-like bioactivity for biomedical applications.


Peptide transport by the multidrug resistance protein MRP1.
Small hydrophobic peptides were studied as possible substrates of the multidrug resistance protein (MRP)-1 (ABCC1) transmembrane transporter molecule. As observed earlier for P-glycoprotein- (Pgp; ABCB1) overexpressing cells, MRP1-overexpressing cells, including cells stably transfected with the MRP1 cDNA, showed distinct resistance to the cytotoxic peptide N-acetyl-Leu-Leu-norleucinal (ALLN). Resistance to this peptide and another toxic peptide derivative, which is based on a Thr-His-Thr-Nle-Glu-Gly backbone conjugated to butyl and benzyl groups (4A6), could be reversed by MRP1 inhibitors. The reduced toxicity of 4A6 in MRP1-overexpressing cells was found to be associated with lower accumulation of a fluorescein-labeled derivative of this peptide. Glutathione (GSH) depletion had a clear effect on resistance to ALLN but hardly affected 4A6 resistance. In a limited structure-activity study using peptides that are analogous to 4A6, MRP1-overexpressing cells were found to be resistant to these peptides as well. Remarkably, when selecting A2780 ovarian cancer cells for resistance to ALLN, even in the absence of Pgp blockers, resulting cell lines had up-regulated MRP1, rather than any of the other currently known multidrug resistance transporter molecules including Pgp, MRP2 (ABCC2), MRP3 (ABCC3), MRP5 (ABCCS), and the breast cancer resistance protein ABCG2. ALLN-resistant, MRP1-overexpressing cells were found to be cross-resistant to 4A6 and the classical multidrug resistance drugs doxorubicin, vincristine, and etoposide. This establishes MRP1 as a transporter for small hydrophobic peptides. More extensive structure-activity relationship studies should allow the identification of clinically useful peptide antagonists of MRP1.


Manipulation of Ovarian Function Significantly Influenced Trabecular and Cortical Bone Volume, Architecture and Density in Mice at Death.
Previously, transplantation of ovaries from young, cycling mice into old, postreproductive-age mice increased life span and decreased cardiomyopathy at death. We anticipated that the same factors that increased life span and decreased cardiomyopathy could also influence the progression of orthopedic disease. At 11 months of age, prepubertally ovariectomized and ovary-intact mice (including reproductively cycling and acyclic mice) received new 60-day-old ovaries. At death, epiphyseal bone in the proximal tibia and the distal femur and mid-shaft tibial and femoral diaphyseal bone was analyzed with micro-computed tomography. For qualitative analysis of osteophytosis, we also included mineralized connective tissue within the stifle joint. Prepubertal ovariectomy had the greatest influence on bone volume, ovarian transplantation had the greatest influence on bone architecture and both treatments influenced bone density. Ovarian transplantation increased cortical, but not trabecular bone density and tended to increase osteophytosis and heterotopic mineralization, except in acyclic recipients. These effects may have been dictated by the timing of the treatments, with ovariectomy appearing to influence early development and ovarian transplantation limited to influencing only the postreproductive period. However, major differences observed between cycling, acyclic and ovariectomized recipients of new ovaries may have been, in part due to differences in the levels of hormone receptors present and the responsiveness of specific bone processes to hormone signaling. Changes that resulted from these treatments may represent a compensatory response to normal age-associated, negative, orthopedic changes. Alternatively, differences between treatments may simply be the 'preservation' of unblemished orthopedic conditions, prior to the influence of negative, age-associated effects. These findings may suggest that in women, tailoring hormone replacement therapy to the patient's current reproductive status may improve therapy effectiveness and that beginning therapy earlier may help preserve trabecular bone mineral density that would otherwise be lost during perimenopause.


The natural virome and pandemic potential: Disease X.
Over the last decade, the emergence of several zoonotic viruses has demonstrated that previously unknown or neglected pathogens have the potential to cause epidemics and therefore to pose a threat to global public health. Even more concerning are the estimated 1.7 million still-undiscovered viruses present in the natural environment or 'global virome', with many of these as-yet uncharacterized viruses predicted to be pathogenic for humans. Thus, in order to mitigate disease emergence and prevent future pandemics, it is crucial to identify the global extent of viral threats to which humans may become exposed. This requires cataloguing the viruses that exist in the environment within their various and diverse host species, and also understanding the viral, host, and environmental factors that dictate the circumstances that result in viral spillover into humans. We also address here which strategies can be implemented as countermeasure initiatives to reduce the risk of emergence of new diseases.


Patient and public understanding and knowledge of antimicrobial resistance and stewardship in a UK hospital: should public campaigns change focus?
The rising global tide of antimicrobial resistance is a well-described phenomenon. Employing effective and innovative antimicrobial stewardship strategies is an essential approach to combat this public health threat. Education of the public and patients is paramount to enable the success of such strategies. A panel of hospital multidisciplinary healthcare professionals was set up and a short quiz containing true/false statements around antimicrobial stewardship and resistance was designed and piloted. An educational leaflet with the correct replies and supporting information was also produced and disseminated. Participants were recruited on a single day (18 November 2015) from the hospital outpatient clinics and the hospital outpatient pharmacy waiting room. One hundred and forty-five completed quizzes were returned, providing a total of 1450 answers. Overall, 934 of 1450 (64%) statements were scored correctly whilst 481 (33%) were scored incorrectly; 35 (3%) statements were left unscored. We speculate that these results may demonstrate that respondents understood the statements, as only a small proportion of statements were left unanswered. The question dealing with the definition of antimicrobial resistance and the question dealing with the definition of antimicrobial stewardship obtained the most incorrect replies (85% and 72%, respectively). However, a specific factual recall question regarding only one microorganism (MRSA) received the most correct responses (99%). We describe a simple, innovative method of engagement with patients and the general public to help educate and disseminate important public health messages around antimicrobial resistance and stewardship. We also identified the need for public health campaigns to address the knowledge gaps found around this topic.


Cell Killing by Reovirus: Mechanisms and Consequences.
Infection of host cells by mammalian reovirus in culture or in tissues of infected animals results in cell death. Cell death of infected neurons and myocytes contributes to the pathogenesis of reovirus-induced encephalitis and myocarditis in a newborn mouse model. Thus, reovirus-induced cell death has been used to investigate the basis of viral disease. Depending on the cell type, infection of host cells by reovirus results in one of two forms of cell death-apoptosis and necroptosis. In addition to the obvious differences in how these two forms of cell death are executed, the mechanisms by which reovirus infection initiates and transduces signals that lead to each of these types of cell death are distinct. In this review, we discuss how apoptosis and necroptosis are triggered by events at different stages of infection. We also describe how innate immune recognition of reovirus genomic material and type I interferon signaling pathways connect with the core components of the apoptosis and necroptosis machinery. The impact of different cell death mediators on viral pathogenesis and the potential of reovirus as an oncolytic vector are also outlined.


[Initial analysis of the results of the genome-wide search for the genes responsive to interferon alpha].
The paper is focused on the primary analysis of the potential genes of primary response to IFNalpha, predicted by the program COTRASIF elaborated in our laboratory. Two web-instruments FatiGO and GOTM were applied for this purpose; the literature search was carried out using IHOP; genes were classified according to the conceptual diagram "Gene Ontology" (GO) and were ranked according to their first priority for the experimental validation. On the basis of conducted analysis 162 genes of potential primary response to IFNalpha were subdivided into two groups--61 genes, for which the experimental data of their responsibility to IFNalpha were found and 101 genes for which such data are absent. We have performed the functional analysis of the "unknown" genes according to GO. The enriched category "Synapse part" encountering three genes of Rattus norvegicus: PRKCA-binding of protein, NMDAR-L and GABAA--receptor subunit alpha-2 were revealed. Among 162 genes the enriched category "Immune response" was detected with Mbl1 gene, which is reckoned among "unknown" genes. The four designated genes are chosen as the candidates for the prior validation.


On the Constitutionality of Hard State Border Closures in Response to the COVID-19 Pandemic.
I investigate the constitutionality of hard state border closures in the United States as a prophylactic response to a pandemic. This type of border closure prevents people from entering a State, except for exempt travelers, a category that includes, for example, military, judicial and government officers, and people granted entry on compassionate grounds. Those allowed to enter usually have to then go through a quarantine regime before being released into the community. During the COVID-19 pandemic, no State has attempted such closures. However, epidemiological experts suggest that, in comparison to other border and non-border measures, such closures are more effective. Given the World Health Organization prediction of more pandemics in the foreseeable future, it is imperative that the constitutionality of such hard closures is investigated. I use structural analysis to argue that a recent challenge to hard border closures in Australia suggests that, under a strict scrutiny review, the Supreme Court is likely to uphold such closures in the United States. While actual implementation requires investigating issues that go beyond a constitutional analysis, these findings highlight the need for a wider conversation around a federal goldilocks zone when responding to the next pandemic.


IDH1 Targeting as a New Potential Option for Intrahepatic Cholangiocarcinoma Treatment-Current State and Future Perspectives.
Cholangiocarcinoma is a primary malignancy of the biliary tract characterized by late and unspecific symptoms, unfavorable prognosis, and few treatment options. The advent of next-generation sequencing has revealed potential targetable or actionable molecular alterations in biliary tumors. Among several identified genetic alterations, the IDH1 mutation is arousing interest due to its role in epigenetic and metabolic remodeling. Indeed, some IDH1 point mutations induce widespread epigenetic alterations by means of a gain-of-function of the enzyme, which becomes able to produce the oncometabolite 2-hydroxyglutarate, with inhibitory activity on α-ketoglutarate-dependent enzymes, such as DNA and histone demethylases. Thus, its accumulation produces changes in the expression of several key genes involved in cell differentiation and survival. At present, small-molecule inhibitors of IDH1 mutated enzyme are under investigation in preclinical and clinical phases as promising innovative treatments for IDH1-mutated intrahepatic cholangiocarcinomas. This review examines the molecular rationale and the results of preclinical and early-phase studies on novel pharmacological agents targeting mutant IDH1 in cholangiocarcinoma patients. Contextually, it will offer a starting point for discussion on combined therapies with metabolic and epigenetic drugs, to provide molecular support to target the interplay between metabolism and epigenetics, two hallmarks of cancer onset and progression.


FLOWERING LOCUS C EXPRESSOR family proteins regulate FLOWERING LOCUS C expression in both winter-annual and rapid-cycling Arabidopsis.
Many naturally occurring Arabidopsis (Arabidopsis thaliana) are very late flowering, unless flowering is promoted by a prolonged period of cold (e.g. winter) known as vernalization. In these winter-annual strains, flowering prior to winter is blocked by the synergistic interaction of FRIGIDA (FRI) and FLOWERING LOCUS C (FLC). FLC acts as a strong floral inhibitor, and FRI is required for high levels of FLC expression. Vernalization, in turn, leads to an epigenetic down-regulation of FLC expression. Most rapid-cycling Arabidopsis carry loss-of-function mutations in FRI, leading to low levels of FLC and rapid flowering in the absence of vernalization. Recent work has shown that FRI acts as a scaffolding protein for the assembly of a FRI complex (FRI-C) that includes both general transcription and chromatin-modifying factors, as well as FRI-specific components such as FRI-LIKE1, FRI ESSENTIAL1 (FES1), SUPPRESSOR OF FRI4 (SUF4), and FLC EXPRESSOR (FLX). Here, we show that FLX-LIKE4 (FLX4) is a novel component of the FRI-C and is essential for the activation of FLC by FRI. Both FLX and FLX4 contain leucine zipper domains that facilitate interaction with FRI. In addition, FLX and FLX4 interact with each other and show synergistic transcription activation activity. Interestingly, we show that FLX, FLX4, FES1, and SUF4 are required for basal levels of FLC expression in the absence of FRI. Thus, components of the FRI-C play a role in the regulation of FLC expression in both FRI-containing winter annuals, as well as fri-null rapid-cycling strains.


Retrograde ejaculation, painful ejaculation and hematospermia.
Although there has been an increased interest on premature ejaculation in the recent years, our understanding regarding the disorders of retrograde ejaculation, painful ejaculation and hematospermia remain limited. All three of these conditions require a keen clinical acumen and willingness to engage in thinking outside of the standard established treatment paradigm. The development of novel investigational techniques and treatments has led to progress in the management of these conditions symptoms; however, the literature almost uniformly is limited to small series and rare randomised trials. Further investigation and randomised controlled trials are needed for progress in these often challenging cases.


A point mutation in the extracellular domain of CD4 completely abolishes CD4 T cell development in C57BL/6 mouse.
In this study, we performed ENU mutagenesis and multi-parameter flow cytometric analysis in C57BL/6 mice to uncover novel genes or alleles regulating immune cell development. We identified a novel mutant allele of Cd4 gene which completely blocked development of a major subset of T cells named CD4 T cell. Our data for the first time showed experimentally in mice the critical role of the first extracellular domain, by obtaining mice with a loss of function mutation from Ile to Asn at the position 99 of CD4 (I99N). Interestingly, such CD4I99N mutant protein can be expressed on the surface of human cells, and the mRNA stability could be also affected by this point mutation, suggesting that absence of CD4 T cells in mice rooted in the deficiency in function and expression of CD4. In addition, we used this novel CD4 T cell deficient model as recipient mice for adoptive transfer experiment, and showed that it could be an optimal model for study of CD4 T cells.


Use of multiplanar computed tomography to aid in endosseous implant placement through hydroxylapatite overlay grafts.
Since the mid-1980's, mandibular ridge augmentation has been carried out with nonresorbable hydroxylapatite, with various results. This article describes the use of multiplanar reformated computed tomography to aid in the placement of endosseous fixtures through such augmentation. Displaced or migrated hydroxylapatite can obscure normal panographic and periapical radiographic landmarks. With the aid of computed tomography, landmarks can be identified, measured, and related to pre-treatment models, facilitating pre-surgical planning and surgical placement of endosseous fixtures.


Mental health and global well-being.
A range of evidence-based, cost-effective interventions can be implemented in parenting, at schools, at the workplace and in older age to promote mental health and well-being. Such programmes need to be supplemented with actions to build mental health capital and promote resilience to manage and cope with the global risks that face humankind over the coming years. Actions need to connect mental and physical health and individuals need to be connected through health-promoting social networks; living environments need to be designed to support mental health and well-being; well-being indicators that include material living conditions, quality of life and sustainability can help drive healthy public policy. There is an urgent need to invest in skills training in decision-making, social interactions, building trust and cooperative behaviour that support the family of humanity as a whole as it faces the unprecedented stressors resulting from climate change.


Trunk sway analysis to quantify the warm-up phenomenon in myotonia congenita patients.
Patients with autosomal recessive myotonia congenita display myotonia and transient paresis that diminish with repetitive muscle contractions (warm-up phenomenon). A new approach is presented to quantify this warm-up phenomenon under clinically relevant gait and balance tasks. Ten patients with DNA proven autosomal recessive myotonia congenita and 14 age-matched controls participated. Subjects performed six everyday gait and balance tasks. Balance control during these tasks was monitored using two angular velocity transducers that measured trunk movements in anterior-posterior (pitch) and medio-lateral (roll) directions at the level of the lumbar vertebral column. Tasks were performed under two conditions in randomised order: after a 10-minute seated rest period ("rested") and after having consecutively repeated the task five times ("warm-up"). Controls were also tested twice. "Rested" patients showed the greatest abnormalities (increased sway in pitch and roll) for tandem walking and walking stairs. Balance impairment was also evident for all other tasks. After "warm-up," balance was markedly improved in patients, as reflected by decreased trunk sway (especially during tandem walking) and reduced task duration for all tasks. These results were not only evident at the group level but also clearly present in individual patients. The results show that trunk sway analysis detects postural instability in myotonia congenita patients during everyday gait and balance tasks. Moreover, this technique provides a useful tool to quantify the warm-up phenomenon, suggesting a potential use as clinical endpoint in future clinical trials.


Voice or choice? Patient and public involvement in the National Health Service in England under New Labour.
Involving patients, caregivers, and citizens in health care and health policy has been recommended by international organizations for over a decade. This article focuses on developments in England under New Labour, places them in the context of broader health policy, and assesses them in the light of the limited empirical evidence. The authors consider a range of possible explanations for these developments. They suggest that we need to distinguish between individual and collective forms of patient involvement, and they chart patient and public involvement in England before New Labour and in three distinct phases under New Labour. There has been a significant extension of opportunities for individual patients and the public to communicate their views, albeit with twists and turns in the policy over time. The authors explain these developments in terms of New Labour's ideological attachment to pragmatism and the Third Way, political calculations about the need to reinvigorate political culture, and attempts to enhance cost-effectiveness. Patient and public involvement seems to be here to stay, but whether this will result in greater equity and a real shift in power away from professionals to citizens and patients is another matter.


Retention mucocele on the lower lip associated with inadequate use of pacifier.
Mucocele forms because of salivary gland mucous extravasation or retention and is usually related to trauma in the area of the lower lips. Ruptured ducts release the mucous that accumulates into adjacent tissues, leading to swelling. This report describes a large mucocele involving the lower lip, which was produced in a child by incorrect use of a pacifier. A few important concepts are discussed to help clinicians in the diagnosis and treatment of this pathology.


Molecular cloning and expression of the cDNA for the hamster alpha 1-adrenergic receptor.
The cDNA for the Syrian hamster alpha 1-adrenergic receptor has been cloned with oligonucleotides corresponding to the partial amino acid sequence of the receptor protein purified from DDT1MF-2 smooth muscle cells. The deduced amino acid sequence encodes a 515-residue polypeptide that shows the most sequence identity with the other adrenergic receptors and the putative protein product of the related clone G-21. Similarities with the muscarinic cholinergic receptors are also evident. Expression studies in COS-7 cells confirm that we have cloned the alpha 1-adrenergic receptor that couples to inositol phospholipid metabolism.


Bone marrow transplantation for chronic lymphocytic leukemia.
The use of bone marrow transplant (BMT) for patients with chronic lymphocytic leukemia (CLL) is a relatively new approach when compared to the use of this therapy for other hematologic malignancies. The long natural history of CLL and significant procedure-related mortality initially deterred the use of BMT for CLL. However, improvement in transplantation procedures and supportive care paved the way for the use of BMT in patients with CLL. There are now multiple reports showing complete remissions with both autologous and allogeneic BMT. The follow-up remains short, therefore, the durability of these remissions remains to be determined. Several innovative approaches to BMT for CLL are now being explored and include non-myeloablative preparative regimens for allogeneic transplants, purging autologous grafts, and induction of graft-versus-leukemia effects. Ultimately, however, long-term disease-free and overall survival will need to be shown before BMT for CLL will gain widespread acceptance.


Sustainability of performance improvements after 26 Kaizen events in a large academic hospital system: a mixed methods study.
Implementing Kaizen can improve productivity in healthcare but maintaining long-term results has proven challenging. This study aimed to assess improved performance achieved and sustained by Kaizen events and find explanatory factors for the persistence or decline of long-term results. Kaizen events were conducted in 26 specialised healthcare units in a large academic hospital system in southern Finland. Primary data for mixed methods analysis was collected from each unit with 21 semi-structured interviews, Kaizen report files and performance metrics. Fifteen explanatory factors were found in this study. Work culture and motivation for continuous improvement stood out as the most important explanatory factor for the persistence of long-term results-lack of time for improvement activities and high workload for the decline. Success in preparation and follow-up was associated with sustained long-term results. Thirteen units achieved long-term results, three units could not sustain the performance improvements and five units struggled to make any improvements. This study explains the long-term sustainability of performance improvements, bringing new insights to Kaizen research. Our findings can guide organising successful Kaizen events. The events can be worth organising even though long-term performance improvements are not guaranteed. Units with supportive working culture and motivation for the Kaizen event will likely succeed. A unit should aim to create a supportive foundation for Kaizen before organising a Kaizen event. Units that lack the foundation can be identified, trained and guided to increase their chances of success. Pitfalls like high workload and insufficient follow-up should be proactively identified and appropriately managed by allocating the required time and resources for the development work.


Intracavitary therapy with radioactive yttrium: results of animal experiments.
The distribution of radioactive yttrium was studied in guinea-pigs. The injection of 1,5 mci of radioactive yttrium leads to a mean surface dose of 20 Gy which can be obtained within 5 days. An extrapolation of these findings to the peritoneal surface of human beings should lead to a corresponding dose of 64 mci.


The mate choice brain: comparing gene profiles between female choice and male coercive poeciliids.
Genes that mediate mate preferences potentially play a key role in promoting and maintaining biological diversity. In this study, we compare mate preference behavior in two related poeciliid fishes with contrasting behavioral phenotypes and relate these behavioral differences to gene profiles in the brain. Results reveal that one poeciliid fish, the Northern swordtail, exhibits robust mate preference as compared to the Western mosquitofish, which utilizes a coercive mating system. Female swordtails display no significant difference in association time between male- and female-exposure trials, whereas female mosquitofish spend significantly less time associating with males relative to females. Furthermore, the preference strength for large males is significantly lower in female mosquitofish relative to swordtails. We then examine expression of three candidate genes previously shown to be associated with mate preference behavior in female swordtails and linked to neural plasticity in other vertebrates: neuroserpin (NS), neuroligin-3 (NLG-3) and N-methyl-d-aspartate receptor (NMDA-R). Whole brain gene expression patterns reveal that two genes (NS and NLG-3) are positively associated with mate preference behavior in female swordtails, a pattern opposing that of the mosquitofish. In mosquitofish females, these genes are downregulated when females express biases toward males yet are elevated in association with total motor activity patterns under asocial conditions, suggesting that the presence of males in mosquitofish species may inhibit expression of these genes. Both gene expression and female behavioral responses to males exhibit opposing patterns between these species, suggesting that this genetic pathway may potentially act as a substrate for the evolution of mate preference behavior.


Role of diet and nutritional management in non-alcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD) encompasses a spectrum ranging from simple steatosis to non-alcoholic steatohepatitis, which causes an increased risk of cirrhosis, type 2 diabetes, and cardiovascular complications. With the worldwide growing incidence of obesity, sedentary lifestyle, and unhealthy dietary pattern, NAFLD has currently been recognized as a major health burden. Dietary patterns and nutrients are the important contributors to the development, progression, and treatment of NAFLD and associated metabolic comorbidities. Generally, hypercaloric diet, especially rich in trans/saturated fat and cholesterol, and fructose-sweetened beverages seem to increase visceral adiposity and stimulate hepatic lipid accumulation and progression into non-alcoholic steatohepatitis, whereas reducing caloric intake, increasing soy protein and whey consumption, and supplement of monounsaturated fatty acids, omega-3 fatty acids, and probiotics have preventive and therapeutic effects. In addition, choline, fiber, coffee, green tea, and light alcohol drinking might be protective factors for NAFLD. Based on available data, at least 3-5% of weight loss, achieved by hypocaloric diet alone or in conjunction with exercise and behavioral modification, generally reduces hepatic steatosis, and up to 10% weight loss may be needed to improve hepatic necroinflammation. A sustained adherence to diet rather than the actual diet type is a major predictor of successful weight loss. Moreover, a healthy diet has benefits beyond weight reduction on NAFLD patients whether obese or of normal weight. Therefore, nutrition serves as a major route of prevention and treatment of NAFLD, and patients with NAFLD should have an individualized diet recommendation.


Granulocyte colony stimulating activity derived from human keratinocytes.
Human keratinocytes are known to produce a variety of soluble mediators including epidermal cell-derived thymocyte activating factor (ETAF), which is closely related, if not identical, to interleukin (IL)-1. In the present studies, crude supernatants derived from normal keratinocytes or from the skin squamous cell carcinoma line, COLO-16, were harvested after 5 days of culture or were purified sequentially by ultrafiltration, Sephadex G-100 chromatography, and high performance liquid chromatography (HPLC), using a Synchropak AX 300 ion exchange column, followed by a TSK 4,000 size exclusion column, and their effects tested on normal human peripheral blood hemopoietic progenitors at 14 days in methylcellulose cultures. Crude and fractionated COLO-16 or normal human keratinocyte supernatants stimulated human granulocyte progenitor growth and differentiation in a dose response fashion, including both neutrophil/macrophage (GM)-type and basophil/eosinophil (Eo)-type colony-forming cells (CFU-c). The supernatants also contained an interleukin-3 (IL-3)-like activity when assayed in a proliferation assay using a subcloned, murine IL-3-dependent cell line. After two-step HPLC separation, a linear stimulatory dose-response curve for colony stimulation was obtained for GM- and Eo-type colony growth in methylcellulose. In addition, a dissociation between ETAF and both GM- or Eo-colony-stimulating activity (CSF) was found after HPLC separation. The derivation of hemopoietic CSF's from human keratinocytes gives further support to the hypothesis that these cells and their products contribute to localized inflammatory or neoplastic skin responses.


Predictive capacity of the International Prognostic Factor Index in patients with peripheral T-cell lymphoma.
The International Prognostic Factor Index has been shown to predict the outcome of patients with predominantly B-cell lymphomas classified using traditional classifications, including the Working Formulation, but its prognostic importance has not been tested in a cohort of patients with exclusively T-cell lymphomas. This study was conducted to evaluate the prognostic significance of the International Prognostic Factor Index in patients with peripheral T-cell lymphoma (PTCL). Seventy-eight patients (48 men and 30 women) with PTCL seen at a single institution between 1985 and 1995 were included in the analysis. The morphology and immunocytochemistry of all the original biopsy specimens were reviewed by a single pathologist and classified using the Revised European-American Lymphoma (REAL) classification. The International Prognostic Factor Index, as well as clinical and biochemical parameters, were evaluated by univariate and multivariate analyses to determine their association with patient outcome. The International Prognostic Factor Index strongly predicted survival when all patients were included in the analysis (P < .001). For patients < or = 60 years, the age-adjusted International Index significantly predicted long-term survival (P = .0218). For patients older than 60 years, the age-adjusted International Index also significantly predicted survival (P = .002). Liver involvement (P = .006) and bone marrow involvement (P = .014) were also significant prognostic factors in the univariate analysis, but only the International Index remained significant in the multivariate analysis (P = .001). The International Prognostic Factor Index, which significantly predicts outcome in patients with aggressive/intermediate-grade B-cell lymphomas, has similar prognostic importance in patients with PTCL.


The response of two arctic tundra plant communities to human trampling disturbance.
A 4-year study was conducted to evaluate the consequences of human trampling on dryas and tussock tundra plant communities. Treatments of 25, 75, 200 and 500 trampling passes were applied in 0.75 m2 vegetation plots at a time of approximately peak seasonal biomass. Immediately after and 1 and 4 years after trampling, plots were evaluated on the basis of plant species cover, percent bare ground, vegetation height, and soil penetration resistance. One year after trampling, soils were collected for nitrogen analysis in highly disturbed and control plots. Immediately after trampling, 500 trampling passes resulted in approximately 50% cover loss in the dryas tundra and 70% cover loss in tussock tundra, but both communities showed a substantial capacity for regrowth. Plots where low and moderate levels of trampling were applied returned to pre-disturbance conditions by 4 years after trampling, but impact was still evident in plots subjected to high levels of disturbance. These results suggest that these tundra communities can tolerate moderate levels of hiking and camping provided that use is maintained below disturbance thresholds and that visitors employ appropriate minimum-impact techniques. By utilizing this information in a visitor education program combined with impact monitoring and management, it is possible to allow dispersed camping and still maintain these vegetation communities with a minimum of observable impact.


Skeletal muscle contractility is preserved in COPD patients with normal fat-free mass.
Peripheral muscle dysfunction often occurs in patients with chronic obstructive pulmonary disease (COPD). The muscle dysfunction may be caused by a loss of force-generating capacity, resulting from a loss of muscle mass, as well as by other alterations in contractile properties of skeletal muscle. The maximal isometric voluntary strength and fatigability were determined in hand-grip and quadriceps muscles from nine male COPD patients (FEV(1) 30-50% predicted) and control subjects matched for fat-free mass (FFM), physical activity level and age. Contractile properties and fatigability of the quadriceps muscle were also studied with electrically evoked isometric contractions. The maximal voluntary force (MVC) and fatigability of the handgrip muscle did not differ between the COPD patients and control subjects. Also the MVC of the quadriceps muscle and the rate of force rise, contraction time, force-frequency relationship and fatigability, as determined with electrically evoked contractions, were similar in patients with COPD and control subjects. Skeletal muscle strength, contractile properties and fatigability are preserved in patients with moderate COPD and a normal FFM and activity level. This suggests that skeletal muscle dysfunction does not take place during moderate COPD until cachexia and/or a decline in physical activity occur.


Psychobiography: Theory and method.
Psychobiography is best understood as a branch of applied personality science in which experimental findings are used to explore the lives and personalities of public figures, from artists to politicians. This article introduces the field, addresses questions commonly raised about it, and explores, among other subjects, what psychobiographical research might offer to the field of psychology. Two recent model psychobiographies are described in detail-McAdams's (2010) profile of George W. Bush and Kasser's (2013) analysis of John Lennon-both of which are oriented scientifically and rooted in experimentally derived concepts. Lastly, criteria for evaluating psychobiographical studies are summarized, as well as attributes of effective psychobiography that tend to include the components of fact, fit, and function. (PsycINFO Database Record


Design, synthesis and biological evaluation of biaryl amide derivatives against SARS-CoV-2 with dual-target mechanism.
The COVID-19 pandemic highlights the urgent need to develop effective small-molecule antivirals. Thirty-three novel biaryl amide derivatives were synthesized and evaluated for anti-coronaviral activity. Some significant SARs were uncovered and the intensive structure modifications led to the most active compounds 8b and 8h. The broad-spectrum anti-coronaviral effects of 8h were validated at RNA and protein levels. 8h inhibits coronavirus replication at multiple stages, from virus entry to virus dsRNA synthesis. The mechanism of action showed that 8h may simultaneously act on 3CLpro and TMPRSS2 to display anti-coronaviral effects. 8h combined with RdRp inhibitor showed synergistic inhibitory activity against coronavirus. This study confirmed that biaryl amide derivatives may be a new class of potential therapeutic agents against coronavirus with multiple target effect, worthy of further investigation.


Interobserver reproducibility of RENAL nephrometry score in comparison to simplified PADUA Renal nephrometry score.
Nephron-sparing surgery (NSS) is the standard of care for renal tumors, especially in the early stages. RENAL Nephrometry scores provide a comprehensive presurgical predictive module for the choice of NSS or Radical Nephrectomy. The validity and reliability of Nephrometry scores is being tested continuously with advancement in the surgical techniques. The Simplified PADUA Nephrometry score (SPARE NS) is a newer proposed score which aims to better the reproducibility of the previously established nephrometry scores. The retrospective observational study studied the comparative inter-observer reliability of RENAL (RENAL NS) and SPARE nephrometry scoring systems amongst two radiologists while assessing solid renal tumors in contrast-enhanced computed tomography scans of 42 patients. Interobserver reliability for all components of both scores, final scores and risk grading was done by Kendall's Concordance Coefficient (Tau). Both RENAL NS and SPARE NS showed strong to excellent agreement (RENAL NS = 78.57% and SPARE NS = 88.09%) among observers with comparable correlation co-efficient (RENAL NS = 0.944 and SPARE NS = 0.935). Lesion radius and exophytic/endophytic properties were the most reproducible components of RENAL NS with 97.61% and 92.85% agreement, respectively. Location across polar lines was the least reproducible component with 85.71% agreement among observers. Exophytic rate (97.61%) and Rim location were the most reproducible components of SPARE NS. The final lesion risk stratification by both observers for both was concordant in 92.85% of cases. The SPARE system of scoring matches up to the RENAL NS in total score and risk stratification reproducibility. However, the individual components of the SPARE score are more reproducible than those of RENAL NS, bringing about better compliance among radiology consultants. Comparable reproducibility with the RENAL NS, lesser number of variables, and ease of doing make SPARE NS a plausible option for the customary preoperative assessment of renal tumors.


Immunohistochemical and molecular markers in the diagnosis of hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) has distinct morphologic features and can be identified in the majority of cases by routine hematoxylin and eosin (H&E)-stained formalin-fixed paraffin-embedded sections. However, distinguishing a well-differentiated HCC from normal or regenerative tissue may be very difficult in some cases, particularly in small needle aspiration or core biopsies. Furthermore, some of the unusual morphologic variants, including clear-cell, pleomorphic, and sarcomatoid variants, may be mistaken for metastases. Similarly, metastases from various primary tumors to the liver may be mistaken for primary hepatic tumors. In this overview, we summarize the immunohistochemical and molecular markers that have been developed to address these diagnostic challenges. Among the numerous diagnostic markers studied, pCEA, HepPar 1, CD34, CK 7, CK 19, CK 20, and albumin in situ (ISH) have been found to be valuable in distinguishing HCC from metastatic neoplasms of extrahepatic sites.


Dialogue as skill: training a health professions workforce that can talk about race and racism.
Efforts in the field of multicultural education for the health professions have focused on increasing trainees' knowledge base and awareness of other cultures, and on teaching technical communication skills in cross-cultural encounters. Yet to be adequately addressed in training are profound issues of racial bias and the often awkward challenge of cross-racial dialogue, both of which likely play some part in well-documented racial disparities in health care encounters. We seek to establish the need for the skill of dialoguing explicitly with patients, colleagues, and others about race and racism and its implications for patient well-being, for clinical practice, and for the ongoing personal and professional development of health care professionals. We present evidence establishing the need to go beyond training in interview skills that efficiently "extract" relevant cultural and clinical information from patients. This evidence includes concepts from social psychology that include implicit bias, explicit bias, and aversive racism. Aiming to connect the dots of diverse literatures, we believe health professions educators and institutional leaders can play a pivotal role in reducing racial disparities in health care encounters by actively promoting, nurturing, and participating in this dialogue, modeling its value as an indispensable skill and institutional priority.


Sulfur-Mediated Electron Shuttling Sustains Microbial Long-Distance Extracellular Electron Transfer with the Aid of Metallic Iron Sulfides.
In addition to serving as an energy source for microbial growth, iron sulfides are proposed to act as naturally occurring electrical wires that mediate long-distance extracellular electron transfer (EET) and bridge spatially discrete redox environments. These hypothetical EET reactions stand on the abilities of microbes to use the interfacial electrochemistry of metallic/semiconductive iron sulfides to maintain metabolisms; however, the mechanisms of these phenomena remain unexplored. To obtain insight into EET to iron sulfides, we monitored EET at the interface between Shewanella oneidensis MR-1 cells and biomineralized iron sulfides in an electrochemical cell. Respiratory current steeply increased with the concomitant formation of poorly crystalline mackinawite (FeS) minerals, indicating that S. oneidensis has the ability to exploit extracellularly formed metallic FeS for long-distance EET. Deletion of major proteins of the metal-reduction (Mtr) pathway (OmcA, MtrC, CymA, and PilD) caused only subtle effects on the EET efficiency, a finding that sharply contrasts the majority of studies that report that the Mtr pathway is indispensable for the reduction of metal oxides and electrodes. The gene expression analyses of polysulfide and thiosulfate reductase suggest the existence of a sulfur-mediated electron-shuttling mechanism by which HS(-) ions and water-soluble polysulfides (HS(n)(-), where n ≥ 2) generated in the periplasmic space deliver electrons from cellular metabolic processes to cell surface-associated FeS. The finding of this Mtr-independent pathway indicates that polysulfide reductases complement the function of outer-membrane cytochromes in EET reactions and, thus, significantly expand the number of microbial species potentially capable of long-distance EET in sulfur-rich anoxic environments.


Effector caspases are dispensable for the early nuclear morphological changes during chemical-induced apoptosis.
Nuclear morphological changes during apoptosis are very distinct and effector caspases have been implicated to play a central role in these processes. To investigate this in greater detail we examined the effect of blocking caspase activity and its activation on the nuclear morphological change in Jurkat T cells undergoing apoptosis after staurosporine treatment. In the presence of caspase inhibitors, like benzyloxycarbonyl-Val-Ala-Asp fluoro-methylketone (z-VAD-FMK), N-acetyl Tyr-Val-Ala-Asp chloromethylketone (Ac-YVAD-CMK) and benzyloxy-carbonyl-Asp-Glu-Val-Asp (OMe) fluoromethylketone (z-DEVD-FMK), staurosporine-treated Jurkat cells displayed a nuclear morphological change distinct from that of normal and apoptotic cells. This nuclear morphological change is an early event, characterised by convoluted nuclei with cavitations, and clumps of chromatin abutting to inner regions of the nuclear envelope between the nuclear pores. Both the nuclear envelope and endoplasmic reticulum were grossly dilated. This pre-apoptotic nuclear change precedes the externalisation of phosphatidylserine, chromatin condensation and DNA laddering, and can be dissociated from the formation of high molecular weight DNA fragments and cell shrinkage. Although cytochrome c efflux from the mitochondria and the processing of caspase-3 were observed in Jurkat cells with pre-apoptotic nuclear morphology, caspase-2, -6, -7 and -8 were not activated. In the presence of z-DEVD-FMK or Ac-YVAD-CMK, caspase-3 was processed to both the p17 and p20 fragments in staurosporine-treated cells, but only to p20 fragment in the presence of z-VAD-FMK. However, the caspase-3 substrate, poly(ADP ribose) polymerase was not cleaved in the presence of z-VAD-FMK, despite >70% of the cells have pre-apoptotic nuclei. In addition, caspase-3 null MCF-7 cells also undergo pre-apoptotic nuclear change when treated with staurosporine in the presence of caspase inhibitors, indicating that caspase-3 is not required for the early nuclear morphological change in cells undergoing apoptosis. Although cell death in staurosporine-treated Jurkat cells was markedly delayed, they eventually die without discernible downstream apoptotic features. Other apoptotic stimuli like etoposide and the heavy metal chelator, N,N,N',N'-tetrakis (2-pyridylmethyl) ethylenediamine also induced this nuclear morphological change in Jurkat cells in the presence of z-VAD-FMK. In summary, the effector caspases are not involved in early nuclear morphological change, which precedes the conventional hallmark morphological changes associated with chemical-induced apoptosis.


Arthroscopic Vastus Elevation (AVE) for Arthrofibrosis of the Knee: Surgical Technique and Literature Review.
Arthrofibrosis of the knee continues to challenge Orthopaedic surgeons. With a wide etiology, lack of knee motion can be debilitating. Its surgical management has several complications. The purpose of this study is to describe a modification of previously described techniques to aid in the management of knee arthrofibrosis. Arthroscopic vastus elevation in conjunction with adjuvant hemostatic agents allows for a controlled quadriceps elevation in the setting of arthrofibrosis. In addition to a thorough intra-articular lysis of adhesions, this appears to improve motion, while minimizing postoperative complications. Minimized postoperative complications include extensor lag, skin necrosis, and bleeding complications. (Journal of Surgical Orthopaedic Advances 29(2):117-120, 2020).


Thyroid hormone regulates the cell cycle inhibitor p27Kip1 in postnatal murine Sertoli cells.
Thyroid hormone regulates early postnatal Sertoli cell proliferation. Transient neonatal hypothyroidism allows prolonged postnatal Sertoli cell mitogenesis and doubles adult Sertoli cell numbers, testis weight, and sperm production. The mechanism of this effect is unknown. Cell proliferation is stimulated by cyclins and cyclin-dependent kinases and inhibited by cyclin-dependent kinase inhibitors (CDKIs). T(3) regulates the CDKI p27(Kip1) in other cell types, and mice lacking p27(Kip1) have increased testis size. To test the hypothesis that T(3) regulates Sertoli cell mitogenesis by acting through p27(Kip1), we compared expression of p27(Kip1) in Sertoli cells of testes from euthyroid, hypothyroid, and hyperthyroid mice. At postnatal d 5-25, testes were collected and immunostained for p27(Kip1) expression, or Sertoli cells were isolated enzymatically and used for p27(Kip1) Western blotting. p27(Kip1) immunostaining was low in rapidly proliferating 5-d-old Sertoli cells but had increased strongly in nonproliferating 25-d-old Sertoli cells. p27(Kip1) immunostaining was reduced in Sertoli cells from hypothyroid mice compared with euthyroid controls at 10 and 16 d, consistent with increased Sertoli cell proliferation in these mice. Western blotting corroborated the p27(Kip1) immunostaining, and p27(Kip1) expression was greater in Sertoli cells from control compared with hypothyroid mice at postnatal d 10 and 16, but p27(Kip1) expression was comparable by d 25. Hyperthyroidism increased p27(Kip1) immunostaining relative to controls, and Western analysis indicated that Sertoli cells from 10-d-old hyperthyroid mice expressed more p27(Kip1) than control mice. These results indicate that thyroid hormone status affects p27(Kip1) expression in neonatal Sertoli cells, suggesting that T(3) effects on Sertoli cell proliferation may be mediated through this CDKI.


Shared and distinct functions of two Gti1/Pac2 family proteins in growth, morphogenesis and pathogenicity of Magnaporthe oryzae.
Gti1/Pac2 are conserved family proteins that regulate morphogenic transition in yeasts such as Schizosaccharomyces pombe and Candida albicans, and they also control toxin production and pathogenicity in filamentous fungus Fusarium graminearum. To test the functions of Gti1/Pac2 paralogues MoGti1 and MoPac2 in the rice blast fungus Magnaporthe oryzae, we generated respective ΔMogti1 and ΔMopac2 mutant strains. We found that MoGti1 and MoPac2 exhibit shared and distinct roles in hyphal growth, conidiation, sexual reproduction, stress responses, surface hydrophobility, invasive hyphal growth and pathogenicity. Consistent with the putative conserved function of MoGti1, we showed that MoGti1-GFP is localized to the nucleus, whereas MoPac2-GFP is mainly found in the cytoplasm. In addition, we provided evidence that the nuclear localization of MoGti1 could be subject to regulation by MoPmk1 mitogen-activated protein kinase. Moreover, we found that the reduced pathogenicity in the ΔMopac2 mutant corresponds with an increased expression of plant defence genes, including PR1a, AOS2, LOX1, PAD4, and CHT1. Taken together, our studies provide a comprehensive analysis of two similar but distinct Gti1/Pac2 family proteins in M. oryzae, which underlines the important yet conserved functions of these family proteins in plant pathogenic fungi.


Duodenal and ileal lipid suppresses postprandial blood glucose and insulin responses in man: possible implications for the dietary management of diabetes mellitus.
Infusion of lipid into the ileum delays the transit of a meal through the stomach and small intestine and could therefore influence the rate and degree of nutrient absorption. Experiments were carried out on human volunteers to investigate the effect of infusion of lipid into either the duodenum or ileum on blood glucose, insulin and gastric emptying after ingestion of a mashed potato meal. Infusions of lipid into either the duodenum or the ileum significantly reduced or abolished the immediate postprandial rises in blood glucose and insulin and significantly delayed gastric emptying. Blood glucose and insulin rose shortly after the lipid infusion terminated. Addition of corn oil to a meal of mashed potato also reduced blood glucose and insulin and delayed gastric emptying. Intestinal lipid can thus modify the glycaemic and insulinaemic responses to a meal, and this modulation probably explains the reduced metabolic responses to a meal containing fat compared with a fat free meal. This principle could be of value in the dietary control of diabetes mellitus.


Primary intravitreal bevacizumab for subfoveal choroidal neovascularization in age-related macular degeneration: results of the Pan-American Collaborative Retina Study Group at 12 months follow-up.
To report the 12-month anatomic and Early Treatment Diabetic Retinopathy Study best-corrected visual acuity (BCVA) response after primary intravitreal bevacizumab (Avastin, Genentech Inc., San Francisco, CA) (1.25 mg or 2.5 mg) in patients with subfoveal choroidal neovascularization secondary to age-related macular degeneration. Sixty-three eyes of 63 consecutive patients with subfoveal choroidal neovascularization secondary to age-related macular degeneration, a mean age of 73.7 +/- 7.5 years and a minimum of 12 months (mean 55.5 +/- 6.2 weeks) of follow-up participated in this interventional retrospective multicenter case series in 7 centers from 6 countries. Patients were treated with at least 1 intravitreal injection of 1.25 mg or 2.5 mg of bevacizumab. Patients underwent Early Treatment Diabetic Retinopathy Study BCVA testing, ophthalmoscopic examination, optical coherence tomography, and fluorescein angiography at baseline and follow-up visits. Repeated measures analysis of variance was used to compare mean values. The mean number of intravitreal bevacizumab injections per eye was 3.5 (range, 1-8). Mean baseline BCVA was 20/320, logarithm of the minimum angle of resolution = 1.2, and mean final BCVA was 20/200, logarithm of the minimum angle of resolution = 1.0 (P < 0.001). Central macular thickness at baseline by optical coherence tomography had a mean of 389.2 +/- 149.6 microm which was significantly reduced to a mean of 281.0 +/- 96.1 microm, 268.2 +/- 82.6 microm, 262.6 +/- 92.3 microm, and 241.3 +/- 76.7 microm at 1, 3, 6, and 12 months after initial treatment, respectively (P < 0.0001). Ocular adverse events included transient increased intraocular pressure in 2 (3.1%) eyes, endophthalmitis in 2 (3.1%) eyes, and transient hypotony in 1 eye (1.1%). No systemic adverse events were observed. Primary intravitreal bevacizumab at doses of 1.25 mg or 2.5 mg seems to provide stability or improvement in BCVA, optical coherence tomography, and fluorescein angiography in subfoveal choroidal neovascularization secondary to age-related macular degeneration at 12 months.


Acetabular distraction hip arthroplasty in neglected transverse and T-shape acetabular fractures : a different application.
Neglected acetabular fractures are often challenging to treat. The aim of this study is to highlight the technique of acetabular distraction with porous metal cup as a viable technique to address such complex cases. This is a prospective study of seven patients who had neglected acetabular fracture or fracture non-union that were managed with total hip arthroplasty using acetabular distraction and porous metal cup with cemented liner. The average age of the patients was 51 years (range 36-58 years). Average time of follow up was 18 months (range 12-36 months). Radiological results at latest follow up and by comparing serial radiographs showed that all cases had a well-fixed implant construct without evidence of loosening or malorientation. Average Harris hip score pre-operatively was 46, which improved to an average of 84 post-operatively. Our study shows that total hip replacement using acetabular distraction with porous metal cup provide early good results in reconstruction of acetabular bony deficiencies due to neglected acetabular fracture.


An ultrastructural study of sporidium formation during infection of a rhabditid nematode by large gun cells of Haptoglossa heteromorpha.
Recently fired gun cells of Haptoglossa heteromorpha, an aplanosporic nematode parasite, were examined ultrastructurally. The everted tubes of the fired cells had penetrated the cuticle of a nematode, and infective sporidia were developing inside the host body. The nematode cuticle was penetrated by the narrow, walled part of the tube below the needle chamber. The lower unwalled part of the tube tail formed the sporidium. The developing sporidium had a multilayered fibrous outer coating and the plasma membrane was separated from the wall in places. Sporidia contained biphasic membrane-bound vesicles that had been generated by the Golgi dictyosome during gun cell development. Immediately following gun cell firing, the nuclear envelope of the sporidium nucleus was not apparent, and the sporidium nucleus contained clusters of electron-dense particles concentrated in the nucleolar region. We compare the structures and organelles found in the mature gun cell with those in the fired cell and attempt to identify the membranous layers around the sporidium.


Splitting of the Mind: When the You I Talk to is Me and Needs Commands.
Self-talk has fascinated scholars for decades but has received little systematic research attention. Three studies examined the conditions under which people talk to themselves as if they are another person, indicating a splitting or fragmentation of the self. Fragmented self-talk, defined by the use of the second person, You, and the imperative, was specifically expected to arise in contexts requiring explicit self-control. Results showed that fragmented self-talk was most prevalent in response to situations requiring direct behavior regulation, such as negative events (Study 1), experiences of autonomy (Study 2), and action as opposed to behavior preparation or behavior evaluation (Study 3). Therefore, people refer to themselves as You and command themselves as if they are another person in situations requiring conscious self-guidance. The implications of these findings for behavior change are discussed.


In-situ measurements of 137Cs in soil by unfolding method.
An improved in-situ spectrometry measurement of 137Cs concentration in soil is introduced. The method uses the information contained in the pulse spectrum in order to forego the need for soil sampling. The approach is based on the unfolding of responses of a collimated and uncollimated HPGe detector to primary 0.662 MeV photons and to photons scattered forward in the soil. The calibration of the in situ equipment has been performed by Monte Carlo calculations and by experiments. For unfolding of experimental detector responses the code SAND II has been found reliable and capable of calculating distribution of 137Cs in soil profile with adequate accuracy for environmental monitoring purposes. The analysis of the spectra indicates that 137Cs concentration in soil 10 y after Chernobyl accident would be measurable using a middle HPGe detector (20-30% relative efficiency) and a counting time on the order of 1 h. Even with smaller detectors, 137Cs concentrations of 5 kBq m(-2) are measurable, and the depth distribution of 137Cs activities above 10 kBq m(-2) in the soil can be estimated by the presented method when a counting time on the order of 3 h is used.


Bayesian semiparametric analysis of developmental toxicology data.
Modeling of developmental toxicity studies often requires simple parametric analyses of the dose-response relationship between exposure and probability of a birth defect but poses challenges because of nonstandard distributions of birth defects for a fixed level of exposure. This article is motivated by two such experiments in which the distribution of the outcome variable is challenging to both the standard logistic model with binomial response and its parametric multistage elaborations. We approach our analysis using a Bayesian semiparametric model that we tailored specifically to developmental toxicology studies. It combines parametric dose-response relationships with a flexible nonparametric specification of the distribution of the response, obtained via a product of Dirichlet process mixtures approach (PDPM). Our formulation achieves three goals: (1) the distribution of the response is modeled in a general way, (2) the degree to which the distribution of the response adapts nonparametrically to the observations is driven by the data, and (3) the marginal posterior distribution of the parameters of interest is available in closed form. The logistic regression model, as well as many of its extensions such as the beta-binomial model and finite mixture models, are special cases. In the context of the two motivating examples and a simulated example, we provide model comparisons, illustrate overdispersion diagnostics that can assist model specification, show how to derive posterior distributions of the effective dose parameters and predictive distributions of response, and discuss the sensitivity of the results to the choice of the prior distribution.


Structural determinants of phosphatidylinositol 4,5-bisphosphate (PIP2) regulation of BK channel activity through the RCK1 Ca2+ coordination site.
Big or high conductance potassium (BK) channels are activated by voltage and intracellular calcium (Ca(2+)). Phosphatidylinositol 4,5-bisphosphate (PIP2), a ubiquitous modulator of ion channel activity, has been reported to enhance Ca(2+)-driven gating of BK channels, but a molecular understanding of this interplay or even of the PIP2 regulation of this channel's activity remains elusive. Here, we identify structural determinants in the KDRDD loop (which follows the αA helix in the RCK1 domain) to be responsible for the coupling between Ca(2+) and PIP2 in regulating BK channel activity. In the absence of Ca(2+), RCK1 structural elements limit channel activation through a decrease in the channel's PIP2 apparent affinity. This inhibitory influence of BK channel activation can be relieved by mutation of residues that (a) connect either the RCK1 Ca(2+) coordination site (Asp(367) or its flanking basic residues in the KDRDD loop) to the PIP2-interacting residues (Lys(392) and Arg(393)) found in the αB helix or (b) are involved in hydrophobic interactions between the αA and αB helix of the RCK1 domain. In the presence of Ca(2+), the RCK1-inhibitory influence of channel-PIP2 interactions and channel activity is relieved by Ca(2+) engaging Asp(367). Our results demonstrate that, along with Ca(2+) and voltage, PIP2 is a third factor critical to the integral control of BK channel activity.


Role of the Strength of Drug-Polymer Interactions on the Molecular Mobility and Crystallization Inhibition in Ketoconazole Solid Dispersions.
The effects of specific drug-polymer interactions (ionic or hydrogen-bonding) on the molecular mobility of model amorphous solid dispersions (ASDs) were investigated. ASDs of ketoconazole (KTZ), a weakly basic drug, with each of poly(acrylic acid) (PAA), poly(2-hydroxyethyl methacrylate) (PHEMA), and polyvinylpyrrolidone (PVP) were prepared. Drug-polymer interactions in the ASDs were evaluated by infrared and solid-state NMR, the molecular mobility quantified by dielectric spectroscopy, and crystallization onset monitored by differential scanning calorimetry (DSC) and variable temperature X-ray diffractometry (VTXRD). KTZ likely exhibited ionic interactions with PAA, hydrogen-bonding with PHEMA, and weaker dipole-dipole interactions with PVP. On the basis of dielectric spectroscopy, the α-relaxation times of the ASDs followed the order: PAA > PHEMA > PVP. In addition, the presence of ionic interactions also translated to a dramatic and disproportionate decrease in mobility as a function of polymer concentration. On the basis of both DSC and VTXRD, an increase in strength of interaction translated to higher crystallization onset temperature and a decrease in extent of crystallization. Stronger drug-polymer interactions, by reducing the molecular mobility, can potentially delay the crystallization onset temperature as well as crystallization extent.


The Effect of Signaling Latencies and Node Refractory States on the Dynamics of Networks.
We describe the construction and theoretical analysis of a framework derived from canonical neurophysiological principles that model the competing dynamics of incident signals into nodes along directed edges in a network. The framework describes the dynamics between the offset in the latencies of propagating signals, which reflect the geometry of the edges and conduction velocities, and the internal refractory dynamics and processing times of the downstream node receiving the signals. This framework naturally extends to the construction of a perceptron model that takes into account such dynamic geometric considerations. We first describe the model in detail, culminating with the model of a geometric dynamic perceptron. We then derive upper and lower bounds for a notion of optimal efficient signaling between vertex pairs based on the structure of the framework. Efficient signaling in the context of the framework we develop here means that there needs to be a temporal match between the arrival time of the signals relative to how quickly nodes can internally process signals. These bounds reflect numerical constraints on the compensation of the timing of signaling events of upstream nodes attempting to activate downstream nodes they connect into that preserve this notion of efficiency. When a mismatch between signal arrival times and the internal states of activated nodes occurs, it can cause a breakdown in the signaling dynamics of the network. In contrast to essentially all of the current state of the art in machine learning, this work provides a theoretical foundation for machine learning and intelligence architectures based on the timing of node activations and their abilities to respond rather than necessary changes in synaptic weights. At the same time, the theoretical ideas we developed are guiding the discovery of experimentally testable new structure-function principles in the biological brain.


Atelerix algirus, the North African Hedgehog: Suitable Wild Host for Infected Ticks and Fleas and Reservoir of Vector-Borne Pathogens in Tunisia.
Small wild mammals are an important element in the emergence and transmission of vector-borne pathogens (VBPs). Among these species, hedgehogs have been found to be a reservoir of VBPs and host of arthropod vectors. Surveillance of VBPs in wildlife and their arthropods are crucial in a one health context. We conducted an exploratory study to screen Atelerix algirus hedgehogs and their infesting ticks and fleas for VBPs using a high throughput microfluidic real-time PCR system. Tested biopsies from hedgehogs were found to be naturally infected by Theileria youngi, Hepatozoon sp., Ehrlichia ewingii, Coxiella burnetii, and Candidatus Ehrlichia shimanensis. Similarly, Haemaphysalis erinacei and Rhipicephalus sanguineus tick species were infected by Ehrlichia ewingii, Rickettsia spp., Rickettsia massiliae, Borrelia sp., Coxiella burnetii, Rickettsia lusitaniae and Anaplasma sp. Archaeopsylla erinacei fleas were infected by Rickettsia asembonensis, Coxiella burnetii, and Rickettsia massiliae. Co-infections by two and three pathogens were detected in hedgehogs and infesting ticks and fleas. The microfluidic real-time PCR system enabled us not only to detect new and unexpected pathogens, but also to identify co-infections in hedgehogs, ticks, and fleas. We suggest that hedgehogs may play a reservoir role for VBPs in Tunisia and contribute to maintaining enzootic pathogen cycles via arthropod vectors.


Group II intron in Bacillus cereus has an unusual 3' extension and splices 56 nucleotides downstream of the predicted site.
All group II introns known to date fold into six functional domains. However, we recently identified an intron in Bacillus cereus ATCC 10987, B.c.I4, that splices 56 nt downstream of the expected 3' splice site in vivo (Tourasse et al. 2005, J. Bacteriol., 187, 5437-5451). In this study, we confirmed by ribonuclease protection assay that the 56-bp segment is part of the intron RNA molecule, and computational prediction suggests that it might form a stable stem-loop structure downstream of domain VI. The splicing of B.c.I4 was further investigated both in vivo and in vitro. Lariat formation proceeded primarily by branching at the ordinary bulged adenosine in domain VI without affecting the fidelity of splicing. In addition, the splicing efficiency of the wild-type intron was better than that of a mutant construct deleted of the 56-bp 3' extension. These results indicate that the intron has apparently adapted to the extra segment, possibly through conformational adjustments. The extraordinary group II intron B.c.I4 harboring an unprecedented extra 3' segment constitutes a dramatic example of the flexibility and adaptability of group II introns.


The developmental biology of brain tumors.
Tumors of the central nervous system (CNS) can be devastating because they often affect children, are difficult to treat, and frequently cause mental impairment or death. New insights into the causes and potential treatment of CNS tumors have come from discovering connections with genes that control cell growth, differentiation, and death during normal development. Links between tumorigenesis and normal development are illustrated by three common CNS tumors: retinoblastoma, glioblastoma, and medulloblastoma. For example, the retinoblastoma (Rb) tumor suppressor protein is crucial for control of normal neuronal differentiation and apoptosis. Excessive activity of the epidermal growth factor receptor and loss of the phosphatase PTEN are associated with glioblastoma, and both genes are required for normal growth and development. The membrane protein Patched1 (Ptc1), which controls cell fate in many tissues, regulates cell growth in the cerebellum, and reduced Ptc1 function contributes to medulloblastoma. Just as elucidating the mechanisms that control normal development can lead to the identification of new cancer-related genes and signaling pathways, studies of tumor biology can increase our understanding of normal development. Learning that Ptc1 is a medulloblastoma tumor suppressor led directly to the identification of the Ptc1 ligand, Sonic hedgehog, as a powerful mitogen for cerebellar granule cell precursors. Much remains to be learned about the genetic events that lead to brain tumors and how each event regulates cell cycle progression, apoptosis, and differentiation. The prospects for beneficial work at the boundary between oncology and developmental biology are great.


New, rapid method to measure dissolved silver concentration in silver nanoparticle suspensions by aggregation combined with centrifugation.
It is unclear whether the antimicrobial activities of silver nanoparticles (AgNPs) are exclusively mediated by the release of silver ions (Ag+) or, instead, are due to combined nanoparticle and silver ion effects. Therefore, it is essential to quantify dissolved Ag in nanosilver suspensions for investigations of nanoparticle toxicity. We developed a method to measure dissolved Ag in Ag+/AgNPs mixtures by combining aggregation of AgNPs with centrifugation. We also describe the reproducible synthesis of stable, uncoated AgNPs. Uncoated AgNPs were quickly aggregated by 2 mM Ca2+, forming large clusters that could be sedimented in a low-speed centrifuge. At 20,100g, the sedimentation time of AgNPs was markedly reduced to 30 min due to Ca2+-mediated aggregation, confirmed by the measurements of Ag content in supernatants with graphite furnace atomic absorption spectrometry. No AgNPs were detected in the supernatant by UV-Vis absorption spectra after centrifuging the aggregates. Our approach provides a convenient and inexpensive way to separate dissolved Ag from AgNPs, avoiding long ultracentrifugation times or Ag+ adsorption to ultrafiltration membranes.


Managing menopausal symptoms after cancer.
The joint burden of cancer and menopause impacts millions of women globally. This review provides an approach to management of menopausal symptoms after cancer in all settings. This includes an overview of current evidence for both hormonal and non-hormonal treatments for vasomotor symptoms and vaginal dryness after cancer. Systemic menopausal hormone therapy provides symptom control and may be used after most cancers but should be avoided after estrogen receptor-positive breast cancer and after some other estrogen-dependent cancers. Non-hormonal therapies have been minimally studied in women after a cancer diagnosis and, where they have been studied, it is usually in women with breast cancer. Non-hormonal methods to manage vasomotor symptoms include cognitive behavioral therapy, hypnosis, selective serotonin reuptake inhibitors, serotonin noradrenaline reuptake inhibitors, clonidine, and gabapentin. Vaginal estrogen may be useful to address vaginal dryness. However, safety data in breast cancer patients are still lacking and there is currently no consensus. Lubricants may also help with pain with sexual activity. Management of menopausal symptoms after cancer may be challenging and should include information about induced menopause and possible symptoms as well as available treatments. Management then requires a holistic and multidisciplinary approach with individualized care.


Chloramine mutagenesis in Bacillus subtilis.
Chloramine (which occurs widely as a by-product of sanitary chlorination of water supplies) is shown to be a weak mutagen, when reversion of trpC to trpC in Bacillus subtilis is used as an assay. Some DNA-repair mutants appear to be more sensitive to chloramine, suggesting the involvement of DNA targets in bactericide. The influence of plating media on survival of cells treated with chloramine suggests a bacterial repair system acting upon potentially lethal lesions induced by chloramine.


Sociodemographic disparities in COVID-19 burden: changing patterns over four pandemic waves in Israel.
Worldwide, the COVID-19 pandemic hit weakest populations hardest, with socioeconomic (SE), racial and ethnic disparities in COVID-19 burden. The study aimed to analyse patterns of SE and ethnic disparities in morbidity, hospitalisation, and vaccination throughout four pandemic waves. A retrospective-archive study was conducted in Israel from 11 March 2020 to 1 December 2021, with data on confirmed cases, hospitalisations, mortality and vaccinations (three doses), obtained from the Israeli Ministry of Health's open COVID-19 database, covering 98.8% of the population, by SE and ethnic characteristics of localities. At the outbreak of the pandemic, there was a higher caseload in Jewish, compared with Arab localities. In the second and third waves, low SE and Arab minority populations suffered 2-3 times higher morbidity, with a similar but attenuated pattern in the fourth wave. A similar trend was observed in hospitalisation of confirmed patients. COVID-19-associated mortality did not demonstrate a clear SE gradient.A strong social gradient in vaccine uptake was demonstrated throughout the period, with 71% and 74% double vaccinated in the two highest SE clusters, and 43% and 27% in the two lowest clusters by December 2021. Uptake of the third dose was 57%-60% in the highest SE clusters and 31%-25% in the lowest clusters. SE disparities in vaccine uptake were larger among the younger age groups and gradually increased from first to third doses. Israel was among the first to lead a rapid vaccination drive, as well as to experience a fourth wave fuelled by diminishing immunity and the delta variant. SE and ethnic disparities were evident throughout most of the pandemic months, though less so for mortality. Despite higher COVID-19 burden, vaccine uptake was lower in disadvantaged groups, with greater disparities in the younger population which widened with subsequent doses.


Cardamonin induces G2/M arrest and apoptosis via activation of the JNK-FOXO3a pathway in breast cancer cells.
Cardamonin (CD), a naturally occurring chalcone isolated from large black cardamom, was previously reported to suppress the proliferation of breast cancer cells. However, its precise molecular anti-tumor mechanisms have not been well elucidated. In this study, we found that CD markedly inhibited the proliferation of MDA-MB 231 and MCF-7 breast cancer cells through the induction of G2/M arrest and apoptosis. Reactive oxygen species (ROS) plays a pivotal role in the inhibition of CD-induced cell proliferation. Treatment with N-acetyl-cysteine (NAC), an ROS scavenger, blocked CD-induced G2/M arrest and apoptosis in this study. Quenching of ROS by overexpression of catalase also blocked CD-induced cell cycle arrest and apoptosis. We showed that CD enhanced the expression and nuclear translocation of Forkhead box O3 (FOXO3a) via upstream c-Jun N-terminal kinase, inducing the expression of FOXO3a and its target genes, including p21, p27, and Bim. This process led to the reduction of cyclin D1 and enhancement of activated caspase-3 expression. The addition of NAC markedly reversed these effects, knockdown of FOXO3a using small interfering RNA also decreased CD-induced G2/M arrest and apoptosis. In vivo, CD efficiently suppressed the growth of MDA-MB 231 breast cancer xenograft tumors. Taken together, our data provide a molecular mechanistic rationale for CD-induced cell cycle arrest and apoptosis in breast cancer cells.


Psychiatric history and subthreshold symptoms as predictors of the occurrence of depressive or anxiety disorder within 2 years.
Past episodes of depressive or anxiety disorders and subthreshold symptoms have both been reported to predict the occurrence of depressive or anxiety disorders. It is unclear to what extent the two factors interact or predict these disorders independently. To examine the extent to which history, subthreshold symptoms and their combination predict the occurrence of depressive (major depressive disorder, dysthymia) or anxiety disorders (social phobia, panic disorder, agoraphobia, generalised anxiety disorder) over a 2-year period. This was a prospective cohort study with 1167 participants: the Netherlands Study of Depression and Anxiety. Anxiety and depressive disorders were determined with the Composite International Diagnostic Interview, subthreshold symptoms were determined with the Inventory of Depressive Symptomatology-Self Report and the Beck Anxiety Inventory. Occurrence of depressive disorder was best predicted by a combination of a history of depression and subthreshold symptoms, followed by either one alone. Occurrence of anxiety disorder was best predicted by both a combination of a history of anxiety disorder and subthreshold symptoms and a combination of a history of depression and subthreshold symptoms, followed by any subthreshold symptoms or a history of any disorder alone. A history and subthreshold symptoms independently predicted the subsequent occurrence of depressive or anxiety disorder. Together these two characteristics provide reasonable discriminative value. Whereas anxiety predicted the occurrence of an anxiety disorder only, depression predicted the occurrence of both depressive and anxiety disorders.


Development and Psychometric Assessment of a Spirituality Questionnaire for Indian Palliative Care Patients.
There are only a few studies on spirituality among palliative care patients in India. This gap in research may be caused by the absence of relevant questionnaires and scales specifically designed for Indian palliative care populations. In this study, we describe the development of such a questionnaire and explain its psychometric characteristics. We designed a questionnaire on the basis of a systematic review of the literature. After a review of the questionnaire by specialists and a subsequent pilot study, the questionnaire was amended. The final questionnaire consisted of a list of 36 spirituality items. It was administered to a sample of 300 cancer patients attending the pain clinic of a tertiary hospital in New Delhi. A factor analysis led to four factors explaining 54.6% of variance: Shifting moral and religious values (Factor 1), support from religious relationship (Factor 2), existential blame (Factor 3), and spiritual trust (Factor 4). The skewness and kurtosis for Factors 1, 3, and 4 were within a tolerable range for assuming a normal distribution, but Factor 2 was skewed. The alphas showed that the four factors have an acceptable internal consistency. Statistically significant associations were observed for age and Factor 3 (P = 0.004), gender and Factor 4 (P = 0.014), marital status and Factors 3 (P = 0.002) and 4 (P = 0.001), educational level and Factors 3 (P < 0.001) and 4 (P < 0.001), and pain scores and Factors 1 (P < 0.001), 2 (P < 0.001), and 3 (P = 0.001). The questionnaire offers promising prospects for the study of spirituality among palliative care patients in India.


The several ways to authentically cure Alzheimer's dementia.
Although drugs may slow its progression, authentic cure of AD has never been accomplished. Here, six approaches are suggested that might achieve genuine cure. The six therapies include: 1) treatments addressing levels of TGF-β and Wnt/β-catenin, that become significantly reduced after MCI transitions to AD, and addressing also the impaired epithelial-to-mesenchymal transition (EMT) in AD's pathogenesis; 2) and 3) are two formulations that address the inadequate counter-responses to initial loss of cognition; 4) treatments addressing the brain cells whose impaired functions result in MCI and dementia; 5) the need for using partner drugs even when a particular drug addresses a single pathogenetic cause such as amyloid deposition; 6) enhancing the likelihood of genuine cure by using combinations of approaches chosen from the foregoing. Briefly, genuine cure of AD is possible; however, since AD denotes not one but multiple, phenotypically similar conditions, no one therapy can be generalized to all cases.


Identification of a novel synaptic protein, TMTC3, involved in periventricular nodular heterotopia with intellectual disability and epilepsy.
Defects in neuronal migration cause brain malformations, which are associated with intellectual disability (ID) and epilepsy. Using exome sequencing, we identified compound heterozygous variants (p.Arg71His and p. Leu729ThrfsTer6) in TMTC3, encoding transmembrane and tetratricopeptide repeat containing 3, in four siblings with nocturnal seizures and ID. Three of the four siblings have periventricular nodular heterotopia (PVNH), a common brain malformation caused by failure of neurons to migrate from the ventricular zone to the cortex. Expression analysis using patient-derived cells confirmed reduced TMTC3 transcript levels and loss of the TMTC3 protein compared to parental and control cells. As TMTC3 function is currently unexplored in the brain, we gathered support for a neurobiological role for TMTC3 by generating flies with post-mitotic neuron-specific knockdown of the highly conserved Drosophila melanogaster TMTC3 ortholog, CG4050/tmtc3. Neuron-specific knockdown of tmtc3 in flies resulted in increased susceptibility to induced seizures. Importantly, this phenotype was rescued by neuron-specific expression of human TMTC3, suggesting a role for TMTC3 in seizure biology. In addition, we observed co-localization of TMTC3 in the rat brain with vesicular GABA transporter (VGAT), a presynaptic marker for inhibitory synapses. TMTC3 is localized at VGAT positive pre-synaptic terminals and boutons in the rat hypothalamus and piriform cortex, suggesting a role for TMTC3 in the regulation of GABAergic inhibitory synapses. TMTC3 did not co-localize with Vglut2, a presynaptic marker for excitatory neurons. Our data identified TMTC3 as a synaptic protein that is involved in PVNH with ID and epilepsy, in addition to its previously described association with cobblestone lissencephaly.


A nonhuman primate aerosol deposition model for toxicological and pharmaceutical studies.
Nonhuman primates may be used as human surrogates in inhalation exposure studies to assess either the (1) adverse health effects of airborne particulate matter or (2) therapeutic effects of aerosolized drugs and proteins. Mathematical models describing the behavior and fate of inhaled aerosols may be used to complement such laboratory investigations. For example, the optimal conditions, in terms of ventilatory parameters (e.g., breathing frequency and tidal volume) and aerosol characteristics (e.g., geometric size and density), necessary to target drug delivery to specific sites within the respiratory tract may be estimated a priori with models. In this work a mathematical description of the rhesus monkey (Macaca mulatta) lung is presented for use with an aerosol deposition model. Deposition patterns of 0.01- to 5-microm-diameter monodisperse aerosols within lungs were calculated for 3 monkey lung models (using different descriptions of alveolated regions) and compared to human lung results obtained using a previously validated mathematical model of deposition physics. Our findings suggest that there are significant differences between deposition patterns in monkeys and humans. The nonhuman primates had greater exposures to inhaled substances, particularly on the basis of deposition per unit airway surface area. However, the different alveolar volumes in the rhesus monkey models had only minor effects on aerosol dosimetry within those lungs. By being aware of such quantitative differences, investigators can employ the respective primate models (human and nonhuman) to more effectively design and interpret the results of future inhalation exposure experiments.


Generation of cytokines in human visceral leishmaniasis: dissociation of endogenous TNF-alpha and IL-1 beta production.
The role of TNF-alpha in visceral leishmaniasis is ambivalent, the eventual outcome of this infection, cure or generalization, being determined by the relative amounts of cytokines produced in vivo. Since release, by monocytes/macrophages, of TNF-alpha and interleukins 1 beta (IL-1 beta) and IL-6 is important in both the induction and effector phases of the immune responses, these mediators were determined in sera and cell culture supernatants of seventeen L. donovani infected patients in Brazil. The results are compared to those of a local control group. Circulating immunoreactive TNF-alpha in patients (median, 140 pg ml-1) was increased ten-fold over controls (median 16 pg ml-1, p less than or equal to 0.0001). In contrast, serum IL-1 beta was less than 20 pg ml-1 in all patients, although detectable in sera of 3/16 Brazilian controls (chi 2 = 3.5, p less than 0.1). Mitogen induced in vitro release of IL-1 beta and IL-6 by patients' circulating mononuclear cells was significantly reduced, and the capacity of patients' peripheral monocytes for H2O2 generation in response to opsonized zymosan was significantly diminished. In the patients, serum TNF-alpha levels were inversely related to IL-1 beta release in vitro (rho = -0.57, p less than or equal to 0.01).


Influence of Brewer's Spent Grain Compounds on Glucose Metabolism Enzymes.
With a yearly production of about 39 million tons, brewer's spent grain (BSG) is the most abundant brewing industry byproduct. Because it is rich in fiber and protein, it is commonly used as cattle feed but could also be used within the human diet. Additionally, it contains many bioactive substances such as hydroxycinnamic acids that are known to be antioxidants and potent inhibitors of enzymes of glucose metabolism. Therefore, our study aim was to prepare different extracts-A1-A7 (solid-liquid extraction with 60% acetone); HE1-HE6 (alkaline hydrolysis followed by ethyl acetate extraction) and HA1-HA3 (60% acetone extraction of alkaline residue)-from various BSGs which were characterized for their total phenolic (TPC) and total flavonoid (TFC) contents, before conducting in vitro studies on their effects on the glucose metabolism enzymes α-amylase, α-glucosidase, dipeptidyl peptidase IV (DPP IV), and glycogen phosphorylase α (GPα). Depending on the extraction procedures, TPCs ranged from 20-350 µg gallic acid equivalents/mg extract and TFCs were as high as 94 µg catechin equivalents/mg extract. Strong inhibition of glucose metabolism enzymes was also observed: the IC50 values for α-glucosidase inhibition ranged from 67.4 ± 8.1 µg/mL to 268.1 ± 29.4 µg/mL, for DPP IV inhibition they ranged from 290.6 ± 97.4 to 778.4 ± 95.5 µg/mL and for GPα enzyme inhibition from 12.6 ± 1.1 to 261 ± 6 µg/mL. However, the extracts did not strongly inhibit α-amylase. In general, the A extracts from solid-liquid extraction with 60% acetone showed stronger inhibitory potential towards a-glucosidase and GPα than other extracts whereby no correlation with TPC or TFC were observed. Additionally, DPP IV was mainly inhibited by HE extracts but the effect was not of biological relevance. Our results show that BSG is a potent source of α-glucosidase and GPα inhibitors, but further research is needed to identify these bioactive compounds within BSG extracts focusing on extracts from solid-liquid extraction with 60% acetone.


Structure-function relationships among highly diverse T cells that recognize a determinant from influenza virus hemagglutinin.
We have analyzed the structural and genetic basis for T cell recognition of the complex formed between antigen and class II products of the major histocompatibility complex by performing sequence analysis of T cell receptors (TCRs) induced in response to the helper T cell site 1 of the influenza virus hemagglutinin. The results demonstrate, first, that structurally highly diverse TCRs can be utilized in recognition of the same antigen/I-Ed complex: 12 of 13 TCRs utilize unique V alpha/V beta gene segment combinations, suggesting that approximately 70 different V alpha/V beta combinations are available to BALB/c mice in response to this determinant. Second, comparison of these sequences with the ability of each hybridoma to recognize a panel of peptide analogues suggests that alpha and beta chains of these TCRs frequently determine specificity for the NH2-terminal and the COOH terminal portions, respectively, of the site 1 determinant.


Martian surface materials: effect of particle size on spectral behavior.
The presence of abundant limonite on Mars has long been the subject of controversy. Some advocates of abundant limonite also suggest that the albedo differences between Martian light and dark areas are caused by different sizes of particles in those areas. We show that the relative albedo is reversed from the blue to the red for samples of limonite with particles of different sizes. Observations of Mars reveal no blue-red albedo reversal between the light and dark areas. Consequently, the hypothesis of particle size control of albedo is incompatible with the presence of abundant limonite on Mars.


Amylolytic fungi in starter cakes for rice beer production.
Two types of starter cakes, viz. amou and perok-kushi, used in the production of rice beer in Assam, India, by the Bodo and Deori communities, respectively, were used for the isolation of amylolytic fungi. Based on the sequencing of their internal transcribed spacer (ITS) regions the fungi were identified as Amylomyces rouxii and Rhizopus oryzae, and given the strain names TU460 and TU465, respectively. Both the strains showed the ability to degrade and saccharify starch. The glucoamylase activity was considerably high in A. rouxii TU460 (14.92 μmol/min) as compared to R. oryzae TU465 (1.41 μmol/min), whereas α-amylase activity was found to be closely related, i.e. 7.02 and 6.09 unit mL-1, respectively. SDS PAGE for the determination of the molecular size of the glucoamylase enzymes revealed the production of two distinct units of 59 kDa and 31 kDa by A. rouxii TU460, and one unit of 72 kDa by R. oryzae TU465. LC MS/MS analysis revealed that no mycotoxins were produced by either of the strains. The overall study indicated a good amylolytic property of both strains and a potential for application in the starch processing industries.