Spleen contributes to chronic restraint stress-induced lung injury through splenic CD11b+ cells.
Chronic stress can induce lung injury. The spleen, as the largest peripheral immune organ, plays a crucial role in various lung diseases. Our previous study found that the spleen underwent significant changes during chronic restraint stress (CRS). However, the exact role of the spleen in CRS-induced lung injury remains unclear. In this study, we found that CRS could increase lung index. CRS could lead to alterations of the lungs such as destruction of alveolar wall, thickening of alveolar septa, dilation of pulmonary capillaries, and increased inflammatory cell infiltration. CRS increases the concentration of malondialdehyde (MDA), decreases the level of surfactant protein A (SP-A), and elevates the levels of pro-inflammatory factors (TNF-α, IL-6, and IL-1β) in the lungs. Additionally, CRS could increase the proportions and numbers of CD11b+Ly6ChiLy6G- monocytes in the lung, while cannot alter proportions and numbers of CD3-NK1.1+ NK cells, CD3+CD4+ T cells, CD3+CD8+ T cells, and CD11b+Ly6G+ neutrophils. Moreover, the levels of inflammatory markers in lung tissues were positively correlated with the proportion of CD11b+Ly6ChiLy6G- monocytes. Interestingly, splenectomy inhibited CRS-induced lung injury and attenuated the alteration in the proportion of CD11b+Ly6ChiLy6G- monocytes in the lungs induced by CRS. Moreover, splenic CD11b+ cells, rather than splenic CD11b- cells, transfused into splenectomized mice, and subsequently exposed to CRS, can cause lung injury. These results suggest that CRS could induce lung injury and CD11b+Ly6ChiLy6G- monocytes aggregation in the lung. The spleen could contribute to CRS-induced lung injury. Furthermore, splenic CD11b+ cells might play an important role in CRS-induced lung injury.


MicroRNA 223 is upregulated in the multistep progression of Barrett's esophagus and modulates sensitivity to chemotherapy by targeting PARP1.
Recent microarray and RNA-sequencing studies have uncovered aberrantly expressed microRNAs (miRNA) in Barrett's esophagus-associated esophageal adenocarcinoma. The functional significance of these miRNAs in esophageal adenocarcinoma initiation and progression is largely unknown. Expression levels of miR-199a/b-3p, -199a-5p, -199b-5p, -200b, -200c, -223, and -375 were determined in microdissected tissues from cardiac mucosa, Barrett's esophagus, dysplastic Barrett's esophagus, and esophageal adenocarcinoma using quantitative real-time PCR. miR-223 expression was validated in precursors and esophageal adenocarcinomas from 95 patients with esophageal adenocarcinoma by in situ hybridization (ISH). miR-223 was transfected into two esophageal adenocarcinoma cell lines, and in vitro assays were conducted. Target genes were identified using Illumina microarray, and results were validated in cell lines and human specimens. miR-199 family members and miR-223 were significantly overexpressed in esophageal adenocarcinoma, however, only miR-223 showed a stepwise increase during esophageal adenocarcinoma carcinogenesis. A similar trend was observed by ISH, which additionally showed that miR-223 is exclusively expressed by the epithelial compartment. miR-223-overexpressing cells had statistically significantly more migratory and invasive potential than scramble sequence-transfected cells. PARP1 was identified as a direct target gene of miR-223 in esophageal adenocarcinoma cells. Increased sensitivity to chemotherapy was observed in cells with enforced miR-223 expression and reduced PARP1. miR-223 is significantly upregulated during the Barrett's esophagus-dysplasia-esophageal adenocarcinoma sequence. Although high miR-223 levels might contribute to an aggressive phenotype, our results also suggest that patients with esophageal adenocarcinoma with high miR-223 levels might benefit from treatment with DNA-damaging agents.


Dissection of the Effects of JAK and BTK Inhibitors on the Functionality of Healthy and Malignant Lymphocytes.
Despite the emergence of small molecule inhibitors, current treatment strategies for chronic lymphocytic leukemia (CLL) are not curative, and the search for new therapeutic modalities continues. Prosurvival signaling derived from the microenvironment is often mediated via JAK signaling. However, whether JAK inhibitors are useful in CLL therapy has not been studied extensively. JAK inhibitors are valuable therapeutic agents in myelofibrosis and show promising results in graft-versus-host-disease. However, JAK inhibition is associated with an increased infection risk, presumably because of the effect on other immune cells, a feature shared with other kinase inhibitors used for CLL treatment, such as the BTK inhibitor ibrutinib and the PI3Kδ inhibitor idelalisib. We compared functional effects of the JAK1/2 inhibitors momelotinib and ruxolitinib, the BTK inhibitors ibrutinib and tirabrutinib, and PI3Kδ inhibitor idelalisib on malignant CLL cells but also on healthy human T, B, and NK lymphocytes. We found several interesting differences among the inhibitors, apart from expected and well-known effects. Momelotinib but not ruxolitinib blocked cytokine-induced proliferation of CLL cells. Momelotinib also reduced BCR signaling, in contrast to ruxolitinib, indicating that these JAK inhibitors in fact have a distinct target spectrum. In contrast to tirabrutinib, ibrutinib had inhibitory effects on T cell activation, probably because of ITK inhibition. Remarkably, both BTK inhibitors stimulated IFN-γ production in a mixed lymphocyte reaction. Collectively, our results demonstrate that kinase inhibitors directed at identical targets may have differential effects on lymphocyte function. Their unique profile could be strategically employed to balance desired versus unwanted lymphocyte inhibition.


Report of the First International Symposium on NUT Carcinoma.
NUT carcinoma is a rare, aggressive cancer defined by rearrangements of the NUTM1 gene. No routinely effective treatments of NUT carcinoma exist, despite harboring a targetable oncoprotein, most commonly BRD4-NUT. The vast majority of cases are fatal. Poor awareness of the disease is a major obstacle to progress in the treatment of NUT carcinoma. While the incidence likely exceeds that of Ewing sarcoma, and BRD4-NUT heralded the bromodomain and extra-terminal domain (BET) inhibitor class of selective epigenetic modulators, NUT carcinoma is incorrectly perceived as "impossibly rare," and therefore receives comparatively little private or governmental funding or prioritization by pharma. To raise awareness, propagate scientific knowledge, and initiate a consensus on standard and targeted treatment of NUT carcinoma, we held the First International Symposium on NUT Carcinoma on March 3, 2021. This virtual event had more than eighty attendees from the Americas, Europe, Asia, and Australia. Patients with NUT carcinoma and family members were represented and shared perspectives. Broadly, the four areas discussed by experts in the field included (1) the biology of NUT carcinoma; (2) standard approaches to the treatment of NUT carcinoma; (3) results of clinical trials using BET inhibitors; and (4) future directions, including novel BET bromodomain inhibitors, combinatorial approaches, and immunotherapy. It was concluded that standard chemotherapeutic approaches and first-generation BET bromodomain inhibitors, the latter complicated by a narrow therapeutic window, are only modestly effective in a minority of cases. Nonetheless, emerging second-generation targeted inhibitors, novel rational synergistic combinations, and the incorporation of immuno-oncology approaches hold promise to improve the prognosis of this disease.


Atlantic salmon skin and fillet color changes effected by perimortem handling stress, rigor mortis, and ice storage.
The changes in skin and fillet color of anesthetized and exhausted Atlantic salmon were determined immediately after killing, during rigor mortis, and after ice storage for 7 d. Skin color (CIE L*, a*, b*, and related values) was determined by a Minolta Chroma Meter. Roche SalmoFan Lineal and Roche Color Card values were determined by a computer vision method and a sensory panel. Before color assessment, the stress levels of the 2 fish groups were characterized in terms of white muscle parameters (pH, rigor mortis, and core temperature). The results showed that perimortem handling stress initially significantly affected several color parameters of skin and fillets. Significant transient fillet color changes also occurred in the prerigor phase and during the development of rigor mortis. Our results suggested that fillet color was affected by postmortem glycolysis (pH drop, particularly in anesthetized fillets), then by onset and development of rigor mortis. The color change patterns during storage were different for the 2 groups of fish. The computer vision method was considered suitable for automated (online) quality control and grading of salmonid fillets according to color.


Binding of methyltrienolone to glucocorticoid receptors in rat muscle cytosol.
Androgen receptor measurements with [3H]-methyltrienolone [17 beta-hydroxy-17 alpha-methyl-estra-4,9,11-trien-3-one (R1881)] or [3H]testosterone in vastus lateralis muscle cytosol of intact male rats yielded similar numbers of binding sites. Gonadectomy significantly increased the androgen receptor values, and both steroids gave similar results. The binding of [3H]-testosterone was not affected by adrenalectomy. In contrast, [3H]R1881 binding increased 2-fold after adrenalectomy. Competition experiments as well as sucrose density gradient analysis indicated that R1881, in addition to its binding of the androgen receptor, was also bound to glucocorticoid receptor. These results show that the synthetic androgen R1881 must be used with caution in tissues suspected to contain glucocorticoid receptors.


The treatment of early Parkinson's disease: levodopa rehabilitated.
Many clinicians regard levodopa as a last resort in the symptomatic treatment of Parkinson's disease. Here we critically review the arguments that are typically used to postpone the start of levodopa for as long as possible. We will point out that most concerns are invalid. Levodopa remains the most effective and best tolerated Parkinson's drug to date, and should have an important role in all therapeutic strategies, both as monotherapy in early Parkinson's disease and as part of combination therapy in advanced disease. Regardless of disease stage, the choice of a particular drug should not be driven by fear of long term complications but by the clinical condition of the patient at the time, with an emphasis on functioning in everyday life and any comorbidity. A 'phobia' for levodopa—or, indeed, for any other antiparkinsonian medication—is unacceptable according to current evidence.


Association between childhood maltreatment and adult emotional dysregulation in a low-income, urban, African American sample: moderation by oxytocin receptor gene.
The ability to effectively regulate emotions and a secure attachment style are critical for maintaining mental health across the life span. The experience of childhood maltreatment interferes with normal development of emotional regulation and dramatically increases risk for a wide range of psychiatric disorders in adulthood. The central nervous system oxytocin systems are critically involved in mediating social attachment and buffering psychophysiological responses to stress. We therefore investigated the impact of childhood maltreatment and an oxytocin receptor (OXTR) single nucleotide polymorphism (rs53576) and their interaction on emotional dysregulation and attachment style in adulthood in a sample of low-income, African American men and women recruited from primary care clinics of an urban, public hospital. Consistent with prior research, we found that the severity of childhood maltreatment was associated with increased levels of emotional dysregulation in adulthood. Childhood maltreatment was also positively associated with ratings of disorganized/unresolved adult attachment style and negatively associated with ratings of secure adult attachment style. There was no direct association between rs53576 and emotional dysregulation or ratings of adult attachment style. However, there were significant interactions between rs53576 and childhood maltreatment in predicting level of adult emotional dysregulation and attachment style. Specifically, G/G genotype carriers were at risk for increased emotional dysregulation when exposed to three or more categories of childhood abuse. In addition, G/G genotype carriers exhibited enhanced disorganized adult attachment style when exposed to severe childhood abuse compared to A/A and A/G carriers. Our findings suggest that A allele carriers of OXTR rs53576 are resilient against the effects of severe childhood adversity, by protection against emotional dysregulation and disorganized attachment.


Leveraging community engaged research partnerships for crisis and emergency risk communication to vulnerable populations in the COVID-19 pandemic.
Community engagement is important for reaching vulnerable populations in the coronavirus disease 2019 (COVID-19) pandemic. A risk communication framework was implemented by a community-engaged research (CEnR) partnership in Southeast Minnesota to address COVID-19 prevention, testing, and socioeconomic impacts. Bidirectional communication between Communication Leaders and community members within their social networks was used by the partnership to refine messages, leverage resources, and advise policy makers. Over 14 days, messages were delivered by 24 Communication Leaders in 6 languages across 9 electronic platforms to 9882 individuals within their networks. CEnR partnerships may effectively implement crisis and emergency risk communication to vulnerable populations in a pandemic.


Towards proteome standards: the use of absolute quantitation in high-throughput biomarker discovery.
The use of proteomics to profile biological fluids and identify therein biomarkers for cancer and other diseases was initially received with considerable excitement. However, results have fallen short of the expectations. Traditionally, protein biomarkers have been identified by measurement of relative expression changes between case and control samples from which differentially expressed proteins are then considered to represent biomarker candidates. We argue that current individual proteomics-based biomarker discovery studies lack the statistical strength for the identification of high-confidence biomarkers. Instead, multi-group efforts are necessary to facilitate the generation of sufficient sample sizes. This is contingent on the ability to collate and cross-compare data from different studies, which will require the use of a common metric or standards. Though profound, the technical challenges for absolute protein quantification can be overcome. The use of matrix specific, shared standards for absolute quantitation presents an opportunity to facilitate the much needed, but currently impossible, comparisons of different studies. In addition to community-wide approaches to standardize pre-analytical biomarker research studies, it is also important to establish means to integrate experimental data from different studies in order to assess the usefulness of proposed biomarkers with sufficient statistical certainty.


Six1 Overexpression Promotes Glucose Metabolism and Invasion Through Regulation of GLUT3, MMP2 and Snail in Thyroid Cancer Cells.
Sineoculis homeobox homolog 1 (Six1) overexpression has been implicated in several human cancers. To date, its clinical significance and potential function in human thyroid cancer remain unclear. Immunohistochemistry was used to examine the protein expression of BCAT1 in 89 cases of thyroid cancer tissues. We overexpressed and knockdown Six1 in TPC-1 and B-CPAP thyroid cancer cell lines. Biological roles and potential mechanisms of Six1 were examined using CCK-8, colony formation assay, Matrigel invasion assay, Western blot, PCR, ATP assay, and 2-NBDG uptake assay. We showed that Six1 protein was upregulated in thyroid cancers and was associated with tumor size and nodal metastasis. Analysis of TCGA dataset indicated that Six1 mRNA was higher in thyroid cancers compared with normal thyroid. CCK-8, colony formation and Matrigel invasion assays demonstrated that Six1 overexpression promoted proliferation, colony number and invasion while Six1 siRNA knockdown inhibited the growth rate, colony formation ability and invasive ability in both cell lines. Notably, Six1 upregulated glucose consumption, lactate production level and ATP level. 2-NBDG uptake analysis showed that Six1 overexpression upregulated glucose uptake while Six1 knockdown inhibited glucose uptake. Further analysis revealed that Six1 overexpression upregulated Snail, MMP2 and GLUT3 at both mRNA and protein levels. TCGA analysis demonstrated positive associations between Six1 and Snail, MMP2 and GLUT3 at the mRNA levels. Taken together, our data demonstrated that Six1 was upregulated in human thyroid cancers and promoted cell proliferation and invasion. Our data also revealed new roles of Six1 in thyroid cancer development by modulating glucose metabolism and invasion, possibly through regulation of Snail, MMP2 and GLUT3.


Bisphenol A emission factors from industrial sources and elimination rates in a sewage treatment plant.
Bisphenol A (BPA) is widely used for the production of epoxy resins and polycarbonate plastics and is considered an endocrine disruptor. Special in vitro test systems and animal experiments showed a weak estrogenic activity. Aquatic wildlife especially could be endangered by waste water discharges. To manage possible risks arising from BPA emissions the major fluxes need to be investigated and the sources of the contamination of municipal treatment plants need to be determined. In this study, five major industrial point sources, two different household areas and the influent and effluent of the corresponding treatment plant (WWTP) were monitored simultaneously at a plant serving 120,000 population equivalents. A paper producing plant was the major BPA contributor to the influent load of the wastewater treatment plant. All the other emissions from point sources, including the two household areas, were considerably lower. The minimum elimination rate in the WTTP could be determined at 78% with an average of 89% of the total BPA-load. For a possible pollution-forecast, or for a comparison between different point sources, emission factors based on COD-emissions were calculated for industrial and household point sources at BPA/COD-ratios between 1.4 x 10(-8) - 125 x 10(-8) and 1.3 x 10(-6) - 6.3 x 10(-6), respectively.


Network-based Identification of novel cancer genes.
Genes involved in cancer susceptibility and progression can serve as templates for searching protein networks for novel cancer genes. To this end, we introduce a general network searching method, MaxLink, and apply it to find and rank cancer gene candidates by their connectivity to known cancer genes. Using a comprehensive protein interaction network, we searched for genes connected to known cancer genes. First, we compiled a new set of 812 genes involved in cancer, more than twice the number in the Cancer Gene Census. Their network neighbors were then extracted. This candidate list was refined by selecting genes with unexpectedly high levels of connectivity to cancer genes and without previous association to cancer. This produced a list of 1891 new cancer candidates with up to 55 connections to known cancer genes. We validated our method by cross-validation, Gene Ontology term bias, and differential expression in cancer versus normal tissue. An example novel cancer gene candidate is presented with detailed analysis of the local network and neighbor annotation. Our study provides a ranked list of high priority targets for further studies in cancer research. Supplemental material is included.


[Nursing absenteeism: experience with workers at a teaching hospital].
The nursing work conditions are surrounded by physical and chemical risks that result in a high number of absences as well as leaves. The purpose of this study was to characterize the absences of nursing workers from a University Hospital located in the city of São José do Rio Preto-SP during the year of 1999. This is a census epidemiological investigation. Data were collected from registries of the Hospital Workers' Attendance Center. Results showed that 333 workers were involved in 662 episodes of absences during this period. Sick leaves of nurses, nursing technicians and auxiliaries occurred mainly due to genitourinary tract disorders and other diseases. With respect to nursing aids, the causes of absences were: disturbances in sensory organs, infectious parasitic and respiratory diseases. These data enabled a better analysis of that hospital's environment as well as the improvement of these professionals' work conditions.


MANAGEMENT OF ENDOCRINE DISEASE: Congenital adrenal hyperplasia due to 21-hydroxylase deficiency: update on the management of adult patients and prenatal treatment.
Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is characterized by cortisol and in some cases aldosterone deficiency associated with androgen excess. Goals of treatment are to replace deficient hormones and control androgen excess, while avoiding the adverse effects of exogenous glucocorticoid. Over the last 5 years, cohorts of adults with CAH due to 21-hydroxylase deficiency from Europe and the United States have been described, allowing us to have a better knowledge of long-term complications of the disease and its treatment. Patients with CAH have increased mortality, morbidity and risk for infertility and metabolic disorders. These comorbidities are due in part to the drawbacks of the currently available glucocorticoid therapy. Consequently, novel therapies are being developed and studied in an attempt to improve patient outcomes. New management strategies in the care of pregnancies at risk for congenital adrenal hyperplasia using fetal sex determination and dexamethasone have also been described, but remain a subject of debate. We focused the present overview on the data published in the last 5 years, concentrating on studies dealing with cardiovascular risk, fertility, treatment and prenatal management in adults with classic CAH to provide the reader with an updated review on this rapidly evolving field of knowledge.


AI-driven molecular generation of not-patented pharmaceutical compounds using world open patent data.
Developing compounds with novel structures is important for the production of new drugs. From an intellectual perspective, confirming the patent status of newly developed compounds is essential, particularly for pharmaceutical companies. The generation of a large number of compounds has been made possible because of the recent advances in artificial intelligence (AI). However, confirming the patent status of these generated molecules has been a challenge because there are no free and easy-to-use tools that can be used to determine the novelty of the generated compounds in terms of patents in a timely manner; additionally, there are no appropriate reference databases for pharmaceutical patents in the world. In this study, two public databases, SureChEMBL and Google Patents Public Datasets, were used to create a reference database of drug-related patented compounds using international patent classification. An exact structure search system was constructed using InChIKey and a relational database system to rapidly search for compounds in the reference database. Because drug-related patented compounds are a good source for generative AI to learn useful chemical structures, they were used as the training data. Furthermore, molecule generation was successfully directed by increasing and decreasing the number of generated patented compounds through incorporation of patent status (i.e., patented or not) into learning. The use of patent status enabled generation of novel molecules with high drug-likeness. The generation using generative AI with patent information would help efficiently propose novel compounds in terms of pharmaceutical patents. Scientific contribution: In this study, a new molecule-generation method that takes into account the patent status of molecules, which has rarely been considered but is an important feature in drug discovery, was developed. The method enables the generation of novel molecules based on pharmaceutical patents with high drug-likeness and will help in the efficient development of effective drug compounds.


Heterogeneity of sodium-dependent D-glucose transport sites along the proximal tubule: evidence from vesicle studies.
The glucose transport properties of brush border membrane vesicles from the outer cortex (early proximal tubule) and outer medulla (late proximal tubule) of rabbit kidney were studied. In the outer cortical preparation the behavior of the sodium-dependent component of D-glucose flux indicated the presence of a low-affinity transport system with Km congruent to 6 mM and Vmax congruent to 10 nmol.min-1.mg protein-1 as measured under zero trans conditions at 40 mM NaCl and 17 degrees C. By contrast, in the outer medullary preparation this component of flux behaved as a high-affinity system with Km congruent to 0.35 mM and Vmax congruent to 4 nmol.min-1.mg protein-1. Differences in transport specificity between the two preparations were also indicated and glucose uptake by the outer cortical vesicles was significantly more sensitive to inhibition by phlorizin. These results suggest the existence of two distinct sodium-dependent D-glucose transport systems in the renal proximal tubule brush border membrane. The kinetic studies presented here were done under zero trans sodium and glucose conditions. The rationale and methodology for carrying out these measurements reliably are discussed in detail.


Sampling-dependent up-regulation of gene expression in sequential samples of human airway epithelial cells.
As part of a study of in vivo gene expression levels in the human airway epithelium in response to chronic cigarette smoking, we have identified a number of genes whose expression levels are altered in a time-dependent fashion resulting from the procedure used to sample epithelial cells. Fiberoptic bronchoscopy and airway epithelium brushing were used to obtain independent samples from a single individual, 1st from the right lung, followed by sampling of the left lung. We observed that a specific subset of early response genes encoding proteins involved in transcription, signal transduction, cell cycle/growth, and apoptosis were significantly up-regulated in the left lung samples (the 2nd region to be sampled) compared with the right lung samples (the 1st region to be sampled). This response was due to the temporal nature of the sampling procedure and not to inherent gene expression differences between airway epithelium of the right and left lungs. When the order of sampling was reversed, with the left airway epithelium sampled 1st, the same subset of genes were up-regulated in the samples obtained from the right airway epithelium. The time-dependent up-regulation of these genes was likely in response to the stress of the procedure and/or the anesthesia used. Sampling-dependent uncertainty of gene expression is likely a general phenomenon relevant to the procedures used for obtaining biological samples, particularly in humans where the sampling procedures are dependent on ensuring comfort and safety.


Combined effects of Er: YAG laser and casein phosphopeptide-amorphous calcium phosphate on the inhibition of enamel demineralization: An in vitro study.
Development of white spot lesions on enamel is a significant and common problem during the fixed orthodontic treatment. Various preventive methods have been suggested. The purpose of this study was to evaluate the preventive potential of MI Paste Plus, Er: YAG Laser and combined under similar in vitro conditions against demineralization. In this experimental in vitro study, 60 extracted premolars were randomly allocated to four groups (n = 15) of control, MI Paste Plus, Laser and MI + Laser (MIL). Enamel surface of each group was treated with one of above materials before and during the pH cycling for 12 days through a daily procedure of demineralization and remineralization for 3 h and 20 h, respectively. Teeth were sectioned and evaluated quantitatively by cross-sectional microhardness testing at 20 μm intervals from the outer enamel surface toward dentinoenamel junction up to 160 μm and data were analyzed using the one-way analysis of variance and Tukey test. P < 0.05 was considered as significant. MIL group had the least amount of demineralization (P < 0.001). Control group (C group) had the greatest relative mineral loss and the laser group (L group) had 45% less mineral loss than the C group and there was no significant difference between the MI Paste Plus and L group (P = 0.154). BASED ON THESE RESULTS, ER: YAG laser was able to decrease demineralization and was a potential alternative to preventive dentistry and was more effective when combined with casein phosphopeptide-amorphous calcium phosphate products.


Ventral brain stem compression in pediatric and young adult patients with Chiari I malformations.
The purposes of this study were as follows: 1) to determine the incidence and degree of ventral brain stem compression (VBSC) in pediatric and young adult patients with Chiari I malformations, and 2) to correlate VBSC with other imaging and clinical factors to help determine what amount of VBSC is successfully treated with a posterior decompressive procedure alone. The magnetic resonance images and clinical histories of 40 pediatric and young adult patients with Chiari I malformations were analyzed for subjective grade of VBSC, distance of tonsillar descent, odontoid's relation to Chamberlain's and Wackenheim's line, clival length, foramen magnum diameter, syringomyelia, scoliosis, hydrocephalus, presenting clinical status, treatment, and outcome. To objectively measure the amount of ventral cervicomedullary encroachment by the odontoid and its investing tissues into the rostral spinal canal, a line (B-C2) was drawn between the basion and posteroinferior aspect of the C2 body on a sagittal magnetic resonance image. A line perpendicular to this line, pB-C2, was drawn through the odontoid tip to the ventral dura, and a distance (representing the amount of ventral canal encroachment) was measured. Flattening and distortion of the ventral brain stem were present in 48 and 28% of the patients, respectively. Only two patients had basilar invagination by traditional definitions. pB-C2 measurements correlated with the subjective grade of VBSC (P < 0.05), age, and distance of tonsillar descent (P < 0.05). Eye motion abnormalities and upper cervical osseous anomalies were associated with higher pB-C2 measurements. All patients with a pB-C2 measurement of less than 9 mm were treated successfully with posterior fossa decompression alone despite any subjective VBSC. Some patients with pB-C2 measurements greater than 9 mm had either preoperative neurological deficits or neurological worsening after posterior fossa decompression referable to VBSC. Patients with a pB-C2 measurement of less than 9 mm do not require treatment directed at VBSC. In select patients with pB-C2 measurements of 9 mm or greater, reduction of VBSC may be prudent before posterior fossa decompression.


Hippocampal synaptic plasticity: role in spatial learning or the automatic recording of attended experience?
Allocentric spatial learning can sometimes occur in one trial. The incorporation of information into a spatial representation may, therefore, obey a one-trial correlational learning rule rather than a multi-trial error-correcting rule. It has been suggested that physiological implementation of such a rule could be mediated by N-methyl-D-aspartate (NMDA) receptor-dependent long-term potentiation (LTP) in the hippocampus, as its induction obeys a correlational type of synaptic learning rule. Support for this idea came originally from the finding that intracerebral infusion of the NMDA antagonist AP5 impairs spatial learning, but studies summarized in the first part of this paper have called it into question. First, rats previously given experience of spatial learning in a watermaze can learn a new spatial reference memory task at a normal rate despite an appreciable NMDA receptor blockade. Second, the classical phenomenon of 'blocking' occurs in spatial learning. The latter finding implies that spatial learning can also be sensitive to an animal's expectations about reward and so depend on more than the detection of simple spatial correlations. In this paper a new hypothesis is proposed about the function of hippocampal LTP. This hypothesis retains the idea that LTP subserves rapid one-trial memory, but abandons the notion that it serves any specific role in the geometric aspects of spatial learning. It is suggested that LTP participates in the automatic recording of attended experience': a subsystem of episodic memory in which events are temporarily remembered in association with the contexts in which they occur. An automatic correlational form of synaptic plasticity is ideally suited to the online registration of context event associations. In support, it is reported that the ability of rats to remember the most recent place they have visited in a familiar environment is exquisitely sensitive to AP5 in a delay-dependent manner. Moreover, new studies of the lasting persistence of NMDA-dependent LTP, known to require protein synthesis, point to intracellular mechanisms that enable transient synaptic changes to be stabilized if they occur in close temporal proximity to important events. This new property of hippocampal LTP is a desirable characteristic of an event memory system.


The effect of postural change on the coefficient of diffusion in sarcoidosis patients.
Granulomatous angiitis is common in sarcoidosis. To test the hypothesis that patients with sarcoidosis have an impaired ability to recruit or to distend the pulmonary vascular bed, the postural change in the coefficient of diffusion (ratio of DCO to alveolar volume) for carbon monoxide was measured in 20 patients with sarcoidosis and in 10 normal subjects. The normal control group increased the coefficient of diffusion (KCO) by 12.8% (p, 0.05), when values at sitting position were compared to values at supine position. The sarcoidosis group did not significantly increase the coefficient with change in position. The absence of an increase in the coefficient of diffusion (KCO) in sarcoidosis with change in position may indicate vascular involvement.


Cluster definition by the optimization of simple measures.
We adopt the following measures of clustering based on simple edge counts in an undirected loop-free graph. Let S be a subset of the points of the graph. The compactness of S is measured by the number of edges connecting points in S to other points in S. The isolation or separation of S is measured by the negative of the number of edges connecting points in S to points not in S. The subset S is a cluster if it is compact and isolated. We study the cluster search problem: find a subset S which maximizes a linear combination of the compactness and (negative) isolation edge counts. We show that a closely related decision problem is NP-complete. We develop a pruned search tree algorithm which is much faster than complete search, especially for graphs which are derived from points embedded in a space of low dimensionality.


Dating violence: outcomes following a brief motivational interviewing intervention among at-risk adolescents in an urban emergency department.
A recent study demonstrated the efficacy of the SafERteens intervention in reducing peer violence among adolescents presenting to the emergency department (ED). The objective of this study was to determine the efficacy of this ED-based brief intervention (BI) on dating violence 1 year following the ED visit among the subsample of adolescents in the original randomized controlled trial reporting past-year dating violence. Patients (aged 14 to 18 years) at an ED were eligible for inclusion if they had past-year violence and alcohol use. Participants were randomized to one of three conditions (BI delivered by a computer [CBI], BI delivered by a therapist and a computer (T+CBI), or control) and completed follow-ups at 3, 6, and 12 months. In addition to content on alcohol misuse and peer violence, adolescents reporting dating violence received a tailored module on dating violence. The outcome of interest was frequency of moderate and severe dating violence victimization and aggression (baseline and 3, 6, and 12 months after ED visit). Among eligible adolescents, 55% (n = 397) reported dating violence and were included in these analyses. Compared to the control group (who received a resource brochure only), participants in the CBI showed reductions in moderate dating victimization at 3 months (inter-rater reliability [IRR] = 0.71; 95% confidence interval [CI] = 0.51 to 0.99; p < 0.05) and 6 months (IRR = 0.56; 95% CI = 0.38 to 0.83; p < 0.01). Models examining interaction effects were significant for the CBI on moderate dating victimization at 3 months (IRR = 0.81; 95% CI = 0.67 to 0.98; p < 0.05) and 6 months (IRR = 0.81; 95% CI = 0.66 to 0.99; p < 0.05). Significant interaction effects were found for the T+CBI on moderate dating violence victimization at 6 months (IRR = 0.81; 95% CI = 0.69 to 0.96; p < 0.01) and 12 months (IRR = 0.76; 95% CI = 0.63 to 0.90; p < 0.001) and severe dating violence victimization at 3 months (IRR = 0.76; 95% CI = 0.59 to 0.96; p < 0.05). ED-based BIs tailored to address multiple risk behaviors (i.e., peer violence, alcohol use, and dating violence) show promise for reducing moderate and severe dating victimization for up to 1 year following an ED visit.


Cellular magnetic resonance imaging: in vivo tracking of gastric cancer cells and detecting of lymph node metastases using microparticles of iron oxide in mice.
Monitoring the fate of implanted cells over time in an experimental animal may provide a new way to track the metastatic process. Lymph node metastase is of extremely importance for the prognostic prediction of gastric carcinoma. The aim of this study was to assess the feasibility of magnetic resonance imaging (MRI), using micron-sized superparamagnetic iron oxide particles (MPIO), for monitoring of the fate of gastric cancer cells and detecting the migration of gastric cancer cells through the lymphatic system in a mouse model. SGC-7901 gastric cancer cells were labeled with green fluorescent MPIO. The cells were monitored in vitro at multiple time points by staining for iron-labeled cells and by flow cytometric detection of the fluorescent MPIO. MPIO-labeled cells were implanted subcutaneously into nude mice, and cellular MRI was performed at different time points until 35 days postinjection. The potential for retention of the iron particles in vitro was evaluated. Our results showed that the labeling and uptake efficiency of MPIO reached 90.0% after 24 hrs of incubation, and a small percentage of cells that retained MPIO could be examined until 16 days after labeling. In vivo MRI-based tracking over several weeks in mice revealed regions of signal loss in the primary tumors for up to 5 weeks. Furthermore, small regions of signal void were detected in images of the inguinal lymph nodes in three mice at day 28 postinjection or later, and histological assays confirmed the presence of iron-labeled cancer cells. This study supports MPIO-based cell tracking is a useful tool for monitoring the fate of gastric cancer cells in mice over time, which may facilitate progress in understanding the mechanisms of early regional lymph node micrometastases.


Violent behavior in children and youth: preventive intervention from a psychiatric perspective. Group for the Advancement of Psychiatry, Committee on Preventive Psychiatry.
To outline causative factors for the epidemic of violence among children and youth in North America and suggest roles for child and adolescent psychiatry in preventive intervention. The committee used literature searches to identify biological, psychological, and sociocultural factors associated with violent behavior. Children and youth are both victims and perpetrators of violence. Risk factors include socioeconomic status, difficult temperament, chronic illness, psychiatric comorbidity, and parental psychopathology. Access to firearms in a culture of violence presents a particularly serious risk. Protective factors include intact family structures, prosocial peer groups, and supportive communities. Preventive interventions include the following: universal, addressed to total population groups; selective, for at-risk populations; and indicated, for children and youth developing violent behavior. Universal interventions including gun control and improved perinatal care are helpful, and selective interventions such as gun-free zones around schools may be successful. Indicated programs such as gun confiscation and conflict resolution for youth at serious risk may be useful, but only when embedded within well-funded, clinically based, and community-focused programs. Single-emphasis programs such as "Boot Camps" have intuitive appeal, but their utility is doubtful. Violent behavior can be prevented, and child and adolescent psychiatrists must be more active in community preventive interventions.


Affected pathways and transcriptional regulators in gene expression response to an ultra-marathon trail: Global and independent activity approaches.
Gene expression (GE) analyses on blood samples from marathon and half-marathon runners have reported significant impacts on the immune and inflammatory systems. An ultra-marathon trail (UMT) represents a greater effort due to its more testing conditions. For the first time, we report the genome-wide GE profiling in a group of 16 runners participating in an 82 km UMT competition. We quantified their differential GE profile before and after the race using HuGene2.0st microarrays (Affymetrix Inc., California, US). The results obtained were decomposed by means of an independent component analysis (ICA) targeting independent expression modes. We observed significant differences in the expression levels of 5,084 protein coding genes resulting in an overrepresentation of 14% of the human biological pathways from the Kyoto Encyclopedia of Genes and Genomes database. These were mainly clustered on terms related with protein synthesis repression, altered immune system and infectious diseases related mechanisms. In a second analysis, 27 out of the 196 transcriptional regulators (TRs) included in the Open Regulatory Annotation database were overrepresented. Among these TRs, we identified transcription factors from the hypoxia-inducible factors (HIF) family EPAS1 (p< 0.01) and HIF1A (p<0.001), and others jointly described in the gluconeogenesis program such as HNF4 (p< 0.001), EGR1 (p<0.001), CEBPA (p< 0.001) and a highly specific TR, YY1 (p<0.01). The five independent components, obtained from ICA, further revealed a down-regulation of 10 genes distributed in the complex I, III and V from the electron transport chain. This mitochondrial activity reduction is compatible with HIF-1 system activation. The vascular endothelial growth factor (VEGF) pathway, known to be regulated by HIF, also emerged (p<0.05). Additionally, and related to the brain rewarding circuit, the endocannabinoid signalling pathway was overrepresented (p<0.05).


Design of an image restoration algorithm for the TOMBO imaging system.
The TOMBO system (thin observation module by bound optics) is a multichannel subimaging system over a single electronic imaging device. Each subsystem provides a low-resolution (LR) image from a unique lateral point of view. By estimating the image's lateral position, a high-resolution (HR) image is restored from the series of the LR images. This paper proposes an multistage algorithm comprised of successive stages, improving difficulties in previous suggested schemes. First, the registration algorithm estimates the subchannel shift parameters and eliminates bias. Second, we introduce a fast image fusion, overcoming visual blockiness artifacts that characterized previously suggested schemes. The algorithm fuses the set of sampled subchannel images into a single image, providing the reconstruction initial estimate. Third, an edge-sensitive quadratic upper bound term to the total variation regulator is suggested. The complete algorithm allows the reconstruction of a clean, HR image, in linear computation time, by the use of the linear conjugate gradient optimization. Finally, we present a simulated comparison between the proposed method and a previously suggested image restoration method. The results show that the proposed method yields better reconstruction fidelity while eliminating spatial speckle artifacts associated with the previously suggested method.


Elimination of symbiotic Aeromonas spp. from the intestinal tract of the medicinal leech, Hirudo medicinalis, using ciprofloxacin feeding.
The use of the medicinal leech (Hirudo medicinalis) in promoting venous drainage in tissues whose vitality is threatened by venous congestion and obstruction, especially in plastic and reconstructive surgery, has been complicated by infections caused by Aeromonas spp. These are leech endosymbionts for which patients undergoing hirudotherapy frequently receive systemic chemoprophylaxis. In order to evaluate the possibility of rendering leeches safe for use on patients, H. medicinalis were fed artificially with a 2 g/L arginine solution (used as a phagostimulant) supplemented with ciprofloxacin (100 mg/L). Aeromonads were detected in 57 out of 80 control leeches (71.3%), but in none of the 56 leeches treated with ciprofloxacin (p <0.001). Treated leeches survived for up to 4 months. Tested weekly, 61% of these leeches took human blood for at least 4 weeks after treatment and all remained negative for aeromonads. All water samples in which leeches were kept before treatment were contaminated with Aeromonas spp.; none were detected in any of the NaCl/arginine solutions with which treated animals were fed. Molecular characterization of two phenotypically distinct isolates using gyrB sequencing showed that one clustered tightly with A. veronii and the other was closely related to A. media. Other environmental bacteria and fungi were isolated from 26.5% of treated leeches that had taken a blood meal 1-4 weeks after treatment. Ciprofloxacin reduced the number of leech-associated aeromonads to undetectable levels for extended periods. Most treated leeches were ready to take a blood meal after treatment, suggesting the possibility of using ciprofloxacin-treated leeches instead of chemoprophylaxis in patients undergoing hirudotherapy.


Molecular Effects of the CTG Repeats in Mutant Dystrophia Myotonica Protein Kinase Gene.
Myotonic Dystrophy type 1 (DM1) is a multi-system disorder characterized by muscle wasting, myotonia, cardiac conduction defects, cataracts, and neuropsychological dysfunction. DM1 is caused by expansion of a CTG repeat in the 3 untranslated region (UTR) of the Dystrophia Myotonica Protein Kinase (DMPK) gene. A body of work demonstrates that DMPK mRNAs containing abnormally expanded CUG repeats are toxic to several cell types. A core mechanism underlying symptoms of DM1 is that mutant DMPK RNA interferes with the developmentally regulated alternative splicing of defined pre-mRNAs. Expanded CUG repeats fold into ds(CUG) hairpins that sequester nuclear proteins including human Muscleblind-like (MBNL) and hnRNP H alternative splicing factors. DM1 cells activate CELF family member CUG-BP1 protein through hyperphosphorylation and stabilization in the cell nucleus. CUG-BP1 and MBNL1 proteins act antagonistically in exon selection in several pre-mRNA transcripts, thus MBNL1 sequestration and increase in nuclear activity of CUG-BP1 both act synergistically to missplice defined transcripts. Mutant DMPK-mediated effect on subcellular localization, and defective phosphorylation of cytoplasmic CUG-BP1, have additionally been linked to defective translation of p21 and MEF2A in DM1, possibly explaining delayed differentiation of DM1 muscle cells. Mutant DMPK transcripts bind and sequester transcription factors such as Specificity protein 1 leading to reduced transcription of selected genes. Recently, transcripts containing long hairpin structures of CUG repeats have been shown to be a Dicer ribonuclease target and Dicer-induced downregulation of the mutant DMPK transcripts triggers silencing effects on RNAs containing long complementary repeats. In summary, mutant DMPK transcripts alter gene transcription, alternative splicing, and translation of specific gene transcripts, and have the ability to trigger gene-specific silencing effects in DM1 cells. Therapies aimed at reversing these gene expression alterations should prove effective ways to treat DM1.


Outreach nurses critical for delivery of HIV care to women in western Victoria.
Women living with HIV in regional Victoria face barriers accessing care. We evaluated the care cascade and outreach nurse support required for women attending our service between 2005 and 2020. A total of 33 women attended; 97% (32/33) were on antiretroviral therapy; 67% (22/33) retained in care, 27% (9/33) transferred and 6% (2/33) lost to follow up. Of women retained in care, 95% (21/22) were on antiretroviral therapy and 91% (20/22) had virological suppression. A total of 91% (30/33) required outreach nurse care (median care episodes 100/woman; IQR 44-179) - most frequently (87%; 26/30) liaising with pharmacies and prescribers. Outreach nurses are critical in achieving UNAIDS targets for women in western Victoria.


National surveillance of Staphylococcus epidermidis recovered from bloodstream infections in Belgian hospitals.
The objectives of this study were: (i) to determine the species diversity of CoNS isolated from bloodstream infections collected during a national surveillance study; and (ii) to examine the antimicrobial resistance and genomic diversity among Staphylococcus epidermidis isolates. Eighty CoNS were identified by MALDI-TOF. Antimicrobial resistance determination, molecular characterization of resistance and virulence genes, and molecular typing were performed for S. epidermidis isolates. The majority (76%) of CoNS were identified as S. epidermidis. Among these S. epidermidis, 77% were resistant to methicillin [methicillin-resistant S. epidermidis (MRSE)] and showed multiresistance to other antimicrobials. Genes implicated in resistance were erm(C), erm(A) and msr(A) for erythromycin, aacA-aphD and aadC for aminoglycosides, tet(K) for tetracycline and mupA for high-level resistance to mupirocin. Molecular typing showed that 34/40 MRSE isolates (85%) belonged to clonal complex (CC) 2 that could be subdivided into CC2-I (ST2) and CC2-II (ST5, ST59 and ST88). In contrast, methicillin-susceptible S. epidermidis displayed high genomic diversity. The majority (70%) of S. epidermidis isolates contained an icaA or arcA gene. The icaA gene was found in the CC2-I subgroup, whereas arcA was more common in methicillin-susceptible S. epidermidis. S. epidermidis was frequently recovered among CoNS isolated from bloodstream infections with a high proportion of MRSE being multiresistant. A large number of S. epidermidis belonged to CC2, a clone that is disseminated worldwide. More studies are needed to understand its clonal evolutionary success.


Studies on the optimization of growth and indole alkaloid production by hairy root cultures of Catharanthus roseus.
Catharanthus roseus hairy root cultures, genetically transformed with Agrobacterium rhizogenes, produce a wide variety of indole alkaloids. The effect of sucrose, phosphate, nitrate, and ammonia concentrations on growth and indole alkaloid production of C. roseus hairy root cultures were studied by using statistical experimental designs and linear regression analysis. Contradictory effects of these nutrients on growth and indole alkaloid production were found. The maximal growth was obtained by having 77. 8 mg NaH(2)PO(4) . H(2)O/L and 1. 311 g KNO(3)/L in the medium, whereas the specific production of alkaloids was highest at the lowest levels of all the nutrients studied. The maximal dry weight was obtained with high values of sucrose and ammonia, but clear optimum concentrations could not be found. When having enough nutrients to support reasonable growth, it appeared difficult to affect the specific alkaloid production rates considerably. The growth (dry wt.) with the optimized nutrient concentrations in the medium was more than 50% better than in the control medium with about the same alkaloid production.


Clinical efficacy and safety of a light mask for prevention of dark adaptation in treating and preventing progression of early diabetic macular oedema at 24 months (CLEOPATRA): a multicentre, phase 3, randomised controlled trial.
We aimed to assess 24-month outcomes of wearing an organic light-emitting sleep mask as an intervention to treat and prevent progression of non-central diabetic macular oedema. CLEOPATRA was a phase 3, single-blind, parallel-group, randomised controlled trial undertaken at 15 ophthalmic centres in the UK. Adults with non-centre-involving diabetic macular oedema were randomly assigned (1:1) to wearing either a light mask during sleep (Noctura 400 Sleep Mask, PolyPhotonix Medical, Sedgefield, UK) or a sham (non-light) mask, for 24 months. Randomisation was by minimisation generated by a central web-based computer system. Outcome assessors were masked technicians and optometrists. The primary outcome was the change in maximum retinal thickness on optical coherence tomography (OCT) at 24 months, analysed using a linear mixed-effects model incorporating 4-monthly measurements and baseline adjustment. Analysis was done using the intention-to-treat principle in all randomised patients with OCT data. Safety was assessed in all patients. This trial is registered with Controlled-Trials.com, number ISRCTN85596558. Between April 10, 2014, and June 15, 2015, 308 patients were randomly assigned to wearing the light mask (n=155) or a sham mask (n=153). 277 patients (144 assigned the light mask and 133 the sham mask) contributed to the mixed-effects model over time, including 246 patients with OCT data at 24 months. The change in maximum retinal thickness at 24 months did not differ between treatment groups (mean change -9·2 μm [SE 2·5] for the light mask vs -12·9 μm [SE 2·9] for the sham mask; adjusted mean difference -0·65 μm, 95% CI -6·90 to 5·59; p=0·84). Median compliance with wearing the light mask at 24 months was 19·5% (IQR 1·9-51·6). No serious adverse events were related to either mask. The most frequent adverse events related to the assigned treatment were discomfort on the eyes (14 with the light mask vs seven with the sham mask), painful, sticky, or watery eyes (14 vs six), and sleep disturbance (seven vs one). The light mask as used in this study did not confer long-term therapeutic benefit on non-centre-involving diabetic macular oedema and the study does not support its use for this indication. The Efficacy and Mechanism Evaluation Programme, a Medical Research Council and National Institute for Health Research partnership.


Use of a controlled-nutrient experiment to test heterosis hypotheses.
A controlled-nutrient (CN) experiment was conducted to test three heterosis hypotheses with reference to tomato yield, and its components, for a set of two inbred lines and their hybrid that had previously exhibited considerable heterosis under field conditions. The CN treatments consisted of periodic applications of differential doses of nutrient solution to plants reared individually in containers filled with vermiculite. Ripe fruit were harvested, counted and weighed over a period of 340 days. The data permitted the partitioning of yield into a closed system of five component variables. Heterosis was not exhibited by yield, nor yield components, at any of the four nutrient levels. Hence the total heterosis phenomenon was classified as nutrient-dependent: heterosis occurring under field conditions, but not under the nutritional restrictions of the CN experiment. Three heterosis hypotheses were examined for their ability to explain all of the nutrient-dependent aspects of the heterosis phenomenon. Hypothesis 1: Heterosis is a consequence of a more efficient hybrid metabolic system in that it can produce more product with equal input. Hypothesis 2: Heterosis is a consequence of the somatic multiplication of additive component traits. Hypothesis 3: Heterosis is a consequence of a faster hybrid growth rate. Although none of the hypotheses are rejected by the field data, the first two are rejected by the CN experimental results. The third hypothesis fits all aspects of the nutrient-dependent heterosis phenomenon remarkably well. It is speculated that the indeterminate pattern of plant development responsible for yield and its components is due to two major gene systems: genes that determine morphogenetic, and genes that determine growth rate manifestations of growth. Under this hypothesis, the CN technique permits separation of the responses due to these two gene systems.


Coordinated control of cholesterol catabolism to bile acids and of gluconeogenesis via a novel mechanism of transcription regulation linked to the fasted-to-fed cycle.
Bile acid metabolism plays an essential role in cholesterol homeostasis and is critical for the initiation of atherosclerotic disease. However, despite the recent advances, the molecular mechanisms whereby bile acids regulate gene transcription and cholesterol homeostasis in mammals still need further investigations. Here, we show that bile acids suppress transcription of the gene (CYP7A1) encoding cholesterol 7alpha-hydroxylase, the rate-limiting enzyme in bile acid biosynthesis, also through an unusual mechanism not involving the bile acid nuclear receptor, farnesoid X receptor. By performing cell-based reporter assays, protein/protein interaction, and chromatin immunoprecipitation assays, we demonstrate that bile acids impair the recruitment of peroxisome proliferator-activated receptor-gamma coactivator-1alpha and cAMP response element-binding protein-binding protein by hepatocyte nuclear factor-4alpha, a master regulator of CYP7A1. We also show for the first time that bile acids inhibit transcription of the gene (PEPCK) encoding phosphoenolpyruvate carboxykinase, the rate-limiting enzyme in gluconeogenesis, through the same farnesoid X receptor-independent mechanism. Chromatin immunoprecipitation assay revealed that bile acid-induced dissociation of coactivators from hepatocyte nuclear factor-4alpha decreased the recruitment of RNA polymerase II to the core promoter and downstream in the 3'-untranslated regions of these two genes, reflecting the reduction of gene transcription. Finally, we found that Cyp7a1 expression was stimulated in fasted mice in parallel to Pepck, whereas the same genes were repressed by bile acids. Collectively, these results reveal a novel regulatory mechanism that controls gene transcription in response to extracellular stimuli and argue that the transcription regulation by bile acids of genes central to cholesterol and glucose metabolism should be viewed dynamically in the context of the fasted-to-fed cycle.


The Development and Piloting of Parallel Scales Measuring External and Internal HIV and Tuberculosis Stigma Among Healthcare Workers in the Free State Province, South Africa.
The dual burden of tuberculosis and human immunodeficiency virus (HIV) is severely impacting the South African healthcare workforce. However, the use of on-site occupational health services is hampered by stigma among the healthcare workforce. The success of stigma-reduction interventions is difficult to evaluate because of a dearth of appropriate scientific tools to measure stigma in this specific professional setting. The current pilot study aimed to develop and test a range of scales measuring different aspects of stigma-internal and external stigma toward tuberculosis as well as HIV-in a South African healthcare setting. The study employed data of a sample of 200 staff members of a large hospital in Bloemfontein, South Africa. Confirmatory factor analysis produced 7 scales, displaying internal construct validity: (1) colleagues' external HIV stigma, (2) colleagues' actions against external HIV stigma, (3) respondent's external HIV stigma, (4) respondent's internal HIV stigma, (5) colleagues' external tuberculosis stigma, (6) respondent's external tuberculosis stigma, and (7) respondent's internal tuberculosis stigma. Subsequent analyses (reliability analysis, structural equation modeling) demonstrated that the scales displayed good psychometric properties in terms of reliability and external construct validity. The study outcomes support the use of the developed scales as a valid and reliable means to measure levels of tuberculosis- and HIV-related stigma among the healthcare workforce in a resource-limited context. Future studies should build on these findings to fine-tune the instruments and apply them to larger study populations across a range of different resource-limited healthcare settings with high HIV and tuberculosis prevalence.


Epistatic interactions between neuraminidase mutations facilitated the emergence of the oseltamivir-resistant H1N1 influenza viruses.
Oseltamivir-resistant H1N1 influenza viruses carrying the H275Y neuraminidase mutation predominated worldwide during the 2007-2009 seasons. Although several neuraminidase substitutions were found to be necessary to counteract the adverse effects of H275Y, the order and impact of evolutionary events involved remain elusive. Here we reconstruct H1N1 neuraminidase phylogeny during 1999-2009, estimate the timing and order of crucial amino acid changes and evaluate their impact on the biological outcome of the H275Y mutation. Of the 12 neuraminidase substitutions that occurred during 1999-2009, 5 (chronologically, V234M, R222Q, K329E, D344N, H275Y and D354G) are necessary for maintaining full neuraminidase function in the presence of the H275Y mutation by altering protein accumulation or enzyme affinity/activity. The sequential emergence and cumulative effects of these mutations clearly illustrate a role for epistasis in shaping the emergence and subsequent evolution of a drug-resistant virus population, which can be useful in understanding emergence of novel viral phenotypes of influenza.


[Exposure to endotoxins in the environment. Occurrence and health hazards].
Endotoxins are lipopolysaccharides from the outer cell wall of Gram-negative bacteria. Exposure to endotoxins can take place in industries where organic material is handled, in agriculture, in garbage handling, and sewage treatment. Byssinosis defined as Monday chest tightness and slight dyspnoea in the work place has been related to endotoxin exposure in cotton mills, but studies indicate that similar symptoms may be found in other work places. Other symptoms are: Headache, nausea, gastrointestinal symptoms and influenza-like symptoms. Several studies have shown a decrease in FEV1 following exposure to endotoxins. The relationship between exposure to organic dust, microorganisms, endotoxins and other chemicals in the work place and disease needs further research.


Evidence that glucagon-mediated inhibition of acetyl-CoA carboxylase in isolated adipocytes involves increased phosphorylation of the enzyme by cyclic AMP-dependent protein kinase.
The kinetic parameters and phosphorylation state of acetyl-CoA carboxylase were analysed after purification of the enzyme by avidin--Sepharose chromatography from extracts of isolated adipocytes treated with glucagon or adrenaline. The results provide evidence that the mechanism of inhibition of acetyl-CoA carboxylase in adipocytes treated with glucagon [Zammit & Corstorphine (1982) Biochem. J. 208, 783-788] involves increased phosphorylation of the enzyme. Hormone treatment had effects on the kinetic parameters of the enzyme similar to those of phosphorylation of the enzyme in vitro by cyclic AMP-dependent protein kinase. Glucagon treatment of adipocytes led to increased phosphorylation of acetyl-CoA carboxylase in the same chymotryptic peptide as that containing the major site phosphorylated on the enzyme by purified cyclic AMP-dependent protein kinase in vitro [Munday & Hardie (1984) Eur. J. Biochem. 141, 617-627]. The dose--response curves for inhibition of enzyme activity and increased phosphorylation of the enzyme were very similar, with half-maximal effects occurring at concentrations of glucagon (0.5-1 nM) which are close to the physiological range. In general, the patterns of increased 32P-labelling of chymotryptic peptides induced by glucagon or adrenaline were similar, although there were quantitative differences between the effects of the two hormones on individual peptides. The results are discussed in terms of the possible roles of cyclic AMP-dependent and -independent protein kinases in the regulation of acetyl-CoA carboxylase activity and of lipogenesis in white adipose tissue.


Comparison of five glucose meters for self-monitoring of blood glucose by diabetic patients.
This study collected data regarding the accuracy of five currently marketed meters for home use of self-monitoring of blood glucose (SMBG). Data regarding ease of operation of each glucose meter as well as cost and availability of meters and necessary materials were compiled. Fasting blood glucose (BG) samples were drawn from patients in the adult and pediatric diabetes clinics at the University of Kansas College of Health Sciences and Hospital. Specimens were tested by a clinical laboratory as well as by each meter. Unadjusted meter readings of whole BG were plotted against laboratory readings of serum glucose and displayed in scattergrams. Scattergrams showed decreased machine accuracy in upper and lower BG ranges. Whole BG values (meter readings) were adjusted to serum glucose levels by multiplication by 1.12. Adjusted values were compared with laboratory serum values using ANOVA for repeated measures and Dunnett's Multiple Comparisons Test: ANOVA and Dunnett's Multiple Comparisons Test showed only Glucochek II (Medistron, Ltd., West Sussex, England; distributed in U.S. by Larken Industries, Lenexa, Indiana) using Dextrostix reagent strips (Ames Division, Miles Laboratories, Elkhart, Indiana) having a statistically significant difference from laboratory values (P less than 0.01). This meter was recently revised and may show changed accuracy readings. Each meter exhibited inherent advantages and disadvantages regarding price, calibration, strip utilization, and ease of operation, which must be considered before recommendation by the health care provider.


Immune parameters in male atheletes after a lacto-ovo vegetarian diet and a mixed Western diet.
The influence of a lacto-ovo vegetarian diet versus a meat-rich Western diet on in vitro measures of immune function was studied in eight male endurance athletes. Subjects consumed two different diets for 2 x 6 wk, separated by 4 wk on an ad libitum diet, in a cross-over design. Both diets consisted of 57 energy % (E%) carbohydrates, 14 E% protein and 29 E% fat. One diet was a mixed meat-rich diet (M) prepared with 69% animal protein sources, whereas the other diet (V) was a lacto-ovo vegetarian diet prepared with 82% vegetable protein sources. Blood for determination of leukocyte subpopulations and in vitro function was collected at the end of each diet period 36 h after the last training bout. Fiber content and P/S ratio of fatty acids were twice as high on the V diet as on the M diet. Training volume was similar on the two diets, and maximal aerobic capacity did not change during diet periods. The number of CD3+ (pan T-cells), CD8+ (mainly T suppressor cells), CD4+ (mainly T helper cells), CD16+ (natural killer cells), and CD14+ (monocytes) was similar after the two different diets. Similarly, proliferations of mononuclear cells after stimulation with interleukin-2 (IL-2), phytohemagglutinin, and purified derivative of tuberculin (PPD), as well as activity of natural killer cells in the unstimulated state and after stimulation with IL-2, indomethacin, and interferon-alpha (IFN-alpha), were identical after the two diet periods.(ABSTRACT TRUNCATED AT 250 WORDS)


Cytotoxic effect of hyperthermia and chemotherapy with platinum salt on ovarian cancer cells: results of an in vitro study.
Hyperthermic intraperitoneal chemotherapy is continuously under evaluation in ovarian cancer. The purpose of the present study was to evaluate the effect of chemotherapy, drug concentration and temperature. A human ovarian carcinoma cell line was used. The effect of hyperthermia combined with chemotherapy was analyzed. When hyperthermia was combined with chemotherapy, the 50% lethal dose (LD(50)) decreased with the duration of exposure. The effect of temperature was similar between 39 and 43 °C for a 30-min exposure. For a 60- to 90-min exposure, the LD(50) was equivalent between 38 and 43 °C. Beyond 40 °C, an increase in platinum salt concentration was necessary to obtain similar results according to the duration of exposure. The cytotoxic effect of the combination seemed to be potentiated and limited at 40 °C.


Acute recovery from patterns in stroke patients: neuropsychological factors.
Acute functional recovery after a stroke was evaluated in 31 subjects over a three-month period. Assessments were made at four, eight and 12 weeks after insult in the areas of activities of daily living, mobility, functional movement and cognitive abilities. Maximum recovery was achieved by eight weeks, although a trend towards recovery was continued to 12 weeks. Significant recovery was noted in activities of daily living, gait and functional movement, but not in cognitive abilities. A hemispheric difference in recovery rate was looked for, but not found, although unilateral spatial neglect, which, in this study, was exclusively found in right hemisphere lesions, emerged as a factor associated with poor outcome. The present results need to be confirmed by long term follow-up studies.


A study of clinicopathologic profile of 15 cases of hypopigmented mycosis fungoides.
Mycosis fungoides (MF) is cutaneous lymphoma of the T-cell lineage. Hypopigmented MF is a clinical variant of MF, described mainly in Asians. This is a retrospective clinicopathologic analysis of hypopigmented MF at a tertiary care center. To describe the clinicopathologic profile of hypopigmented MF. Records of clinicopathologic notes over a 5-year period ranging from January 2005 up to December 2009 were reviewed over a period of 3 months, of which 15 cases were diagnosed with hypopigmented MF based on clinicopathologic correlation. Hypopigmented MF was found to be more common in males, and between second and fourth decades of life. The latent period between onset and diagnosis was around 3.83 years. Most of the patients were asymptomatic 80% (12/15), with skin changes of subtle atrophy in 46.66% (7/15), scaling in 20% (3/15) and focal changes of poikiloderma in 26.66% (4/15) patients. Most common sites of distribution of the lesions were the trunk and extremities. Many of the cases had been clinically mistaken for Hansen's disease prior to correct diagnosis. Marked epidermotropism and tagging of epidermis by large lymphocytes characterizes the condition histopathologically. Of the 15 cases, immunohistochemistry was possible in 10 cases, of which 8 showed predominant CD8 positive epidermotropic infiltrates and two cases showed absence of CD8 positive and CD4 positive lymphocytic infiltrate in the epidermis. Hypopigmented MF presents as hypopigmented asymptomatic patches without any erythema or infiltration in its early stage and mimics Hansen's disease. Skin biopsy clinches the diagnosis.


[Attitude of medical personnel to the health care reform].
A total of 953 managers and doctors, specializing in various spheres at therapeutic-and-prevention facilities, were polled on the basis of questionnaires for the purpose of assessing the healthcare personnel attitude towards the changes occurring in society and inside their branch. The thus obtained results are presented in the article.


Neurotoxicity of N-methyl-D-aspartate is markedly enhanced in developing rat central nervous system.
The neurotoxic lesion produced by direct injection of 25 nmol of N-methyl-D-aspartate (NMDA) into the corpus striatum of 7-day-old rats was compared to the effects of injecting 75 nmol into the striatum or hippocampus of adults. The area of histopathology in the immature striatum was 21 X larger than the striatal lesion in adults. Damage from NMDA injected into the immature striatum also extended into the dorsal hippocampus and produced an area of destruction which was 16 X larger than observed after direct injection into the adult hippocampus. Several studies have implicated excessive N-methyl-D-aspartate receptor activation in the pathogenesis of hypoxic-ischemic and hypoglycemic injury and our results suggest that this neurotoxic mechanism is extremely active in the immature brain.


The human toll and humanitarian crisis of the Russia-Ukraine war: the first 162 days.
We examined the human toll and subsequent humanitarian crisis resulting from the Russian invasion of Ukraine, which began on 24 February 2022. We extracted and analysed data resulting from Russian military attacks on Ukrainians between 24 February and 4 August 2022. The data tracked direct deaths and injuries, damage to healthcare infrastructure and the impact on health, the destruction of residences, infrastructure, communication systems, and utility services - all of which disrupted the lives of Ukrainians. As of 4 August 2022, 5552 civilians were killed outright and 8513 injured in Ukraine as a result of Russian attacks. Local officials estimate as many as 24 328 people were also killed in mass atrocities, with Mariupol being the largest (n=22 000) such example. Aside from wide swaths of homes, schools, roads, and bridges destroyed, hospitals and health facilities from 21 cities across Ukraine came under attack. The disruption to water, gas, electricity, and internet services also extended to affect supplies of medications and other supplies owing to destroyed facilities or production that ceased due to the war. The data also show that Ukraine saw an increase in cases of HIV/AIDS, tuberculosis, and Coronavirus (COVID-19). The 2022 Russia-Ukraine War not only resulted in deaths and injuries but also impacted the lives and safety of Ukrainians through destruction of healthcare facilities and disrupted delivery of healthcare and supplies. The war is an ongoing humanitarian crisis given the continuing destruction of infrastructure and services that directly impact the well-being of human lives. The devastation, trauma and human cost of war will impact generations of Ukrainians to come.


Supercapacitors in Motion: Autonomous Microswimmers for Natural-Resource Recovery.
An electroadsorption technique similar to the ultrafast charging mechanism in supercapacitors is utilized to remove metals with different sizes and hydrophilicities from contaminated water using self-propelled microswimmers. The swimmers carry graphite fibre or bismuth with a layered crystal structure providing high electrostatic double-layer capacitances. Unlike previous methods, this electrochemical technique does not only utilize the surface of the swimmers, but due to the interlayer spacing of the graphite and bismuth, it is able to store metals in ≈400 layers, allowing removal and recovery of >50 ppm lithium in only 5 min. A larger interlayer distance between bismuth sheets allows the removal of bigger cations (sodium and calcium), expanding the application of this method to a large variety of natural elements. Finally, magnetic navigation of charged swimmers to an oxygen-saturated media causes oxidation and thus immediate release of the metal ions from the swimmers.


Nuclear matrix bound fibroblast growth factor receptor is associated with splicing factor rich and transcriptionally active nuclear speckles.
We have used confocal microscopy combined with computer image analysis to evaluate the functional significance of a constitutively expressed form of the receptor tyrosine kinase FGFR1 (fibroblast growth factor receptor 1) in the nucleus of rapidly proliferating serum stimulated TE 671 cells, a medullobastoma human cell line. Our results demonstrate a limited number of large sites and numerous smaller sites of FGFR1 in the nuclear interior. The larger sites showed virtually complete colocalization (>90%) with splicing factor rich nuclear speckles while the smaller sites showed very limited overlap (<20%). Similar results were found for several other proliferating cell lines grown in culture. An in situ transcription assay was used to determine colocalization with transcription sites by incorporating 5-bromouridine triphosphate (BrUTP) followed by dual staining for BrUTP and FGFR1. These results combined with those from using an antibody against the large subunit of RNA polymerase II suggest a significant degree of colocalization (26-38%) over both the large and small sites. No colocalization was detected with sites of DNA replication. The spatial arrangements of FGFR1 sites and colocalization with nuclear speckles were maintained following extraction for nuclear matrix. Moreover, immunoblots indicated a significant enrichment of FGFR1 in the nuclear matrix fraction. Our findings suggest an involvement of a nuclear matrix bound FGFR1 in transcriptional and RNA processing events in the cell nucleus. We further propose that nuclear speckles, aside from a role in transcriptional/RNA processing events, may serve as fundamental regulatory factories for the integration of diverse signaling and regulatory factors that impact transcription and cellular regulation.


Disability inclusion in higher education in Uganda: Status and strategies.
Uganda has embraced inclusive education and evidently committed itself to bringing about disability inclusion at every level of education. Both legal and non-legal frameworks have been adopted and arguably are in line with the intent of the Convention on the Rights of Persons with Disabilities (CRPD) on education. The CRPD, in Article 24, requires states to attain a right to education for persons with disabilities without discrimination and on the basis of equal opportunities at all levels of education. Despite Uganda's robust disability legal and policy framework on education, there is evidence of exclusion and discrimination of students with disabilities in the higher education institutions. The main objective of this article is to explore the status of disability inclusion in higher education and strategies for its realisation, using evidence from Emong's study, workshop proceedings where the authors facilitated and additional individual interviews with four students with disabilities by the authors. The results show that there are discrimination and exclusion tendencies in matters related to admissions, access to lectures, assessment and examinations, access to library services, halls of residence and other disability support services. The article recommends that institutional policies and guidelines on support services for students with disabilities and special needs in higher education be developed, data on students with disabilities collected to help planning, collaboration between Disabled Peoples Organisations (DPO's) strengthened to ensure disability inclusion and the establishment of disability support centres.


Food Insecurity and Depression among US Adults: NHANES 2005-2016.
A growing body of evidence suggests that food insecurity is associated with adverse mental health outcomes such as depression and anxiety. In this study, the relationship between food insecurity and depression was examined using data from the 2005−2016 National Health and Nutrition Examination Survey (NHANES). Food insecurity was assessed with the 18-item United States Food Security Survey Module with zero affirmative responses indicating high food security, 1 or 2 affirmative responses indicating marginal food security, and ≥3 affirmative responses indicating food insecurity. Depression was assessed with the Patient Health Questionnaire-9 with scores ≥10 indicating depression. Data were analyzed from 28,448 adult participants aged 20 or older. Food insecurity was present in 19.2% of the sample population (n = 5452). Food security status was significantly associated with gender, race, education level, marital status, smoking status, and BMI (Rao-Scott chi-square, p < 0.05). Fully food secure and very low food security adults experienced depression at a rate of 5.1% and 25.8%, respectively (Rao-Scott chi-square, p < 0.0001). Participants with very low food security had a significantly greater odds of depression than food secure adults, OR = 3.50 (95% CI: 2.98, 4.12). These findings suggest that food insecurity is a significant risk factors for depression in US adults over 20 years of age. To address this issue in our citizenry, police initiatives and public health interventions addressing both food access and mental health should be prioritized.


Coupling between Land⁻Ocean⁻Atmosphere and Pronounced Changes in Atmospheric/Meteorological Parameters Associated with the Hudhud Cyclone of October 2014.
India is vulnerable to all kinds of natural hazards associated with land, ocean, biosphere, atmosphere, and snow/glaciers. These natural hazards impact large areas and the population living in the affected regions. India is surrounded by ocean on three sides and is vulnerable to cyclonic activities. Every year cyclones hit the east and west coasts of India, affecting the population living along the coasts and infrastructure and inland areas. The extent of the affected inland areas depends on the intensity of the cyclone. On 12 October 2014, a strong cyclone "Hudhud" hit the east coast of India that caused a high degree of devastation along the coast. The impact of this cyclone was seen up to the Himalayan region. Detailed analysis of satellite and ground data show a strong coupling between land-ocean-atmosphere associated with the Hudhud cyclone. The contrast between land and ocean temperature was found to be closely related with the formation of the cyclone in the ocean and its movements towards land. Pronounced changes in the ocean, land, atmospheric, and meteorological parameters with the development of the cyclone and its landfall have been observed. Changes in total column ozone (TCO), relative humidity (RH), and volume mixing ratio of CO (CO VMR), water mixing ratio (H₂O MMR), surface latent heat flux (SLHF), and aerosol optical properties derived from satellite data show characteristic behavior of the Hudhud cyclone.


Mechanical characterization of tissue-engineered cartilage using microscopic magnetic resonance elastography.
Knowledge of mechanical properties of tissue-engineered cartilage is essential for the optimization of cartilage tissue engineering strategies. Microscopic magnetic resonance elastography (μMRE) is a recently developed MR-based technique that can nondestructively visualize shear wave motion. From the observed wave pattern in MR phase images the tissue mechanical properties (e.g., shear modulus or stiffness) can be extracted. For quantification of the dynamic shear properties of small and stiff tissue-engineered cartilage, μMRE needs to be performed at frequencies in the kilohertz range. However, at frequencies greater than 1 kHz shear waves are rapidly attenuated in soft tissues. In this study μMRE, with geometric focusing, was used to overcome the rapid wave attenuation at high frequencies, enabling the measurement of the shear modulus of tissue-engineered cartilage. This methodology was first tested at a frequency of 5 kHz using a model system composed of alginate beads embedded in agarose, and then applied to evaluate extracellular matrix development in a chondrocyte pellet over a 3-week culture period. The shear stiffness in the pellet was found to increase over time (from 6.4 to 16.4 kPa), and the increase was correlated with both the proteoglycan content and the collagen content of the chondrocyte pellets (R(2)=0.776 and 0.724, respectively). Our study demonstrates that μMRE when performed with geometric focusing can be used to calculate and map the shear properties within tissue-engineered cartilage during its development.


Accuracy of an intimate partner violence screening tool for female VHA patients: a replication and extension.
The 4-item Hurt/Insult/Threaten/Scream (HITS) tool accurately detects past-year intimate partner violence (IPV) among female Veterans Health Administration (VHA) patients; however, it lacks a sexual IPV item. This study evaluated the accuracy of an extended HITS (E-HITS), which adds a sexual IPV item, in female VHA patients. A sample of 80 female U.S. veteran VHA patients in New England completed a mail survey (50.0% response rate) that included the 5-item E-HITS and the Revised Conflict Tactics Scales (CTS-2). Women were included if they were in an intimate relationship in the past year. The women averaged 49 years of age and 86.0% of the sample was White. Accuracy of the 4-item HITS was compared to the 5-item E-HITS, using the CTS-2 as the reference. There were 20 women (25.0%) who reported past-year IPV on the CTS-2. The receiver operator characteristic curves demonstrated that the HITS and E-HITS performed nearly identically at their optimal cutoff scores of 6 and 7, respectively. At these cutoff scores, the sensitivity of both tools was .75, 95% CI [.55, .95]. The specificities were similar; .83 for the HITS, 95% CI [.73, .92], and .82 for the E-HITS, 95% CI [.72, .90]. Including a sexual IPV item may be clinically beneficial; it also attains the same accuracy of case identification as the HITS.


Serum antibody levels to retinal S-antigen determined by ELISA in experimental and human uveitis.
Retinal S-antigen is thought to play an important role in the immunopathogenesis of uveitis. To investigate the role of S-antigen in human uveitis, an immunopotent bioactive S-antigen and a sensitive enzyme-linked immunosorbent assay (ELISA) were developed to determine the anti-S-antigen antibody contents in experimental autoimmune uveitis (EAU) and human uveitis. The bovine S-antigen was isolated by ammonium sulfate precipitation, Fractogel TSK HW-55 gel filtration and hydroxylapatite absorption chromatography. A test by SDS polyacrylamide electrophoresis showed that it was a highly purified protein. The purified S-antigen evoked EAU in 60% of eyes of guinea pigs even at the dose of 1 microgram. ELISA was developed to determine the contents of serum immunoglobulin G antibody against S-antigen. In the serum of S-antigen immunized rabbit, the antibody was measurable up to 1:160,000 in dilution. The antibody content appeared on the 10th day following inoculation and rose gradually until the 22nd day when the maximal antibody content was observed. Regarding human uveitis, the anti-S-antigen antibody was higher in 31 patients with Behçet's disease and 8 with Harada's disease than in normal controls although statistical significance existed only in the former (p less than 0.02). In this study, the method to purify the bovine retinal S-antigen proved relatively rapid and efficient. The ELISA developed herein appeared sensitive and well reproducible in the detection of anti-S-antigen antibody. The results showed that retinal autoimmunity may play a role in the pathogenesis in EAU and human uveitis.


Up-regulation of tissue factor expression by platelet-derived growth factor in human vascular smooth muscle cells in culture--role of mitogen-activated protein kinase and effects of intracellular cyclic AMP.
Human vascular smooth muscle cells (SMCs) isolated from normal adult arteries were investigated for the expression of Tissue Factor (TF) procoagulant activity at their membrane surface and TF antigen in lysed cells. In growing conditions, SMCs expressed high levels of TF activity and antigen. When cells were made quiescent by 72 h of subculture in serum-poor medium, these levels fell to about one third of the initial values. Platelet-derived growth factor BB (PDGF) up-regulated in a dose-dependent manner TF expression with a significant increase (1.8 fold) within five hours. PD 98059, a specific inhibitor of mitogen-activated protein kinase (MAPK) pathway involved in cell response to PDGF, also prevented TF up-regulation. Short-term treatment by Pentoxifylline and dibutyryl cAMP also completely prevented induced TF up-regulation, but remained without effect on baseline levels of quiescent, unstimulated cells. At their effective concentrations, pentoxifylline and dibutyryl cAMP also inhibited the activation of MAPK induced by PDGF. The rapid induction of TF upon growth factor stimulation may be important in circumstances where SMCs proliferate, such as vascular events or vessel development.


Topography of Cholinergic Changes in Dementia With Lewy Bodies and Key Neural Network Hubs.
The authors investigated the topography of cholinergic vulnerability in patients with dementia with Lewy bodies (DLB) using positron emission tomography (PET) imaging with the vesicular acetylcholine transporter (VAChT) [18F]-fluoroethoxybenzovesamicol ([18F]-FEOBV) radioligand. Five elderly participants with DLB (mean age, 77.8 years [SD=4.2]) and 21 elderly healthy control subjects (mean age, 73.62 years [SD=8.37]) underwent clinical assessment and [18F]-FEOBV PET. Compared with the healthy control group, reduced VAChT binding in patients with DLB demonstrated nondiffuse regionally distinct and prominent reductions in bilateral opercula and anterior cingulate to mid-cingulate cortices, bilateral insula, right (more than left) lateral geniculate nuclei, pulvinar, right proximal optic radiation, bilateral anterior and superior thalami, and posterior hippocampal fimbria and fornices. The topography of cholinergic vulnerability in DLB comprises key neural hubs involved in tonic alertness (cingulo-opercular), saliency (insula), visual attention (visual thalamus), and spatial navigation (fimbria/fornix) networks. The distinct denervation pattern suggests an important cholinergic role in specific clinical disease-defining features, such as cognitive fluctuations, visuoperceptual abnormalities causing visual hallucinations, visuospatial changes, and loss of balance caused by DLB.


Glucagon-like peptide (GLP)-2 reduces chemotherapy-associated mortality and enhances cell survival in cells expressing a transfected GLP-2 receptor.
Chemotherapeutic agents produce cytotoxicity via induction of apoptosis and cell cycle arrest. Rapidly proliferating cells in the bone marrow and intestinal crypts are highly susceptible to chemotherapy, and damage to these cellular compartments may preclude maximally effective chemotherapy administration. Glucagon-like peptide (GLP)-2 is an enteroendocrine-derived regulatory peptide that inhibits crypt cell apoptosis after administration of agents that damage the intestinal epithelium. We report here that a human degradation-resistant GLP-2 analogue, h[Gly2]-GLP-2 significantly improves survival, reduces bacteremia, attenuates epithelial injury, and inhibits crypt apoptosis in the murine gastrointestinal tract after administration of topoisomerase I inhibitor irinotecan hydrochloride or the antimetabolite 5-fluorouracil. h[Gly2]-GLP-2 significantly improved survival and reduced weight loss but did not impair chemotherapy effectiveness in tumor-bearing mice treated with cyclical irinotecan. Furthermore, h[Gly2]-GLP-2 reduced chemotherapy-induced apoptosis, decreased activation of caspase-8 and -3, and inhibited poly(ADP-ribose) polymerase cleavage in heterologous cells transfected with the GLP-2 receptor. These observations demonstrate that the antiapoptotic effects of GLP-2 on intestinal crypt cells may be useful for the attenuation of chemotherapy-induced intestinal mucositis.


Aneuploidogenic effects and DNA oxidation induced in vitro by differently sized gold nanoparticles.
Gold nanoparticles (Au NPs) are used in many fields, including biomedical applications; however, no conclusive information on their potential cytotoxicity and genotoxicity mechanisms is available. For this reason, experiments in human primary lymphocytes and murine macrophages (Raw264.7) were performed exposing cells to spherical citrate-capped Au NPs with two different nominal diameters (5 nm and 15 nm). The proliferative activity, mitotic, apoptotic, and necrotic markers, as well as chromosomal damage were assessed by the cytokinesis-block micronucleus cytome assay. Fluorescence in situ hybridization with human and murine pancentromeric probes was applied to distinguish between clastogenic and aneuploidogenic effects. Our results indicate that 5 nm and 15 nm Au NPs are able to inhibit cell proliferation by apoptosis and to induce chromosomal damage, in particular chromosome mis-segregation. DNA strand breaks were detected by comet assay, and the modified protocol using endonuclease-III and formamidopyrimidine-DNA glycosylase restriction enzymes showed that pyrimidines and purines were oxidatively damaged by Au NPs. Moreover, we show a size-independent correlation between the cytotoxicity of Au NPs and their tested mass concentration or absolute number, and genotoxic effects which were more severe for Au NP 15 nm compared to Au NP 5 nm. Results indicate that apoptosis, aneuploidy, and DNA oxidation play a pivotal role in the cytotoxicity and genotoxicity exerted by Au NPs in our cell models.


Hydration Status, Executive Function, and Response to Orthostatism After a 118-km Mountain Race: Are They Interrelated?
Martínez-Navarro, I, Chiva-Bartoll, O, Hernando, B, Collado, E, Porcar, V, and Hernando, C. Hydration status, executive function and response to orthostatism after a 118-km mountain race: are they interrelated? J Strength Cond Res 32(2): 441-449, 2018-The present study aimed to explore whether blood pressure (BP) and heart rate (HR) variability (HRV) responsiveness to orthostatism, jointly with executive function (EF) performance, was diminished after an ultra-endurance mountain race. Besides, we wanted to assess whether hydration status was related to either performance or the abovementioned alterations. Fifty recreational ultra-endurance athletes participating in the Penyagolosa Trails CSP115 race (118 km and a total positive elevation of 5,439 m) were evaluated before and after the competition. The HRV and BP were measured in response to an orthostatic challenge. The EF was evaluated using the color-word interference task of the Stroop test. Body mass (BM) and urine specific gravity (USG) changes were used to assess hydration status. The HRV and BP responsiveness to orthostatism was diminished after the race. Besides, a significant BM loss of 3.51 ± 2.03% was recorded. Conversely, EF and USG showed no significant changes from prerace to postrace. Eventually, BM loss was inversely related to finishing time (r = -0.34) and postrace orthostatic HR and EF were positively associated (r = 0.60). The USG and BM loss appear to provide different insights into hydration status, and our results challenge the well-established criteria that BM losses >2% are detrimental to performance. Coaches are advised to consider athletes' performance level when interpreting their BM changes during an ultra-endurance competition. Similarly, coaches should be aware that increased vulnerability to orthostatism is a common phenomenon after ultra-endurance races, and diminished HR responsiveness to orthostatism could constitute a practical indicator of EF worsening.


High efficiency polyoma DNA transfection of chloroquine treated cells.
Chloroquine treatment of rodent cells during the first hours of polyoma DNA transfection increase the fraction of cells expressing viral functions. The effect has been observed after DNA absorption using both the DEAE-dextran and calcium phosphate coprecipitation methods. Exposure to chloroquine increased the proportion of transfected mouse cells to approximately 40%. From a culture of one million such cells, microgram quantities of newly synthesized viral DNA could be isolated. Similarly, the transformation frequency of rat cells following polyoma DNA transfection was approximately 6-fold increased by chloroquine treatment. The effect of the compound was even more pronounced in transfections with linear forms of polyoma DNA, suggesting that chloroquine inhibits degradation of DNA absorbed by the cells.