An investigation of PRESAGE® 3D dosimetry for IMRT and VMAT radiation therapy treatment verification.
The purpose of this work was to characterize three formulations of PRESAGE(®) dosimeters (DEA-1, DEA-2, and DX) and to identify optimal readout timing and procedures for accurate in-house 3D dosimetry. The optimal formulation and procedure was then applied for the verification of an intensity modulated radiation therapy (IMRT) and a volumetric modulated arc therapy (VMAT) treatment technique. PRESAGE(®) formulations were studied for their temporal stability post-irradiation, sensitivity, and linearity of dose response. Dosimeters were read out using a high-resolution optical-CT scanner. Small volumes of PRESAGE(®) were irradiated to investigate possible differences in sensitivity for large and small volumes ('volume effect'). The optimal formulation and read-out technique was applied to the verification of two patient treatments: an IMRT plan and a VMAT plan. A gradual decrease in post-irradiation optical-density was observed in all formulations with DEA-1 exhibiting the best temporal stability with less than 4% variation between 2-22 h post-irradiation. A linear dose response at the 4 h time point was observed for all formulations with an R(2) value >0.99. A large volume effect was observed for DEA-1 with sensitivity of the large dosimeter being ~63% less than the sensitivity of the cuvettes. For the IMRT and VMAT treatments, the 3D gamma passing rates for 3%/3 mm criteria using absolute measured dose were 99.6 and 94.5% for the IMRT and VMAT treatments, respectively. In summary, this work shows that accurate 3D dosimetry is possible with all three PRESAGE(®) formulations. The optimal imaging windows post-irradiation were 3-24 h, 2-6 h, and immediately for the DEA-1, DEA-2, and DX formulations, respectively. Because of the large volume effect, small volume cuvettes are not yet a reliable method for calibration of larger dosimeters to absolute dose. Finally, PRESAGE(®) is observed to be a useful method of 3D verification when careful consideration is given to the temporal stability and imaging protocols for the specific formulation used.


Practising in a post-truth world: Pandemic ethics can inform patient autonomy and clinical communication.
The COVID-19 pandemic posed an unprecedented challenge to modern bioethical frameworks in the clinical setting. Now, as the pandemic stabilises and we learn to 'live with COVID', the medical community has a duty to evaluate its response to the challenge, and reassess our ethical reasoning, considering how we practise in the future. This article considers a number of clinical and bioethical challenges encountered by the author team and colleagues during the most severe waves of the pandemic. We argue that the changed clinical context may require reframing our ethical thought in such a manner as to adequately accommodate all parties in the clinical interaction. We argue that clinicians have become relatively disempowered by the 'infodemic', and do not necessarily have adequate skills or training to assess the scientific literature being published at an unprecedented rate. Conversely, we acknowledge that patients and families are more empowered by the infodemic, and bring this empowerment to bear on the clinical consultation. Sometimes these interactions can be unpleasant and threatening, and involve inviting clinicians to practise against best evidence or even illegally. Generally, these requests are framed within 'patient autonomy' (which some patients or families perceive to be unlimited), and several factors may prevent clinicians from adequately navigating these requests. In this article, we conclude that embracing a framework of shared decision-making (SDM), which openly acknowledges clinical expertise and in which patient and family autonomy is carefully balanced against other bioethics principles, could serve us well going forward. One such principle is the recognition of clinician expertise as holding weight in the clinical encounter, when framed in terms of non-maleficence and beneficence. Such a framework incorporates much of our learning and experience from advising and treating patients during the pandemic.


Quantitative proteomic profiling of prostate cancer reveals a role for miR-128 in prostate cancer.
Multiple, complex molecular events characterize cancer development and progression. Deciphering the molecular networks that distinguish organ-confined disease from metastatic disease may lead to the identification of biomarkers of cancer invasion and disease aggressiveness. Although alterations in gene expression have been extensively quantified during neoplastic progression, complementary analyses of proteomic changes have been limited. Here we interrogate the proteomic alterations in a cohort of 15 prostate-derived tissues that included five each from adjacent benign prostate, clinically localized prostate cancer, and metastatic disease from distant sites. The experimental strategy couples isobaric tags for relative and absolute quantitation with multidimensional liquid phase peptide fractionation followed by tandem mass spectrometry. Over 1000 proteins were quantified across the specimens and delineated into clinically localized and metastatic prostate cancer-specific signatures. Included in these class-specific profiles were both proteins that were known to be dysregulated during prostate cancer progression and new ones defined by this study. Enrichment analysis of the prostate cancer-specific proteomic signature, to gain insight into the functional consequences of these alterations, revealed involvement of miR-128-a/b regulation during prostate cancer progression. This finding was validated using real time PCR analysis for microRNA transcript levels in an independent set of 15 clinical specimens. miR-128 levels were elevated in benign prostate epithelial cell lines compared with invasive prostate cancer cells. Knockdown of miR-128 induced invasion in benign prostate epithelial cells, whereas its overexpression attenuated invasion in prostate cancer cells. Taken together, our profiles of the proteomic alterations of prostate cancer progression revealed miR-128 as a potentially important negative regulator of prostate cancer cell invasion.


Mechanism of an Exceptional Class of photostabilizers: a seam of conical intersection parallel to excited state intramolecular proton transfer (ESIPT) in o-hydroxyphenyl-(1,3,5)-triazine.
We present a detailed CASSCF study of the mechanism of excited-state intramolecular proton transfer (ESIPT) in the o-hydroxyphenyl triazine class of photostabilizers. The valence-bond analysis of the ground state and the two pipi* excited states permits a simple chemical interpretation of the mechanistic information. Our results show that the barrier to enol-keto tautomerism on the ground-state adiabatic surface is high. Following photoexcitation to the charge-transfer state, the ESIPT is predicted to take place without a barrier. Radiationless decay to the ground state is associated with an extended seam of conical intersection, with a sloped topology lying parallel to the ESIPT path, which can be accessed at any point along the reaction path. Our results show that the triazine class of photostabilizers has the photochemical and photophysical qualities associated with exceptional photostability.


Everyday discrimination, daily affect, and physical symptoms during the COVID-19 pandemic.
Abundant evidence has linked everyday discrimination with health risks. Because the COVID-19 pandemic has increased exposure to discrimination (e.g., based on age and race), it is important to understand the day-to-day implications of discrimination experiences for well-being. Furthermore, daily positive events were examined as a moderator due to their potential for mitigating the associations between everyday discrimination and well-being. From March to August 2020, 1,212 participants aged 18-91 in the United States and Canada (84% women, 75% White) completed surveys for seven consecutive evenings about everyday discrimination, positive events, physical health symptoms, and positive and negative affect. Data were analyzed using multilevel models and controlled for sociodemographic factors. Everyday discrimination was reported on 9.3% of days when in-person or remote social interactions occurred. Within-persons, positive affect was lower and negative affect and physical symptoms were higher on days when discrimination occurred versus on days without discrimination. Positive events mitigated the within-person association between everyday discrimination and same-day negative affect, but not for positive affect or physical symptoms. Discrimination perceived to be due to age was associated with higher negative affect and lower positive affect within-persons. Positive events did not moderate the associations between age-based discrimination and same-day outcomes. Everyday discrimination was related to lower daily positive affect and higher negative affect and physical symptoms during the COVID-19 pandemic. This study provides initial evidence that daily positive events partially offset the increased negative affect associated with same-day discrimination. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Mucosal damage in the esophageal remnant after esophagectomy and gastric transposition.
To assess development of mucosal damage in the esophageal remnant in regard to the level of the esophagogastrostomy reconstruction either in a right chest or in a left neck position. Esophagectomy with gastric interposition creates an in vivo human model of pathologic esophageal reflux with the potential for long-term reflux disease complications. Eighty-four esophagectomy patients were assessed over time by symptoms, endoscopy and biopsies of their esophageal remnant after the operation. The anastomosis was in the right upper chest (n = 36) or in a left cervical position (n = 48). Visual quantification of damage, details of histopathology, and time period since surgery were recorded. Twenty-nine patients (81%) with a right chest reconstruction had reflux symptoms when compared with 25 patients (53%) with a neck reconstruction (P = 0.007). Visualized reflux esophagitis was observed in 31 patients (81%) with chest anastomoses and in 22 patients (46%) with cervical anastomoses (P = 0.006). Documented mucosal damage and columnar lined metaplasia were significantly more frequent in the chest anastomosis group than the cervical group. The median of all mucosal damage and columnar lined metaplastic-free evolution were 13 +/- 3 and 20.5 +/- 6 months for the intrathoracic anastomosis, and 22 +/- 6 months and 40 +/- 8 months for the cervical anastomosis (P = 0.087). Two factors affecting the development of metaplasia were included in the multivariate analysis: an intrathoracic anastomosis (P = 0.018) and the presence of a previous Barrett esophagus (P = 0.064). When a gastric transplant is used after esophagectomy, a high prevalence of mucosal damage is observed in the esophageal remnant independently of the level of reconstruction. A left cervical anastomosis favors less reflux symptoms, less visualized damage, and delays the development of mucosal damage over time.


Polyglandular autoimmune syndrome type I - a novel AIRE mutation in a North American patient.
Autoimmune polyglandular syndrome type 1 (APS-1), also referred to as autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED), is a rare autoimmune disease that results from autosomal recessive mutations of the human autoimmune regulatory (AIRE) gene. We present the case of a 17-year-old North American girl of primarily Norwegian descent with a novel AIRE gene mutation causing APS-1. In addition to the classic triad of chronic candidiasis, hypoparathyoidism and autoimmune adrenocortical insufficiency, she also has vitiligo, intestinal malabsorption, autoimmune hepatitis, autoimmune hypothyroidism, myositis, myalgias, chronic fatigue, and failure to thrive. Genetic testing revealed heterozygosity for c.20_115de196 and c.967_979del13 mutations in the AIRE gene. The AIRE gene c.20_115de196 mutation has not been previously reported.


Determination of optimized protocols for the extraction of anticholinesterasic compounds in environmental samples containing cyanobacteria species.
The effects of temperature and pH during extraction of toxin(s) from the cyanobacterial genus Anabaena were studied. Different extracts were compared for their anticholinesterase potency against purified eel acetylcholinesterase. A lyophilized powder from Anabaeba spiroides cells was dissolved in ethanol, sonicated, and filtered. This solution was then evaporated at three different temperatures (40, 60, or 80 degrees C), resuspended in chloroform, and washed in deionized water at three different pH values (3.3, 5.0, or 8.5). At 40 degrees C, the estimated 50% inhibition concentration (IC50) was 2.06, 2.19, and 8.64 mg lyophilized A. spiroides cells ml(-1) for the extraction pH of 3.3, 5.0, and 8.5, respectively. At 60 degrees C, the values were 4.82, 3.06, and 12.0 mg/ml at increasing pH values. Finally, at 80 degrees C, the IC50 values were 3.52, 4.35, and 30.1 mg/ml at the same pH values. These results indicate that pH 3.3 and a temperature of 40 degrees C should be employed to preserve toxin content. The optimized protocol was employed to evaluate the anticholinesterase potency of environmental samples of water containing different numbers of filaments of cyanobacterial species. Registered inhibition ranged from 5.11 to 11.1%, supporting the feasibility of the method.


Growing unculturable bacteria.
The bacteria that can be grown in the laboratory are only a small fraction of the total diversity that exists in nature. At all levels of bacterial phylogeny, uncultured clades that do not grow on standard media are playing critical roles in cycling carbon, nitrogen, and other elements, synthesizing novel natural products, and impacting the surrounding organisms and environment. While molecular techniques, such as metagenomic sequencing, can provide some information independent of our ability to culture these organisms, it is essentially impossible to learn new gene and pathway functions from pure sequence data. A true understanding of the physiology of these bacteria and their roles in ecology, host health, and natural product production requires their cultivation in the laboratory. Recent advances in growing these species include coculture with other bacteria, recreating the environment in the laboratory, and combining these approaches with microcultivation technology to increase throughput and access rare species. These studies are unraveling the molecular mechanisms of unculturability and are identifying growth factors that promote the growth of previously unculturable organisms. This minireview summarizes the recent discoveries in this area and discusses the potential future of the field.


Chronic Respiratory Disorders and Their Treatment among Older People with Intellectual Disability and/or Autism Spectrum Disorder in Comparison with the General Population.
Respiratory disorders are common among people with intellectual disabilities (ID). However, few studies have investigated these disorders among older people with ID. We identified 7936 people, aged 55+ years, with ID and a reference cohort from the general population. Data on diagnoses of chronic respiratory disorders, with a focus on asthma and chronic obstructive pulmonary disease (COPD), were collected, as was information on health care visits due to such disorders. We also added data on the prescription of drugs for obstructive airway diseases. Whereas the risk of having at least one diagnosis of asthma during the study period was similar in the two cohorts, people with ID were less likely than the general population to have been diagnosed with COPD. The same was found for health care visits due to asthma and COPD, respectively. The patterns of drug prescription were similar among people with ID and the general population, with the exception of adrenergics for systemic use, which were more commonly prescribed to people with ID. Thus, older people with ID do not seem to have an increased risk of asthma or COPD. Moreover, the indications are that when diagnosed with any of these disorders, they receive treatment adapted to their particular needs.


To be funny or not to be funny: Gender differences in student perceptions of instructor humor in college science courses.
For over 50 years instructor humor has been recognized as a way to positively impact student cognitive and affective learning. However, no study has explored humor exclusively in the context of college science courses, which have the reputation of being difficult and boring. The majority of studies that explore humor have assumed that students perceive instructor humor to be funny, yet students likely perceive some instructor humor as unfunny or offensive. Further, evidence suggests that women perceive certain subjects to be more offensive than men, yet we do not know what impact this may have on the experience of women in the classroom. To address these gaps in the literature, we surveyed students across 25 different college science courses about their perceptions of instructor humor in college science classes, which yielded 1637 student responses. Open-coding methods were used to analyze student responses to a question about why students appreciate humor. Multinomial regression was used to identify whether there are gender differences in the extent to which funny, unfunny, and offensive humor influenced student attention to course content, instructor relatability, and student sense of belonging. Logistic regression was used to examine gender differences in what subjects students find funny and offensive when joked about by college science instructors. Nearly 99% of students reported that they appreciate instructor humor and reported that it positively changes the classroom atmosphere, improves student experiences during class, and enhances the student-instructor relationship. We found that funny humor tends to increase student attention to course content, instructor relatability, and student sense of belonging. Conversely, offensive humor tends to decrease instructor relatability and student sense of belonging. Lastly, we identified subjects that males were more likely to find funny and females were more likely to find offensive if a college science instructor were to joke about them.


Characterisation of methicillin-resistant Staphylococcus aureus isolates by restriction endonuclease digestion of chromosomal DNA.
Clinical isolates of Staphylococcus aureus from the London Hospital were characterised by genetic analysis of antibiotic-resistance determinants and by restriction endonuclease digestion of chromosomal DNA and compared with isolates from elsewhere in the UK. Restriction enzyme digestion of chromosomal DNA confirmed that a single strain of methicillin-resistant S. aureus (MRSA) persists at the London Hospital, although its antibiotic-resistance profile and plasmid carriage are not constant. Methicillin-sensitive isolates, on the other hand, each had readily distinguishable and unique DNA restriction patterns. The DNA restriction digest pattern of the London Hospital MRSA isolates was identical to that of "epidemic" (E) MRSA isolates from the Thames regions. By contrast, other MRSA isolates had DNA restriction patterns which differed from those of EMRSA isolates and from each other. These results confirm the discriminatory value of restriction pattern analysis as a typing method.


Leprosy reactions: humoral and cellular immune responses to M. leprae, 65kDa, 28kDa, and 18 kDa antigens.
This study examines the immune responses against some stress proteins of Mycobacterium leprae in leprosy patients with and without leprosy reactions. Leprosy patients showed a higher level of antibodies to all antigens compared to healthy controls. The antibody response to 18kDa antigen was significantly higher in patients with Type 1 reaction compared to those of TT or borderline patients without Type 1 reaction, or those with Type 2 reaction. Borderline (BT/BL), lepromatous (LL) and patients with reactions (Type 1 and Type 2) had higher levels of antibodies to M. leprae soluble extract (MLSE) and 65kDa than those of the tuberculoid (TT) group. LL, borderline patients, and patients with Type 1 reaction had a higher level of antibody to 28kDa than those of healthy controls. However, no significant differences could be observed in antibody response to these antigens (MLSE, 65kDa, and 28kDa) between patients with reaction and without reaction. A significant proportion of TT/BT patients showed positive lymphoproliferative response to MLSE compared to BL/LL patients. In addition, the lymphoproliferative response to MLSE was significantly greater in patients with Type 1 reaction compared to patients without reaction. No difference in proliferative response to 65kDa could be observed in any of these groups. The finding of high levels of antibodies against stress proteins in patients with Type 1 reactions, especially to 18 kDa antigen, along with a heightened lymphoproliferative response to MLSE is suggestive of a coexistence of cell mediated and humoral immunity in leprosy patients during Type 1 reactions. On the other hand, in Type 2 reactions no significant role of stress proteins could be demonstrated except a heightened lymphoproliferative response to the 28 kDa antigen.


Short versus long cephalomedullary nailing of intertrochanteric fractures: a meta-analysis of 3208 patients.
The purpose of the study was to compare treatment outcomes after short or long cephalomedullary nailing for intertrochanteric femur fractures. A systematic review of perioperative outcomes after short or long cephalomedullary nailing for intertrochanteric femur fractures was performed. The following databases were used: using the Cochrane Database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, PubMed (1980-2019), and MEDLINE (1980-2019). The queries were performed in June 2019. The following search term query was used: "Intramedullary Nail AND Intertrochanteric Fracture OR "Long OR Short Nail AND intertrochanteric Fracture." Studies were excluded if they were "single-arm" studies (i.e., reporting on either long or short CMN but not both), or did not report at least one of the outcomes being meta-analyzed. Furthermore, cadaveric studies, animal studies, basic science articles, editorial articles, surveys and studies were excluded. Two investigators independently reviewed abstracts from all identified articles. Full-text articles were obtained for review if necessary, to allow further assessment of inclusion and exclusion criteria. Additionally, all references from the included studies were reviewed and reconciled to verify that no relevant articles were missing from the systematic review. Short nails were associated with statistically significantly less estimated blood loss and operative time compared to long nails. There were no significant differences in transfusion rates, implant failures or overall re-operation rates between implant lengths. Similarly, there was no significant difference in peri-implant fracture between implant lengths. Overall, the available clinical evidence supports the use of short cephalomedullary nails for the majority of intertrochanteric femur fractures. Meta-analysis; Level III, therapeutic.


Quantitative OCT Angiography of the Retinal Microvasculature and the Choriocapillaris in Myopic Eyes.
To study the retinal capillary microvasculature and the choriocapillaris (CC) in myopic eyes using quantitative optical coherence tomography angiography (OCTA) analysis. Macular OCTA images of 3 × 3 mm were obtained using the RTVue-XR Avanti with AngioVue. Quantitative measurements of the retinal capillary microvascular layers and the CC were analyzed using en face projection images. Vessel density and fractal dimension of the superficial and deep retinal capillary plexus, and area and density of flow reduction in the CC were analyzed, quantified, and compared with an age-matched control group. Fifty eyes with myopia and 34 age-matched healthy eyes were included in this study. The vessel density and the vessel branching complexity using fractal dimension of the retinal capillary microvasculature were significantly lower in myopic eyes (P < 0.001 and P = 0.001). The total number of flow voids in the CC was lower (108.93 vs. 138.63, P = 0.001) but the total and average flow void area was significantly higher (total area 3.715 ± 0.257 vs. 3.596 ± 0.194 mm2, P = 0.026; average area 0.044 ± 0.029 vs. 0.028 ± 0.010 mm2, P = 0.002) compared with the healthy control group. Average choroidal thickness was lower in the myopic group versus the normal control cohort (123.538 ± 73.477 vs. 246.97 ± 41.745 μm, P < 0.05) and significantly reduced in eyes with lacquer cracks (LC) compared with myopic eyes without LC formation (P = 0.003). There was no correlation between choroidal thickness and quantitative parameters of the CC in the myopic eyes. The density of the retinal capillary microvasculature is reduced and the area of flow deficit in the CC is increased in eyes with greater myopia. The relevance of microvascular alterations in the setting of myopia warrants further study.


Long-term neurocognitive effects of olanzapine or low-dose haloperidol in first-episode psychosis.
Neurocognitive deficits are severe in first-episode psychosis. Patients (N = 263) with first-episode psychosis (schizophrenia, schizoaffective, or schizophreniform disorders) were randomly assigned to double-blind treatment with olanzapine (mean 11.30 mg/day) or haloperidol (mean 4.87 mg/day) for 104 weeks. A neurocognitive battery was administered at baseline (n = 246) and 12 (n = 167), 24 (n = 126), 52 (n = 89), and 104 (n = 46) weeks during treatment. Weighted principal component and unweighted composite scores were derived from individual tests. Both treatment groups demonstrated significant improvement on both composite scores. On the basis of the weighted composite score, olanzapine had greater improvement than haloperidol only at 12 (p = .014) and 24 (p = .029) weeks. For the unweighted composite, olanzapine had significantly better improvement compared with haloperidol only at week 12 (p = .044). At week 12 only, olanzapine improved performance on the Digit Symbol and Continuous Performance Test significantly more than haloperidol. Both antipsychotic agents appeared to improve neurocognitive functioning among first-episode psychosis patients with schizophrenia. A significantly greater benefit in terms of neurocognitive improvement was found with olanzapine than with haloperidol at weeks 12 and 24.


Synthesis of Heterocyclic Compounds Derived From Dimedone and their Anti-tumor and Tyrosine Kinase Inhibitions.
The reaction of dimedone with arylaldehydes gave the benzylidene derivatives 3a-c, the latter underwent a series of heterocyclization reactions to give fused thiophene, pyrazole isoxazole and pyridazine derivatives. The synthesized compounds were evaluated against different kinds of cancer cell lines together tyrosine kinases and Pim-1 kinase inhibitions. All the synthesized compounds were assessed for the inhibitory activities against A549 (non-small cell lung cancer), H460 (human lung cancer), HT-29 (human colon cancer) and MKN-45 (human gastric cancer) cancer cell lines together with foretinib as the positive control by a MTT assay. The promising compounds were 3c, 5b, 5e, 5f, 7c, 7f, 9c, 11b, 12c, 12d, 13b, 13d, 14b, 16c and 16d among the tested compounds. On the other hand, compounds 5b, 5e, 5f, 7c, 11b, 12c, 12d, 13d, 14b, 16c and 16d were the most effective inhibitors against tyrosine kinases and compounds 5b, 11b, 12d, 13d, 14b and 16c were the most potent against Pim-1 kinase.


Prevention of renal carriage of leptospirosis in dogs by vaccination.
Dogs vaccinated with a Leptospira interrogans vaccine containing serogroups Canicola and Icterohaemorrhagiae and prepared from cultures grown in a protein-free medium resisted challenge with heterologous representatives of these serogroups. In contrast, control dogs were pyrexic and leptospires were isolated from the blood for nine days following Canicola challenge and six days after Icterohaemorrhagiae challenge. Leptospires were isolated from the urine of controls throughout the post challenge period and from kidneys at post mortem examination of six out of six and four out of six Canicola and Icterohaemorrhagiae challenged controls, respectively. There have been no reports of hypersensitivity reactions to this vaccine since its introduction in 1980.


Transcriptional targets of TWIST1 in the cranial mesoderm regulate cell-matrix interactions and mesenchyme maintenance.
TWIST1, a basic helix-loop-helix transcription factor is essential for the development of cranial mesoderm and cranial neural crest-derived craniofacial structures. We have previously shown that, in the absence of TWIST1, cells within the cranial mesoderm adopt an abnormal epithelial configuration via a process reminiscent of a mesenchymal to epithelial transition (MET). Here, we show by gene expression analysis that loss of TWIST1 in the cranial mesoderm is accompanied by a reduction in the expression of genes that are associated with cell-extracellular matrix interactions and the acquisition of mesenchymal characteristics. By comparing the transcriptional profiles of cranial mesoderm-specific Twist1 loss-of-function mutant and control mouse embryos, we identified a set of genes that are both TWIST1-dependent and predominantly expressed in the mesoderm. ChIP-seq was used to identify TWIST1-binding sites in an in vitro model of a TWIST1-dependent mesenchymal cell state, and the data were combined with the transcriptome data to identify potential target genes. Three direct transcriptional targets of TWIST1 (Ddr2, Pcolce and Tgfbi) were validated by ChIP-PCR using mouse embryonic tissues and by luciferase assays. Our findings reveal that the mesenchymal properties of the cranial mesoderm are likely to be regulated by a network of TWIST1 targets that influences the extracellular matrix and cell-matrix interactions, and collectively they are required for the morphogenesis of the craniofacial structures.


Repetition of deliberate self-harm and subsequent suicide risk: long-term follow-up study of 11,583 patients.
Repetition of deliberate self-harm (DSH) is a risk factor for suicide. Little information is available on the risk for specific groups of people who deliberately harm themselves repeatedly. To investigate the long-term risk of suicide associated with repetition of DSH by gender, age and frequency of repetition. A mortality follow-up study to the year 2000 was conducted on 11 583 people who presented to the general hospital in Oxford between 1978 and 1997. Repetition of DSH was determined from reported episodes prior to the index episode and episodes presenting to the same hospital during the follow-up period. Deaths were identified through national registers. Thirty-nine per cent of patients repeated the DSH. They were at greater relative risk of suicide than the single-episode DSH group (2.24; 95% CI 1.77-2.84). The relative risk of suicide in the repeated DSH group compared with the single-episode DSH group was greater in females (3.5; 95% CI 1.3-2.4) than males (1.8; 95% CI 2.3-5.3) and was inversely related to age (up to 54 years). Suicide risk increased further with multiple repeat episodes of DSH in females. Repetition of DSH is associated with an increased risk of suicide in males and females. Repetition may be a better indicator of risk in females, especially young females.


Decreased synaptic plasticity in the medial prefrontal cortex underlies short-term memory deficits in 6-OHDA-lesioned rats.
Parkinson's disease (PD) is characterized by motor dysfunction associated with dopaminergic degeneration in the dorsolateral striatum (DLS). However, motor symptoms in PD are often preceded by short-term memory deficits, which have been argued to involve deregulation of medial prefrontal cortex (mPFC). We now used a 6-hydroxydopamine (6-OHDA) rat PD model to explore if alterations of synaptic plasticity in DLS and mPFC underlie short-term memory impairments in PD prodrome. The bilateral injection of 6-OHDA (20μg/hemisphere) in the DLS caused a marked loss of dopaminergic neurons in the substantia nigra (>80%) and decreased monoamine levels in the striatum and PFC, accompanied by motor deficits evaluated after 21 days in the open field and accelerated rotarod. A lower dose of 6-OHDA (10μg/hemisphere) only induced a partial degeneration (about 60%) of dopaminergic neurons in the substantia nigra with no gross motor impairments, thus mimicking an early premotor stage of PD. Notably, 6-OHDA (10μg)-lesioned rats displayed decreased monoamine levels in the PFC as well as short-term memory deficits evaluated in the novel object discrimination and in the modified Y-maze tasks; this was accompanied by a selective decrease in the amplitude of long-term potentiation in the mPFC, but not in DLS, without changes of synaptic transmission in either brain regions. These results indicate that the short-term memory dysfunction predating the motor alterations in the 6-OHDA model of PD is associated with selective changes of information processing in PFC circuits, typified by persistent changes of synaptic plasticity.


Interactions of gene expression, alternative splicing, and DNA methylation in determining nodule identity.
Soybean nodulation is a highly controlled process that involves complex gene regulation at both transcriptional and post-transcriptional levels. In the present study, we profiled gene expression changes, alternative splicing events, and DNA methylation patterns during nodule formation, development, and senescence. The transcriptome data uncovered key transcription patterns of nodule development that included 9669 core genes and 7302 stage-specific genes. Alternative splicing analysis uncovered a total of 2323 genes that undergo alternative splicing events in at least one nodule developmental stage, with activation of exon skipping and repression of intron retention being the most common splicing events in nodules compared to roots. Approximately 40% of the differentially spliced genes were also differentially expressed at the same nodule developmental stage, implying a substantial association between gene expression and alternative splicing. Genome-wide-DNA methylation analysis revealed dynamic changes in nodule methylomes that were specific to each nodule stage, occurred in a sequence-specific manner, and impacted the expression of 1864 genes. An attractive hypothesis raised by our data is that increased DNA methylation may contribute to the efficiency of alternative splicing. Together, our results provide intriguing insights into the associations between gene expression, alternative splicing, and DNA methylation that may shape transcriptome complexity and proteome specificity in developing soybean nodules.


Detailed delineation of an interferon-gamma-responsive element important in human HLA-DRA gene expression in a glioblastoma multiform line.
In this report, we determined that induction of the DR alpha-chain by recombinant human interferon-gamma (IFN-gamma) in a human glioblastoma multiform cell line is transcriptionally regulated and showed that protein synthesis is not necessary for this to occur. The regions of the DR alpha-chain gene that are responsible for basal and recombinant IFN-gamma-induced gene transcription have been determined by gene transfer of a series of 5' deletion mutants in which the upstream region of the DR alpha chain was linked to a reporter gene, chloramphenicol acetyltransferase. Chloramphenicol acetyltransferase transcript and protein levels were determined by S1 nuclease protection and chloramphenicol acetyltransferase enzyme assays, respectively. By using these deletion mutants, we were able to draw the following conclusions. (i) One hundred and nine base pairs of upstream sequence contains the basic DR alpha-chain gene promoter and represents the minimal amount of sequence necessary for basal gene expression. (ii) An additional 9 base pairs of upstream sequence can mediate recombinant IFN-gamma induction. (iii) Maximal recombinant IFN-gamma induction requires at most an additional 23 base pairs of upstream sequence. (iv) The sequence between positions -267 and -141 does not appear to contain any additional positive or negative regulatory elements. These results suggest that the region between positions -141 and -109 contains a critical IFN-gamma-responsive element. Substitution mutagenesis was performed to confirm this suggestion.


Obstructive Sleep Apnea in Pregnancy: Early Lessons From Our Sleep Pregnancy Clinic.
Obstructive sleep apnea (OSA) is underdiagnosed during pregnancy, but there is strong theoretical and some empiric evidence that treatment may improve obstetric outcomes. Barriers to screening, testing, and treatment are common during pregnancy. The goal of this described intervention was to reduce these barriers and improve detection of OSA in pregnancy. Representatives from sleep medicine and perinatology established a cross-disciplinary, collaborative Sleep Pregnancy Clinic offering a streamlined referral process for multimodal screening, testing, and treatment of OSA during pregnancy. This is a retrospective analysis of data from the clinic's first 19 months. Between June 2017 and December 2018, 134 pregnant women were referred for OSA testing. Sixty-three (47.0%) completed objective sleep testing, and 38 (60.3%) of the women who completed testing met diagnostic criteria for OSA. This intervention resulted in a statistically significant increase in the number of diagnostic sleep apnea tests performed (average 22.4 tests per year pre-intervention, 77 per year post-intervention [P = 0.0012]). Despite a streamlined referral pipeline, completion rates of OSA testing in pregnant women remained below 50%. However, the overall number of women referred and who completed testing increased significantly during this time period. Of those who completed testing, the majority were diagnosed with OSA. Since starting this clinic, we have created resources to familiarize patients with the equipment and worked to reduce other barriers. Assessment of these interventions and the impact of treatment on obstetric outcomes is ongoing, as is assessment of reasons women do not complete diagnostic testing.


On the Stimulation Artifact Reduction during Electrophysiological Recording of Compound Nerve Action Potentials.
Recording neuronal activity triggered by electrical impulses is a powerful tool in neuroscience research and neural engineering. It is often applied in acute electrophysiological experimental settings to record compound nerve action potentials. However, the elicited neural response is often distorted by electrical stimulus artifacts, complicating subsequent analysis. In this work, we present a model to better understand the effect of the selected amplifier configuration and the location of the ground electrode in a practical electrophysiological nerve setup. Simulation results show that the stimulus artifact can be reduced by more than an order of magnitude if the placement of the ground electrode, its impedance, and the amplifier configuration are optimized. We experimentally demonstrate the effects in three different settings, in-vivo and in-vitro.


Falls are a fact of life for some patients after discharge from a rehabilitation programme.
This paper reports on a 3-month post-discharge falls profile of a cohort of patients discharged home from an acute rehabilitation setting. A consecutively-sampled cohort of consenting patients was prospectively followed for 3 months after discharge from a large Australian metropolitan acute rehabilitation facility. Patients were diagnostically classified by the reason for admission. All patients completed a monthly falls diary. Each month, falls were described by severity, location, frequency and outcome. Patients also completed the World Health Organisation's quality of life (QoL-BREF) at Months 1 and 3. Almost a third of the study participants fell during the study [105 fallers of 342 total (31%)]. Of these, 21 fell more than once (20%). There were significant differences in falls reported per diagnostic group, including frequency, impact, severity and ability to get up. Irrespective of diagnostic group, fallers had lower QoL scores compared to non-fallers. Both the neurological and orthopaedic trauma groups were most at risk of falling in Months 1 and 3, whereas in Month 2, only the neurological group was most at risk of falling. Falls are a fact of life for some patients during their community rehabilitation phase. Patients from different diagnostic groups have different over-time risk profiles for falling. Thus, there is no 'one-size fits all' solution for falls prevention. This study provides useful data to assist in formulating appropriate falls prevention strategies for recently ill people.


Mevalonate Pathway Blockade, Mitochondrial Dysfunction and Autophagy: A Possible Link.
The mevalonate pathway, crucial for cholesterol synthesis, plays a key role in multiple cellular processes. Deregulation of this pathway is also correlated with diminished protein prenylation, an important post-translational modification necessary to localize certain proteins, such as small GTPases, to membranes. Mevalonate pathway blockade has been linked to mitochondrial dysfunction: especially involving lower mitochondrial membrane potential and increased release of pro-apoptotic factors in cytosol. Furthermore a severe reduction of protein prenylation has also been associated with defective autophagy, possibly causing inflammasome activation and subsequent cell death. So, it is tempting to hypothesize a mechanism in which defective autophagy fails to remove damaged mitochondria, resulting in increased cell death. This mechanism could play a significant role in Mevalonate Kinase Deficiency, an autoinflammatory disease characterized by a defect in Mevalonate Kinase, a key enzyme of the mevalonate pathway. Patients carrying mutations in the MVK gene, encoding this enzyme, show increased inflammation and lower protein prenylation levels. This review aims at analysing the correlation between mevalonate pathway defects, mitochondrial dysfunction and defective autophagy, as well as inflammation, using Mevalonate Kinase Deficiency as a model to clarify the current pathogenetic hypothesis as the basis of the disease.


Healthcare seeking behavior among subjects with irritable bowel syndrome.
The behaviors exhibited by individuals who seek healthcare for irritable bowel syndrome (IBS) are numerous, varied, and sometimes controversial, and/or inconsistent. This study aims to review the factors associated with these healthcare seeking behaviors. Through a selective review of the literature, articles were identified by using the following search terms: IBS, healthcare seeking, conventional and non-conventional treatment, cultural factors, and gender differences. The roles of different factors associated with healthcare seeking are discussed.


Telomere Maintenance Variants and Survival after Colorectal Cancer: Smoking- and Sex-Specific Associations.
Telomeres play an important role in colorectal cancer prognosis. Variation in telomere maintenance genes may be associated with survival after colorectal cancer diagnosis, but evidence is limited. In addition, possible interactions between telomere maintenance genes and prognostic factors, such as smoking and sex, also remain to be investigated. We conducted gene-wide analyses of colorectal cancer prognosis in 4,896 invasive colorectal cancer cases from the Genetics and Epidemiology of Colorectal Cancer Consortium (GECCO); 1,871 common variants within 13 telomere maintenance genes were included. Cox models were fit to estimate associations of these variants individually with overall and colorectal cancer-specific survival. Likelihood ratio tests were used to test for interaction by smoking and sex. P values were adjusted using Bonferroni correction. The association between minor allele of rs7200950 (ACD) with colorectal cancer-specific survival varied significantly by smoking pack-years (corrected P = 0.049), but no significant trend was observed. By sex, minor alleles for rs2975843 (TERF1), rs75676021 (POT1), and rs74429678 (POT1) were associated with decreased overall and/or colorectal cancer-specific survival in women but not in men. Our study reported a gene-wide statistically significant interaction with sex (TERF1, POT1). Although significant interaction by smoking pack-years (ACD) was observed, there was no evidence of a dose response. Validation of these findings in other large studies and further functional annotation on these SNPs are warranted. Our study found a gene-smoking and gene-sex interaction on survival after colorectal cancer diagnosis, providing new insights into the role of genetic polymorphisms in telomere maintenance on colorectal cancer prognosis.


[Diagnosis and therapy of myofascial trigger points].
Myofascial trigger points (MTrPs) are hyperirritable tender spots in palpable tense bands of skeletal muscle. Muscle is an orphan organ, no medical specialty claims muscle as its organ. The article aims at filling some of the gaps in the current knowledge of MTrPs. The presented findings were partly obtained in experiments on anesthetised rabbits, partly they are the result of ample experience with patients suffering from MTrPs. Each muscle has a characteristic elicited referred pain pattern that, for active MTrPs, is familiar to the patient. Without a laboratory test or imaging method, diagnosis of MTrPs depends entirely on history and physical examination. MTrP symptoms follow muscle overload, are activated acutely by sudden overload, or develop gradually with prolonged contractions or repetitive activity. The diagnostic skill required depends on considerable innate palpation ability, authoritative training, and extensive clinical experience. Effective treatment methods include manual stretching by trigger-point pressure release, contract-relax, vapo coolant spray-and-stretch, and dry needling or injection of MTrPs. The integrated hypothesis presents an explanation for the pathophysiology of MTrPs and begins with excessive release of acetylcholine from involved motor endplates. It depends on a new understanding of the abnormality of endplate noise. Biopsies demonstrate segmental shortening of groups of sarcomeres in individual muscle fibres and possibly waves of contracted sarcomeres to account for palpable taut bands.


Comments on Sabariego et al. Measuring Disability: Comparing the Impact of Two Data Collection Approaches on Disability Rates. Int. J. Environ. Res. Public Health, 2015, 12, 10329-10351.
In the article, Measuring Disability: Comparing the Impact of Two Data Collection Approaches on Disability Rates, in Volume 12 of the Journal International Journal of Environmental Research and Public Health, Carla Sabariego et al. [1] raise several issues regarding the use of the short set of questions developed by the Washington Group on Disability Statistics (WG) as compared with the approach to disability measurement proposed through the Model Disability Survey (MDS). We address these below. [...].


A Dual-Scale Lead-Separated Transformer for ECG Classification.
This work proposes a novel dual-scale lead-separated transformer for the auxiliary diagnosis of 12-lead electrocardiograms (ECGs). We added a new structure design on the basis of traditional ECG signal processing, which led to our model with only 2.6M parameters. The output of the system is the classification results. The fixed 0.5 second ECG segments of each lead are interpreted as independent patches. Together with the reduced dimension signal, patches form a dual-scale representation. As a method to reduce interference from segments with low correlation, a lead-orthogonal attention module is proposed. Experimental results show the effectiveness and scalability of our model.Clinical relevance- Our method improves the scores of clinical 12-lead ECG classification and shows generalization ability. Our model is suitable for single-label and multi-label classification tasks on clinical 12-lead ECG and is compatible with single lead classification. The integration of clinical information can further improve the effectiveness of the model.


Batch experiment on H2S degradation by bacteria immobilised on activated carbons.
Biological treatments of odorous compounds, as compared to chemical or physical technologies, are in general ecologically and environmentally favourable. However, there are some inefficiencies relative to the media used in biofiltration processes, such as the need for an adequate residence time; the limited lifetime, and pore blockage of media, which at present render the technology economically non-viable. The aim of the study is to develop novel active media to be used in performance-enhanced biofiltration processes, by achieving an optimum balance and combination of the media adsorption capacity with the biodegradation of H2S through the bacteria immobilised on the media. An enrichment culture was obtained from activated sludges in order to metabolise thiosulphate. Batch-wise experiments were conducted to optimise the bacteria immobilisation on activated carbon, so as to develop a novel "biocarbon". Biofilm was mostly developed through culturing the bacteria with the presence of carbons in mineral media. SEM and BET tests of the carbon along with the culturing process were used to identify, respectively, the biofilm development and biocarbon porosity. Breakthrough tests evaluated the biocarbon performance with varying gas resistance time, inlet H2S concentration, and type of support materials. Fundamental issues were discussed, including type of support material, mode of bacteria immobilisation, pore blockages, and biodegradation kinetics, etc. This batch-wise study provides a basis for our future research on optimisation of the biofiltration process using a bio-trickling reactor.


A case of reactive plasmacytosis mimicking multiple myeloma in a patient with primary Sjogren's syndrome.
Primary Sjogren's syndrome (pSS) is a chronic autoimmune disease with well-documented association of lymphoid malignancies during the progress of the disease. Although several types of malignancy and pseudomalignancy have been reported in pSS, low-grade non-Hodgkin's lymphomas are the most frequently observed. Reactive plasmacytosis mimicking myeloma is a very rare condition in association with pSS. We describe a 72-yr-old woman with pSS who presented with hypergammaglobulinemia, and extensive bone marrow and lymph node plasmacytosis, which mimicked multiple myeloma. In this patient, there was an abnormal differentiation of memory B cells to plasma cells in the peripheral blood suggesting underlying pathogenetic mechanism for this condition.


Comparison between Field Effect Transistors and Bipolar Junction Transistors as Transducers in Electrochemical Sensors.
Field effect transistors (FET) have been widely used as transducers in electrochemical sensors for over 40 years. In this report, a FET transducer is compared with the recently proposed bipolar junction transistor (BJT) transducer. Measurements are performed on two chloride electrochemical sensors that are identical in all details except for the transducer device type. Comparative measurements show that the transducer choice significantly impacts the electrochemical sensor characteristics. Signal to noise ratio is 20 to 2 times greater for the BJT sensor. Sensitivity is also enhanced: BJT sensing signal changes by 10 times per pCl, whereas the FET signal changes by 8 or less times. Also, sensor calibration curves are impacted by the transducer choice. Unlike a FET sensor, the calibration curve of the BJT sensor is independent of applied voltages. Hence, a BJT sensor can make quantitative sensing measurements with minimal calibration requirements, an important characteristic for mobile sensing applications. As a demonstration for mobile applications, these BJT sensors are further investigated by measuring chloride levels in artificial human sweat for potential cystic fibrosis diagnostic use. In summary, the BJT device is demonstrated to be a superior transducer in comparison to a FET in an electrochemical sensor.


Implications of Dual Practice among Health Workers: A Systematic Review.
Mixed health care systems to work simultaneously on both public and private facilities, is common today. This phenomenon referred to as dual practice (DP), has potential implications for access, quality, cost and equity of health services. This paper aimed to review systematically studies that assess the implications of DP among health workers. MEDLINE, EMBASE, and The Cochrane library were searched for obtaining published literature between Feb 1990 and May 2014. Google and Google Scholars, organizational websites, and reference lists of relevant papers searched to get grey literature. Only studies concentrated on consequences and impacts of DP among health professionals and conducted using "randomized controlled trials", "non-randomized controlled trials", "controlled before and after studies", or "interrupted time series" were eligible for inclusion. From 3242 records, we focused on 19 studies, which aimed to assess effects and impacts of dual practice. After that, the current understanding of DP positive and negative implications was categorized and discussed based on two perspectives. There has been a propensity to over-reliance on theoretical methods in predicting the implications of this phenomenon. Almost all of the mentioned implications are based on theoretical predictions undermined in the broader literature. Furthermore, assessing the current literature showed positive and negative impacts of DP on different parts of the health system and various dimensions of service delivery. These implications are contexted specific and may vary from system to system.


Left atrial appendage closure versus medical therapy in patients with atrial fibrillation: the APPLY study.
Left atrial appendage closure (LAAC) with AMPLATZER occluders is used for stroke prevention in atrial fibrillation (AF). Net clinical benefit compared to medical therapy has not been tested. The aim of this study was to test whether long-term clinical outcome after LAAC with AMPLATZER occluders may be similar to medical therapy. Five hundred consecutive patients who underwent LAAC with AMPLATZER occluders were compared to 500 patients with medical therapy by propensity score matching. The primary efficacy endpoint was a composite of stroke, systemic embolism and cardiovascular/unexplained death. The primary safety endpoint consisted of major procedural adverse events and major bleedings. For assessment of net clinical benefit, all of the above-mentioned hazards were combined. After 2,645 patient-years at a mean follow-up of 2.7±1.5 years, the primary efficacy endpoint was reached by 75/1,342, 5.6% in the LAAC group versus 102/1,303, 7.8% per 100 patient-years (hazard ratio [HR] 0.70, 95% confidence interval [CI]: 0.53-0.95, p=0.026). The primary safety endpoint occurred in 48/1,342, 3.6% versus 60/1,303, 4.6% per 100 patient-years (HR 0.80, 95% CI: 0.55-1.18, p=0.21), and the combined hazard endpoint in 109/1,342, 8.1% versus 142/1,303, 10.9% per 100 patient-years (HR 0.76, 95% CI: 0.60-0.97, p=0.018). Patients receiving LAAC demonstrated lower rates of both all-cause and cardiovascular mortality (111/1,342, 8.3% vs 151/1,303, 11.6% per 100 patient-years [HR 0.72, 95% CI: 0.56-0.92, p=0.005] and 54/1,342, 4.0% vs 84/1,303, 6.5% per 100 patient-years [HR 0.64, 95% CI: 0.46-0.89, p=0.007]). LAAC with AMPLATZER devices showed a net clinical benefit over medical therapy by superior efficacy, similar safety and a benefit in all-cause and cardiovascular mortality.


Novel Pd_{2}Se_{3} Two-Dimensional Phase Driven by Interlayer Fusion in Layered PdSe_{2}.
Two-dimensional (2D) materials are easily fabricated when their bulk form has a layered structure. The monolayer form in layered transition-metal dichalcogenides is typically the same as a single layer of the bulk material. However, PdSe_{2} presents a puzzle. Its monolayer form has been theoretically shown to be stable, but there have been no reports that monolayer PdSe_{2} has been fabricated. Here, combining atomic-scale imaging in a scanning transmission electron microscope and density functional theory, we demonstrate that the preferred monolayer form of this material amounts to a melding of two bulk monolayers accompanied by the emission of Se atoms so that the resulting stoichiometry is Pd_{2}Se_{3}. We further verify the interlayer melding mechanism by creating Se vacancies in situ in the layered PdSe_{2} matrix using electron irradiation. The discovery that strong interlayer interactions can be induced by defects and lead to the formation of new 2D materials opens a new venue for the exploration of defect engineering and novel 2D structures.


Neuropsychological assessment and telemedicine: a preliminary study examining the reliability of neuropsychology services performed via telecommunication.
Within the last decade, various applications of two-way audio-visual telecommunication have been implemented in the Norwegian health care system to facilitate the delivery of medical services to patients located in rural areas away from the hospitals. This technology may also be useful to extend neuropsychological services to underserved areas and thereby reduce the patients' travel time and costs. In the current study, a total of 12 visual, verbal and performance tests were administered face-to-face and via videophones to 32 volunteer participants to examine the consistency and stability of test-scores when delivered via different formats. The obtained reliability coefficients ranged from .37 to .86 with a median value of .74. When testing for mean differences, the measures on verbal learning (WMS-Logical Memory I) and auditory attention (Seashore Rhythm Test) differed significantly due to administration format (t = 2.34, p = .024 and t = 2.37, p = .025, respectively). The findings imply that administration format does not appear to affect the reliability of measurement but neuropsychological test performance is significantly higher for the measures of attention and memory when delivered via videophone. Additional research on these cognitive domains is required, and if the observed differences due to testing format persist, separate normative data via telecommunication will be required before assessments are carried out. Also, since only normal participants were included in this study, the findings may have external validity for normal populations, but similar controlled trials with patients need to be completed before remote assessments can be implemented in regular clinical practice.


Regulatory organization of the staphylococcal sae locus.
This paper describes an investigation of the complex internal regulatory circuitry of the staphylococcal sae locus and the impact of modifying this circuitry on the expression of external genes in the sae regulon. The sae locus contains four genes, the saeR and S two-component signalling module (TCS), and saeP and Q, two upstream genes of hitherto unknown function. It is expressed from two promoters, P(A)sae, which transcribes only the TCS, and P(C)sae, which transcribes the entire locus. A bursa aurealis (bursa) transposon insertion in saeP in a derivative of Staphylococcus aureus NCTC 8325 has a profound effect on sae function. It modifies the activity of the TCS, changing the expression of many genes in the sae regulon, even though transcription of the TCS (from P(A)sae) is not interrupted. Moreover, these effects are not due to disruption of saeP since an in-frame deletion in saeP has essentially no phenotype. The phenotype of S. aureus strain Newman is remarkably similar to that of the saeP : : bursa and this similarity is explained by an amino acid substitution in the Newman saeS gene that is predicted to modify profoundly the signalling function of the protein. This concurrence suggests that the saeP : : bursa insertion affects the signalling function of saeS, a suggestion that is supported by the ability of an saeQR clone, but not an saeR clone, to complement the effects of the saeP : : bursa insertion.


Claims-based Prevalence of Disease Progression among Patients with Fibrosing Interstitial Lung Disease Other than Idiopathic Pulmonary Fibrosis in the United States.
Rationale: Chronic fibrosing interstitial lung disease (ILD) with a progressive phenotype is a clinical concept describing the broad group of ILDs characterized by progressive pulmonary fibrosis. The prevalence of progressive fibrotic ILDs other than idiopathic pulmonary fibrosis (IPF) is not well understood. Objectives: We used a novel algorithm to estimate the prevalence range of disease progression among patients with non-IPF fibrotic ILD in a U.S. claims database. Methods: This was a retrospective study including adults with commercial or Medicare Advantage with Part D (MAPD) insurance using administrative claims data from October 2015 to September 2019. Patients likely to have non-IPF fibrosing ILD with a progressive phenotype were identified via an algorithm that incorporated ILD-related diagnosis codes (excluding IPF) and claims-based proxies for fibrotic ILD progression, including pulmonary function tests, chest imaging, oral corticosteroid (OCS) medications, immunosuppressive medications, lung transplant, oxygen therapy, palliative care, and respiratory hospitalization. The prevalence range of non-IPF fibrotic ILD with progressive disease behavior was calculated using strict and lenient case definitions to account for potential imprecision in the progression proxies. Results: Of nearly 9 million study-eligible patients, 17,136 were identified with non-IPF fibrosing ILD. The prevalence of disease progression per 10,000 (95% confidence interval) ranged from 12.14 (11.74-12.54) to 29.05 (28.43-29.67) over a mean observation time of 1.44 years for MAPD enrollees (n = 14,686), and from 0.89 (0.81-0.97) to 2.36 (2.24-2.48) over a mean observation time of 1.29 years for commercial enrollees (n = 2,450). Prevalence estimates increased with age for both insurance types. Among patients with progression, 4,097 met at least two progression proxies not considering OCS (strict case definition) and 9,946 met at least one progression proxy (lenient case definition). The mean (standard deviation) number of proxies met was 2.1 (1.3), and the most common individual proxies met (alone or in combination with other proxies) were OCS use (48.9%), respiratory hospitalization (44.2%), and oxygen therapy (44.1%). Conclusions: This is among the first claims-based estimates of the prevalence of non-IPF chronic fibrosing ILD with a progressive phenotype. Our analysis indicates that this phenotype is rare in the overall population but increases substantially with increasing age.


Ischemic tolerance of the heart by adaptation to chronic hypoxia is suppressed by high subchronic carbon monoxide exposure.
This study was designed to investigate whether exposure to carbon monoxide (CO) could alter the ischemic tolerance induced by chronic hypoxia. We aimed to determine whether chronic hypoxia-induced cardiovascular adaptation was modified during the return to normoxia or by subchronic CO exposure. The degree of resistance to an in vitro transient ischemia was measured, using the Langendorff method, in hearts from rats previously exposed to chronic hypoxic hypoxia and/or subchronic CO exposure to 600 ppm. Chronic hypoxia decreased ischemic contracture (15.6 +/- 04.9 vs. 60.8 +/- 07.7%) and improved both contractile recovery (59.6 +/- 07.3 vs. 21.8 +/- 06.8%) and ventricular arrhythmia during reperfusion (0 vs. 45%) compared to a control normoxic group. However, in our chronic hypoxia regression model many parameters returned near to control values except for the persistence of cardiomegaly, a significant decrease in both ischemic contracture (22.0 +/- 04.9 vs. 60.8 +/- 07.7%), and ventricular tachycardia (25 vs. 45%). CO exposure alone increased the coronary flow and improved both contractile recovery (42.6 +/- 7.2 vs. 21.8 +/- 6.8%) and ventricular arrhythmia (16.7 vs. 45%) without altering the action potential shape. These two models causing tissue hypoxia induced the same degree of polycythemia or cardiomegaly and provided similar ischemic tolerance. CO exposure after chronic hypoxia exacerbated ischemic contracture (69.3 +/- 10.5 vs. 22.0 +/- 14.5%) and ventricular tachycardia incidence (100 vs. 50%) but with significant alteration in contractile recovery (12.7 +/- 10.5%) compared to the chronic hypoxia or CO exposure. Thus, CO exposure completely suppressed the chronic hypoxia-induced ischemic tolerance.


Can We Improve Diosmetin Activity? The State-of-the-Art and Promising Research Directions.
Diosmetin is a natural substance widely distributed in nature, with documented multidirectional biological effects. The wide spectrum of biological activity of diosmetin gives hope that derivatives of this flavonoid may also be used as drugs or dietary supplements used in many diseases. Modification of the structure may, on the one hand, lead to an increase in biological potency, new biological activity, or an increase in solubility and thus bioavailability. This is an important direction of research because the use of pure diosmetin is limited due to its low bioavailability. This work is an attempt to collect information on the possibility of modifying the structure of diosmetin and its impact on biological activity.


Genotyping for severe drug hypersensitivity.
Over the past decade, there have been significant advances in our understanding of the immunopathogenesis and pharmacogenomics of severe immunologically-mediated adverse drug reactions. Such T-cell-mediated adverse drug reactions such as Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN), drug-induced liver disease (DILI) and other drug hypersensitivity syndromes have more recently been shown to be mediated through interactions with various class I and II HLA alleles. Key examples have included the associations of HLA-B*15:02 and carbamazepine induced SJS/TEN in Southeast Asian populations and HLA-B*57:01 and abacavir hypersensitivity. HLA-B*57:01 screening to prevent abacavir hypersensitivity exemplifies a successful translational roadmap from pharmacogenomic discovery through to widespread clinical implementation. Ultimately, our increased understanding of the interaction between drugs and the MHC could be used to inform drug design and drive pre-clinical toxicity programs to improve drug safety.


The purine analog fludarabine acts as a cytosolic 5'-nucleotidase II inhibitor.
For several years the IMP/GMP-preferring cytosolic 5'-nucleotidase II (cN-II) has been considered as a therapeutic target in oncology. Indeed, various reports have indicated associations between cN-II expression level and resistance to anticancer agents in several cancer cell lines and in patients affected with neoplasia, mainly by hematologic malignancies. In this paper we present evidence showing that, among the commonly used cytotoxic nucleoside analogs, fludarabine can act as a cN-II inhibitor. In vitro studies using the wild type recombinant cN-II demonstrated that fludarabine inhibited enzymatic activity in a mixed manner (Ki 0.5 mM and Ki' 9 mM), whereas no inhibition was observed with clofarabine and cladribine. Additional experiments with mutant recombinant proteins and an in silico molecular docking indicated that this inhibition is due to an interaction with a regulatory site of cN-II known to interact with adenylic compounds. Moreover, synergy experiments between fludarabine and 6-mercaptopurine in human follicular lymphoma (RL) and human acute promyelocytic leukemia (HL-60) cells transfected with control or cN-II-targeting shRNA-encoding plasmids, showed synergy in control cells and antagonism in cells with decreased cN-II expression. This is in line with the hypothesis that fludarabine acts as a cN-II inhibitor and supports the idea of using cN-II inhibitors in association with other drugs to increase their therapeutic effect and decrease their resistance.


Far-infrared radiation alleviates cisplatin-induced vascular damage and impaired circulation via activation of HIF-1α.
Severe vascular damage and complications are often observed in cancer patients during treatment with chemotherapeutic drugs such as cisplatin. Thus, development of potential options to ameliorate the vascular side effects is urgently needed. In this study, the effects and the underlying mechanisms of far-infrared radiation (FIR) on cisplatin-induced vascular injury and endothelial cytotoxicity/dysfunction in mice and human umbilical vein endothelial cells (HUVECs) were investigated. An important finding is that the severe vascular stenosis and poor blood flow seen in cisplatin-treated mice were greatly mitigated by FIR irradiation (30 minutes/day) for 1-3 days. Moreover, FIR markedly increased the levels of phosphorylation of PI3K and Akt, and VEGF secretion, as well as the expression and the activity of hypoxia-inducible factor 1α (HIF-1α) in cisplatin-treated HUVECs in a promyelocytic leukemia zinc finger protein (PLZF)-dependent manner. However, FIR-stimulated endothelial angiogenesis and VEGF release were significantly diminished by transfection with HIF-1α siRNA. We also confirmed that HIF-1α, PI3K, and PLZF contribute to the inhibitory effect of FIR on cisplatin-induced apoptosis in HUVECs. Notably, FIR did not affect the anticancer activity and the HIF-1α/VEGF cascade in cisplatin-treated cancer cells under normoxic or hypoxic condition, indicating that the actions of FIR may specifically target endothelial cells. It is the first study to demonstrate that FIR effectively attenuates cisplatin-induced vascular damage and impaired angiogenesis through activation of HIF-1α-dependent processes via regulation of PLZF and PI3K/Akt. Taken together, cotreatment with the noninvasive and easily performed FIR has a therapeutic potential to prevent the pathogenesis of vascular complications in cancer patients during cisplatin treatment.


Lactate and Immunosuppression in Sepsis.
Serum lactate levels are traditionally interpreted as a marker of tissue hypoxia and often used clinically as an indicator of severity and outcome of sepsis/septic shock. Interestingly, recent studies involving the effects of tumor-derived lactate suggest that lactate itself may have an immunosuppressive effect in its local environment. This finding adds to the recent advances in immunometabolism that shed light on the importance of metabolism and metabolic intermediates in the regulation of innate immune and inflammatory responses in sepsis. In this article, we summarize recent studies, showing that the activation of immune cells requires aerobic glycolytic metabolism and that lactate produced by aerobic glycolysis may play an immunosuppressive role in sepsis.


Phosphorylation of neurofilament proteins in intact neurons: demonstration of phosphorylation in cell bodies and axons.
The principal subunits of neurofilaments (NFs) of immature cultured sympathetic neurons have apparent Mr of 68,000 and 145,000; a 200,000 Mr subunit is also present, but at comparatively low levels. These subunits are referred to as the low (NFL), middle (NFM), and high (NFH) Mr subunits, respectively. We studied the phosphorylation of NFL and NFM in these neurons in order to characterize the NFL and NFM isoforms generated by this important posttranslational modification. NFL resolved into a single spot in 2-dimensional gels, although 2 spots were occasionally observed. NFM typically resolved into 3 variants, termed NFM a, b, and c, in order of increasing mobility, but as many as 6 variants were detected in some gels. NFL and, to a much greater degree, NFM became labeled following incubation of intact neurons with 32P-PO4. Although all 3 major NFM variants became labeled, NFM a was the most heavily labeled, followed by NFM b, and then NFM c. Two observations suggest that the generation of these 3 NFM variants is due to their phosphorylation. First, treatment of NFs with phosphatase prior to analysis reduced NFM to a single spot or band that comigrated with NFM c; NFM a and b were completely eliminated. However, NFM c was not fully dephosphorylated because it still reacted with a monoclonal antibody (mAb) specific for a phosphate-dependent epitope on NFM. Second, NFM was recognized by 4 mAbs to distinctly different phosphorylated epitopes of NFM, which suggested that at least 4 distinct sites on NFM can be phosphorylated in cultured neurons. Explant cultures were used to study the phosphorylation of NFL and NFM in cell bodies and axons. In these cultures, a central cell body mass (CBM) 0.5 mm in diameter contains all of the cell bodies, while peripheral to the CBM is a halo of pure axons that extends for 4-6 mm. These cultures were incubated with 32P-PO4 and CBM and axon regions were analyzed separately. NFL became phosphorylated to a greater extent in the CBM than in axons. NFM also became labeled in the CBM and axons, although the relative labeling of NFM a, b, and c in these regions differed considerably from each other and also from the pattern observed in whole neurons (cell bodies plus neurites, see above).(ABSTRACT TRUNCATED AT 400 WORDS)


Soluble dominant-negative receptor uncovers essential roles for fibroblast growth factors in multi-organ induction and patterning.
Despite a wealth of experimental data implicating fibroblast growth factor (FGF) signaling in various developmental processes, genetic inactivation of individual genes encoding specific FGFs or their receptors (FGFRs) has generally failed to demonstrate their role in vertebrate organogenesis due to early embryonic lethality or functional redundancy. Here we show that broad mid-gestational expression of a novel secreted kinase-deficient receptor, specific for a defined subset of the FGF superfamily, caused agenesis or severe dysgenesis of kidney, lung, specific cutaneous structures, exocrine and endocrine glands, and craniofacial and limb abnormalities reminiscent of human skeletal disorders associated with FGFR mutations. Analysis of diagnostic molecular markers revealed that this soluble dominant-negative mutant disrupted early inductive signaling in affected tissues, indicating that FGF signaling is required for growth and patterning in a broad array of organs and in limbs. In contrast, transgenic mice expressing a membrane-tethered kinase-deficient FGFR were viable. Our results demonstrate that secreted FGFR mutants are uniquely effective as dominant-negative agents in vivo, and suggest that related soluble receptor isoforms expressed in wild-type mouse embryos may help regulate FGF activity during normal development.


Identification of new drugs to counteract anti-spike IgG-induced hyperinflammation in severe COVID-19.
Previously, we and others have shown that SARS-CoV-2 spike-specific IgG antibodies play a major role in disease severity in COVID-19 by triggering macrophage hyperactivation, disrupting endothelial barrier integrity, and inducing thrombus formation. This hyperinflammation is dependent on high levels of anti-spike IgG with aberrant Fc tail glycosylation, leading to Fcγ receptor hyperactivation. For development of immune-regulatory therapeutics, drug specificity is crucial to counteract excessive inflammation whereas simultaneously minimizing the inhibition of antiviral immunity. We here developed an in vitro activation assay to screen for small molecule drugs that specifically counteract antibody-induced pathology. We identified that anti-spike-induced inflammation is specifically blocked by small molecule inhibitors against SYK and PI3K. We identified SYK inhibitor entospletinib as the most promising candidate drug, which also counteracted anti-spike-induced endothelial dysfunction and thrombus formation. Moreover, entospletinib blocked inflammation by different SARS-CoV-2 variants of concern. Combined, these data identify entospletinib as a promising treatment for severe COVID-19.


Magic angle effect in MR imaging of ankle tendons: influence of foot positioning on prevalence and site in asymptomatic subjects and cadaveric tendons.
The influence of foot positioning on prevalence of the magic angle effect (MAE) in ankle tendons was investigated. In 30 asymptomatic volunteers and five cadaveric feet, MR imaging of the ankle was performed in the supine (neutral position of the foot) and prone (plantar-flexed foot) position. MAE was considered if increased T1-weighted signal at a certain site was seen in one position only. Histological correlation was obtained at 25 sites of the cadaveric posterior tibialis tendons (PTT). MAE occurred in 6/30 vs 1/30 (supine vs prone) anterior tibialis tendons (ATT), 30/30 vs 0/30 extensor hallucis longus and 27/30 vs 0/30 extensor digitorum longus tendons, 29/30 vs 0/30 PTTs, 30/30 vs 0/30 flexor digitorum and flexor hallucis longus tendons, 30/30 vs 1/30 peroneus brevis and 23/30 vs 1/30 peroneus longus tendons. At 12/25 cadaveric PTT sites where MAE was exclusively responsible for the increased signal, histology revealed normal tissue (11/12) or minimal degeneration (1/12). In conclusion, the supine body position with neutral position of the foot, a high prevalence (77-100%) of MAE in ankle tendons except for the ATT (20%) is seen. MAE is almost absent in the prone body position with plantar flexion of the foot.


In Vitro Inhibitory Potential of Different Anthocyanin-Rich Berry Extracts in Murine CT26 Colon Cancer Cells.
Anti-oxidant, -inflammatory, and -carcinogenic activities of bioactive plant constituents, such as anthocyanins, have been widely discussed in literature. However, the potential interaction of anthocyanin-rich extracts with routinely used chemotherapeutics is still not fully elucidated. In the present study, anthocyanin-rich polyphenol extracts of blackberry (BB), bilberry (Bil), black currant (BC), elderberry (EB), and their respective main anthocyanins (cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, cyanidin-3-O-rutinoside, and cyanidin-3-O-sambubioside) were investigated concerning their cytotoxic and DNA-damaging properties in murine CT26 cells either alone or in combination with the chemotherapeutic agent SN-38. BB exerted potent cytotoxic effects, while Bil, BC, and EB only had marginal effects on cell viability. Single anthocyanins comprised of the extracts could not induce comparable effects. Even though the BB extract further pronounced SN-38-induced cytotoxicity and inhibited cell adhesion at 100-200 µg/mL, no effect on DNA damage was observed. In conclusion, anti-carcinogenic properties of the extracts on CT26 cells could be ranked BB >> BC ≥ Bil ≈ EB. Mechanisms underlying the potent cytotoxic effects are still to be elucidated since the induction of DNA damage does not play a role.


Effects of short-wave therapy in patients with knee osteoarthritis: a systematic review and meta-analysis.
To evaluate the efficacy and safety of short-wave therapy with sham or no intervention for the management of patients with knee osteoarthritis. We searched the following databases from their inception up to 26 October 2016: MEDLINE, CENTRAL, EMBASE, Physiotherapy Evidence Database, CINAHL and OpenGrey. Studies included randomized controlled trials compared with a sham or no intervention in patients with knee osteoarthritis. The results were calculated via standardized mean difference (SMD) and risk ratio for continuous variables outcomes as well as dichotomous variables, respectively. Heterogeneity was explored by the I2 test and inverse-variance random effects analysis was applied to all studies. Eight trials (542 patients) met the inclusion criteria. The effect of short-wave therapy on pain was found positive (SMD, -0.53; 95% CI, -0.84 to -0.21). The pain subgroup showed that patients received pulse modality achieved clinical improvement (SMD, -0.83; 95% CI, -1.14 to -0.52) and the pain scale in female patients decreased (SMD, -0.53; 95% CI, -0.98 to -0.08). In terms of extensor strength, short-wave therapy was superior to the control group ( p < 0.05, I2 = 0%). There was no significant difference in the physical function (SMD, -0.16; 95% CI, -0.36 to 0.05). For adverse effects, there was no significant difference between the treatment and control group. Short-wave therapy is beneficial for relieving pain caused by knee osteoarthritis (the pulse modality seems superior to the continuous modality), and knee extensor muscle combining with isokinetic strength. Function is not improved.


hedgehog signaling independent of engrailed and wingless required for post-S1 neuroblast formation in Drosophila CNS.
The hedgehog gene product, secreted from engrailed-expressing neuroectoderm, is required for the formation of post-S1 neuroblasts in rows 2, 5 and 6. The hedgehog protein functions not only as a paracrine but also as an autocrine factor and its transient action on the neuroectoderm 1-2 hours (at 18 degrees C) prior to neuroblast delamination is necessary and sufficient to form normal neuroblasts. In contrast to epidermal development, hedgehog expression required for neuroblast formation is regulated by neither engrailed nor wingless. hedgehog and wingless bestow composite positional cues on the neuroectodermal regions for S2-S4 neuroblasts at virtually the same time and, consequently, post-S1 neuroblasts in different rows can acquire different positional values along the anterior-posterior axis. The average number of proneural cells for each of three eagle-positive S4-S5 neuroblasts was found to be 5-9, the same for S1 NBs. As with wingless (Chu-LaGraff et al., Neuron 15, 1041-1051, 1995), huckebein expression in putative proneural regions for certain post-S1 neuroblasts is under the control of hedgehog. hedgehog and wingless are involved in separate, parallel pathways and loss of either is compensated for by the other in NB 7-3 formation. NBs 6-4 and 7-3, arising from the engrailed domain, were also found to be specified by the differential expression of two homeobox genes, gooseberry-distal and engrailed.


Relationship of psychosocial work factors and health-related quality of life in male automotive assembly workers in Malaysia.
The present study investigates the relationship between psychosocial work factors and health-related quality of life (HRQOL) in male automotive assembly plant workers in Malaysia. A total of 728 male workers were recruited in March-July 2005 from 2 major automotive assembly plants in Selangor and Pahang. In this cross-sectional study, information on socio-demography, psychosocial work factors using the 97-item Job Content Questionnaire (JCQ) and an abbreviated 26-item version of the World Health Organization Quality of Life-Brief Version (WHOQOL-BREF) questionnaire containing 4 domains (physical health, psychological, social relationship, and environment) was self-administered to all workers involved. The prevalence of reported good or very good overall HRQOL and general health was 64.9% and 53.7%, respectively. Multiple linear regression analysis indicated that created skill was positively associated with physical health and psychological domains; whilst, skill discretion was positively associated with social relationship and environment domains. Social support was positively associated with physical health and environment domains; whilst, co-worker support was positively associated with psychological and social relationship domains. Job insecurity and hazardous condition were negatively associated with all domains, whilst psychological job demands was negatively associated with the environment domain of HRQOL.


Quality of Life after post-prostatectomy intensity modulated radiation therapy to the prostate bed with or without the use of gold fiducial markers for image guidance or higher total radiotherapy doses.
To evaluate quality of life (QoL) after post-prostatectomy intensity modulated radiation therapy (IMRT) in the "adjuvant" setting starting within 4 months of radical prostatectomy for adverse features; and "salvage" setting for a PSA≥0.2ng/mL. Retrospective review of 130 patients who underwent IMRT to the prostate bed±gold fiducial marker placement for image guidance to 64.8-72.0Gy (median, 70.2Gy) between 2004 and 2013. Higher doses were defined as 70.2-72.0Gy and lower doses were defined as 64.8-68.4Gy. Androgen deprivation therapy (ADT) was given to 4/48 (8%) adjuvant patients and 9/82 (11%) salvage patients. International Prostate Symptom Score (IPSS), Sexual Health Inventory for Men (SHIM), and Expanded Prostate Cancer Index Composite-26-bowel (EPIC-26-bowel) questionnaires were used to assess urinary, sexual, and bowel QoL, respectively. Median follow-up was 46 months. There were better urinary (p=0.03) and sexual (p=0.002) QoL scores with adjuvant IMRT relative to salvage IMRT. The use of prostate bed fiducial markers did not significantly affect urinary, sexual, or bowel QoL (p=0.39, p=0.49, and p=0.40, respectively). Higher total radiotherapy doses did not significantly affect urinary, sexual, or bowel QoL (p=0.21, p=0.61, and p=0.36, respectively). There was no significant change in urinary, sexual, and bowel sexual QoL with post-prostatectomy IMRT regardless of whether prostate bed fiducial markers or higher total radiotherapy doses were used. QoL with IMRT in the present study compares favorably with prior reports for three-dimensional conformal radiation therapy.


The effects of lidocaine and fentanyl on airway irritability during inhalation induction with desflurane.
Inhalation induction with desflurane can cause airway irritability and sympathetic stimulation. The aim of this study was to investigate whether lidocaine and fentanyl could reduce these unwanted reactions. Seventy-five patients who had premedication with midazolam were randomly allocated to one of three groups to receive intravenous saline (S group), lidocaine 1.5 mg/kg (L group), fentanyl 1 microgram/kg (F group), respectively, before tidal volume induction with desflurane in oxygen and nitrous oxide. We recorded airway irritability such as cough, apnea, laryngospasm and excitatory movement and hemodynamic changes. Airway irritability was not significantly different between the groups. In F group, mean blood pressure at LOC ver and LOC BIS and heart rate at LOC ver, LOC BIS and just before intubation were lower than those of S group (P < 0.05). Other results were not significantly different. The results of the study showed that intravenous fentanyl and lidocaine had no beneficial effects to reduce airway irritability, but intravenous fentanyl could significantly reduce hemodynamic stimulation during inhalation induction with desflurane in the patients who were premedicated with midazolam.


Knockdown of GALNT1 suppresses malignant phenotype of hepatocellular carcinoma by suppressing EGFR signaling.
O-glycosylation is a common protein modification. Aberrant O-glycosylation is associated with many cancers. GALNT1 is a GalNAc-transferase that initiates protein O-glycosylation. We found that GALNT1 is frequently up-regulated in hepatocellular carcinoma (HCC) and is associated with poor patient survival. Overexpression of GALNT1 increased and knockdown decreased HCC cell migration and invasion. Knockdown of GALNT1 inhibited EGF-induced migration and invasion. Knockdown of GALNT1 decreased EGFR activation and increased EGFR degradation, by decreasing EGFR O-glycosylation. This study demonstrates that down-regulation of GALNT1 is sufficient to suppress malignant phenotype of HCC cells by decreasing EGFR signaling. Thus, GALNT1 is a potential target in HCC.


Rop GTPase-dependent dynamics of tip-localized F-actin controls tip growth in pollen tubes.
Tip-growing pollen tubes provide a useful model system to study polar growth. Although roles for tip-focused calcium gradient and tip-localized Rho-family GTPase in pollen tube growth is established, the existence and function of tip-localized F-actin have been controversial. Using the green fluorescent protein-tagged actin-binding domain of mouse talin, we found a dynamic form of tip-localized F-actin in tobacco pollen tubes, termed short actin bundles (SABs). The dynamics of SABs during polar growth in pollen tubes is regulated by Rop1At, a Rop GTPase belonging to the Rho family. When overexpressed, Rop1At transformed SAB into a network of fine filaments and induced a transverse actin band behind the tip, leading to depolarized growth. These changes were due to ectopic Rop1At localization to the apical region of the plasma membrane and were suppressed by guanine dissociation inhibitor overexpression, which removed ectopically localized Rop1At. Rop GTPase-activating protein (RopGAP1) overexpression, or Latrunculin B treatments, also recovered normal actin organization and tip growth in Rop1At-overexpressing tubes. Moreover, overexpression of RopGAP1 alone disrupted SABs and inhibited growth. Finally, SAB oscillates and appears at the tip before growth. Together, these results indicate that the dynamics of tip actin are essential for tip growth and provide the first direct evidence to link Rho GTPase to actin organization in controlling cell polarity and polar growth in plants.


Effects of aging on the dynamics of lymphocyte organ distribution in mice: use of a radioiodinated cell membrane probe.
We have employed a derivatized aminostyrylpyridinium dye, [125I]I2P-Di-6-ASP, to provide a relatively stable tag on mixed mouse splenocytes and purified B and T cells for the purpose of tracking the distribution of those cells among the organs of normal young (4 months) and aged (> 26 months) recipient mice. Cells from both young and aged donor spleens were studied. Special emphasis was placed on localization of donor cells in the spleens of the recipients because the majority of circulating lymphocytes localize in the spleen and the spleen is the principal organ of primary immune response. There was a profound difference in the efficiency of splenic acquisition of donor cells between young and aged recipients, a difference not found in the liver, lungs, kidneys or heart. In contrast young and old donor lymphocytes lodged equally well in the spleens of recipients of the same age. It was clear that the competence of the splenic microenvironment to serve as a lodging site for circulating lymphocytes deteriorated with age. Such a change could contribute significantly to the deficient immune response of aged subjects. We suggest that aging results in significant change in the splenic extracellular matrix to serve as an adhesive substratum for lymphocytes. Our data point to a need for detailed studies on age-related changes in components of the extracellular matrix within lymphoid tissues. The novel compound which we employed for cell labeling is both radioactive and fluorescent and should be quite suitable for such studies.


Work-related injuries in a state trauma registry: relationship between industry and drug screening.
Work-related injuries exert a great financial and economic burden on the US population. The study objectives were to identify the industries and occupations associated with worker injuries and to determine the predictors for injured worker drug screening in trauma centers. Work-related injury cases were selected using three criteria (expected payer source of workers' compensation, industry-related e-codes, and work-related indicator) from the Kentucky Trauma Registry data set for years 2008 to 2012. Descriptive analyses and multiple logistic regression were performed on the work-related injury cases. The "other services" and construction industry sectors accounted for the highest number of work-related cases. Drugs were detected in 55% of all drug-screened work-related trauma cases. Higher percentages of injured workers tested positive for drugs in the natural resources and mining, transportation and public utilities, and construction industries. In comparison, higher percentages of injured workers in the other services as well as transportation and public utilities industries were drug screened. Treatment at Level I trauma centers and Glasgow Coma Scale (GCS) scores indicating a coma or severe brain injury were both significant independent predictors for being screened for drugs; industry was not a significant predictor for being drug screened. The injured worker was more likely to be drug screened if the worker had a greater than mild injury, regardless of whether the worker was an interfacility transfer. These findings indicate that there may be elevated drug use or abuse in natural resources and mining, transportation and public utilities, as well as construction industry workers; improved identification of the specific drug types in positive drug screen results of injured workers is needed to better target prevention efforts. Epidemiologic study, level III.


Characterization of the N-methoxyindole-3-carbinol (NI3C)--induced cell cycle arrest in human colon cancer cell lines.
Recent results have shown that indole-3-carbinol (I3C) inhibits the cellular growth of human cancer cell lines. In some cruciferous vegetables, another indole, N-methoxyindole-3-carbinol (NI3C), is found beside I3C. Knowledge about the biological effects of NI3C is limited. The aim of the present study was to show the effect of NI3C on cell growth of two human colon cancer cell lines, DLD-1 and HCT-116. For the first time it is shown that NI3C inhibits cellular growth of DLD-1 and HCT-116 and that NI3C is a more potent inhibitor of cell proliferation than I3C. In addition to the inhibition of cellular proliferation, NI3C caused an accumulation of HCT-116 cells in the G2/M phase, in contrast to I3C, which led to an accumulation of the colon cells in G0/G1 phase. Furthermore, NI3C delays the G1-S phase transition of synchronized HCT-116 cells. The indole-mediated cell-cycle arrest may be related to the increased levels of the CDK-inhibitors p21 and p27 (only induced by NI3C). Only an initial increase of cdc2 protein was observed, whereas prolonged treatment with NI3C or I3C downregulates the mRNA and proteins of cyclin-dependent kinases and cyclins. These results indicate that both NI3C and I3C inhibit the proliferation of human colon cells but via different mechanisms.


Early communicative gestures prospectively predict language development and executive function in early childhood.
Using an epidemiological sample (N = 1,117) and a prospective longitudinal design, this study tested the direct and indirect effects of preverbal and verbal communication (15 months to 3 years) on executive function (EF) at age 4 years. Results indicated that whereas gestures (15 months), as well as language (2 and 3 years), were correlated with later EF (φs ≥ .44), the effect was entirely mediated through later language. In contrast, language had significant direct and indirect effects on later EF. Exploratory analyses indicated that the pattern of results was comparable for low- and not-low-income families. The results were consistent with theoretical accounts of language as a precursor of EF ability, and highlighted gesture as an early indicator of EF.


Association of risk factor variables and coronary artery disease documented with angiography.
Stepwise linear discrimination was used to analyze risk factors in 431 consecutive patients who underwent coronary angiography to determine which variables were most closely associated with coronary artery disease. Twenty-one risk factors were considered: total plasma cholesterol and triglycerides; the cholesterol and triglyceride content of high-density lipoproteins (HDL), low-density lipoproteins (LDL) and very low density lipoproteins (VLDL); and the percentage of total cholesterol and triglycerides in each fraction. Age, smoking history, family history, hypertension, diabetes mellitus and relative weight were also considered. Coronary artery disease was assessed using three standard grading scores. There were significant differences in risk factors between males and females. In males, LDL cholesterol and age were selected by multivariate analysis. In females, the ratio of HDL cholesterol to total cholesterol, as well as relative weight, family history, age and smoking were selected. The discriminating value of HDL cholesterol as the percentage of total cholesterol was significantly greater than that of HDL cholesterol itself. Despite highly significant associations between risk factors and the presence of coronary artery disease, the discrimination did not provide sufficient separation of the groups to give results that are useful diagnostically in individual patients.


[Identification of endocrine cells in the gastrointestinal tract].
The gastric and duodenal endocrine cells (apudocytes) of children and immature rats were studied by electron microscopy. It is shown that every cell type has its own morphological peculiarities of secretory granules formation, maturation and disintegration. This feature together with the size and morphology of mature granules is recommended to be used as a classification criterion for the identification of apudocyte types.


Determination of bacterial cell concentrations by electrical measurements.
An instrument was developed to measure the concentration of bacterial suspensions by their electrical characteristics. It employed a square-wave signal generator, a tetra-polar electrode probe, and a voltage detector in the form of an oscilloscope. When electrical measurements were made on washed bacteria obtained from cultures or urine specimens, there was a direct relationship between the concentration of the cells and the electrical characteristics of the system as reflected by voltage changes. As little as 10(3) organisms per ml could be detected. The resolution between readings taken on samples containing 10(3) to 10(9) cells per ml was found to be a function of the input frequency. The maximal resolution between concentration readings was obtained at a input frequency of 10 Hz. Thus, with relatively simple instrumentation, bacterial concentrations could be determined within a few minutes. This technique, therefore, eliminates the more lengthy laboratory procedures as plate counts or the accumulation of measurable metabolic changes (such as the utilization of radioactive or other substrates). This method can efficiently monitor clinical urine specimens when a bacteriuria is suspected.