Resistance to local anaesthetics administered via epidural, intrathecal and pudendal injections.
A primigravida presented to our institution in established labour. Her past medical history included joint hypermobility, postural orthostatic tachycardia syndrome, Raynaud's syndrome, fibromyalgia and gastroparesis. Two technically uneventful lumbar epidurals with bupivacaine and fentanyl provided no analgesia. The spinal element of a planned combined spinal and epidural was also ineffective, so alternative analgesia was offered. While this was being prepared, the obstetric team recommended an instrumental delivery. An attempted pudendal nerve block with lidocaine had no effect, and general anaesthesia was therefore provided for a lower segment caesarean delivery. We believe this is the first report of local anaesthetic resistance via three distinct routes of administration in a single patient. Resistance to local anaesthetics is unusual and is more common in patients with hypermobility spectrum disorders. This case demonstrates the unique experience of a patient with a hypermobility condition who had failed epidural, spinal and pudendal local anaesthetic. We suggest that patients with a hypermobility condition should be specifically assessed for local anaesthetic resistance as part of anaesthetic pre-assessment, to enable early planning.


Maximum carotid intima-media thickness and NT-pro BNP in association with retinal vein occlusion.
The purpose is to clarify the relationship between patients with retinal vein occlusion (RVO), maximal intima-media complex thickness (Max IMT), and N-terminal pro-brain natriuretic peptide (NT-proBNP), which is useful in assessing atherosclerosis. This was a retrospective observation, single center study. The patients were 86 RVO patients (male: female = 43:43, mean age 63.3 years), 25 with central retinal vein occlusion (CRVO) and 61 with branch retinal vein occlusion (BRVO), classified as ≧50 years old and <50 years old, Max IMT≧1.1 and less, NT-pro BNP≧55 and less. Results showed that Max IMT ≧1.1 was significantly more common in both the CRVO and BRVO groups at ≧50 years, and NT-pro BNP ≧55 was significantly more common in the CRVO group. Max IMT≧1.1 was seen in 80% of the BRVO group and in 85% of patients aged ≧50 years. Sixty-eight percent of patients in the CRVO group had Max IMT≧1.1, but none of those < 50 years had Max IMT≧1.1. Forty-eight percent of RVO patients had NT-pro BNP≧55, and significantly more patients had Max IMT≧1.1 than those who did not have NT-pro BNP more than 55 (p = 0.02). Multiple regression analysis with Max IMT as the dependent variable showed that age and NT-pro BNP were significantly associated with RVO (p = 0.015, 0.022). RVO patients were more likely to have a Max IMT≧1.1, which was associated with atherosclerosis. Max IMT and NT-pro BNP were also associated with RVO patients, so NT-pro BNP may be a marker of RVO.


Lumpfish (Cyclopterus lumpus, Linnaeus) is susceptible to viral nervous necrosis: Result of an experimental infection with different genotypes of Betanodavirus.
In recent years, the use of cleaner fish for biological control of sea lice has increased considerably. Along with this, a number of infectious diseases have emerged. The aim of this study was to investigate the susceptibility of lumpfish (Cyclopterus lumpus) to Betanodavirus since it was detected in asymptomatic wild wrasses in Norway and Sweden. Three betanodaviruses were used to challenge lumpfish: one RGNNV genotype and two BFNNV genotypes. Fish were injected and monitored for 4 weeks. Brain samples from clinically affected specimens, from weekly randomly selected fish and survivors were subjected to molecular testing, viral isolation, histopathology and immunohistochemistry. Reduced survival was observed but was attributed to tail-biting behaviour, since no nervous signs were observed throughout the study. Betanodavirus RNA was detected in all samples, additionally suggesting an active replication of the virus in the brain. Viral isolation confirmed molecular biology results and revealed a high viral titre in BFNNV-infected groups associated with typical lesions in brains and eyes of survivor fish. We concluded that lumpfish are susceptible to Betanodavirus, as proven by the high viral titre and brain lesions detected, but further studies are necessary to understand if Betanodavirus can cause clinical disease in this species.


Case report: Persistent shedding of a live vaccine-derived rubella virus in a young man with severe combined immunodeficiency and cutaneous granuloma.
A young man with X-linked severe combined immunodeficiency developed a persistent vaccine-derived rubella virus (VDRV) infection, with the emergence of cutaneous granulomas more than fifteen years after receipt of two doses of measles-mumps-rubella (MMR) vaccine. Following nasopharyngeal swab (NP) collection, VDRV was detected by real-time polymerase chain reaction (RT-qPCR) and sequencing, and live, replication-competent VDRV was isolated in cell culture. To assess duration and intensity of viral shedding, sequential respiratory samples, one cerebrospinal fluid sample, and two urine samples were collected over 15 months, and VDRV RNA was detected in all samples by RT-qPCR. Live VDRV was cultured from nine of the eleven respiratory specimens and from one urine specimen. To our knowledge, this was the first reported instance of VDRV cultured from respiratory specimens or from urine. To assess potential transmission to close contacts, NP specimens and sera were collected from all household contacts, all of whom were immunocompetent and previously vaccinated with MMR. VDRV RNA was not detected in any NP swabs from the contacts, nor did serologic investigations suggest VDRV transmission to any contacts. This report highlights the need to understand the prevalence and duration of VDRV shedding in granuloma patients and to estimate the risk of VDRV transmission to immune and non-immune contacts.


Inhibition of gene expression in plant cells by expression of antisense RNA.
Due to the paucity of mutations in biochemical pathways in plants, an alternative approach to classical genetics was tested by expressing antisense RNA in plant cells. A series of plasmids was constructed with the bacterial gene for chloramphenicol acetyltransferase (EC 126.96.36.199) linked in either the sense or antisense orientation to several different plant gene promoters. Various ratios of sense and antisense chloramphenicol acetyltransferase gene plasmids were introduced into plant protoplasts by electric field-mediated DNA transfer ("electroporation") and the level of expression in each combination was monitored by chloramphenicol acetyltransferase assays. Transcription of antisense RNA was found to effectively block the expression of target genes. Thus, the observation that antisense RNA inhibits gene expression in bacteria and animal systems has been extended to the plant kingdom. Antisense RNA techniques have immediate practical applications in both basic research and in plant genetic engineering.


Prenatal oxidative balance and risk of asthma and allergic disease in adolescence.
Fetal oxidative balance (achieved when protective prenatal factors counteract sources of oxidative stress) might be critical for preventing asthma and allergic disease. We examined prenatal intakes of hypothesized protective nutrients (including antioxidants) in conjunction with potential sources of oxidative stress in models of adolescent asthma and allergic disease. We used data from 996 mother-child pairs in Project Viva. Exposures of interest were maternal prepregnancy body mass index and prenatal nutrients (energy-adjusted intakes of vitamins D, C, and E; β-carotene; folate; choline; and n-3 and n-6 polyunsaturated fatty acids [PUFAs]), air pollutant exposures (residence-specific third-trimester black carbon or particulate matter with a diameter of less than 2.5 μm [PM2.5]), acetaminophen, and smoking. Outcomes were offspring's current asthma, allergic rhinitis, and allergen sensitization at a median age of 12.9 years. We performed logistic regression. Continuous exposures were log-transformed and modeled as z scores. We observed protective associations for vitamin D (odds ratio [OR], 0.69 [95% CI, 0.53-0.89] for allergic rhinitis), the sum of the n-3 PUFAs eicosapentaenoic acid and docosahexaenoic acid (OR, 0.81 [95% CI, 0.66-0.99] for current asthma), and the n-3 PUFA α-linolenic acid (OR, 0.78 [95% CI, 0.64-0.95] for allergen sensitization and OR, 0.80 [95% CI 0.65-0.99] for current asthma). Black carbon and PM2.5 were associated with an approximately 30% increased risk for allergen sensitization. No multiplicative interactions were observed for protective nutrient intakes with sources of oxidative stress. We identified potential protective prenatal nutrients (vitamin D and n-3 PUFAs), as well as adverse prenatal pro-oxidant exposures that might alter the risk of asthma and allergic disease into adolescence.


Effects of dietary chloride and magnesium on the incidence of tibial dyschondroplasia in chickens fed on Chinese practical diets.
1. The effects of varying the concentrations of dietary magnesium and chloride on the performance, incidence of tibial dyschondroplasia and tissue element contents of broilers fed on Chinese practical maize soya-bean meal diets was assessed. 2. Two concentrations of dietary chloride (1.5 and 3.5 g/kg) and 3 of dietary magnesium (2.0, 4.0 and 6.0 g/kg) were used to form 6 dietary treatments for a 21-d experimental period. 3. Results from this study indicated that the dietary ingredients of maize and soya-bean meals produced in the northeast of China may predispose or influence the susceptibility of chickens to tibial dyschondroplasia. 4. High dietary chloride induces a high incidence of tibial dyschondroplasia (P less than 0.05) which could be ameliorated by dietary magnesium supplementation. 5. Dietary magnesium plays an anti-rachitic role by improving the deposition of copper and zinc in the bone tissue. 6. Supplementation with magnesium to 4.0 g/kg is recommended for broiler diets in northeast China.


DPAGT1 myasthenia and myopathy: genetic, phenotypic, and expression studies.
To investigate patients with DPAGT1 (UDP-N-acetylglucosamine-dolichyl-phosphate N-acetylglucosaminephosphotransferase 1)-associated myasthenic syndrome. We performed exome and Sanger sequencing, determined glycoprotein expression in patient muscles, assessed pathogenicity of the mutant proteins by examining their expression and enzymatic activity in transfected cells, evaluated structural changes in muscle and the neuromuscular junction, and examined electrophysiologic aspects of neuromuscular transmission in vitro. Patients 1 and 2, 16 and 14 years of age, had progressive fatigable weakness since infancy and are intellectually disabled. Patient 3, a less severely affected brother of patient 1, also has autistic features. Each patient harbors 2 novel heteroallelic mutations in DPAGT1, an enzyme subserving protein N-glycosylation. Patients 1 and 3 harbor Met1Leu, which reduces protein expression, and His375Tyr, which decreases enzyme activity. Patient 2 carries Val264Met, which abolishes enzyme activity, and a synonymous Leu120Leu mutation that markedly augments exon skipping, resulting in some skipped and infrequent nonskipped alleles. Therefore, the nonskipped allele rescues the phenotype. Intracellular microelectrode studies indicate combined pre- and postsynaptic defects of neuromuscular transmission with evidence for somatic mosaicism in patient 2. Structural studies reveal hypoplastic endplates, fiber-type disproportion, tubular aggregates, and degeneration of muscle fiber organelles resulting in autophagocytosis. DPAGT1 myasthenia affects multiple parameters of neuromuscular transmission, causes fiber-type disproportion and an autophagic myopathy, and can be associated with intellectual disability. We speculate that hypoglycosylation of synapse-specific proteins causes defects in central as well as motor synapses.


The Hombres Saludables Physical Activity Web-Based and Mobile Phone Intervention: Pilot Randomized Controlled Trial With Latino Men.
Owing to structural-level, interpersonal-level, and individual-level barriers, Latino men have disproportionately high rates of physical inactivity and experience related chronic diseases. Despite these disparities, few physical activity (PA) interventions are culturally targeted for Latino men. This study reported the feasibility and acceptability of Hombres Saludables PA intervention for Latino men. We also reported the preliminary efficacy of the intervention on PA change and provided the results of the exploratory moderator and mediator analysis. We completed a 6-month, single-blind, pilot randomized controlled trial of Hombres Saludables with Latino men aged between 18 and 65 years. Men were randomized to either (1) a theory-driven, individually tailored, internet-based and SMS text message-based, Spanish-language PA intervention arm or (2) a nutrition and wellness attention contact control arm that was also delivered via the web and SMS text message. We assessed the primary study outcomes of feasibility using participant retention and acceptability using postintervention survey and open-ended interview questions. We measured the preliminary efficacy via change in minutes of moderate to vigorous PA per week using ActiGraph wGT3X-BT accelerometry (primary measure) and self-reported minutes per week using 7-day Physical Activity Recall. Participants completed the assessments at study enrollment and after 6 months. The 38 participants were predominantly Dominican (n=8, 21%) or Guatemalan (n=5, 13%), and the mean age was 38.6 (SD 12.43) years. Retention rates were 91% (21/23) for the PA intervention arm and 100% (15/15) for the control arm. Overall, 95% (19/20) of the intervention arm participants reported that the Hombres study was somewhat to very helpful in getting them to be more physically active. Accelerometry results indicated that participants in the intervention group increased their PA from a median of 13 minutes per week at study enrollment to 34 minutes per week at 6 months, whereas the control group participants showed no increases. On the basis of self-reports, the intervention group was more likely to meet the US PA guidelines of 150 minutes per week of moderate to vigorous PA at 6-month follow-up, with 42% (8/19) of the intervention participants meeting the PA guidelines versus 27% (4/15) of the control participants (odds ratio 3.22, 95% CI 0.95-13.69). Exploratory analyses suggested conditional effects on PA outcomes based on baseline stage of motivational readiness, employment, and neighborhood safety. The PA intervention demonstrated feasibility and acceptability. Results of this pilot study indicate that the Hombres Saludables intervention is promising for increasing PA in Latino men and suggest that a fully powered trial is warranted. Our technology-based PA intervention provides a potentially scalable approach that can improve health in a population that is disproportionately affected by low PA and related chronic disease. ClinicalTrials.gov NCT03196570; https://classic.clinicaltrials.gov/ct2/show/NCT03196570. RR2-10.2196/23690.


Regulation of cancer stem cells by p53.
The hypothesis that cancer stem cells are responsible for the chemoresistant and metastatic phenotypes of many breast cancers has gained support using cell-sorting strategies to enrich the tumor-initiating population of cells. The mechanisms regulating the cancer stem cell pool, however, are less clear. Two recent publications suggest that loss of p53 permits expansion of presumptive cancer stem cells in mouse mammary tumors and in human breast cell lines. These results add restriction of cancer stem cells as a new tumor suppressor activity attributed to p53.


JNK1 stress signaling is hyper-activated in high breast density and the tumor stroma: connecting fibrosis, inflammation, and stemness for cancer prevention.
Mammography is an important screening modality for the early detection of DCIS and breast cancer lesions. More specifically, high mammographic density is associated with an increased risk of breast cancer. However, the biological processes underlying this phenomenon remain largely unknown. Here, we re-interrogated genome-wide transcriptional profiling data obtained from low-density (LD) mammary fibroblasts (n = 6 patients) and high-density (HD) mammary fibroblasts (n = 7 patients) derived from a series of 13 female patients. We used these raw data to generate a "breast density" gene signature consisting of>1250 transcripts that were significantly increased in HD fibroblasts, relative to LD fibroblasts. We then focused on the genes that were increased by ≥ 1.5-fold (P<0.05) and performed gene set enrichment analysis (GSEA), using the molecular signatures database (MSigDB). Our results indicate that HD fibroblasts show the upregulation and/or hyper-activation of several key cellular processes, including the stress response, inflammation, stemness, and signal transduction. The transcriptional profiles of HD fibroblasts also showed striking similarities to human tumors, including head and neck, liver, thyroid, lung, and breast cancers. This may reflect functional similarities between cancer-associated fibroblasts (CAFs) and HD fibroblasts. This is consistent with the idea that the presence of HD fibroblasts may be a hallmark of a pre-cancerous phenotype. In these biological processes, GSEA predicts that several key signaling pathways may be involved, including JNK1, iNOS, Rho GTPase(s), FGF-R, EGF-R, and PDGF-R-mediated signal transduction, thereby creating a pro-inflammatory, pro-proliferative, cytokine, and chemokine-rich microenvironment. HD fibroblasts also showed significant overlap with gene profiles derived from smooth muscle cells under stress (JNK1) and activated/infected macrophages (iNOS). Thus, HD fibroblasts may behave like activated myofibroblasts and macrophages, to create and maintain a fibrotic and inflammatory microenvironment. Finally, comparisons between the HD fibroblast gene signature and breast cancer tumor stroma revealed that JNK1 stress signaling is the single most significant biological process that is shared between these 2 data sets (with P values between 5.40E-09 and 1.02E-14), and is specifically associated with tumor recurrence. These results implicate "stromal JNK1 signaling" in the pathogenesis of human breast cancers and the transition to malignancy. Augmented TGF-β signaling also emerged as a common feature linking high breast density with tumor stroma and breast cancer recurrence (P = 5.23E-05). Similarities between the HD fibroblast gene signature, wound healing, and the cancer-associated fibroblast phenotype were also noted. Thus, this unbiased informatics analysis of high breast density provides a novel framework for additional experimental exploration and new hypothesis-driven breast cancer research, with a focus on cancer prevention and personalized medicine.


Effects of lesions of the nigrostriatal pathway and of nigral grafts on striatal serotonergic innervation in adult rats.
Neonatal destruction of the nigrostriatal dopaminergic system leads to serotonergic hyperinnervation of the striatum. However, it is not clear whether this occurs in adult animals. We investigated whether serotonergic sprouting occurs in adult animals, and also studied the effects of prior or subsequent implantation of dopamine-rich intrastriatal grafts. One group of adult rats received maximal 6-hydroxydopamine lesions. Other rats received maximal lesions and intrastriatal grafts 2 months later, or vice versa. The lesioned non-grafted rats showed clear serotonergic hyperinnervation throughout the striatum ipsilateral to the lesion. Intrastriatal grafts did not prevent or revert this serotonergic hyperinnervation, and were themselves densely innervated by serotonergic fibers. Serotonergic neurons usually present in the grafted cell suspension also contributed to the serotonergic innervation of the graft and the surrounding striatum.


Negative reactivity and attachment: Down syndrome's contribution to the attachment-temperament debate.
This longitudinal study investigated the relation between negative reactivity and attachment in children with Down syndrome (DS). Specifically, we examined whether extremely low negative reactivity is associated with the enactment and organization of attachment behaviors. To address this issue 30 infants with DS (13 female, 17 male) were observed in Ainsworth's Strange Situation at 19 and 27 months. The intensity and duration of distress reactions were coded in 15-s intervals throughout each Strange Situation episode. Children's distress intensity during separations from caregivers was used to place children into Low versus Moderate/High Negative Reactivity groups. In addition, ratings of infants' social interactive behaviors (contact maintenance, proximity seeking, distance interaction, resistance, and avoidance) were made for each episode and used to assess the quality of infants' attachment relationships. The data indicated that there are high rates of Type A and D insecure attachments within the DS population at each age studied. However, low negative reactivity was not consistently related to the activation of the attachment behavioral system, or associated with insecure attachment relationships. Alternative explanations for the high rate of insecure attachments are discussed.


Molecular Detection of Trypanosoma spp. in Questing and Feeding Ticks (Ixodidae) Collected from an Endemic Region of South-West Australia.
A growing number of indigenous trypanosomes have been reported to naturally infect a variety of Australian wildlife with some species of Trypanosoma implicated in the population decline of critically endangered marsupials. However, the mode of transmission of Australian trypanosomes is unknown since their vectors remain unidentified. Here we aimed to fill this current knowledge gap about the occurrence and identity of indigenous trypanosomes in Australian invertebrates by conducting molecular screening for the presence of Trypanosoma spp. in native ticks collected from south-west Australia. A total of 231 ticks (148 collected from vegetation and 83 retrieved directly from 76 marsupial hosts) were screened for Trypanosoma using a High-Resolution Melt (HRM) qPCR assay. An overall Trypanosoma qPCR positivity of 37% (46/125) and 34% (26/76) was detected in questing ticks and host-collected (i.e., feeding) ticks, respectively. Of these, sequencing revealed 28% (35/125) of questing and 28% (21/76) of feeding ticks were infected with one or more of the five species of trypanosome previously reported in this region (T. copemani, T. noyesi, T. vegrandis, T. gilletti, Trypanosoma sp. ANU2). This work has confirmed that Australian ticks are capable of harbouring several species of indigenous trypanosome and likely serve as their vectors.


The Thirteenth International Conference on Endothelin (ET-13), Tokyo, 2013.
The Thirteenth International Conference on Endothelin (ET-13) was held from September 8-11, 2013 in Tokyo, Japan chaired by Noriaki Emoto, Kobe Pharmaceutical University, Japan, and Takashi Miyauchi, University of Tsukuba, Japan and held on the Tokyo Campus of Tsukuba University. The International Conferences on Endothelin were launched in December of 1988 shortly after the discovery of endothelin and organized by Sir John Vane, laureate of the Nobel Prize in Physiology or Medicine 1982, as the Conference Chair at The William Harvey Research Institute, London. Since then, the conference has been held every two years alternating between North America, Europe and Asia. In 2013, the conference was again held in Asia and also marked the 25th anniversary of the discovery of endothelin at University of Tsukuba. The 25th anniversary of the discovery of endothelin was celebrated by almost 300 attendees from 25 different countries, the largest number of delegates in the recent history of the conference. Conference delegates who traveled to Japan were from Argentina, Australia, Austria, Brazil, Canada, China, Czech Republic, Denmark, France, Germany, Greece, Hong Kong, Hungary, Indonesia, Italy, Japan, Korean Republic, Netherlands, Sweden, Switzerland, Taiwan, Turkey, United Kingdom, United States, and from Vietnam. In this article we summarize the conference highlights, its speakers, and some of the festivities related to the celebration of the 25th anniversary of the discovery of endothelin.


Second-site compensatory mutations of HIV-1 capsid mutations.
The human immunodeficiency virus (HIV) capsid (CA) protein assembles into a hexameric lattice that forms the mature virus core. Contacts between the CA N-terminal domain (NTD) of one monomer and the C-terminal domain (CTD) of the adjacent monomer are important for the assembly of this core. In this study, we have examined the effects of mutations in the NTD region associated with this interaction. We have found that such mutations yielded modest reductions of virus release but major effects on viral infectivity. Cell culture and in vitro assays indicate that the infectivity defects relate to abnormalities in the viral cores. We have selected second-site compensatory mutations that partially restored HIV infectivity. These mutations map to the CA CTD and to spacer peptide 1 (SP1), the portion of the precursor Gag protein immediately C terminal to the CTD. The compensatory mutations do not locate to the molecularly modeled intermolecular NTD-CTD interface. Rather, the compensatory mutations appear to act indirectly, possibly by realignment of the C-terminal helix of the CA CTD, which participates in the NTD-CTD interface and has been shown to serve an important role in the assembly of infectious virus.


Complementary and Integrative Approaches for Pediatric Headache.
In this article, the use of complementary and integrative medicine for the management of pediatric headache is reviewed. Despite limited numbers of studies for pediatric headaches, children and families seek these services. Integrative medicine focuses on treating the whole person, integrating conventional medicine with mind-body-spirit methods. Nutriceuticals include dietary supplements in the form of vitamins (vitamin D), minerals (magnesium), coenzyme Q, butterbur, and melatonin. Acupuncture, stimulation, physical therapy and Transcutaneous Electrical Nerve Stimulations (TENS) or Transcranial Magnetic Stimulation (TMS) may also be useful in selected patients. The efficacy of all these therapeutic alternatives in pediatric headache is presented here. Primary care providers, neurologists, and headache specialists alike need to be informed of such interventions and integrate these approaches, when appropriate, in the management of children with headaches.


[Achievements in the study of the development and ultrastructure of spermatozoa (on the 300th anniversary of the discovery of spermatozoa)].
A review of principal advances in studying spermiogenesis and fine structure of spermatozoa is given on the occasion of the 3rd centennary of the discovery of spermatozoa by Loevenhuk. The most important achievements are the discovery of acrosome and its role in fertilization, the discovery of centriole and its role in the formation of flagellum, the discovery of axial complex and the role of its components in the flagellum movement. A study of spermatogenesis has revealed the syncytial nature of spermatogenic epithelium, synaptinemal complexes and their role in chromosome conjugation. The role of nuclear membrane in chromosome conjugation was established, the ultrastructural and biochemical transformation of chromatin was investigated, as well as that of microtubules in spermatogenesis. Some other discoveries were made which are of great importance for diagnosis of sterility.


Inferior olivary neurons in the awake cat: detection of contact and passive body displacement.
We have recorded from 306 neurons in the inferior olive of six alert cats. Most of the cats were trained to perform a simple task with the forelimb. We observed the neural responses to a wide variety of cutaneous and proprioceptive stimuli, as well as responses during spontaneous and learned active movements. Neurons responsive to somatosensory stimulation were found in all parts of the inferior olive, and they were roughly evenly divided between those responsive to cutaneous stimulation and those responsive to proprioceptive stimulation. In the dorsal accessory olive all neurons were responsive to somatosensory stimulation. In the medial accessory nucleus 88% and in the principal olive 74% of cells were responsive to somatosensory stimulation. Cells responsive to cutaneous stimulation usually had small receptive fields, commonly on the paw. These cells had low-threshold responses to one or more forms of cutaneous stimulation and typically fired one spike at the onset of the stimulus on 80% or more of stimulus applications. Cells responsive to proprioceptive stimulation most commonly responded to passive displacements of a limb. These cells were often very sensitive, responding to linear displacements of less than 1 cm in one specific direction. No cells in our sample responded reliably during active movement by the animal. Only 21% of cells responding to passive proprioceptive stimulation showed any modulation during active movement, and the modulation was weak. Likewise, cells responsive to cutaneous stimulation generally failed to respond when a similar stimulus was produced by an active movement by the animal. Exceptions to this were stimuli produced during exploratory movements or when the receptive field unexpectedly made contact with an object during active movement. Electrical stimulation applied in the inferior olive failed to evoke movements or to modify ongoing movement. Our results are consistent with the hypothesis that inferior olivary neurons function as somatic event detectors responding particularly reliably to unexpected stimuli.


Correlation of geomagnetic activity with implantable cardioverter defibrillator shocks and antitachycardia pacing.
To investigate a potential relationship between implantable cardioverter defibrillator (ICD) therapies and daily geomagnetic activity (GMA) recorded in a large database. The ALTITUDE database, derived from the Boston Scientific LATITUDE remote monitoring system, was retrospectively analyzed for the frequency of ICD therapies. Daily GMA was expressed as the planetary K-index and the integrated A-index and was graded as levels I (quiet), II (unsettled), III (active), and IV (storm). A daily mean ± SD of 59,468±11,397 patients were monitored between January 1, 2009, and May 15, 2012. The distribution of days according to GMA was as follows: level I, 924/1231 (75%); level II, 226/1231 (18%); level III, 60/1231 (5%); and level IV, 21/1231 (2%). The daily mean ± SD numbers of ICD shocks received per 1000 active patients in the database were 1.29±0.47, 1.17±0.46, 1.03±0.37, and 0.94±0.29 on level I, II, III, and IV days, respectively; the daily mean ± SD sums of shocks and antitachycardia pacing therapies were 9.29±2.86, 8.46±2.45, 7.92±1.80, and 7.83±2.28 on quiet, unsettled, active, and storm days, respectively. A significant inverse relationship between GMA and frequency of ICD therapies was identified, with the most pronounced difference between level I and level IV days (P<.001 for shocks; P=.008 for shocks + antitachycardia pacing). In a large-scale cohort analysis, ICD therapies were delivered less frequently on days of higher GMA, confirming the previous pilot data and suggesting that higher GMA does not pose an increased risk of arrhythmias using ICD therapies as a surrogate marker. Further studies are needed to gain an in-depth understanding of the underlying mechanisms.


Shock waves in weakly compressed granular media.
We experimentally probe nonlinear wave propagation in weakly compressed granular media and observe a crossover from quasilinear sound waves at low impact to shock waves at high impact. We show that this crossover impact grows with the confining pressure P0, whereas the shock wave speed is independent of P0-two hallmarks of granular shocks predicted recently. The shocks exhibit surprising power law attenuation, which we model with a logarithmic law implying that shock dissipation is weak and qualitatively different from other granular dissipation mechanisms. We show that elastic and potential energy balance in the leading part of the shocks.


Developmental and oncogenic radiation effects on neural stem cells and their differentiating progeny in mouse cerebellum.
Neural stem cells are highly susceptible to radiogenic DNA damage, however, little is known about their mechanisms of DNA damage response (DDR) and the long-term consequences of genotoxic exposure. Patched1 heterozygous mice (Ptc1(+/-)) provide a powerful model of medulloblastoma (MB), a frequent pediatric tumor of the cerebellum. Irradiation of newborn Ptc1(+/-) mice dramatically increases the frequency and shortens the latency of MB. In this model, we investigated the mechanisms through which multipotent neural progenitors (NSCs) and fate-restricted progenitor cells (PCs) of the cerebellum respond to DNA damage induced by radiation, and the long-term developmental and oncogenic consequences. These responses were assessed in mice exposed to low (0.25 Gy) or high (3 Gy) radiation doses at embryonic day 13.5 (E13.5), when NSCs giving rise to the cerebellum are specified but the external granule layer (EGL) has not yet formed, or at E16.5, during the expansion of granule PCs to form the EGL. We found crucial differences in DDR and apoptosis between NSCs and fate-restricted PCs, including lack of p21 expression in NSCs. NSCs also appear to be resistant to oncogenesis from low-dose radiation exposure but more vulnerable at higher doses. In addition, the pathway to DNA repair and the pattern of oncogenic alterations were strongly dependent on age at exposure, highlighting a differentiation-stage specificity of DNA repair pathways in NSCs and PCs. These findings shed light on the mechanisms used by NSCs and PCs to maintain genome integrity during neurogenesis and may have important implications for radiation risk assessment and for development of targeted therapies against brain tumors.


Does unilateral transtibial amputation lead to greater metabolic demand during walking?
Previous literature reports greater metabolic demand of walking following transtibial amputation. However, most research focuses on relatively older, less active, and often dysvascular amputees. Servicemembers with traumatic amputation are typically young, fit, and highly active before and often following surgical amputation of their lower limb. This study compared the metabolic demand of walking in young, active individuals with traumatic unilateral transtibial amputation (TTA) and nondisabled controls. Heart rate (HR), rate of oxygen consumption, and rating of perceived exertion (RPE) were calculated as subjects walked at a self-selected velocity and at five standardized velocities based on leg length. The TTA group completed a Prosthetics Evaluation Questionnaire. Oxygen consumption (p = 0.89), net oxygen consumption (p = 0.32), and RPE (p = 0.14) did not differ between groups. Compared with controls, HR was greater in the TTA group and increased to a greater extent with velocity (p < 0.001). Overall, the TTA group rated their walking abilities as high (mean: 93% out of 100%). This is the first study to report equivalent metabolic demand between persons with amputation and controls walking at the same velocity. These results may reflect the physical fitness of the young servicemembers with traumatic amputations and may serve to guide outcome expectations in the future.


[Correlation between adipocytokines with iodine deficiency criteria in patients with metabolic syndrome and type 2 diabetes mellitus in Carpathians region].
69 patients with metabolic syndrome (MS) and type 2 diabetes mellitus (DM) have been examined. It is certain that activation of the cytokines system plays important part in formation of the insulin resistance syndrome. The results of our studies testify that insulin resistance in patients with MS and type 2 DM is associated by the decrease in the level of adiponectin and increase in the level of resistin. Lower level of adiponectin and higher level of resistin in blood serum can be examined as markers of the metabolic syndrome. Content of adiponectin in blood serum negatively correlates with an insulin resistance index HOMA IR, the level of TSH, proinflammatory cytokines-tumor necrosis factor alpha (TNFa) and cortisol. The author showed direct correlations between the TSH level and proinflammatory cytokines--TNFalpha, IL-6, CRP and negative correlations between HOMA IR indexes and iodinuria. It indicates on the connection of insulin resistance with the iodine deficiency in biosphere.


Suramin and the suramin analogue NF307 discriminate among calmodulin-binding sites.
Calmodulin-binding sites on target proteins show considerable variation in primary sequence; hence compounds that block the access of calmodulin to these binding sites may be more selective than compounds that inactivate calmodulin. Suramin and its analogue NF307 inhibit the interaction of calmodulin with the ryanodine receptor. We have investigated whether inhibition of calmodulin binding to target proteins is a general property of these compounds. Suramin inhibited binding of [(125)I]calmodulin to porcine brain membranes and to sarcoplasmic reticulum from skeletal muscle (IC(50)=4.9+/-1.2 microM and 19.9+/-1.8 microM, respectively) and blocked the cross-linking of [(125)I]calmodulin to some, but not all, target proteins in brain membranes by [(125)I]calmodulin. Four calmodulin-binding proteins were purified [ryanodine receptor-1 (RyR1) from rabbit skeletal muscle, neuronal NO synthase (nNOS) from Sf9 cells, G-protein betagamma dimers (Gbetagamma) from porcine brain and a glutathione S-transferase-fusion protein comprising the C-terminal calmodulin-binding domain of the metabotropic glutamate receptor 7A (GST-CmGluR7A) from bacterial lysates]. Three of the proteins employed (Gbetagamma, GST-CmGluR7A and RyR1) display a comparable affinity for calmodulin (in the range of 50-70 nM). Nevertheless, suramin and NF307 only blocked the binding of Gbetagamma and RyR1 to calmodulin-Sepharose. In contrast, the association of GST-CmGluR7A and nNOS was not impaired, whereas excess calmodulin uniformly displaced all proteins from the matrix. Thus suramin and NF307 are prototypes of a new class of calmodulin antagonists that do not interact directly with calmodulin but with calmodulin-recognition sites. In addition, these compounds discriminate among calmodulin-binding motifs.


How to survive a new CEO.
A new CEO means everyone in the organization is now a new hire--an unknown entity that must prove he or she can and will enthusiastically, tirelessly support the CEO's agenda. Those who want to stay on the payroll must avoid a business-as-usual attitude and demonstrate that they share the CEO's dedication to change. If you're in a staff job, it's important to show you're essential to the new regime, quickly and convincingly. Some important steps to consider as you plan for the transition include: Do your home-work; anticipate the CEO's needs; be supportive and nonpolitical; be realistic with your direct reports; and appear poised to leave willingly and quietly--and you may not have to.


Interocular suppression in strabismic amblyopia results in an attenuated and delayed hemodynamic response function in early visual cortex.
Factors such as strabismus or anisometropia during infancy can disrupt normal visual development and result in amblyopia, characterized by reduced visual function in an otherwise healthy eye and often associated with persistent suppression of inputs from the amblyopic eye by those from the dominant eye. It has become evident from fMRI studies that the cortical response to stimulation of the amblyopic eye is also affected. We were interested to compare the hemodynamic response function (HRF) of early visual cortex to amblyopic vs. dominant eye stimulation. In the first experiment, we found that stimulation of the amblyopic eye resulted in a signal that was both attenuated and delayed in its time to peak. We postulated that this delay may be due to suppressive effects of the dominant eye and, in our second experiment, measured the cortical response of amblyopic eye stimulation under two conditions--where the dominant eye was open and seeing a static pattern (high suppression) or where the dominant eye was patched and closed (low suppression). We found that the HRF in response to amblyopic eye stimulation depended on whether the dominant eye was open. This effect was manifested as both a delayed HRF under the suppressed condition and an amplitude reduction.


Sustained cardiac beta adrenoceptor blockade in vitro and increased vulnerability to aconitine-induced arrhythmias in vivo after propranolol withdrawal in rats.
The present study was undertaken in order to investigate the possibility of cardiac hypersensitivity to norepinephrine (NE) after propranolol withdrawal in rats. The effect of NE was studied on heart rate and left intraventricular pressure development (maximal dP/dt) in the isolated perfused heart at various time periods after termination of the propranolol feeding. Also, the influence of propranolol withdrawal on vulnerability to aconitine-induced arrhythmias in vivo was evaluated in anesthetized rats. No hypersensitivity to NE was seen in the perfused rat heart 1 and 3 days after propranolol withdrawal. Rather, a depressed response to NE was registered suggesting a long half-life for the disappearance of beta adrenoceptor blockade. Increased vulnerability to aconitine-induced arrhythmias was observed 1 and 3 days after treatment with propranolol had ceased. Rats receiving propranolol continuously until experimentation exhibited an elevated threshold to aconitine-induced arrhythmias. These data indicate that a hypothetical rebound phenomenon after withdrawal of propranolol is not associated with an increase in mechanical performance of the heart in response to NE. A decreased threshold for development of arrhythmias appears, however, to be at hand, perhaps due to an inbalance in the transmembrane sodium flux in the heart in the early time course after propranolol withdrawal.


Mercury-Based Cuprate High-Transition Temperature Grain-Boundary Junctions and SQUIDs Operating Above 110 Kelvin.
The superconducting transport characteristics of HgBa(2) CaCu(2)O(6+delta) (Hg-1212) films and grain-boundary junctions grown on (100)-oriented SrTiO(3) bicrystal substrates have been investigated. The films exhibit a zero-resistance temperature of approximately 120 kelvin and sustain large critical current densities, with values as high as 10(6) amperes per square centimeter at around 100 kelvin. On the other hand, the grain boundaries behave as weak links, with substantially lower critical currents, as is observed for other cuprate superconductors. A reduction of three orders of magnitude in critical current was observed for transport across a 36.8 degrees grain boundary. The current-voltage characteristics of bridges across such a grain boundary show weak-link behavior qualitatively resembling that of a resistively shunted junction. Single-level direct-current superconducting quantum interference devices (SQUIDs) have been fabricated with such bicrystal junctions. These SQUIDs show clear periodic voltage modulations when subjected to applied magnetic fields. The SQUIDs operate at temperatures as high as 111.8 kelvin, which makes them attractive for operation in portable sensors and devices that utilize nonconventional cooling methods.


[Adherence of patients with stable angina to treatment with trimetazidine MR and frequency of emergency medical care: results of the EFFECT study].
The adherence of patients with stable angina to antianginal therapy is the key factor of controlling the disease. The purpose of the study was to evaluate the relationship of adherence of patients with stable angina to treatment with trimetazidine modified release (MR) with frequency (risk) of emergency medical care. We consistently included in the study patients with stable angina in primary health care. The results of treatment for 16 weeks were monitored at patients with angina attacks three times per week or more, use of short nitrate and treatment with generic trimetazidine. To strengthen the antianginal therapy generic was replaced with original trimetazidine MR. Adherence is considered relatively high while taking 80-120% of the recommended dose of the drug (70 mg/day). The effectiveness of treatment evaluated by the frequency of emergency hospitalizations and/or ambulance calls because of the pain, discomfort, tightness in the chest or ischemic changes on the electrocardiogram. 870 patients were included in the study, the results of treatment in 185 were assessed. Patients with a relatively high adherence to trimetazidine MR (n=151) were used (median) 99% (98, 104), with low (<80%, n=34) adherence - 67% (49, 76) of the recommended dose of the drug. During the study period, the primary end point is fixed in 7 (21%) patients with low and in 18 (12%) - with relatively high adherence (p=0.182). The number of angina attacks, having necessitated taking short-nitrate, decreased in the groups, respectively, with 5 (3; 10) and 6 (4; 10) to 2 (1; 3) per week (p=0.791). Thus, replacing generic trimetazidine with original trimetazidine MR in patients with a high frequency of angina attacks can achieve significant antianginal effect. Adherence of patients to the reception of the drug by an average 1/3 below the recommended amount does not affect the risk of emergency hospitalizations and/or ambulance calls for 16 weeks.


NetPyNE, a tool for data-driven multiscale modeling of brain circuits.
Biophysical modeling of neuronal networks helps to integrate and interpret rapidly growing and disparate experimental datasets at multiple scales. The NetPyNE tool (www.netpyne.org) provides both programmatic and graphical interfaces to develop data-driven multiscale network models in NEURON. NetPyNE clearly separates model parameters from implementation code. Users provide specifications at a high level via a standardized declarative language, for example connectivity rules, to create millions of cell-to-cell connections. NetPyNE then enables users to generate the NEURON network, run efficiently parallelized simulations, optimize and explore network parameters through automated batch runs, and use built-in functions for visualization and analysis - connectivity matrices, voltage traces, spike raster plots, local field potentials, and information theoretic measures. NetPyNE also facilitates model sharing by exporting and importing standardized formats (NeuroML and SONATA). NetPyNE is already being used to teach computational neuroscience students and by modelers to investigate brain regions and phenomena.


[Arterial chemoembolization for metastatic colorectal cancer in the liver].
Transcatheter hepatic artery chemoembolization (THACE) for inoperable colorectal carcinoma metastatic to the liver has been evaluated. Cytostatics were given to 67 patients (Dioxadet 10-30 mg--23; Doxorubicin 40-100 mg--44) in 183 courses using oil and gelatin sponge (1986-1999). Response to treatment (partial decrease in tumor size or growth stabilization) for Dioxadet and Doxorubicin was 47 and 66%, respectively; major complications--17% (fatalities--9%) and 11%, respectively. Mean survival rate was 10 and 11 months; median survival--9 and 10 months, respectively. Repeated THACE-assisted treatment with Doxorubicin was followed by mean survival of 14 months, median--12; one-year survival increased by 77%. THACE-mediated treatment with Dioxadet proved less effective and involved more complications. Further research in modifications of endovascular intervention is needed, including their combinations with systemic treatment of metastatic cancer of the liver.


Enhanced intrinsic fluorescence from carboxidized nano-sculptured thin films of silver and their application for label free dual detection of glycated hemoglobin.
Enhanced intrinsic fluorescence (~x103) from novel carboxidized nanosculptured thin films (CO-nSTFs) of silver is reported. The sources of intrinsic fluorescence, confirmed by X-ray photoelectron spectroscopy, are Ag2O grains and residual carbon formed on the outer layer of silver nSTFs when exposed to air, while the localized surface plasmons on silver nSTFs enhance this intrinsic fluorescence. The CO-nSTFs are optimized with respect to porosity for the maximum enhancement. A sensor developed by using the self-assembled monolayer technique on optimized CO-nSTF is used for the label free detection of glycated hemoglobin, performed by simultaneously using fluorescence imaging and spectroscopy. The specificity of the sensor is established from control experiments on hemoglobin. These novel nanorod like intrinsically fluorescent CO-nSTFs pose huge potential in label free biosensing, light sources, imaging and many more applications.


What are the Benefits and Harms of Ureteroscopy Compared with Shock-wave Lithotripsy in the Treatment of Upper Ureteral Stones? A Systematic Review.
Extracorporeal shock wave lithotripsy (SWL) and ureteroscopy (URS), with or without intracorporeal lithotripsy, are the most common treatments for upper ureteric stones. With advances in technology, it is unclear which treatment is most effective and/or safest. To systematically review literature reporting benefits and harms of SWL and URS in the management of upper ureteric stones. Databases including Medline, Embase, and the Cochrane library were searched from January 2000 to November 2014. All randomised controlled trials (RCTs), quasi-randomised controlled trials, and nonrandomised studies comparing any subtype or variation of URS and SWL were included. The primary benefit outcome was stone-free rate (SFR). The primary harm outcome was complications. Secondary outcomes included retreatment rate, need for secondary, and/or adjunctive procedures. The Cochrane risk of bias tool was used to assess RCTs, and an extended version was used to assess nonrandomised studies. Grading of Recommendations Assessment, Development, and Evaluation was used to assess the quality of evidence. Five thousand-three hundred and eighty abstracts and 387 full-text articles were screened. Forty-seven studies met inclusion criteria; 19 (39.6%) were RCTs. No studies on children met inclusion criteria. URS and SWL were compared in 22 studies (4 RCTs, 1 quasi-randomised controlled trial, and 17 nonrandomised studies). Meta-analyses were inappropriate due to data heterogeneity. SFR favoured URS in 9/22 studies. Retreatment rates were higher for SWL compared with URS in all studies but one. Longer hospital stay and adjunctive procedures (most commonly the insertion of a JJ stent) were more common when primary treatment was URS. Complications were reported in 11 out of 22 studies. In eight studies, it was possible to report this as a Clavien-Dindo Grade. Higher complication rates across all grades were reported for URS compared with SWL. For intragroup (intra-SWL and intra-URS) comparative studies, 25 met the inclusion criteria. These studies varied greatly in outcomes measured with data being heterogeneous. Compared with SWL, URS was associated with a significantly greater SFR up to 4 wk but the difference was not significant at 3 mo in the included studies. URS was associated with fewer retreatments and need for secondary procedures, but with a higher need for adjunctive procedures, greater complication rates, and longer hospital stay. In this paper, the relative benefits and harms of the two most commonly offered treatment options for urinary stones located in the upper ureter were reviewed. We found that both treatments are safe and effective options that should be offered based on individual patient circumstances and preferences.


Behavioural activation for family dementia caregivers: A systematic review and meta-analysis.
The current study aims to investigate the effectiveness of behavioral activation (BA) for family dementia caregivers. A systematic literature search was conducted through PubMed, Medline, CINAHL, Cochrane, Embase and PsycINFO for studies published from March 1988 to September 2019. Standardized mean differences (SMDs) were combined to synthesize pooled effect measures using random effects. The review was based on ten randomized controlled trials. Depression was significantly reduced after participants received BA (n = 9; 786 participants; SMD = -0.69; 95% CI: -1.12 to -0.25; p = 0.002). BA also has a positive impact on other areas of psychological health (e.g., self-efficacy, anxiety, and distress) and physical health (e.g., interleukin-6) in family dementia caregivers. In conclusion, BA not only has a moderate effect on reducing depression but also has the potential to improve other areas of psychological and cardiovascular health in family caregivers. However, more interventional studies of BA for family dementia caregivers are needed.


Measurement of O6-methyldeoxyguanosine in DNA methylated by the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone using a biotin-avidin enzyme-linked immunosorbent assay.
Rabbit antibodies raised against O6-methyl-2'-deoxyguanosine (O6-MedGuo) were used to develop a competitive Biotin-Avidin enzyme-linked immunosorbent assay (BA-ELISA) for the quantitation of this adduct in DNA from animals exposed to tobacco-specific N-nitrosamines. The assay was able to detect as little as 30 mumol O6-MedGuo per mol deoxyguanosine. The specificity of the assay was at least 10 000-fold greater for O6-MedGuo than for unmodified nucleosides or 7-methyl-2'-deoxyguanosine. The sensitivity of the assay was increased to 1 mumol O6-MedGuo per mol deoxyguanosine by the prior separation of O6-MedGuo from unmodified nucleosides by h.p.l.c. This assay was used to study the methylating ability of the tobacco-specific nitrosamine 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in F344 rats. Four hours after exposure to a single i.v. dose of NNK O6-MedGuo was detected in the target tissues nasal mucosa, lung and liver (219, 13.2 and 34.5 mumol/mol deoxyguanosine) but not in the non-target tissues esophagus, spleen, heart or kidney. Chronic exposure by 14 daily i.p. injections of NNK also resulted in detectable amounts of O6-MedGuo in the lung and nasal mucosa (11.4 and 7.9 mumol/mol deoxyguanosine). These results demonstrate that NNK is capable of methylating DNA in vivo. The methodology developed in these studies will allow us to examine in detail the role of DNA methylation in NNK-mediated carcinogenesis.


Low peripheral blood chemokine (C-C motif) ligand 5 and tumor necrosis factor α gene expression is associated with unfavorable progression of respiratory syncytial virus bronchiolitis in infants.
We aimed to analyze whether the expression of inflammatory and antiviral genes in respiratory syncytial virus (RSV)-infected infants' peripheral blood is associated with bronchiolitis progression. We conducted a prospective study on 117 infants between 2015 and 2023. The expression levels of nine genes were quantified by quantitative polymerase chain reaction. Infants were classified according to their clinical evolution during hospital admission: (i) non-progression (n = 74), when the RSV bronchiolitis severity remained stable or improved; (ii) unfavorable progression (n = 43), when the RSV bronchiolitis severity increased. The association analysis was performed by logistic regression, adjusted by age, gender, prematurity, and RSV bronchiolitis severity in the emergency room. Infants were 57.3% male, and the median age of the study population was 61 days. Thirty-five infants (30.7%) were admitted to the intensive care unit after hospital admission. Univariate logistic models showed that tumor necrosis factor (TNFα) and chemokine (C-C motif) ligand (CCL5) gene expression at baseline were inversely associated with unfavorable progression, which was confirmed by multivariate analyses: TNFα (adjusted odds ratio = 0.8 [95% confidence interval = 0.64-0.99], P-value = 0.038) and CCL5 (adjusted odds ratio = 0.76 [95% confidence interval = 0.62-0.93], P-value = 0.007). An inadequate immune response to RSV, characterized by reduced gene expression levels of CCL5 and TNFα in peripheral blood, was associated with an unfavorable progression of RSV bronchiolitis.


[Steps to improve the quality of urinary incontinence care].
Urinary incontinence is a frequent symptom rarely spontaneously reported. This emphasizes the need to address it in every patient. Once disclosed, its origin needs to be specified so that therapeutic issues may be reached. These being often efficient, patient quality of life will improve significantly. In order to improve urinary incontinence management, a multidisciplinary group of healthcare professionals of our institution have proposed to spread basic knowledge, screening tools and evaluation scales of this bothersome worldwide symptom. This team enables healthcare providers to unite themselves in a network which will help to orientate patients in order to fulfill their needs.


Surgical management of symptomatic T8 vertebral hemangioma: case report and review of the literature.
Vertebral hemangiomas are benign vascular lesions of the vertebral column; only 0.9-1.2% of all vertebral hemangiomas cause spinal cord compression. We report a 34-year-old female who was admitted to the neurosurgery clinic with a history of back pain, poor quality of life and easy fatigability for 1.5 years. Her medical history revealed a fall from a height of 2 meters 1.5 years ago. Neurology examination revealed bilateral hypoesthesia below the T8 level and hyperactive deep tendon reflexes in her left leg. Computed tomography scan of the thoracic spine showed T8 vertebral hemangioma, and magnetic resonance imaging showed a T8 hemangioma compressing the spinal cord. Surgical intervention was planned and T8 total laminectomy was performed. The tumor extending into the anterior spinal cord was resected, and T8 vertebroplasty with short segment posterior stabilization and fusion was performed. We aimed to present a new treatment approach for symptomatic vertebral hemangiomas and reviewed the relevant literature.


Epidermolysis bullosa pruriginosa masquerading as psychogenic pruritus.
Epidermolysis bullosa pruriginosa is a rare clinical subtype of dystrophic epidermolysis bullosa characterized by intense pruritus, secondary scratching-induced lesions, and pronounced scarring. We describe a patient with epidermolysis bullosa pruriginosa who was misdiagnosed as having psychogenic pruritus for several years. Except for nail (toenail) dystrophy, no features of the disease were evident among his immediate family members. An underlying new heterozygous donor splice-site mutation in the type VII collagen gene (IVS55 + 1G>C) was found in both the patient and his family members with nail dystrophy. Inheritance was autosomal dominant. The patient was treated with cyclosporine and experienced significant reduction in pruritus, with subsequent improvement of the skin condition. Pruritus is an important factor in the development of epidermolysis bullosa pruriginosa and is the focus of management. Patients with this inherited skin disorder can be easily misdiagnosed as having psychogenic pruritus, and this article aims to make physicians aware of this diagnostic pitfall.


The relevance of coagulation factor X protection of adenoviruses in human sera.
Intravenous delivery of adenoviruses is the optimal route for many gene therapy applications. Once in the blood, coagulation factor X (FX) binds to the adenovirus capsid and protects the virion from natural antibody and classical complement-mediated neutralisation in mice. However, to date, no studies have examined the relevance of this FX/viral immune protective mechanism in human samples. In this study, we assessed the effects of blocking FX on adenovirus type 5 (Ad5) activity in the presence of human serum. FX prevented human IgM binding directly to the virus. In individual human sera samples (n=25), approximately half of those screened inhibited adenovirus transduction only when the Ad5-FX interaction was blocked, demonstrating that FX protected the virus from neutralising components in a large proportion of human sera. In contrast, the remainder of sera tested had no inhibitory effects on Ad5 transduction and FX armament was not required for effective gene transfer. In human sera in which FX had a protective role, Ad5 induced lower levels of complement activation in the presence of FX. We therefore demonstrate for the first time the importance of Ad-FX protection in human samples and highlight subject variability and species-specific differences as key considerations for adenoviral gene therapy.


Association Between Respiratory Alkalosis and the Prognosis of COVID-19 Patients.
Aim: The aim of the study was to describe the clinical characteristics of patients with or without respiratory alkalosis, and analyze the relationship of respiratory alkalosis and the outcome of adult coronavirus disease 2019 (COVID-19) patients. Methods: Clinical and laboratory data of adult COVID-19 patients in a single center in China, were retrospectively collected and analyzed. The Kaplan-Meier (KM) curve and cox regression were adopted to analyze the association between respiratory alkalosis and prognosis of COVID-19 patients. Results: Of 230 adult COVID-19 patients, 66 patients (28.7%) had respiratory alkalosis on admission. Of 66 patients, the median age was 53 years old (range, 21-84 years), and 43 (65.2%) were female. Compared with those without respiratory alkalosis, patients with respiratory alkalosis were significantly older (P = 0.002), had a higher proportion of female (P = 0.004), and showed higher ratios of underlying diseases including hypertension (P = 0.023) and cardiovascular disease (P = 0.028). Moreover, they demonstrated higher proportion of severe events (P = 0.001). Patients with respiratory alkalosis had a higher possibility of developing severe events compared with those without respiratory alkalosis (Log Rank P = 0.001). After adjusting for gender, age, and comorbidities, patients with respiratory alkalosis still showed significantly elevated risks of developing to severe cases (HR 2.445, 95% CI 1.307-4.571, P = 0.005) using cox regression analyses. Conclusions: Respiratory alkalosis as a common acid-base disorder in COVID-19 patients, was associated with a higher risk of developing severe event.


Single-pulse electrical stimulation artifact removal using the novel matching pursuit-based artifact reconstruction and removal method (MPARRM).
Single-pulse electrical stimulation (SPES) has been widely used to probe effective connectivity. However, analysis of the neural response is often confounded by stimulation artifacts. We developed a novel matching pursuit-based artifact reconstruction and removal method (MPARRM) capable of removing artifacts from stimulation-artifact-affected electrophysiological signals.

Approach: To validate MPARRM across a wide range of potential stimulation artifact types, we performed a bench-top experiment in which we suspended electrodes in a saline solution to generate 110 types of real-world stimulation artifacts. We then added the generated stimulation artifacts to ground truth signals (stereoelectroencephalography signals from 9 human subjects recorded during a receptive speech task), applied MPARRM to the combined signal, and compared the resultant denoised signal with the ground truth signal. We further applied MPARRM to artifact-affected neural signals recorded from the hippocampus while performing SPES on the ipsilateral basolateral amygdala in 9 human subjects.

Results: MPARRM could remove stimulation artifacts without introducing spectral leakage or temporal spread. It accommodated variable stimulation parameters and recovered the early response to SPES within a wide range of frequency bands. Specifically, in the early response period (5 to 10 ms following stimulation onset), we found that the broadband gamma power (70-170 Hz) of the denoised signal was highly correlated with the ground truth signal (R=0.98±0.02, Pearson), and the broadband gamma activity of the denoised signal faithfully revealed the responses to the auditory stimuli within the ground truth signal with 94±1.47% sensitivity and 99±1.01% specificity. We further found that MPARRM could reveal the expected temporal progression of broadband gamma activity along the anterior-posterior axis of the hippocampus in response to the ipsilateral amygdala stimulation.

Significance: MPARRM could faithfully remove SPES artifacts without confounding the electrophysiological signal components, especially during the early-response period. This method can facilitate the understanding of the neural response mechanisms of SPES.&#xD.


Non-Pharmacological Approaches to Headaches: Non-Invasive Neuromodulation, Nutraceuticals, and Behavioral Approaches.
Significant side effects or drug interactions can make pharmacological management of headache disorders very difficult. Non-conventional and non-pharmacological treatments are becoming increasingly used to overcome these issues. In particular, non-invasive neuromodulation, nutraceuticals, and behavioral approaches are well tolerated and indicated for specific patient categories such as adolescents and pregnant women. This paper aims to present the main approaches reported in the literature in the management of headache disorders. We therefore reviewed the available literature published between 2010 and 2020 and performed a narrative presentation for each of the three categories (non-invasive neuromodulation, nutraceuticals, and behavioral therapies). Regarding non-invasive neuromodulation, we selected transcranial magnetic stimulation, supraorbital nerve stimulation, transcranial direct current stimulation, non-invasive vagal nerve stimulation, and caloric vestibular stimulation. For nutraceuticals, we selected Feverfew, Butterbur, Riboflavin, Magnesium, and Coenzyme Q10. Finally, for behavioral approaches, we selected biofeedback, cognitive behavioral therapy, relaxation techniques, mindfulness-based therapy, and acceptance and commitment therapy. These approaches are increasingly seen as a valid treatment option in headache management, especially for patients with medication overuse or contraindications to drug treatment. However, further investigations are needed to consider the effectiveness of these approaches also with respect to the long-term effects.


Mesmerize is a dynamically adaptable user-friendly analysis platform for 2D and 3D calcium imaging data.
Calcium imaging is an increasingly valuable technique for understanding neural circuits, neuroethology, and cellular mechanisms. The analysis of calcium imaging data presents challenges in image processing, data organization, analysis, and accessibility. Tools have been created to address these problems independently, however a comprehensive user-friendly package does not exist. Here we present Mesmerize, an efficient, expandable and user-friendly analysis platform, which uses a Findable, Accessible, Interoperable and Reproducible (FAIR) system to encapsulate the entire analysis process, from raw data to interactive visualizations for publication. Mesmerize provides a user-friendly graphical interface to state-of-the-art analysis methods for signal extraction & downstream analysis. We demonstrate the broad scientific scope of Mesmerize's applications by analyzing neuronal datasets from mouse and a volumetric zebrafish dataset. We also applied contemporary time-series analysis techniques to analyze a novel dataset comprising neuronal, epidermal, and migratory mesenchymal cells of the protochordate Ciona intestinalis.


Assessment of utilization of provider-initiated HIV testing and counseling as an intervention for prevention of mother to child transmission of HIV and associated factors among pregnant women in Gondar town, North West Ethiopia.
Detection of maternal HIV infection early in pregnancy is critical for prevention of mother to child transmission of HIV/AIDS. Most efforts have focused on VCT as the primary means of encouraging people to become aware of their HIV status. However, its uptake is low in many parts of sub-Saharan Africa including Ethiopia. Provider-initiated HIV testing and counseling provides a critical opportunity to diagnose HIV infection, to begin chronic care, and to prevent mother to child transmission. However, little is known about its acceptance and associated factors among pregnant women in the country and particularly in the present study area. Health institution based cross-sectional quantitative study was conducted in Gondar town from July 22-August 18, 2010. A total of 400 pregnant women were involved in the study using stratified sampling technique and multiple logistic regression analysis was employed using SPSS version 16. A total of 400 pregnant women actively participated in this study and 330 (82.5%) of them accepted provider-initiated HIV testing and counseling to be tested for HIV and 70(17.5%) of them refused. Acceptance of provider-initiated HIV testing and counseling was positively associated with greater number of antenatal care visits [Adj. OR (95%CI) = 2.64(1.17, 5.95)], residing in the urban areas[Adj. OR (95%CI) = 2.85(1.10, 7.41)], having comprehensive knowledge on HIV [Adj. OR (95%CI) = 4.30(1.72, 10.73)], positive partners reaction for HIV positive result [Adj. OR (95%CI) = 8.19(3.57, 18.80)] and having knowledge on prevention of mother to child transmission of HIV[Adj. OR (95%CI) = 3.27(1.34, 7.94)], but negatively associated with increased maternal age and education level. Utilization of provider-initiated HIV testing and counseling during antenatal care was relatively high among pregnant women in Gondar town. Couple counseling and HIV testing should be strengthened to promote provider-initiated HIV testing and counseling among male partners and to reduce HIV related violence of women from their partner and access to and consistent use of antenatal care should be improved to increase the uptake of provider-initiated HIV testing and counseling service.


Efficacy of constraint-induced movement therapy compared with bimanual intensive training in children with unilateral cerebral palsy: a systematic review.
To systematically review the evidence on the effect of constraint-induced movement therapy compared with bimanual intensive training in children with unilateral cerebral palsy. Seven electronic databases (Cinahl, Cochrane Library, EMBASE, Ovid MEDLINE, PEDro, PsycINFO, PubMed) were searched from database inception through December 2016. A systematic review was performed using the American Academy of Cerebral Palsy and Developmental Medicine and Preferred Reporting Items for Systematic Review and Meta-Analysis guidelines. Standardised mean differences (effect sizes) were calculated for each study and outcome. Nine studies met the eligibility criteria. All studies provided level II evidence. Methodological quality was high in two studies, moderate in four studies and low in three studies. The methodology, participant and intervention characteristics were heterogeneous. The participants' ages ranged from 1.5 to 16 years. Their initial hand function ranged from Manual Ability Classification System Level I to Level III. The total intervention dose ranged from 24 to 210 hours and duration from one week to ten weeks. The studies measured outcomes assessing unimanual and bimanual hand and arm function, participation and attainment of individualised goals. Overall, the effect sizes did not favour one of the interventions at short- or long-term follow-up. The 95% confidence intervals were broad, indicating inaccurate precision of the effect sizes. Pooling of the data for a meta-analysis was judged to be of little clinical value owing to heterogeneity. It is not possible to conclude whether constraint-induced movement therapy or bimanual intensive training is more effective than the other in children with unilateral cerebral palsy.


Middle ear fluid lysozyme source in experimental pneumococcal otitis media.
Middle ear infection with Streptococcus pneumoniae is important in the pathogenesis of acute and chronic otitis media, and lysozyme in middle ear fluid (MEF) is an important inflammatory mediator in this disease. To determine the source of lysozyme during the early period of acute pneumococcal otitis media, chinchillas were irradiated to induce neutropenia, and their middle ears were inoculated with heat-killed, encapsulated pneumococci. The number of inflammatory cells and concentration of lysozyme were measured in MEF between 6 and 72 hours after inoculation. In pneumococcus-inoculated ears, the mean number of inflammatory cells but not lysozyme was significantly lower in MEF from irradiated animals than that from nonirradiated animals at 6 hours. Since lysozyme accumulated in MEF before the influx of inflammatory cells in irradiated animals, the initial release of this inflammatory mediator is most likely not from inflammatory cells; and mucosal epithelial cells, the only other known source of lysozyme in the middle ear environment, were the probable source induced by the direct stimulation of pneumococci. Inflammatory cells may contribute lysozyme later in the inflammatory response, since cellular and lysozyme concentrations in irradiated and nonirradiated animals were similar between 24 and 72 hours. These results suggest that future therapeutic interventions to limit middle ear inflammation in acute otitis media may need to recognize the direct action of pneumococcal cells or their envelope components on middle ear epithelium.


Analysis in Cos-1 cells of processing and polyadenylation signals by using derivatives of the herpes simplex virus type 1 thymidine kinase gene.
Bal31 nuclease was used to resect the herpes simplex virus type 1 thymidine kinase (tk) gene from its 3' end, and a plasmid, pTK206, was isolated that lacked the processing and polyadenylation signals normally found at the 3' end of the gene. The wild-type gene, pTK2, and pTK206 were each transferred to pSV010, a plasmid containing the simian virus 40 (SV40) origin of DNA replication, allowing replication and analysis of the patterns of transcription in Cos-1 cells. Fragments of DNA containing processing and polyadenylation signals from SV40 and polyoma virus were inserted into the 3' end of the resected tk gene, pTK206. We found that tk gene expression requires a processing and polyadenylation signal, that signals from SV40 and polyoma virus could substitute for the herpes simplex virus tk signal, and that considerable differences in the levels of tk mRNA were present in Cos-1 cells transfected by these gene constructs. In addition, tk gene expression was restored to a low level after the insertion of an 88-base-pair fragment from the middle of the SV40 early region. Processing and polyadenylation do not occur in the vicinity of this fragment in SV40, even though it contains the hexanucleotide 5'-AAUAAA-3'.


Resistance of Leishmania (Viannia) Panamensis to Meglumine Antimoniate or Miltefosine Modulates Neutrophil Effector Functions.
Leishmania (Viannia) panamensis (L. (V.) p.) is the main causative agent of cutaneous leishmaniasis in Colombia and is usually treated with either meglumine antimoniate (MA) or miltefosine (MIL). In recent years, there has been increasing evidence of the emergence of drug-resistance against these compounds. Neutrophils are known to play an important role in immunity against Leishmania. These cells are rapidly recruited upon infection and are also present in chronic lesions. However, their involvement in the outcome of infection with drug-resistant Leishmania has not been examined. In this study, human and murine neutrophils were infected in vitro with MA or MIL drug-resistant L. (V.) p. lines derived from a parental L. (V.) p. drug-susceptible strain. Neutrophil effector functions were assessed analyzing the production of reactive oxygen species (ROS), the formation of neutrophil extracellular trap (NET) and the expression of cell surface activation markers. Parasite killing by neutrophils was assessed using L. (V.) p. transfected with a luciferase reporter. We show here that MA and MIL-resistant L. (V.) p. lines elicited significantly increased NET formation and MA-resistant L. (V.) p. induced significantly increased ROS production in both murine and human neutrophils, compared to infections with the parental MIL and MA susceptible strain. Furthermore, neutrophils exposed to drug-resistant lines showed increased activation, as revealed by decreased expression of CD62L and increased expression of CD66b in human neutrophils yet presented higher survival within neutrophils than the drug-susceptible strain. These results provide evidence that parasite drug-susceptibility may influences neutrophil activation and function as well as parasite survival within neutrophils. Further investigaton of the inter-relationship of drug susceptibility and neutrophil effector function should contribute to better understanding of the factors involved in susceptibility to anti-Leishmania drugs.


Socioecological aspects of human reproduction in Sub-Saharan Africa.
The purpose of this study was to compare fertility outcome of two populations of northern Namibia, following different ways of subsistence. The total number of offspring, but also the number of dead and surviving offspring was compared between 236 !Kung San (91 females, 145 males) hunter gatherers and 248 Kavango (87 females, 161 males) horticultural pastoralists and a small number of Kavango people living in the urban center of Rundu. While no typical differences in fertility outcome between the study populations could be observed in males, marked differences were found for the female sample. As to be expected traditional Kavango women had given birth to a higher number of children and these children had a higher chance to survive in comparison to those of !Kung San women. On the other hand Kavango females living in urban centers reported a significantly lower number of offspring. It can be concluded that even in recent populations fertility differences according to subsistence patterns are observable.


A Case of Post-radiotherapy Urethral Stricture with Spontaneous Bladder Rupture, Mimicking Obstructive Uropathy due to Cancer Metastasis.
Non-traumatic, spontaneous urinary bladder rupture is a rare complication of urethral stricture. Furthermore, its symptoms are often nonspecific, and misdiagnosis is common. The authors experienced a case of urethral stricture with spontaneous bladder rupture and bilateral hydronephrosis, mimicking obstructive uropathy attributed to cancer metastasis. A 55-year-old woman was admitted with abdominal pain and distension, oliguria, and an elevated serum creatinine level. She had undergone radical hysterectomy for uterine cervical cancer and received post-operative concurrent chemoradiation therapy 13 years previously. Non-contrast enhanced computed tomography showed massive ascites and bilateral hydronephrosis. The initial diagnosis was acute kidney injury due to obstructive uropathy caused by malignant disease. After improvement of her renal function by bilateral percutaneous nephrostomy catheterization, contrast-enhanced computed tomography and a cytologic examination of ascites showed no evidence of malignancy. However, during retrograde pyelography, a severe urethral stricture was found, and subsequent cystography showed leakage of contrast into the peritoneal cavity and cystoscopy revealed a defect of the posterior bladder wall. After urethral dilatation and primary closure of the bladder wall, acute kidney injury and ascites were resolved.


Triadic interactions in families with preterm children: a comparative study with children born at term.
The relationship between parents and infants born preterm is multifaceted and could present some relational patterns which are believed to predict psychological risk more than others. For example, insensitive parenting behavior has been shown to place very preterm children at greater risk of emotional and behavioral dysregulation. The main objective of this study was to compare the quality of family interactions in a sample of families with preterm children with one of the families with at-term children, exploring possible differences and similarities. The second aim of this research was to consider the associations among family interactions and parental empowerment, the child's temperament, parenting stress, and perceived social support. The sample consisted of 52 children and their families: 25 families, one with two preterm brothers with preterm children (mean 22.3 months, SD 12.17), and 26 families with children born at term (mean 22.2 months, SD 14.97). The Lausanne Trilogue Play procedure was administered to the two groups to assess the quality of their family interactions. The preterm group was also administered the Questionari Italiani del Temperamento, the Family Empowerment Scale, the Multidimensional Scale of Perceived Social Support, and the Parenting Stress Index - Short Form. Differences in the quality of family interactions emerged between the preterm and at-term groups. The preterm group showed significantly lower quality of family interactions than the at-term group. The parenting stress of both parents related to their parental empowerment, and maternal stress was also related to the partner's parental empowerment. Social support had a positive influence on parenting stress, with maternal stress also related to perceived social support from the partner, which underscores the protective role of the father on the dyad.


Writing a group practice business plan.
A business plan offers group practices a blueprint to accomplish a variety of goals, such as securing capital, marketing the practice's services, recruiting new employees, developing a strategic plan or a budget, or planning for growth. A business plan should be informative, specific, and visionary. Elements that every business plan should address are a mission statement, strategy, planning, management information, and action scheme. A business plan should include certain information in a prescribed order. By writing a realistic business plan, group practices can work more efficiently and minimize the risk of not meeting their financial projections.


From Melancholia to Depression: Disordered Mood in Nineteenth-Century Psychiatry.
My book From Melancholia to Depression: Disordered Mood in Nineteenth-Century Psychiatry charts how melancholia was reconceptualized in the nineteenth century as a modern mood disorder and a precursor to clinical depression. The book shows how this occurred chiefly in two ways. First, the idea of disordered mood as a medical concept was created through the appropriation of language from experimental physiology into the realm of psychopathology. Second, the interplay of statistical and diagnostic practices formed the basis for modern psychiatric classification and facilitated the standardization of melancholia as a psychiatric diagnosis. These developments were key to the reconceptualization of melancholia and the subsequent emergence of clinical depression, and were foundational to modern psychiatric theory and practice.


Survey of Professionals of the European Public Health Association (EUPHA) towards Direct-to-Consumer Genetic Testing.
The increasing availability of Direct-to-Consumer Genetic Tests (DTC-GTs) has great implications for public health (PH) and requires literate healthcare professionals to address the challenges they pose. We designed and conducted a survey to assess the state of knowledge, attitudes and behaviours of PH professionals members of the European Public Health Association (EUPHA) towards DTC-GTs. EUPHA members were invited to participate and fill in the survey. We performed multivariable logistic regression to evaluate associations between selected covariates and knowledge, attitudes and behaviours of healthcare professionals towards DTC-GT. Three hundred and two professionals completed the survey, 66.9% of whom were not involved in genetics or genomics within their professional activities. Although 74.5% of respondents were aware that DTC-GTs could be purchased on the web, most of them reported a low level of awareness towards DTC-GTs applications and regulatory aspects. The majority did not approve the provision of DTC-GTs without consultation of a healthcare professional (91.4%), were doubtful about the test utility and validity (61%) and did not feel prepared to address citizens' questions (65.6%). Predictors of knowledge on DTC-GT were the involvement in genetics/genomics and receiving training during the studies (P < 0.0001 and P = 0.043). Predictors of attitudes were medical degree and knowledge about DTC-GTs (P = 0.006 and P = 0.027). Our results revealed a high level of awareness of DTC-GT web purchasing and a moderate to low level of awareness towards their applications. Despite the overall positive attitudes, PH professionals reported a high need for strengthening regulatory aspects of DTC-GTs provision process.


Angiogenesis, vasculogenesis, and vasculogenic mimicry in ovarian cancer.
Neovascularization is essential for tumor growth and metastasis. An adequate vasculature feeds tumor growth and enhances the potential of metastasis. For many years, tumor vessels were thought to be lined exclusively by endothelial cells (ECs). However, therapeutic benefits from the promising antiangiogenic strategy targeting genetically stable ECs are frequently limited by the development of resistance, implying an oversimplified view of tumor vasculature. In fact, latest studies have revealed that in addition to ECs, other cells including bone marrow-derived and plastic tumor cells do contribute to tumor vascularization, which is also indicated in ovarian cancer, the most lethal gynecologic malignancy characterized by widespread metastases within the peritoneal cavity upon diagnosis. Given the principle that tumor progression and metastasis are dependent on a persistent blood supply, it is logical that the capability of generating neovessels through diverse mechanisms of ovarian cancer is associated with its malignant potential. This review will discuss the diverse origins of ovarian cancer vascular cells and emphasize their clinical relevance (in the hope of providing insight into the prognostic assessment of women at risk for aggressive disease behavior) and alternative targets for therapeutic intervention.


Hydrogel-based nanocomposites and mesenchymal stem cells: a promising synergistic strategy for neurodegenerative disorders therapy.
Hydrogel-based materials are widely employed in the biomedical field. With regard to central nervous system (CNS) neurodegenerative disorders, the design of injectable nanocomposite hydrogels for in situ drug or cell release represents an interesting and minimally invasive solution that might play a key role in the development of successful treatments. In particular, biocompatible and biodegradable hydrogels can be designed as specific injectable tools and loaded with nanoparticles (NPs), to improve and to tailor their viscoelastic properties upon injection and release profile. An intriguing application is hydrogel loading with mesenchymal stem cells (MSCs) that are a very promising therapeutic tool for neurodegenerative or traumatic disorders of the CNS. This multidisciplinary review will focus on the basic concepts to design acellular and cell-loaded materials with specific and tunable rheological and functional properties. The use of hydrogel-based nanocomposites and mesenchymal stem cells as a synergistic strategy for nervous tissue applications will be then discussed.


Cooperation in oncogenesis between BK virus early region gene and the activated human c-Harvey ras oncogene.
Rapidly growing, undifferentiated brain tumours were induced in newborn Syrian hamsters by intracerebral inoculation of a recombinant DNA (pBK/c-rasA) carrying the BK virus (BKV) early region gene and the activated human c-Harvey-ras (c-Ha-ras) oncogene. Neither of the two genes inoculated alone nor recombinant DNA of the BKV early region gene and the normal human c-Ha-ras proto-oncogene were tumourigenic. Tumour-derived cell lines propagated in culture were immortalized and had growth characteristics consistent with a fully transformed phenotype. Tumours and tumour cell lines contained pBK/c-rasA sequences integrated into cellular DNA and expressed BKV- and c-Ha-ras-specific transcripts as well as BKV T antigen and c-Ha-ras p21. These findings are discussed in relation to a possible cooperation or synergism between BKV and cellular oncogenes in human neoplasia.


Nucleoside Diphosphate Kinase Escalates A-to-C Mutations in MutT-Deficient Strains of Escherichia coli.
The chemical integrity of the nucleotide pool and its homeostasis are crucial for genome stability. Nucleoside diphosphate kinase (NDK) is a crucial enzyme that carries out reversible conversions from nucleoside diphosphate (NDP) to nucleoside triphosphate (NTP) and deoxynucleoside diphosphate (dNDP) to deoxynucleoside triphosphate (dNTP). Guanosine nucleotides (GDP, GTP, dGDP, and dGTP) are highly susceptible to oxidative damage to 8-oxo-GDP (8-O-GDP), 8-O-dGTP, 8-O-GTP, and 8-O-dGTP. MutT proteins in cells hydrolyze 8-O-GTP to 8-O-GMP or 8-O-dGTP to 8-O-dGMP to avoid its incorporation in nucleic acids. In Escherichia coli, 8-O-dGTP is also known to be hydrolyzed by RibA (GTP cyclohydrolase II). In this study, we show that E. coli NDK catalyzes the conversion of 8-O-dGDP to 8-O-dGTP or vice versa. However, the rate of NDK-mediated phosphorylation of 8-O-dGDP to 8-O-dGTP is about thrice as efficient as the rate of dephosphorylation of 8-O-dGTP to 8-O-dGDP, suggesting an additive role of NDK in net production of 8-O-dGTP in cells. Consistent with this observation, the depletion of NDK (Δndk) in E. coli ΔmutT or ΔmutT ΔribA strains results in a decrease of A-to-C mutations. These observations suggest that NDK contributes to the physiological load of MutT in E. coliIMPORTANCE Nucleoside diphosphate kinase (NDK), a ubiquitous enzyme, is known for its critical role in homeostasis of cellular nucleotide pools. However, NDK has now emerged as a molecule with pleiotropic effects in DNA repair, protein phosphorylation, gene expression, tumor metastasis, development, and pathogen virulence and persistence inside the host. In this study, we reveal an unexpected role of NDK in genome instability because of its activity in converting 8-O-dGDP to 8-O-dGTP. This observation has important consequences in escalating A-to-C mutations in Escherichia coli The severity of NDK in enhancing these mutations may be higher in the organisms challenged with high oxidative stress, which promotes 8-O-dGDP/8-O-dGTP production.


Defining the Core Citrus Leaf- and Root-Associated Microbiota: Factors Associated with Community Structure and Implications for Managing Huanglongbing (Citrus Greening) Disease.
Stable associations between plants and microbes are critical to promoting host health and productivity. The objective of this work was to test the hypothesis that restructuring of the core microbiota may be associated with the progression of huanglongbing (HLB), the devastating citrus disease caused by Liberibacter asiaticus, Liberibacter americanus, and Liberibacter africanus The microbial communities of leaves (n = 94) and roots (n = 79) from citrus trees that varied by HLB symptom severity, cultivar, location, and season/time were characterized with Illumina sequencing of 16S rRNA genes. The taxonomically rich communities contained abundant core members (i.e., detected in at least 95% of the respective leaf or root samples), some overrepresented site-specific members, and a diverse community of low-abundance variable taxa. The composition and diversity of the leaf and root microbiota were strongly associated with HLB symptom severity and location; there was also an association with host cultivar. The relative abundance of Liberibacter spp. among leaf microbiota positively correlated with HLB symptom severity and negatively correlated with alpha diversity, suggesting that community diversity decreases as symptoms progress. Network analysis of the microbial community time series identified a mutually exclusive relationship between Liberibacter spp. and members of the Burkholderiaceae, Micromonosporaceae, and Xanthomonadaceae This work confirmed several previously described plant disease-associated bacteria, as well as identified new potential implications for biological control. Our findings advance the understanding of (i) plant microbiota selection across multiple variables and (ii) changes in (core) community structure that may be a precondition to disease establishment and/or may be associated with symptom progression.IMPORTANCE This study provides a comprehensive overview of the core microbial community within the microbiomes of plant hosts that vary in extent of disease symptom progression. With 16S Illumina sequencing analyses, we not only confirmed previously described bacterial associations with plant health (e.g., potentially beneficial bacteria) but also identified new associations and potential interactions between certain bacteria and an economically important phytopathogen. The importance of core taxa within broader plant-associated microbial communities is discussed.


The Bacterial Microbiome Associated With Arid Biocrusts and the Biogeochemical Influence of Biocrusts Upon the Underlying Soil.
Biocrusts are aggregated crusts that exist on the soil surface of arid environments. They are complex microbial communities comprised of cyanobacteria, lichens, mosses, algae and fungi. Recently, biocrusts have gained significant attention due to their ubiquitous distribution and likely important ecological roles, including soil stabilization, soil moisture retention, carbon (C) and nitrogen (N) fixation, as well as microbial engineers for semi-arid ecosystem restoration. Here, we collected three co-occurring types of biocrust (Cyanobacterial crust, Crustose lichen, and Foliose lichen) and their underlying soil from arid zones within Western Australia. Bacterial microbiome composition was determined through 16S rRNA gene amplicon sequencing to assess the extent of microbiome selection within the crusts versus underlying soil and biogeochemical measures performed to determine whether the crusts had significant impact upon the underlying soil for nutrient input. We determined that the bacterial communities of native biocrusts are distinct from those in their underlying soil, where dominant bacterial taxa differed according to crust morphologies. δ15N revealed that N-fixation appeared most evident in Foliose lichen crust (1.73 ± 1.04‰). Consequently, depending upon the crust type, biocrusts contained higher concentrations of organic C (2 to 50 times), total N (4 to 16 times) and available ammonium (2 to 4 times), though this enrichment did not extend to the soils underneath them. These findings demonstrate that biocrust communities are seemingly islands of biological activity in an arid landscape, uniquely different from their surrounding and underlying soil.


LC/LC-MS/MS of an innovative prostate human epithelial cancer (PHEC) in vitro model system.
This work describes the proteomic characterization of a novel in vitro prostate cancer model system, the clonal prostatic human epithelial cancer (PHEC) cell lines. The model is composed of three cell lines representing the three progressive cancer states found in vivo: non-tumorigenic, tumorigenic, and metastatic. The cell lines were evaluated for differential protein expression between states using two dimensional liquid:liquid chromatographic separation followed by mass spectral identification. The proteins from cellular extracts were first separated using liquid:liquid primary separation based on their isoelectric points and hydrophobicity. The resulting peptide fractions were applied to liquid chromatography-mass spectrometry (LC-MS) separation for mass determination and protein identification based on Mascot database inquiry. Over 200 proteins that change expression over the course of progression of this in vitro prostate cancer model were discovered during the comparative analysis of the three cell lines. The importance of these proteins on prostate cancer progression remains to be elucidated with further characterizations. The combination of the two dimensional liquid:liquid separation and mass spectral identifications was used to successfully analyze differential protein expression between multiple cell lines.


Integration of Independent Heat Transfer Mechanisms for Non-Contact Cold Sensation Presentation With Low Residual Heat.
Thermal sensation is crucial to enhancing our comprehension of the world and enhancing our ability to interact with it. Therefore, the development of thermal sensation presentation technologies holds significant potential, providing a novel method of interaction. Traditional technologies often leave residual heat in the system or the skin, affecting subsequent presentations. Our study focuses on presenting thermal sensations with low residual heat, especially cold sensations. To mitigate the impact of residual heat in the presentation system, we opted for a non-contact method, and to address the influence of residual heat on the skin, we present thermal sensations without significantly altering skin temperature. Specifically, we integrated two highly responsive and independent heat transfer mechanisms: convection via cold air and radiation via visible light, providing non-contact thermal stimuli. By rapidly alternating between perceptible decreases and imperceptible increases in temperature on the same skin area, we maintained near-constant skin temperature while presenting continuous cold sensations. In our experiments involving 15 participants, we observed that when the cooling rate was -0.2 to -0.24 °C/s and the cooling time ratio was 30 to 50%, more than 86.67% of the participants perceived only persistent cold without any warmth.