Rapid Internalization of Nanoparticles by Human Cells at the Single Particle Level.
Nanoparticle uptake by cells has been studied for applications both in nanomedicine and in nanosafety. While the majority of studies have focused on the biological mechanisms underlying particle internalization, less attention has been given to questions of a more quantitative nature, such as how many nanoparticles enter cells and how rapidly they do so. To address this, we exposed human embryonic kidney cells to 40-200 nm carboxylated polystyrene nanoparticles and the particles were observed by live-cell confocal and super-resolution stimulated emission depletion fluorescence microscopy. How long a particle remained at the cell membrane after adsorbing onto it was monitored, distinguishing whether the particle ultimately desorbed again or was internalized by the cell. We found that the majority of particles desorb, but interestingly, most of the particles that are internalized do so within seconds, independently of particle size. As this is faster than typical endocytic mechanisms, we interpret this observation as the particles entering via an endocytic event that is already taking place (as opposed to directly triggering their own uptake) or possibly via an as yet uncharacterized endocytic route. Aside from the rapidly internalizing particles, a minority of particles remain at the membrane for tens of seconds to minutes before desorbing or being internalized. We also followed particles after cell internalization, observing particles that appeared to exit the cell, sometimes as rapidly as within tens of seconds. Overall, our results provide quantitative information about nanoparticle cell internalization times and early trafficking.


A potential regulatory region near the EDN3 gene may control both harness racing performance and coat color variation in horses.
The Swedish-Norwegian Coldblooded trotter and the heavier North-Swedish draught horse both descend from the North-Swedish horse, but the Coldblooded trotters have been selected for racing performance while the North-Swedish draught horse is mainly used for agricultural and forestry work. By comparing the genomes of Coldblooded trotters, North-Swedish draught horses and Standardbreds for a large number of single-nucleotide polymorphisms (SNPs), the aim of the study was to identify genetic regions that may be under selection for racing performance. We hypothesized that the selection for racing performance, in combination with unauthorized crossbreeding of Coldblooded trotters and Standardbreds, has created regions in the genome where the Coldblooded trotters and Standardbreds are similar, but differ from the North-Swedish draught horse. A fixation index (Fst) analysis was performed and sliding window Delta Fst values were calculated across the three breeds. Five windows, where the average Fst between Coldblooded trotters and Standardbreds was low and the average Fst between Coldblooded trotters and North-Swedish draught horses was high, were selected for further investigation. Associations between the most highly ranked SNPs and harness racing performance were analyzed in 400 raced Coldblooded trotters with race records. One SNP showed a significant association with racing performance, with the CC genotype appearing to be negatively associated. The SNP identified was genotyped in 1915 horses of 18 different breeds. The frequency of the TT genotype was high in breeds typically used for racing and show jumping while the frequency of the CC genotype was high in most pony breeds and draught horses. The closest gene in this region was the Endothelin3 gene (EDN3), a gene mainly involved in melanocyte and enteric neuron development. Both functional genetic and physiological studies are needed to fully understand the possible impacts of the gene on racing performance.


Raman spectroscopy as a means for the identification of plattnerite (PbO2), of lead pigments and of their degradation products.
The Raman spectra of plattnerite [lead(IV) oxide, PbO2] and of the lead pigments red lead (Pb3O4), lead monoxide [PbO, litharge (tetragonal) and massicot (orthorhombic)], lead white [basic lead carbonate, 2PbCO3.Pb(OH)2] and of their laser-induced degradation products were recorded using a range of different excitation lines, spectrometer systems and experimental conditions. The degradation of PbO2 is more extensive along the pathway PbO2-->Pb3O4-->PbO (litharge)-->PbO (massicot) the shorter the wavelength of the excitation line and the higher its power. The Raman spectrum of PbO2, which is black and of the rutile structure, is particularly difficult to obtain but three bands, at 653, 515 and 424 cm-1, were identified as arising from the b2g, a1g and e(g) modes respectively, by analogy with the corresponding modes of isostructural SnO2 (776, 634 and 475 cm-1). A further oxide was identified, PbO1.55, the Raman spectrum of which does not correspond to that of any of the laser-induced degradation products of PbO2 at any of the wavelengths used. The Raman results are critical to the future use of Raman microscopy for the identification of lead pigments on artworks.


The Wellness Group: a novel intervention for coping with disruptive behavior among [corrected] elderly nursing home residents.
Elderly nursing home residents face the challenge of coping with the loss of independence, reduced physical abilities, feelings of isolation, anxiety, depression, and helplessness. Maintaining one's integrity within the nursing home is a challenge easily lost. Issues of coping with disability, change in body image, the promotion of self-empowerment, and autonomy are critical to preserving the personhood of a nursing home resident. Current emphasis on the management of behavior has focused entirely on the control of disruption rather than the promotion of resident strengths and abilities. We developed a group therapy program designed to enhance self-awareness, self-esteem, and body awareness among the demented elderly. Techniques of meditation, relaxation, sensory awareness and guided imagery were introduced and reinforced. This group is easily reproducible and offers benefits to both residents and interdisciplinary staff.


Cardiac patients' perceptions of neighboring patients' risk: influence on psychological stress in the ED and subsequent posttraumatic stress.
As many as 12% of acute coronary syndrome (ACS) patients screen positive for post-traumatic stress disorder (PTSD) symptoms due to their cardiac event, and emergency department (ED) factors such as overcrowding have been associated with risk for PTSD. We tested the association of patients' perceptions of their proximity to a critically ill patient during ED evaluation for ACS with development of posttraumatic stress symptoms (PSS) in the month after hospital discharge. Participants were enrolled in the REactions to Acute Care and Hospitalization (REACH) study during evaluation for ACS in an urban ED. Participants reported whether they perceived a patient near them was close to death. They also reported their current fear, concern they may die, perceived control, and feelings of vulnerability on an Emergency Room Perceptions questionnaire. One month later, participants reported on PTSD symptoms specific to the cardiac event and ED hospitalization. Of 763 participants, 12% reported perceiving a nearby patient was likely to die. In a multivariate linear regression model [F(9757) = 19.69, p < .001, R2 adjusted = .18] with adjustment for age, sex, GRACE cardiac risk score, discharge ACS diagnosis, Charlson comorbidity index, objective ED crowding, and depression symptoms at baseline, perception of a nearby patients' likely death was associated with a 2.33 point (95% CI, 0.60-4.61) increase in 1 month PTSD score. A post hoc mediation analysis with personal threat perceptions [F(10,756) = 25.28, p < .001, R2 adjusted = .24] showed increased personal threat perceptions during the ED visit, B = 0.71 points on the PCL per point on the personal threat perception questionnaire, β = 0.27, p = .001, fully mediated association of participants' perceptions of nearby patients' likely death with 1-month PTSD score (after adjustment for ED threat perceptions,) B = 0.89 (95% CI, -1.33 to 3.12), β = 0.03, p = .43, accounting for 62% of the adjusted effect and causing the main effect to become statistically nonsignificant. We found patients who perceived a nearby patient was likely to die had significantly greater PTSD symptoms at 1 month. Awareness of this association may be helpful for designing ED patient management procedures to identify and treat patients with an eye to post-ACS psychological care.


Allopurinol desensitization with A 2 weeks modified protocol in an elderly patients with multiple comorbidities: a case report.
Allopurinol is an effective urate-lowering drug that is well tolerated by the majority of patients. Patients with chronic renal insufficiency have an increased risk of hypersensitivity reactions with allopurinol. 75 year old male patient with gout, renal insufficiency, history of metastatic colorectal carcinoma status post-resection was referred to Allergy clinic for a maculopapular eruption that developed 1 week after initiating therapy with allopurinol. The rash resolved with discontinuation of allopurinol. However, his serum urate level rose to 19.9 mg/dl. We initially proposed a slow 4 week oral allopurinol desensitization. The treating nephrologist felt it was critical to lower urate more rapidly. As a result, we modified the dose and standard 4 week protocol down to 2 weeks. A suspension of allopurinol was prepared by the allergy nurse practitioner with a 300 mg allopurinol tablet. The sensitization protocol was modified as a starting dose of 0.3 mg escalating to a final dose of 300 mg/day in 2 weeks. There was no reaction during or after the desensitization. The patient's urate level normalized (6.3 mg/dl) and has continued on 300 mg allopurinol daily without reaction. A 2 week modified allopurinol desensitization protocol is a safe alternative for elderly patients with multiple comorbidities.


Leukocytic cell sources of airway tissue kallikrein.
Lung tissue kallikrein (TK) is a serine proteinase that putatively plays a role in the pathophysiology of asthma by generating kallidin and bradykinin, mediators that contribute to airway hyperresponsiveness. In previous studies we observed biphasic increases in TK activity in bronchoalveolar lavage fluid following airway allergen challenge in allergic sheep. Although glandular TK is likely a major source of the initial increase in TK, the sources of the late increases in TK that are associated with the development of airway hyperresponsiveness may be dependent on activated resident and recruited inflammatory cells including alveolar macrophages (AMs) and neutrophils (PMNs). These cells increase concomitantly with the late increases in TK activity. To test this hypothesis, we obtained AMs from bronchoalveolar lavage fluid and PMNs and monocytes (precursors of AMs) from sheep blood and determined whether these cells contained TK and whether these same cells could release TK upon activation. Using confocal microscopy, immunocytochemical techniques, and enzyme activity assays, we found that all three cell types contained and secreted TK. All three cell types demonstrated basal release of TK, which could be increased after stimulation with zymosan. In addition, PMNs also released TK in the presence of phorbol ester, suggesting multiple secretory pathways in these cells. Furthermore, we showed that human monocytes also contain and secrete TK. We conclude that in the airways, monocytes, PMNs, and AMs may contribute to increased TK activity. Knowing the sources of TK in the airways could be important in understanding the mechanisms of inflammation that contribute to the pathophysiology of asthma and may help in the development of new therapies to control the disease.


The MultiFurcating Neighbor-Joining Algorithm for Reconstructing Polytomic Phylogenetic Trees.
Results from phylogenetic analyses that study the evolution of species according to their biological characteristics are frequently structured as phylogenetic trees. One of the most widely used methods for reconstructing them is the distance-based method known as the neighbor-joining (NJ) algorithm. It is known that the NJ algorithm can produce different phylogenetic trees depending on the order of the taxa in the input matrix of evolutionary distances, because the method only yields bifurcating branches or dichotomies. According to this, results and conclusions published in articles that only calculate one of the possible dichotomic phylogenetic trees are somehow biased. We have generalized the formulas used in the NJ algorithm to cope with Multifurcating branches or polytomies, and we have called this new variant of the method the multifurcating neighbor-joining (MFNJ) algorithm. Instead of the dichotomic phylogenetic trees reconstructed by the NJ algorithm, the MFNJ algorithm produces polytomic phylogenetic trees. The main advantage of using the MFNJ algorithm is that only one phylogenetic tree can be obtained, which makes the experimental section of any study completely reproducible and unbiased to external issues such as the input order of taxa.


Advances in Antibody-Drug Conjugates in the Treatment of HER2-Positive Breast Cancer.
Although targeted drugs improved the therapeutic effect of HER2-positive breast cancer, the long-term prognosis was still poor. In this regard, the research and development of antibody-drug conjugates (ADCs) came into being and made a lot of progress. ADCs had the characteristics of both chemotherapeutic agents and targeted agents by combining chemotherapeutic agents and targeted agents through a linker. It not only had a strong anti-tumor effect on HER2-positive breast cancer, but also had certain anti-tumor effects on HER2-low and even HER2-negative patients. In addition, the clinical researches of ADCs combined with immune checkpoint inhibitors (ICIs) therapy had also made a great breakthrough. This review aimed to summarize the clinical progress of ADCs, in particular the two drugs approved by the US Food and Drug Administration (FDA) for HER2-positive metastatic breast cancer as well as to summarize the current status of ADCs in combination with ICIs.


[Killer systems of Saccharomyces cerevisiae yeasts].
The killer systems of Saccharomyces cerevisiae are a peculiar group of cytoplasmic symbionts of primitive eukaryotes. The genetic material of these symbionts is double-stranded RNA. Their basic properties are linearity of genome, its fragmentation, resulting in two separately replicating major and minor segments, and the ability to control the synthesis of secretory proteins--mycocins which can kill the taxonomically related strains. Secretion of mycocins also confers immunity to their action. The strains containing killer symbionts are toxigenic and resistant to their own toxins, while those with no killer double-stranded RNA are sensitive to mycocins. The killer systems of Saccharomyces cerevisiae possess some properties relevant to viruses and evidently are evolved during the evolution of infectious viruses. Occurrence of such systems in monocellular eucaryotic organisms is an example of genome complication in the course of putting together the virus-like components. The peculiarities of replication and expression of killer systems and their utilization for the construction of vector molecules are discussed.


Amino acid derivative-mediated detoxification and functionalization of dual cure dental restorative material for dental pulp cell mineralization.
Current dental restorative materials are only used to fill the defect of hard tissues, such as dentin and enamel, because of their cytotoxicity. Therefore, exposed dental pulp tissues in deep cavities must be first covered by a pulp capping material like calcium hydroxide to form a layer of mineralized tissue. However, this tissue mineralization is based on pathological reaction and triggers long-lasting inflammation, often causing clinical problems. This study tested the ability of N-acetyl cysteine (NAC), amino acid derivative, to reduce cytotoxicity and induce mineralized tissue conductivity in resin-modified glass ionomer (RMGI), a widely used dental restorative material having dual cure mechanism. Rat dental pulp cells were cultured on untreated or NAC-supplemented RMGI. NAC supplementation substantially increased the percentage of viable cells from 46.7 to 73.3% after 24-h incubation. Cell attachment, spreading, proliferative activity, and odontoblast-related gene and protein expressions increased significantly on NAC-supplemented RMGI. The mineralization capability of cells, which was nearly suppressed on untreated RMGI, was induced on NAC-supplemented RMGI. These improved behaviors and functions of dental pulp cells on NAC-supplemented RMGI were associated with a considerable reduction in the production of intracellular reactive oxygen species and with the increased level of intracellular glutathione reserves. These results demonstrated that NAC could detoxify and functionalize RMGIs via two different mechanisms involving in situ material detoxification and antioxidant cell protection. We believe that this study provides a new approach for developing dental restorative materials that enables mineralized tissue regeneration.


Value of standardised exercise tests and blood biochemistry in the selection and training of breeding stallions.
Stallions selected by the Royal Dutch Warmblood Society were submitted to a standardised lungeing test at the beginning and at the end of a 100-day test of performance and ability. The heart rate, haematology and biochemistry values obtained in the first lungeing test showed no significant differences between the 15 stallions which were rejected by the Royal Dutch Warmblood Society during the first month of the 100-day test, the 15 stallions rejected during the last month and the 11 stallions which were approved for registration in the studbook. The 26 stallions submitted to the second lungeing test had significantly lower heart rates and blood lactate concentrations than in the first test. The standardised lungeing test had no value in predicting the rejection or approval of the stallions, and the fitness of a stallion at the beginning of the 100-day test did not influence its chance of being approved as a breeding stallion. The differences between the results of the first and the second tests suggest that the fitness of the stallions improved during the 100-day test.


CAPS-1 promotes fusion competence of stationary dense-core vesicles in presynaptic terminals of mammalian neurons.
Neuropeptides released from dense-core vesicles (DCVs) modulate neuronal activity, but the molecules driving DCV secretion in mammalian neurons are largely unknown. We studied the role of calcium-activator protein for secretion (CAPS) proteins in neuronal DCV secretion at single vesicle resolution. Endogenous CAPS-1 co-localized with synaptic markers but was not enriched at every synapse. Deletion of CAPS-1 and CAPS-2 did not affect DCV biogenesis, loading, transport or docking, but DCV secretion was reduced by 70% in CAPS-1/CAPS-2 double null mutant (DKO) neurons and remaining fusion events required prolonged stimulation. CAPS deletion specifically reduced secretion of stationary DCVs. CAPS-1-EYFP expression in DKO neurons restored DCV secretion, but CAPS-1-EYFP and DCVs rarely traveled together. Synaptic localization of CAPS-1-EYFP in DKO neurons was calcium dependent and DCV fusion probability correlated with synaptic CAPS-1-EYFP expression. These data indicate that CAPS-1 promotes fusion competence of immobile (tethered) DCVs in presynaptic terminals and that CAPS-1 localization to DCVs is probably not essential for this role.


Effect of xylazine and ketamine on intraocular pressure in horses.
Intraocular pressure was measured with a MacKay-Marg tonometer in eight horses following auriculopalpebral nerve block and topical application of lignocaine. Measurements were recorded before and after xylazine, 1.1 mg/kg intravenously, every two minutes for 16 minutes after administration of ketamine, 2.2 mg/kg intravenously, and after recovery from anaesthesia. Before xylazine, intraocular pressure was 17.1 +/- 3.9 and 18.4 +/- 2.2 mm Hg in the left and right eyes, respectively. Intraocular pressure tended to decrease after administration of xylazine and ketamine, with a significant decrease in one eye six minutes after injection of ketamine.


Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus of D. melanogaster.
We have determined the complete nucleotide sequence of a 3292 bp cloned segment derived from the 63B heat shock cytogenetic locus of D. melanogaster. Within this segment we have positioned the start of transcription and RNA splice sites of the unique gene that encodes the 83,000 d heat shock polypeptide (hsp83 gene) by S1 mapping and synthesis of cDNA from restriction fragment primed mRNA. The sequence begins at a point 879 bp upstream from the transcription start and includes the 149 bp nontranslated first exon, the 1139 bp intron and extends 1125 bp into the protein coding region. These data identify a single open translation reading frame for the first 375 amino acids of the 83,000 d polypeptide, beginning with the first ATG codon located at the 3' intron-exon junction. We discuss and demonstrate the use of E. coli exonuclease III generated single-strand DNA probes as an alternative to strand separation for S1 mapping of mRNA. We also use homology search criteria based upon known protein-DNA binding sites to compare our hsp83 sequence with other sequenced Drosophila heat shock genes. These comparisons indicate that a large region of approximately 80 bp centered around the transcription initiation point of the hsp83 gene shares only a 31% homology with the corresponding region of the hsp70 gene, whereas the hsp22, 23, 26, and 27 genes share a 54% homology with hsp70 in this region. The lower homology of the hsp83 gene is consistent with the deviant nature of this heat shock gene.


Legumes: the most important dietary predictor of survival in older people of different ethnicities.
To identify protective dietary predictors amongst long-lived elderly people (N= 785), the "Food Habits in Later Life "(FHILL) study was undertaken among five cohorts in Japan, Sweden, Greece and Australia. Between 1988 and 1991, baseline data on food intakes were collected. There were 785 participants aged 70 and over that were followed up to seven years. Based on an alternative Cox Proportional Hazard model adjusted to age at enrollment (in 5-year intervals), gender and smoking, the legume food group showed 7-8% reduction in mortality hazard ratio for every 20g increase in daily intake with or without controlling for ethnicity (RR 0.92; 95% CI 0.85-0.99 and RR 0.93; 95% CI 0.87-0.99, respectively). Other food groups were not found to be consistently significant in predicting survival amongst the FHILL cohorts.


A replication study confirms the association of dendritic cell immunoreceptor (DCIR) polymorphisms with ACPA - negative RA in a large Asian cohort.
Dendritic cell immunoreceptor (DCIR) has been implicated in development of autoimmune disorders in rodent and DCIR polymorphisms were associated with anti-citrullinated proteins antibodies (ACPA)-negative rheumatoid arthritis (RA) in Swedish Caucasians. This study was undertaken to further investigate whether DCIR polymorphisms are also risk factors for the development of RA in four Asian populations originated from China and Malaysia. We genotyped two DCIR SNPs rs2377422 and rs10840759 in Han Chinese population (1,193 cases, 1,278 controls), to assess their association with RA. Subsequently, rs2377422 was further genotyped in three independent cohorts of Malaysian-Chinese subjects (MY_Chinese, 254 cases, 206 controls), Malay subjects (MY_ Malay, 515 cases, 986 controls), and Malaysian-Indian subjects (MY_Indian, 378 cases, 285 controls), to seek confirmation of association in various ethnic groups. Meta-analysis was preformed to evaluate the contribution of rs2377422 polymorphisms to the development of ACPA-negative RA in distinct ethnic groups. Finally, we carried out association analysis of rs2377422 polymorphisms with DCIR mRNA expression levels. DCIR rs2377422 was found to be significantly associated with ACPA -negative RA in Han Chinese (OR 1.92, 95% CI 1.27-2.90, P=0.0020). Meta-analysis confirms DCIR rs2377422 as a risk factor for ACPA-negative RA across distinct ethnic groups (OR(overall) =1.17, 95% CI 1.06-1.30, P=0.003). The SNP rs2377422 polymorphism showed significant association with DCIR mRNA expression level, i.e. RA-risk CC genotype exhibit a significant increase in the expression of DCIR (P=0.0023, Kruskal-Wallis). Our data provide evidence for association between DCIR rs2377422 and RA in non-Caucasian populations and confirm the influence of DCIR polymorphisms on RA susceptibility, especially on ACPA-negative RA.


Fully Automated Detection of Isolated Rapid-Eye-Movement Sleep Behavior Disorder Using Actigraphy.
Isolated rapid-eye-movement (REM) sleep behavior disorder (iRBD) is caused by motor disinhibition during REM sleep and is a strong early predictor of Parkinson's disease. However, screening questionnaires for iRBD lack specificity due to other sleep disorders that mimic the symptoms. Nocturnal wrist actigraphy has shown promise in detecting iRBD by measuring sleep-related motor activity, but it relies on sleep diary-defined sleep periods, which are not always available. Our aim was to precisely detect iRBD using actigraphy alone by combining two actigraphy-based markers of iRBD - abnormal nighttime activity and 24-hour rhythm disruption. In a sample of 42 iRBD patients and 42 controls (21 clinical controls with other sleep disorders and 21 community controls) from the Stanford Sleep Clinic, the nighttime actigraphy model was optimized using automated detection of sleep periods. Using a subset of 38 iRBD patients with daytime data and 110 age-, sex-, and body-mass-index-matched controls from the UK Biobank, the 24-hour rhythm actigraphy model was optimized. Both nighttime and 24-hour rhythm features were found to distinguish iRBD from controls. To improve the accuracy of iRBD detection, we fused the nighttime and 24-hour rhythm disruption classifiers using logistic regression, which achieved a sensitivity of 78.9%, a specificity of 96.4%, and an AUC of 0.954. This study preliminarily validates a fully automated method for detecting iRBD using actigraphy in a general population.Clinical relevance- Actigraphy-based iRBD detection has potential for large-scale screening of iRBD in the general population.


Evidence that the epidermal growth factor receptor and non-tyrosine kinase hormone receptors stimulate phosphoinositide hydrolysis by independent pathways.
We have shown previously that exposure of a non-transformed continuous line of rat liver epithelial (WB) cells to epidermal growth factor (EGF), adrenaline, angiotensin II or [Arg8]vasopressin results in an accumulation of the inositol phosphates InsP1, InsP2 and InsP3 [Hepler, Earp & Harden (1988) J. Biol. Chem. 263, 7610-7619]. Studies were carried out with WB cells to determine whether the EGF receptor and other, non-tyrosine kinase, hormone receptors stimulate phosphoinositide hydrolysis by common, overlapping or separate pathways. The time courses for accumulation of inositol phosphates in response to angiotensin II and EGF were markedly different. Whereas angiotensin II stimulated a very rapid accumulation of inositol phosphates (maximal by 30 s), increases in the levels of inositol phosphates in response to EGF were measurable only following a 30 s lag period; maximal levels were attained by 7-8 min. Chelation of extracellular Ca2+ with EGTA did not modify this relative difference between angiotensin II and EGF in the time required to attain maximal phospholipase C activation. Under experimental conditions in which agonist-induced desensitization no longer occurred in these cells, the inositol phosphate responses to EGF and angiotensin II were additive, whereas those to angiotensin II and [Arg8]vasopressin were not additive. In crude WB lysates, angiotensin II, [Arg8]vasopressin and adrenaline each stimulated inositol phosphate formation in a guanine-nucleotide-dependent manner. In contrast, EGF failed to stimulate inositol phosphate formation in WB lysates in the presence or absence of guanosine 5'-[gamma-thio]triphosphate (GTP[S]), even though EGF retained the capacity to bind to and stimulate tyrosine phosphorylation of its own receptor. Pertussis toxin, at concentrations that fully ADP-ribosylate and functionally inactivate the inhibitory guanine-nucleotide regulatory protein of adenylate cyclase (Gi), had no effect on the capacity of EGF or hormones to stimulate inositol phosphate accumulation. In intact WB cells, the capacity of EGF, but not angiotensin II, to stimulate inositol phosphate accumulation was correlated with its capacity to stimulate tyrosine phosphorylation of the 148 kDa isoenzyme of phospholipase C. Taken together, these findings suggest that, whereas angiotensin II, [Arg8]vasopressin and alpha 1-adrenergic receptors are linked to activation of one or more phospholipase(s) C by an unidentified G-protein(s), the EGF receptor stimulates phosphoinositide hydrolysis by a different pathway, perhaps as a result of its capacity to stimulate tyrosine phosphorylation of phospholipase C-gamma.


Distribution of TMEM100 in the mouse and human gastrointestinal tract--a novel marker of enteric nerves.
Identification of markers of enteric neurons has contributed substantially to our understanding of the development, normal physiology, and pathology of the gut. Previously identified markers of the enteric nervous system can be used to label all or most neuronal structures or for examining individual cells by labeling just the nucleus or cell body. Most of these markers are excellent but have some limitations. Transmembrane protein 100 (TMEM100) is a gene at locus 17q32 encoding a 134-amino acid protein with two hypothetical transmembrane domains. TMEM100 expression has not been reported in adult mammalian tissues but does appear in the ventral neural tube of embryonic mice and plays a role in signaling pathways associated with development of the enteric nervous system. We showed that TMEM100 messenger RNA is expressed in the gastrointestinal tract and demonstrated that TMEM100 is a membrane-associated protein. Furthermore TMEM100 immunoreactivity was restricted to enteric neurons and vascular tissue in the muscularis propria of all regions of the mouse and human gastrointestinal tract. TMEM100 immunoreactivity colocalized with labeling for the pan-neuronal marker protein gene product 9.5 (PGP9.5) but not with the glial marker S100ß or Kit, a marker of interstitial cells of Cajal. The signaling molecule, bone morphogenetic protein (BMP) 4, was also expressed in enteric neurons of the human colon and co-localized with TMEM100. TMEM100 is also expressed in neuronal cell bodies and fibers in the mouse brain and dorsal root ganglia. We conclude that TMEM100 is a novel, membrane-associated marker for enteric nerves and is as effective as PGP9.5 for identifying neuronal structures in the gastrointestinal tract. The expression of TMEM100 in the enteric nervous system may reflect a role in the development and differentiation of cells through a transforming growth factor β, BMP or related signaling pathway.


Cyclosporine Reduces Sclerosis in Morphea: A Retrospective Study in 12 Patients and a Literature Review.
The treatment of severe morphea is challenging, and treatment experience concerning the use of immunosuppressive agents for this condition is limited. The purpose of this study is to analyze the use of cyclosporine, its tolerability, and its effect on skin sclerosis. Patients with severe morphea who underwent treatment with cyclosporine were studied retrospectively. Five of 12 patients with morphea showed complete remission and 6 patients had partial remission at the end of therapy (9-46 months, median 14) under a median cyclosporine dose of 2.4 mg/kg. The mean affected body surface area fell from 50% (2-80, median 65) to 17% (0-40, median 18). Side effects were hypertension, elevated transaminases, cholesterol, and weight gain. Cyclosporine can be effective in morphea. The side effects were reversible. However, the duration of treatment with cyclosporine is limited because of its potential permanent side effects. Prospective placebo-controlled studies are needed to establish the superiority of cyclosporine over other immunosuppressive drugs in this setting.


Placement of endosseous implants in grafted alveolar clefts.
The purpose of this study was to determine the optimal timing for placement and to evaluate short- and long-term outcomes of endosseous implants in bone-grafted alveolar clefts. Fourteen patients who underwent alveolar cleft bone grafting (ACBG) and closure of an oronasal fistula followed by restoration of the missing lateral incisor tooth using endosseous implants (EI) were studied. The oronasal fistulae were closed using local flaps, and the alveolus was grafted with fresh autogenous iliac marrow. Endosseous implants were placed a minimum of 4 months following ACBG. The average age at ACBG was 20.35 years (range, 12-65 yr), and at implant placement 22.2 years (range, 15-66 yr). It was necessary to regraft the alveolar cleft region in six patients to provide adequate bone volume for implant placement. Those who required alveolar regrafting had an increased mean interval between the initial ACBG and planned implant placement compared to the patients with adequate bone available for implant placement 26.4 months (range, 4-46 mo) versus 15.75 months (range, 4-36 mo). Twenty-nine implants were placed in 14 patients, 9 outside of the cleft region and 20 in grafted alveolar clefts. Eighteen of 20 (90%) implants in the bone-grafted alveolar clefts have been successfully restored. The mean follow-up after implant placement was 39.1 months (range, 1-54 mo), and after restoration 28.5 months (range, 1-47 mo). Endosseous implants can be placed in bone-grafted alveolar clefts. Consideration should be given to the adequacy of graft volume and ridge morphology at the time of ACBG. The interval between ACBG and implant placement is important. The greater the interval beyond 4 months, the more likely there will be inadequate bone volume to accept an implant.


[The taxonomic aspects of the transmission of the causative agent of Lyme disease].
According to world literature data 17 species of ixodid ticks have been studied for natural infection with the Lyme disease agent. Analysis of the data on the level of the infection, transovarial and transphase transmission has shown that main biological vectors of Borrelia burgdorferi are the species of the subgenus Ixodes s. str. - I. ricinus, I. persulcatus (Eurasia), I. dammini, I. pacificus (North America). Potential vectors are I. scapularis, I. dentatus, Amblyomma americanum, Dermacentor variabilis. Single isolations were registered for I. neotomae, Haemaphysalis leporispalustris, D. occidentalis. Nonidentified spirocheta was isolated from A. americanum, D. variabilis, D. parumapertum, Rhipicephalus sanguineus. No agent was isolated from I. cookey, D. albipictus, R. reticulatus, H. concinna. On the basis of comparative and ontogenetic data the species from a group of main vectors: I. ricinus, I. persulcatus, I. pacificus had been attributed by me to the phyletic group persulcatus before Lyme disease was discovered and its causative agent isolated. The question whether I. scapularis belongs to the group persulcatus was also discussed at that time but left open due to somewhat aberrant structure of gnathosoma at preimaginal phases (Filippova, 1969, 1971, 1973). 6 Palaearctic, 2 Indomalayan and 3 Nearctic species were referred to the group persulcatus at the time. I. dammini was described later, in 1979. Gnathosoma of its preimaginal phases has an intermediate structure between I. scapularis and other species of the group persulcatus. Sexually mature phase and nymph of I. dentatus have much in common with Palaearctic members of the group, I. pavlovskyi, I. kazakstani, I. kashmicus. Preimaginal phases of I. scapularis and nymph of I. dentatus were studied by me on the collection material. Thus, it is possible to speak of the belonging of main vectors of B. burgdorferi to a common phyletic group within the subgenus Ixodes s. str. and, therefore, of common origin of ecological medium for the agent. At the same time each species of the vector is an evolutionally developed difference of ecological medium for B. burgdorferi. Roots of the group persulcatus could originate as far as in Paleocene before the land connection between North America and Europe disappeared. Conditions for the existence of recent species, however, appeared considerably later and their flourishing is dated by Pliocene. The main epidemiological role belongs now to I. ricinus, I. persulcatus, I. dammini, I. pacificus.(ABSTRACT TRUNCATED AT 400 WORDS)


Production and properties of Bordetella pertussis heat-labile toxin.
Twelve selected strains of Bordetella pertussis were compared quantitatively for their ability to produce heat-labile toxin (HLT); all proved to be active producers, with only a three-fold range between the highest and the lowest. Bordet-Gengou agar, charcoal agar, modified Hornibrook medium and Stainer and Schölte (12G) medium differed little in their ability to support toxin production by three B. pertussis strains. However, cells grown on the solid media for 24 h were slightly more toxic than their counterparts grown for 72 h whereas in the liquid media the opposite was true. The concentration of iron in the medium did not influence HLT production, but high concentrations of nicotinic acid significantly reduced the HLT content of the cells. Crude preparations of toxin underwent only a 10% loss of toxicity per annum at -20 degrees C and were stable for up to 2 weeks at 4 degrees C. At 37 degrees C, toxicity was lost within a few days. The toxin was partially purified by a series of mild procedures and had a mol. wt by gel filtration of 89 000 +/- 10%. HLT was toxoided by treatment with formaldehyde to give a product which was immunogenic in rabbits but not in mice. Because anti-HLT could be absorbed out of the rabbit antisera by treatment with intact B. pertussis, it was concluded that some of the HLT in the bacteria is surface-exposed even though the main part may have a cytoplasmic location.


Advancements in the management of spine disorders.
Spinal disorders and especially back and neck pain affect more people and have greater impact on work capacity and health-care costs than any other musculoskeletal condition. One of the difficulties in reducing the burden of spinal disorders is the wide and heterogeneous range of specific diseases and non-specific musculoskeletal disorders that can involve the spinal column, most of which manifest as pain. Despite, or perhaps because of its impact, spinal disorders remain one of the most controversial and difficult conditions for clinicians, patients and policymakers to manage. This paper provides a brief summary of advances in the understanding of back and neck pain over the past decade as evidenced in the current literature. This paper includes the following sections: a classification of spinal disorders; the epidemiology of spine pain in the developed and developing world; key advancements in biological and biomechanical sciences in spine pain; the current status of potential methods for the prevention of back and neck pain; rheumatological and systemic disorders that impact the spine; and evidence-based surgical and non-surgical management of spine pain. The final section of this paper looks to the future and proposes actions and strategies that may be considered by the international Bone and Joint Decade (BJD), by providers, institutions and by policymakers so that we may better address the burden of spine disorders at global and local levels.


Smoking and first acute myocardial infarction: age, mortality and smoking cessation rate.
Cigarette smoking is a major contributor to the risk of acute myocardial infarction and the subsequent morbidity and mortality. Physicians can play an important role in smoking cessation among patients with AMI because of their frequent contact with the patient during the event. To study the prevalence of smoking, age, localization of coronary occlusion, mortality and rate of smoking cessation in consecutive patients who were diagnosed with a first AMI in our center in 1989-93. The study included 1,510 consecutive patients with first AMI: 973 men (512 smokers, 52.6%) and 537 women (215 smokers, 40%), whose mean age was 64.1 +/- 6.7 and 68.6 +/- 5.2 years respectively. The median age at the first AMI in non-smoking and smoking men differed significantly (70.4 +/- 6.8 vs. 56.6 +/- 6.1 years, P < 0.001) while the difference in the women was smaller (70.4 +/- 6.9 vs. 66.8 +/- 7.2). The proportion of smokers/non-smokers among men was greater at a younger age and decreased proportionally with age. The overall mortality was 11.3% with a significant difference in mortality rate in the younger age groups between smokers and non-smokers (1% vs. 0% in the age group 31-40 years, P < 0.05, and 6.1% vs. 0.8% in the 41-50 year age group, P < 0.001). Only 62% of the smokers who survived the AMI declared that they had received anti-smoking advice from a physician during hospitalization. The cessation rate in this group was significantly higher than in smokers who had not been cautioned against smoking (56% vs. 18%). Current smokers sustained their first AMI more than one decade earlier than non-smokers, and the younger smokers had a higher mortality rate. The majority of the smokers who received anti-smoking advice during their hospitalization for AMI quit smoking in the year following the acute event.


Additional Effects of a Physical Therapy Protocol on Headache Frequency, Pressure Pain Threshold, and Improvement Perception in Patients With Migraine and Associated Neck Pain: A Randomized Controlled Trial.
To evaluate the additional effect provided by physical therapy in migraine treatment. Randomized controlled trial. Tertiary university-based hospital. Among the 300 patients approached, 50 women (age range, 18-55y) diagnosed with migraine were randomized into 2 groups: a control group (n=25) and a physiotherapy plus medication group (n=25) (N=50). Both groups received medication for migraine treatment. Additionally, physiotherapy plus medication patients received 8 sessions of physical therapy over 4 weeks, comprised mainly of manual therapy and stretching maneuvers lasting 50 minutes. A blinded examiner assessed the clinical outcomes of headache frequency, intensity, and self-perception of global change and physical outcomes of pressure pain threshold and cervical range of motion. Data were recorded at baseline, posttreatment, and 1-month follow-up. Twenty-three patients experienced side effects from the medication. Both groups reported a significantly reduced frequency of headaches; however, no differences were observed between groups (physiotherapy plus medication patients showed an additional 18% improvement at posttreatment and 12% improvement at follow-up compared with control patients, P>.05). The reduction observed in the physiotherapy plus medication patients was clinically relevant at posttreatment, whereas clinical relevance for control patients was demonstrated only at follow-up. For pain intensity, physiotherapy plus medication patients showed statistical evidence and clinical relevance with reduction posttreatment (P<.05). In addition, they showed better self-perception of global change than control patients (P<.05). The cervical muscle pressure pain threshold increased significantly in the physiotherapy plus medication patients and decreased in the control patients, but statistical differences between groups were observed only in the temporal area (P<.05). No differences were observed between groups regarding cervical range of motion. We cannot assume that physical therapy promotes additional improvement in migraine treatment; however, it can increase the cervical pressure pain threshold, anticipate clinically relevant changes, and enhance patient satisfaction.


Effectiveness of a course designed to teach handoffs to medical students.
Handoffs of patient care are increasingly common and are known to contribute to medical errors. A significant number, if not the large majority, of first-year Internal Medicine residents have not received formal education pertaining to handoffs during medical school. To develop a program designed to teach handoffs to medical students entering their fourth year of training. University of Colorado Denver School of Medicine. Our Handoff Selective was first offered in April 2007 as part of a 2-week Integrated Clinician's Course conducted once yearly between the third and fourth years of medical school. The Selective consisted of a didactic session in which communication theory and elements were discussed and a practicum in which students used faculty-developed case scenarios to practice both giving and receiving handoffs. Sixty (the maximum number of spots available) out of 150 students participated in the course, although many more students chose the course than spots available. Prior to taking the Selective, medical students' confidence in performing handoffs was poor, but it improved after the course (P < 0.001); 92% of students felt the Handoff Selective was "useful" or "extremely useful." While both components of the course were thought to be useful to the large majority of students, the practicum portion was thought to be more useful (P < 0.001). Formal education on handoffs is well received by medical students and improves their self-perceived understanding and performance of handoffs.


Treprostinil for severe inoperable chronic thromboembolic pulmonary hypertension.
Chronic thromboembolic pulmonary hypertension (CTEPH) results from non-resolving pulmonary thromboemboli that are resistant to plasmatic anticoagulation. Because of a secondary pulmonary arteriopathy accompanying major vessel obstruction, the disorder may be a target for vasodilator therapy. In an open-label uncontrolled study, we investigated the prostacyclin analog treprostinil given s.c. in patients with severe inoperable CTEPH. Between September 1999 and September 2005, 25 patients were included if their World Health Organization (WHO) functional class was III or IV, if their six-minute walking distance (6-MWD) <or= 380 m, and if they had undergone at least one hospitalization for right heart decompensation within the prior six months, albeit not within one month before treatment start. Right heart catheterization was performed at baseline and after a minimum of 12 months (range: 12-33 months) of treatment. Treprostinil plasma concentrations were determined after at least six months of treatment. A historical group of 31 patients at our center with inoperable CTEPH matched for disease severity was used for comparative analyses. Treprostinil-treated patients demonstrated significant improvements in 6-MWD (P = 0.01), WHO functional class (P = 0.001), B-type brain natriuretic peptide plasma levels (P = 0.02), cardiac outputs (P = 0.007) and pulmonary vascular resistances (P = 0.01) after 19 +/- 6.3 months. Treprostinil plasma concentrations correlated with drug dose (P < 0.001), indicating stable absorption over time. Long-term survival was significantly better than in controls. Treprostinil improves exercise capacity, hemodynamics and survival in patients with severe inoperable CTEPH. We speculate that the effects may be explained by a combined vasodilatory, platelet-antagonistic and potential antiproliferative action of the drug.


Identification of compounds in the essential oil of nutmeg seeds (Myristica fragrans Houtt.) that inhibit locomotor activity in mice.
The present study was designed to evaluate the inhibitory effect of nutmeg (Myristica fragrans Houtt.) seed essential oil on the locomotor activity of mice in a wheel cage. Active compounds in the essential oil were identified by off-line solid phase extraction (SPE-C18) and GC/MS analysis. The essential oil was administered by inhalation at doses of 0.1, 0.3, and 0.5 mL/cage. The results showed that inhalation of nutmeg seed essential oil at a dose of 0.5 mL/cage decreased locomotion by 68.62%; and inhalation of 0.1 and 0.3 mL/cage inhibited locomotion by 62.81% and 65.33%, respectively. Generally, larger doses and longer administrations of nutmeg seed essential oil exhibited greater locomotor inhibition. Subsequently, the plasma concentrations of essential oil compounds were measured. The most concentrated compound in the plasma was myristicin. Half an hour after the addition of 1 mL/cage of nutmeg seed oil, the plasma concentration of myristicin was 3.7 μg/mL; one and two hours after the addition, the blood levels of myristicin were 5.2 μg/mL and 7.1 μg/mL, respectively. Other essential oil compounds identified in plasma were safrole (two-hour inhalation: 1.28 μg/mL), 4-terpineol (half-hour inhalation: 1.49 μg/mL, one-hour inhalation: 2.95 μg/mL, two-hour inhalation: 6.28 μg/mL) and fatty esters. The concentrations of the essential oil compounds in the blood plasma were relatively low (μg/mL or ppm). In conclusion, the volatile compounds of nutmeg seed essential oil identified in the blood plasma may correlate with the locomotor-inhibiting properties of the oil when administered by inhalation.


Regulation of interferon-alpha responsiveness by the duration of Janus kinase activity.
Daudi B lymphoblastoid cells are highly sensitive to the anti-growth and anti-viral effects of interferon (IFN). Unlike many cell lines, these cells show prolonged transcription of IFN-stimulated genes following treatment with IFN-alpha. This prolonged response correlated with the continued presence of the activated transcription factor, IFN-stimulated gene factor 3 (ISGF3). Pulse-chase labeling experiments indicated that the half-life of the phosphorylation of signal transducers and activators of transcription (Stat)1 and Stat2 was short (<2 h) although the turnover of the proteins themselves was slow (>24 h), indicative of a constitutive phosphatase activity. The administration of protein-tyrosine kinase inhibitors at any time point during IFN stimulation led to rapid inhibition of the response, indicating that tyrosine kinase activity was continuously required. Catalytic activity of Jak1 and Tyk2 kinases remained elevated for prolonged periods following stimulation. Continuous presence of IFN-alpha was necessary for maintaining prolonged activation of ISGF3 and of Janus kinases, an activity that was blocked by antibodies to IFN-alpha or by cycloheximide. Conditioned medium of IFN-alpha-stimulated cells was capable of stimulating STAT activation in naive cells. Taken together, these results suggest that the response to IFN-alpha is controlled by the duration of stimulated Janus kinase activity over the background of constitutive dephosphorylation and that this response can be sustained by autocrine secretion of IFN-alpha.


Role of an estrogen receptor-dependent mechanism in the regulation of estrogen receptor mRNA in MCF-7 cells.
We have previously demonstrated that regulation of estrogen receptor (ER) expression in MCF-7 breast cancer cells is a complex process involving transcriptional and posttranscriptional regulation by estradiol. Treatment of MCF-7 cells with estradiol results in the down-regulation of receptor expression; posttranscriptional suppression of receptor mRNA appears to be the predominant mechanism. To determine whether posttranscriptional regulation of ER gene expression is mediated by an ER-dependent mechanism independent of protein synthesis, we have used the competitive estrogen antagonist, 4-hydroxytamoxifen, and the inhibitor of protein synthesis, cycloheximide, to study regulation of ER mRNA by estradiol. 4-Hydroxytamoxifen had no effect on the steady-state level of receptor mRNA and effectively blocked the suppression of ER mRNA by estradiol. The metabolic inhibitor, cycloheximide, was unable to prevent the estrogen induced decrease in ER mRNA. These data provide evidence that the posttranscriptional suppression of ER expression through estradiol is mediated through the ER independent of protein synthesis. A study of the effects of estradiol on the steady-state levels of nuclear and cytoplasmic receptor mRNA suggest that posttranscriptional suppression is a nuclear event.


[Functional role of vacuolization of the T-system of skeletal muscle fibers].
T-tubules of skeletal muscle fibres easily transform into large vacuoles under the influence of various factors. These include osmotic shock produced by the efflux of small molecular weight molecules (e.g. glycerol), hypertonic shock, muscle fatigue and muscle damage. In most cases, vacuolation is reversible but the molecular mechanisms involved are not clear. Also, the functional role of reversible vacuolation has not been established. However, three possibilities may be considered. (1) Redistribution of ions and water between the cytoplasm and the extracellular space comprised by the T-system. Thus, the formation of large vacuoles may be a mechanisms for rapid osmoregulation that corresponds to regulated volume decrease in other types of cell. However, in our hands, inhibitors of various pathways that participate in volume regulation had no effect on reversible vacuolation. (2) Resealing of mechanical damage of the plasma membrane. This is usually accompanied by the development near the damaged membrane of numerous vacuoles which we have observed by confocal microscopy and use of a hydrophobic dye (RH414), to arise in part from T-tubules. (3) By confocal microscopy, it has also been shown that extracellular fluorescein dextran (Mr = 10,000), and both plasmid DNA (pUC18) and sonicated high molecular weight DNA stained with YOYO, enter vacuoles derived from T-tubules. This finding may indicate that reversible vacuolation, in the absence of membrane damage, could provide a pathway from the extracellular environment to the cytoplasm that is additional or complimentary to endocytosis; it may also be particularly relevant to the ability of muscle to be transfected by the direct injection of DNA. These several observations strongly indicate that the function of the T-system in skeletal muscle fibers is not restricted to excitation-contraction coupling.


The dermoscopic features in infantile hemangioma.
Dermoscopy is useful for evaluating the precise vascular structure. We recently applied dermoscopy to infantile hemangiomas without proliferation and showed polymorphous vascular structures. We now demonstrate two typical dermoscopic figures of proliferative infantile hemangiomas. One was a superficial hemangioma which showed polymorphous vascular structure without obvious red linear vessels and red dilated vessels. The other was a thicker hemangioma with a deeper component. This hemangioma had polymorphous vascular structures with red linear and red dilated vessels. These observations indicates the usefulness of dermoscopy for subclassifying infantile hemangioma.


Preliminary clinical study on perforating prosthokeratoplasty.
To observe the effects of perforating prosthokeratoplasty on patients with leucoma who failed in keratoplasty or were not suitable for keratoplasty, and improved their vision. Five cases with leucoma (4 with chemical burn and 1 with blast) received Yakimenko Style keratoprosthesis implantation. Preoperative examination showed the visual acuity in 4 of the 5 cases was light perception, and that of the other one was FC/20 cm. The light orientation in 3 patients was definite, and that in the other two was indefinite. The vision improved in 4 of 5 patients in the follow-up period of 9 months to 3 years. Their visual acuity showed 0.09 to 0.8. And there was no change of vision in the other 1 case. Prosthokeratoplasty is the first choice for rehabilitation of the blind that have leucoma but are not suitable for or failed in penetrating keratoplasty.


Salivary biomarkers, vital signs and behaviour of pre-school children during their first dental visit.
Anxiety may influence dental treatment unfavourably, thus evaluation of the psychophysiological reactions of the child may reduce its negative effects. This study aimed to evaluate and correlate the levels of cortisol and alpha-amylase, vital signs and behaviour of children during their first dental visit. Study sample consisted of 32 children (11 male and 21 female patients) aged between 4 and 6 years, who would go to the dentist for the first time, with no pain or chronic illness, and without the use of medication. Three saliva samples were collected: before, immediately after and 20 minutes after the dental procedure (dental prophylaxis). Data were analysed using Student's t test and Pearson's correlation test (p = 0.05). In the comparison of the levels of alpha-amylase, the heart rate and blood pressure were higher after the dental visit and the levels of salivary cortisol were significantly higher before. There was no statistically significant difference in the correlation between the Frankl Behaviour Rating Scale and the levels of cortisol and alpha-amylase, but the results showed that the levels of these two salivary biomarkers increased gradually as the child had a less cooperative behaviour. CONCLUSION The sample revealed that the first dental visit generated anxiety, which was manifested through physiological and behavioural alterations.


Initial refinement of data from video-based single-cell tracking.
Video recording of cells offers a straightforward way to gain valuable information from their response to treatments. An indispensable step in obtaining such information involves tracking individual cells from the recorded data. A subsequent step is reducing such data to represent essential biological information. This can help to compare various single-cell tracking data yielding a novel source of information. The vast array of potential data sources highlights the significance of methodologies prioritizing simplicity, robustness, transparency, affordability, sensor independence, and freedom from reliance on specific software or online services. The provided data presents single-cell tracking of clonal (A549) cells as they grow in two-dimensional (2D) monolayers over 94 hours, spanning several cell cycles. The cells are exposed to three different concentrations of yessotoxin (YTX). The data treatments showcase the parametrization of population growth curves, as well as other statistical descriptions. These include the temporal development of cell speed in family trees with and without cell death, correlations between sister cells, single-cell average displacements, and the study of clustering tendencies. Various statistics obtained from single-cell tracking reveal patterns suitable for data compression and parametrization. These statistics encompass essential aspects such as cell division, movements, and mutual information between sister cells. This work presents practical examples that highlight the abundant potential information within large sets of single-cell tracking data. Data reduction is crucial in the process of acquiring such information which can be relevant for phenotypic drug discovery and therapeutics, extending beyond standardized procedures. Conducting meaningful big data analysis typically necessitates a substantial amount of data, which can stem from standalone case studies as an initial foundation.


[A method of examining the seminal fluid].
Clinical trails of a photo-television device for the assessment of the semen by the count of spermatozoa, their mobility and velocity were carried out. Preliminary treatment of the sperm sample TV image, used in this device, helps essentially improve the photomethod resolution. A method for the quantitative estimation of the results of microscopic examination of the semen is described. Use of this device for the assessment of the semen samples from men whose marriage is infertile has shown that gametic velocity and number of their mobile forms are the most informative characteristics of these cells.


Third-trimester fetoscopic ablation therapy for types II and III vasa previa.
Vasa previa is an obstetrical condition in which fetal vessels located near the cervix traverse the fetal membranes unprotected by underlying placenta. Type I vasa previa arises directly from a velamentous cord root, whereas types II and III arise from an accessory lobe or a distal lobe of the same placenta, respectively. Fetoscopic laser ablation for types II and III vasa previa is a novel therapeutic option with benefits that include surgical resolution of the vasa previa, avoidance of prolonged hospitalization, and opportunity for a term vaginal delivery. The potential risks of fetoscopy can be mitigated by delaying laser surgery until a gestational age of 31 to 33 weeks, immediately before anticipated hospitalized surveillance. This study aimed to assess feasibility and outcomes of types II and III vasa previa patients treated via fetoscopic laser ablation in the third trimester. This is a retrospective study of singleton pregnancies with types II and III vasa previa treated with fetoscopic laser ablation at a gestational age ≥31 weeks at a single center between 2006 and 2022. Pregnancy and newborn outcomes were assessed. Continuous variables are expressed as mean±standard deviation. Of 84 patients referred for vasa previa, 57 did not undergo laser ablation: 19 either had no or resolved vasa previa, 25 had type I vasa previa (laser-contraindicated), and 13 had type II or III vasa previa but declined laser treatment. Of the remaining 27 patients who underwent laser ablation, 7 were excluded (laser performed at <31 weeks and/or twins), leaving 20 study patients. The mean gestational age at fetoscopic laser ablation was 32.0±0.6 weeks, and total operative time was 62.1±19.6 minutes. There were no perioperative complications. All patients had successful occlusion of the vasa previa vessels (1 required a second procedure). All patients were subsequently managed as outpatients. The mean gestational age at delivery was 37.2±1.8 weeks, the mean birthweight was 2795±465 g, and 70% delivered vaginally. Neonatal intensive care unit admission occurred in 3 cases: 1 for respiratory distress syndrome and 2 for hyperbilirubinemia requiring phototherapy. There were no cases of neonatal transfusion, intraventricular hemorrhage, sepsis, patent ductus arteriosus, or death. Laser ablation for types II and III vasa previa at 31 to 33 gestational weeks was technically achievable and resulted in favorable outcomes.


[The physically ill patient needs psychotherapy].
Any somatic diseased should be offered (additional) psychotherapy and many patients are able to profit a good deal by it. This accounts generally not only in the defined "psychosomatic illnesses". Specific indications according to target symptoms have to be applied; but no primary exclusion! This is a big challenge for a new generation of psychotherapists, having almost not been regarded in the classical "psychotherapeutic schools" up to now. It wants regarding of hic et nunc during the acute crises of disease, should however be in fluent transition to long term psychotherapy. It accounts specially for the rehabilitation patient, the incurable patient, the geriatric patient in general. Relatives as well as the nursing personal must be included in the sense of a "systemic approach". This shows, that one single psychotherapeutic school cannot be prepared to fulfill these demands. Psychotherapeutic "schools" present an important basis, should however lead towards an "integrated psychotherapy" to be problem-centered, patient-centered and school-pluralistic. Psychotherapy in somatic illness shows this demands very distinctly. We however consider the "integrated approach" a main street for future psychotherapy in general.


Autoimmunity to tetraspanin-7 in type 1 diabetes.
Type 1 diabetes is an autoimmune disease whereby components of insulin-secreting pancreatic beta cells are targeted by the adaptive immune system leading to the destruction of these cells and insulin deficiency. There is much interest in the development of antigen-specific immune intervention as an approach to prevent disease development in individuals identified as being at risk of disease. It is now recognised that there are multiple targets of the autoimmune response in type 1 diabetes, the most recently identified being a member of the tetraspanin family, tetraspanin-7. The heterogeneity of autoimmune responses to different target antigens complicates the assessment of diabetes risk by the detection of autoantibodies, as well as creating challenges for the design of strategies to intervene in the immune response to these autoantigens. This review describes the discovery of tetraspanin-7 as a target of autoantibodies in type 1 diabetes and how the detection of autoantibodies to the protein provides a valuable marker for future loss of pancreatic beta-cell function.


Dissecting embryonic and extraembryonic lineage crosstalk with stem cell co-culture.
Embryogenesis necessitates harmonious coordination between embryonic and extraembryonic tissues. Although stem cells of both embryonic and extraembryonic origins have been generated, they are grown in different culture conditions. In this study, utilizing a unified culture condition that activates the FGF, TGF-β, and WNT pathways, we have successfully derived embryonic stem cells (FTW-ESCs), extraembryonic endoderm stem cells (FTW-XENs), and trophoblast stem cells (FTW-TSCs) from the three foundational tissues of mouse and cynomolgus monkey (Macaca fascicularis) blastocysts. This approach facilitates the co-culture of embryonic and extraembryonic stem cells, revealing a growth inhibition effect exerted by extraembryonic endoderm cells on pluripotent cells, partially through extracellular matrix signaling. Additionally, our cross-species analysis identified both shared and unique transcription factors and pathways regulating FTW-XENs. The embryonic and extraembryonic stem cell co-culture strategy offers promising avenues for developing more faithful embryo models and devising more developmentally pertinent differentiation protocols.


Intra-Day Solar Power Forecasting Strategy for Managing Virtual Power Plants.
Solar energy penetration has been on the rise worldwide during the past decade, attracting a growing interest in solar power forecasting over short time horizons. The increasing integration of these resources without accurate power forecasts hinders the grid operation and discourages the use of this renewable resource. To overcome this problem, Virtual Power Plants (VPPs) provide a solution to centralize the management of several installations to minimize the forecasting error. This paper introduces a method to efficiently produce intra-day accurate Photovoltaic (PV) power forecasts at different locations, by using free and available information. Prediction intervals, which are based on the Mean Absolute Error (MAE), account for the forecast uncertainty which provides additional information about the VPP node power generation. The performance of the forecasting strategy has been verified against the power generated by a real PV installation, and a set of ground-based meteorological stations in geographical proximity have been used to emulate a VPP. The forecasting approach is based on a Long Short-Term Memory (LSTM) network and shows similar errors to those obtained with other deep learning methods published in the literature, offering a MAE performance of 44.19 W/m2 under different lead times and launch times. By applying this technique to 8 VPP nodes, the global error is reduced by 12.37% in terms of the MAE, showing huge potential in this environment.


Evaluation of the protective efficacy of the killed Coccidioides immitis spherule vaccine in humans. The Valley Fever Vaccine Study Group.
A vaccine consisting of formaldehyde-killed spherules of Coccidioides immitis, previously shown to provide protection against development of lethal coccidioidomycosis in laboratory animals, was evaluated in humans. This double blind "Phase 3" study, conducted during the period 1980 to 1985, involved 2,867 healthy subjects with no history of coccidioidomycosis and negative skin tests. Randomized into vaccine (n = 1,436) or placebo (n = 1,431) groups, the former received three intramuscular injections of 1.75 mg (dry weight) of spherules, the latter received three injections of sterile 0.85% NaCl solution. Compatible clinical presentation with cultural or serologic findings permitted detection of coccidioidomycosis. Of those receiving vaccine, nine developed coccidioidomycosis and nine additional were suspected of having the disease. Of the group receiving placebo, 12 developed coccidioidomycosis, and 13 additional were suspected of having the disease. All cases and suspected cases were mild. Under the conditions of this study, a slight but statistically insignificant reduction in the incidence of coccidioidomycosis was noted in the vaccinated group compared with that in the placebo group, and there was no discernible difference in severity of disease between these groups.


Loss of genetic diversity, recovery and allele surfing in a colonizing parasite, Geomydoecus aurei.
Understanding the genetic consequences of changes in species distributions has wide-ranging implications for predicting future outcomes of climate change, for protecting threatened or endangered populations and for understanding the history that has led to current genetic patterns within species. Herein, we examine the genetic consequences of range expansion over a 25-year period in a parasite (Geomydoecus aurei) that is in the process of expanding its geographic range via invasion of a novel host. By sampling the genetics of 1,935 G. aurei lice taken from 64 host individuals collected over this time period using 12 microsatellite markers, we test hypotheses concerning linear spatial expansion, genetic recovery time and allele surfing. We find evidence of decreasing allelic richness (AR) with increasing distance from the source population, supporting a linear, stepping stone model of spatial expansion that emphasizes the effects of repeated bottleneck events during colonization. We provide evidence of post-bottleneck genetic recovery, with average AR of infrapopulations increasing about 30% over the 225-generation span of time observed directly in this study. Our estimates of recovery rate suggest, however, that recovery has plateaued and that this population may not reach genetic diversity levels of the source population without further immigration from the source population. Finally, we employ a grid-based sampling scheme in the region of ongoing population expansion and provide empirical evidence for the power of allele surfing to impart genetic structure on a population, even under conditions of selective neutrality and in a place that lacks strong barriers to gene flow.


The lived experience of fathers of preterm infants in the Neonatal Intensive Care Unit: a systematic review of qualitative studies.
To systematically review the experience of fathers of preterm infants hospitalised in the Neonatal Intensive Care Unit. Family-centred care is more and more acknowledged in Neonatal Intensive Care Units, advocating for active engagement of both parents in the care journey. Nonetheless, fathers' Neonatal Intensive Care Unit experience has received limited research attention. Systematic review of qualitative studies. Four electronic databases (CINHAL, ISI Web of Science, PubMed, Scopus) were explored and studies published between 2000-2014 were included. Preferred Reporting Item for Systematic Reviews and Meta-analysis (PRISMA) and Joanna Briggs Institute (JBI) Critical Appraisal Tool for Qualitative Studies guidelines were adopted. Key themes were extracted and synthesised. Five main themes resuming fathers' experience of preterm birth and Neonatal Intensive Care Unit stay were identified from 14 studies. Themes were: emotional roller-coaster, paternal needs, coping strategies, self-representation and caregiving engagement. These dimensions were found to be dynamically shaped across three critical turning points: preterm birth, Neonatal Intensive Care Unit stay and at home. Neonatal Intensive Care Unit fathers of preterm infants experience ambivalence, a set of different needs and coping strategies. They modify their self-representations along the Neonatal Intensive Care Unit journey and needs specific nursing support and intervention to sustain caregiving engagement and transition to parenthood. A systematic and deepened understanding of preterms' fathers lived experience in Neonatal Intensive Care Unit would be helpful to inform nursing practice. Specific action priorities are suggested within the frame of family-centred care.


[Radial methods of investigation in expert assessment of some liver disease in pilots].
Potentialities of the present-day methods of radial diagnostics of some liver diseases in pilots have been studied. 79 pilots with liver diseases were investigated. A high degree of correlation between the changes observed in laboratory studies and ultrasonic picture of the liver has been revealed. The pattern of these changes is considered to be a result of chronic disorder of the liver blood flow caused by decreasing of pressure in the liver arteries and increasing of the venous pressure during accelerations. The fan-shaped type of blood circulation infers that these changes propagate in the periphery-to-center direction and, therefore, changes in the peripheral parts of the right lobe of the liver, i.e. the most poorly blood supplied regions, are initiated by the developing connective tissue component. The ultrasonic investigation proved to be the major and most efficient diagnostic procedure of radial investigation within the program of medical qualification of flying personnel with liver diseases. Methods of mathematical analysis raised the value of diagnostics in 79.4 per cent of the observations through assessment of the quantitative indices of the severity of changes in the liver parenchyma.


Impact of multiple micronutrient supplementation ("sprinkles") on iron deficiency anemia in Bedouin Arab and Jewish infants.
A high prevalence of iron deficiency anemia persists in Bedouin Arab and Jewish pediatric populations in southern Israel. To compare the effect of daily use of the micronutrient supplementation (MMS), "Sprinkles," a powdered formulation of iron, vitamins A and C, folic acid and zinc, with liquid iron and vitamins A and D on iron deficiency at 12 months of age. The 621 eligible Bedouin and Jewish infants in the study were assigned to the MMS and control arms and received supplementations from age 6 to 12 months. We examined the change in hemoglobin, hematocrit, mean cell volume, red blood cell distribution, serum ferritin and transferrin saturation. In addition, we used the high Iron Deficiency Index (IDI) if two or more of the above six parameters showed abnormal levels. Rates of anemia decreased significantly over the 6 month period, from 58.8% to 40.6% among Bedouin infants (P = 0.037) and from 40.6 to 15.8% among Jewish infants (P = 0.017). In Bedouin infants the prevalence of high IDI decreased significantly from 79.2% to 67.4% (P = 0.010) in the MMS group, but there was no change in the controls. Among Jewish infants, the high IDI prevalence decreased from 67% to 55.6% with no statistically significant difference in the two study arms. In the multivariate analysis in Bedouin infants MMS use was associated with a reduced risk of 67% in high IDI at age 12 months as compared to controls (P = 0.001). Fewer side effects in the intervention groups in both ethnic populations were reported. MMS fortification of home food can be recommended as an effective and safe method for preventing iron deficiency anemia at 12 months of age.


Retinal detachment after silicone oil removal is prevented by 360 degrees laser treatment.
To investigate the effect of 360 degrees laser retinopexy on the incidence of retinal detachment (RD) after silicone oil removal. In a prospective, randomised clinical trial, 303 patients (303 eyes) affected with primary (n = 211) or recurring (n = 92) rhegmatogenous RD treated by vitrectomy with silicone oil (1000 cSt) and endolaser photocoagulation of retinal breaks were randomised to receive 360 degrees laser retinopexy or not. After at least 4 months, in eyes with a fully attached retina, the silicone oil was removed. The incidence of RD after silicone oil removal was evaluated. 151 eyes received 360 degrees laser retinopexy (completed intraoperatively in 93 eyes and postoperatively in 58 eyes, in nine of them after cataract extraction); 152 eyes served as controls. Silicone oil was removed from 139 laser-treated eyes (92.05%) and 129 controls (84.87%; NS). In the first group, 12 eyes (8.63%) developed an RD posterior to laser treatment (including the macula), three eyes (2.16%) had a localised posterior RD (treated by laser), and 14 (10.07%) had an RD anterior to the laser treatment. In the control group, RD occurred in 27 eyes (20.93%; p = 0.007). 360 degrees laser retinopexy reduces the incidence of RD after silicone oil removal; it should be completed intraoperatively.


Genome-wide analysis study of gestational diabetes mellitus and related pathogenic factors in a Chinese Han population.
Gestational diabetes mellitus (GDM) affects the metabolism of both the mother and fetus during and after pregnancy. Genetic factors are important in the pathogenesis of GDM, and associations vary by ethnicity. However, related studies about the relationship between the susceptibility genes and glucose traits remain limited in China. This study aimed to identify genes associated with GDM susceptibility in Chinese Han women and validate those findings using clinical data during pregnancy and postpartum period. A genome-wide association study (GWAS) of 398 Chinese Han women (199 each with and without GDM) was conducted and associations between single nucleotide polymorphisms (SNPs) and glucose metabolism were identified by searching public databases. Relationships between filtered differential SNPs and glucose metabolism were verified using clinical data during pregnancy. The GDM group were followed up postpartum to evaluate the progression of glucose metabolism. We identified five novel SNPs with genome-wide significant associations with GDM: rs62069863 in TRPV3 gene and rs2232016 in PRMT6 gene were positive correlated with 1 h plasma glucose (1hPG) and 2 h plasma glucose (2hPG), rs1112718 in HHEX/EXOC6 gene and rs10460009 in LPIN2 gene were positive associated with fasting plasma glucose, 1hPG and 2hPG, rs927316 in GLIS3 gene was negative correlated with 2hPG. Of the 166 GDM women followed up postpartum, rs62069863 in TRPV3 gene was positively associated with fasting insulin, homoeostasis model assessment of insulin resistance. The variants of rs62069863 in TRPV3 gene, rs2232016 in PRMT6 gene, rs1112718 in HHEX/EXOC6 gene, rs927316 in GLIS3 gene, and rs10460009 in LPIN2 gene were newly-identified susceptibility loci for GDM in the Chinese Han population. TRPV3 was associated with worse insulin resistance postpartum. This study was registered in the Chinese Clinical Trial Registry. ChiCTR2100043762. Date of first registration: 28/02/2021.


Prospective on using fibre mid-infrared supercontinuum laser sources for in vivo spectral discrimination of disease.
Mid-infrared (MIR) fibre-optics may play a future role in in vivo diagnosis of disease, including cancer. Recently, we reported for the first time an optical fibre based broadband supercontinuum (SC) laser source spanning 1.3 to 13.4 μm wavelength to cover the spectral 'fingerprint region' of biological tissue. This work has catalysed the new field of fibre MIR-SC and now very bright sources equivalent to a 'few synchrotrons' have been demonstrated in fibre. In addition, we have made record transparency MIR fibre for routeing the MIR light and reported first-time MIR photoluminescence (with long lifetime) in small-core, rare earth ion doped, MIR fibre - an important step towards MIR fibre lasing at >4 μm wavelength for pumping fibre MIR-SC. First time fibre MIR-SC spectroscopic imaging of colon tissue is described at wavelengths in the 'fingerprint region'.


Ultra-Processing or Oral Processing? A Role for Energy Density and Eating Rate in Moderating Energy Intake from Processed Foods.
Recent observational data and a controlled in-patient crossover feeding trial show that consumption of "ultra-processed foods" (UPFs), as defined by the NOVA classification system, is associated with higher energy intake, adiposity, and at a population level, higher prevalence of obesity. A drawback of the NOVA classification is the lack of evidence supporting a causal mechanism for why UPFs lead to overconsumption of energy. In a recent study by Hall the energy intake rate in the UPF condition (48 kcal/min) was >50% higher than in the unprocessed condition (31 kcal/min). Extensive empirical evidence has shown the impact that higher energy density has on increasing ad libitum energy intake and body weight. A significant body of research has shown that consuming foods at higher eating rates is related to higher energy intake and a higher prevalence of obesity. Energy density can be combined with eating rate to create a measure of energy intake rate (kcal/min), providing an index of a food's potential to promote increased energy intake. The current paper compared the association between measured energy intake rate and level of processing as defined by the NOVA classification. Data were pooled from 5 published studies that measured energy intake rates across a total sample of 327 foods. We show that going from unprocessed, to processed, to UPFs that the average energy intake rate increases from 35.5 ± 4.4, to 53.7 ± 4.3, to 69.4 ± 3.1 kcal/min (P < 0.05). However, within each processing category there is wide variability in the energy intake rate. We conclude that reported relations between UPF consumption and obesity should account for differences in energy intake rates when comparing unprocessed and ultra-processed diets. Future research requires well-controlled human feeding trials to establish the causal mechanisms for why certain UPFs can promote higher energy intake.


[The swimmer's otitis. An up to date and prevention options].
The swimmer's otitis or acute otitis externa, is a pathology that often occurs in those who practice swimming at a competitive level. The same problem often occurs in the summer with the attendance of swimming pools and bathing areas. A survey made in the United States in 2007 confirms the dynamics of the onset of this pathology, because the contamination of fungi and bacteria in the waters of the swimming pools and the sea cause the inflammation of the epithelium of the auditory canal. To face this issue, in addition to a correct diagnosis, and the necessary and appropriate therapies, it may follow the use of these medicinal preparations. The first one (A) is protective-acting, the second one (B) is characterized as a preventive, hygroscopic, moistening, antiseptic and antimycotic-acting solution. Swimmers who have been using the two (pre-and post) preparations daily for about a year have reported sporadic episodes of otitis. However, there is no doubt that these two compounds deserve a meticulous clinical trial in order to confirm their preventive and therapeutic potentials in external acute otitis.


Medical ethics course for undergraduate medical students: a needs assessment study.
Education needs assessment is one of the essential components of curriculum development. In this study, we aimed to assess the educational needs of general physicians for medical ethics. We conducted a three-stage Delphi study of general physicians' views on important ethical issues in their practice. In the item generation stage we retrieved 45 important educational items from a survey of general physicians, patients, well known ethical clinicians, and a review of other universities' curricula and international literature. The questionnaire was designed to ask the importance of each generated item. We then sent the questionnaire to general physicians. Items scored as highly important by more than 80% of the respondents in the first or second consensus development surveys were considered as educational priorities. Four academic medical ethics teachers reviewed and commented on the findings. The response rate to the first consensus development survey was 38%, of whom 77% also responded to the second survey. We developed consensus on 24 medical ethics items for inclusion in medical ethics curriculum. All items were also considered important by medical ethics teachers, and they added four further items to the list. Despite the attention given to ethical issues originating from technological advances, the most important educational needs of general physicians in medical ethics are still the traditional issues concerning the doctor-patient relationship and professionalism.


[Osteoradionecrosis in adults].
Osteoradionecrosis (ORN) is not exceptional, despite advances in irradiation techniques. Six cases are reported, involving the pelvis, coxofemoral joint, mandible and vertebrae; in this last case, the semiologic value of the lucent intrasomatic image seen on plain films and tomographies of the vertebrae is underscored. The irradiation dose (above 3 000 rad) is the chief factor in osteoradionecrosis, which may be precipitated by adjuvant factors and potentiating events such as trauma and infection. Pathologic study shows several lesions whose association is suggestive: cell lesions, osteoporosis, vascular lesions, and foci of necrosis. The pathogenic significance of lesions of bone cells is demonstrated, while the part played by vascular lesions is controversial. Involvement of the pelvis and hips following irradiation of pelvic carcinoma is the most common. The scapular girdle and ribs may be involved in irradiation for breast cancer. In involvement of the mandible, remarkable features are its frequency following irradiation of carcinoma of the mouth, the significant part played by potentiating factors, i.e. infection and trauma, severity of complications, i.e. fistulae and hemorrhage, and lastly difficulties of management. Among infrequent sites, involvement of the vertebrae is of interest as it may mimic collapse due to osteoporosis or metastasis. Diagnosis rests on an association of criteria, and fortunately bone biopsy is usually unnecessary. The clinical features, topographical characteristics and course of the disease allow differentiation from bone metastasis; it may be more difficult to distinguish postirradiation sarcoma, which is exceptional, or a number of benign conditions, such as aseptic necrosis, infectious osteoarthritis, and destructive coxarthrosis.


Therapeutic impact of erythropoietin-encapsulated liposomes targeted to bone marrow on renal anemia.
Bone marrow is a key element in the diagnosis of disorders of erythropoiesis, including anemia, and a potential target in their treatment. However, because efficient delivery of diagnostic and therapeutic agents to bone marrow is difficult, such delivery is achieved by administering drugs in large quantities that often have adverse effects. Here, we achieved selective delivery of recombinant human erythropoietin (rHuEPO) to bone marrow, via its encapsulation in liposomes with l-glutamic acid, N-(3-carboxy-1-oxopropyl)-, 1,5-dihexadecyl ester (SA) (liposome-EPO). The result, in a rabbit model of renal anemia, was a beneficial effect on hematopoiesis, better than with rHuEPO alone. Also, we determined that liposome-EPO delivery to bone marrow depended on specific uptake by bone marrow macrophages because of the presence of SA. These results indicate both that liposome-EPO is a new, promising erythropoietin-stimulating agent and that liposomes with SA have potential for diagnostic and therapeutic applications in diseases originating from bone marrow.


STAT6 expression in glioblastoma promotes invasive growth.
Glioblastoma (GBM) is a highly aggressive malignant primary brain tumor, characterized by rapid growth, diffuse infiltration of cells into both adjacent and remote brain regions, and a generalized resistance to currently available treatment modalities. Recent reports in the literature suggest that Signal Transducers and Activators of Transcription (STATs) play important roles in the regulation of GBM pathophysiology. STAT6 protein expression was analyzed by Western blotting in GBM cell lines and by immunohistochemistry in a tissue microarray (TMA) of glioma patient tissues. We utilized shRNA against STAT6 to investigate the effects of prolonged STAT6 depletion on the growth and invasion of two STAT6-positive GBM cell lines. Cell proliferation was assessed by measuring (3)H-Thymidine uptake over time. Invasion was measured using an in vitro transwell assay in which cells invade through a type IV collagen matrix toward a chemoattractant (Fetal Bovine Serum). Cells were then stained and counted. Kaplan-Meyer survival curves were generated to show the correlation between STAT6 gene expression and patient survival in 343 glioma patients and in a subset of patients with only GBM. Gene expression microarray and clinical data were acquired from the Rembrandt 1 public data depository (https://caintegrator.nci.nih.gov/rembrandt/). Lastly, a genome-wide expression microarray analysis was performed to compare gene expression in wild-type GBM cells to expression in stable STAT6 knockdown clones. STAT6 was expressed in 2 GBM cell lines, U-1242MG and U-87MG, and in normal astrocytes (NHA) but not in the U-251MG GBM cell line. In our TMA study, STAT6 immunostaining was visible in the majority of astrocytomas of all grades (I-IV) but not in normal brain tissue. In positive cells, STAT6 was localized exclusively in the nuclei over 95% of the time. STAT6-deficient GBM cells showed a reduction in (3)H-Thymidine uptake compared to the wild-type. There was some variation among the different shRNA- silenced clones, but all had a reduction in (3)H-Thymidine uptake ranging from 35%- 70% in U-1242MG and 40- 50% in U-87MG cells. Additionally, STAT6- depleted cells were less invasive than controls in our in vitro transmembrane invasion assay. Invasiveness was decreased by 25-40% and 30-75% in U-1242MG and U-87MG cells, respectively. The microarray analysis identified matrix metalloproteinase 1 (MMP-1) and urokinase Plasminogen activator (uPA) as potential STA6 target genes involved in the promotion of GBM cell invasion. In a Kaplan-Meier survival curve based on Rembrandt 1 gene expression microarray and clinical data, there was a significant difference in survival (P < 0.05) between glioma patients with up- and down-regulated STAT6. Decreased STAT6 expression correlated with longer survival times. In two subsets of patients with either grade IV tumors (GBM) or Grade II/III astrocytomas, there was a similar trend that however did not reach statistical significance. Taken together, these findings suggest a role for STAT6 in enhancing cell proliferation and invasion in GBM, which may explain why up-regulation of STAT6 correlates with shorter survival times in glioma patients. This report thus identifies STAT6 as a new and potentially promising therapeutic target.


Indications and Outcomes of Secondary Hip Procedures After Failed Hip Arthroscopy: A Systematic Review.
(1) To identify present indications for secondary procedures in patients with failed hip arthroscopy and (2) to assess patient-reported outcomes (PROs) of the secondary procedures, including revision arthroscopy, periacetabular osteotomy (PAO), and total hip arthroplasty (THA). Study groups included patients who had a secondary procedure after failed previous hip arthroscopy whereas the control groups were patients who had a primary procedure but did not require a secondary procedure. Indications and procedures at the time of the secondary operation were documented for each study. Average PROs were recorded, and standardized mean difference was calculated to estimate effect size. Eighteen studies reporting on patients undergoing a secondary procedure after a previous hip arthroscopy were included. The 3 main secondary procedure groups were revision hip arthroscopy, secondary PAO, and secondary THA. Regarding the revision arthroscopy group, the most common indications were labral tears, cam deformity, and pincer deformity. In addition, the most common procedures were femoroplasty, acetabuloplasty, capsular release, and labral reconstruction. The most common indications for the secondary PAO and THA groups were dysplasia and osteoarthritis respectively. Five of the revision arthroscopy studies found that revision patients had worse outcomes than the primary arthroscopy group. One PAO study found that the previous arthroscopy group had slightly worse outcomes, and 2 studies found no differences in PROs. Two THA studies reported worse outcomes for the prior arthroscopy group, and 2 studies reported no differences in outcomes. The most common indications for revision hip arthroscopy were labral tears and femoroacetabular impingement. Patients undergoing a revision hip arthroscopy demonstrated good postoperative outcomes but to an overall lesser extent than their primary counterparts. The secondary PAO and THA groups also had favorable PROs, but the studies were inconclusive in determining superior outcomes between the primary and secondary groups. IV, Systematic review of Level II-IV investigations.


Interstitial cells of cajal are involved in neurotransmission in the gastrointestinal tract.
Interstitial cells of Cajal (ICC) are important cells which coordinate gastrointestinal motility. ICC express Kit receptor tyrosine kinase, and Kit immunohistochemistry reveals ICC morphology and distribution in the gastrointestinal musculature. ICC show a highly branched morphology and form unique networks. Myenteric ICC (ICC-MY) are located at the layer of the myenteric plexus and serve as electrical pacemakers. Intramuscular ICC (ICC-IM) and ICC in the deep muscular plexus (ICC-DMP) are distributed within the muscular layers, and are densely innervated by excitatory and inhibitory enteric motor neurons and in close contact with nerve terminals. Recent studies combined with morphological and functional techniques directly revealed that ICC-IM and ICC-DMP are mediators of enteric motor neuro-transmission. These types of ICC express several receptors for neurotransmitters such as acetylcholine and substance P and show responses to excitatory nerve stimulations. ICC also express receptive mechanisms for nitric oxide, which is an inhibitory neurotransmitter in the gastrointestinal tract. They can respond to nitrergic nerve stimulation by cyclic GMP production. Kit mutant mice lack ICC-IM and show attenuated postsynaptic responses after intrinsic nerve stimulation. These findings indicate the importance for ICC in neurotransmission in the gastrointestinal tract.


Wildfires disproportionately affected jaguars in the Pantanal.
The Pantanal wetland harbours the second largest population of jaguars in the world. Alongside climate and land-use changes, the recent mega-fires in the Pantanal may pose a threat to the jaguars' long-term survival. To put these growing threats into perspective, we addressed the reach and intensity of fires that have affected jaguar conservation in the Pantanal ecoregion over the last 16 years. The 2020 fires were the most severe in the annual series, burned 31% of the Pantanal and affected 45% of the estimated jaguar population (87% of these in Brazil); 79% of the home range areas, and 54% of the protected areas within home ranges. Fires consumed core habitats and injured several jaguars, the Pantanal's apex predator. Displacement, hunger, dehydration, territorial defence, and lower fecundity are among the impacts that may affect the abundance of the species. These impacts are likely to affect other less mobile species and, therefore, the ecological stability of the region. A solution to prevent the recurrence of mega-fires lies in combating the anthropogenic causes that intensify drought conditions, such as implementing actions to protect springs, increasing the number and area of protected areas, regulating fire use, and allocating fire brigades before dry seasons.


[Radiation block of mitoses and the action of stimulating factors].
Using the HeLa cell culture, the dynamics of the appearance of the mitotic block after irradiation with 600 rads has been studied, in addition to the influence of some proliferating stimulatory factors on this process. It has been shown that in the early periods after irradiation with 600 rads, on the background of the developing G2--M block, the low dose irradiation (20 rads), changes to the medium, and high temperature (41--42 degrees C) effects lead to the increase in the number of mitoses. It is supposed that this increase in the mitotic activity may be due to a heterogeneity of cell populations as to their velocity of passing through the G2-phase.


A Domino Route toward Polysubstituted Pyrroles from 2-Imidazolines and Electron-Deficient Alkynes.
The reaction of 1,2-disubstituted 2-imidazolines with electron-deficient alkynes proceeds as a pseudo-three-component process and forms imidazolidines with an N-vinylpropargylamine fragment. Heating the resulting imidazolidines in xylene on air leads to an effective formation of polysubstituted pyrroles through a domino sequence of aza-Claisen rearrangement/transannular nucleophilic addition/oxidative ring opening reactions. The direct one-pot transformation of 2-imidazolines to pyrroles has been also realized.