External carotid artery-radial artery graft-posterior cerebral artery bypass for complex vertebrobasilar aneurysms: efficacy and analysis of outcome in a single center.
The purpose of this research was to demonstrate the effectiveness and clinical outcome of an external carotid artery-radial artery graft-posterior cerebral artery (ECA-RAG-PCA) bypass in the treatment of complex vertebrobasilar artery aneurysms (VBANs) in a single-center retrospective study. An ECA-RAG-PCA bypass may be a last and very important option in the treatment of complex VBANs when conventional surgical clipping or endovascular interventions fail to achieve the desired outcome. This study retrospectively analyzed the clinical presentation, case characteristics, aneurysm location, size and morphology, choice of surgical strategy, complications, clinical follow-up, and prognosis of the patients enrolled. The data involved were analyzed by the appropriate statistical methods. A total of 24 patients with complex VBANs who met the criteria were included in this study. Eighteen (75.0%) were male and the mean age was 54.1 ± 8.83 years. The aneurysms were located in the vertebral artery, the basilar artery, and in the vertebrobasilar artery with simultaneous involvement. All patients underwent ECA-RAG-PCA bypass surgery via an extended middle cranial fossa approach, with 8 (33.3%) undergoing ECA-RAG-PCA bypass only, 3 (12.5%) undergoing ECA-RAG-PCA bypass combined with aneurysm partial trapping, and 12 (50.0%) undergoing ECA-RAG-PCA bypass combined with proximal occlusion of the parent artery. The average clinical follow-up was 22.0 ± 13.35 months. The patency rate of the high-flow bypass was 100%. At the final follow-up, 15 (62.5%) patients had complete occlusion of the aneurysm, 7 (29.2%) patients had subtotal occlusion of the aneurysm, and 2 (8.3%) patients had stable aneurysms. The rate of complete and subtotal occlusion of the aneurysm at the final follow-up was 91.7%. The clinical prognosis was good in 21 (87.5%) patients and no procedure-related deaths occurred. Analysis of the good and poor prognosis groups revealed a statistically significant difference in aneurysm size (P = 0.034, t-test). Combining the results of this study and the clinical experience of our center, we propose a surgical algorithm and strategy for the treatment of complex VBANs.The technical approach of ECA-RAG-PCA bypass for complex VBANs remains important, even in an era of rapid advances in endovascular intervention. When conventional surgical clipping or endovascular intervention has failed, an ECA-RAG-PCA bypass plays a role that cannot be abandoned and is a very important treatment option of last resort.


Immunohistochemical study of angiogenesis and proliferative activity in epiretinal membranes.
Formation of epiretinal membranes (ERMs) is a serious complication of retinal diseases, the most important being proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). In this study, our goal was to (i) calculate the microvessel density (MVD), (ii) evaluate vascular endothelial growth factor (VEGF) expression and (iii) correlate angiogenesis with the proliferative activity as expressed by the expression of Ki67 marker, in both membrane types. We performed immunohistochemistry in 14 PVR and eight PDR membranes, using antibodies against CD34, VEGF, Ki67 and glial fibrillary acidic protein. PDR membranes presented higher average count of microvessels compared with PVR membranes (p = 0.0015). No differences were observed concerning VEGF expression (p = 0.1). The expression of Ki67 was not correlated with microvessel number or VEGF expression. Our study confirms the presence of vascularisation in PDR membranes, as well as the presence of VEGF even in avascular PVR membranes, suggesting that immunoreactivity for VEGF may not be accompanied by angiogenesis.


Age, aging and physiological dysregulation in safety-critical work: a retrospective longitudinal study of helicopter emergency medical services pilots.
Concurrent progressive physiological dysregulation in different organ systems may be a driver of phenotypical frailty and is associated with morbidity and performance declines. In aging societies, health and performance of older workers are given increased scrutiny. However, there are few studies researching the effect of age on physiological dysregulation specifically in the working population and none for high-risk occupations with close medical supervision. Here, we aim to investigate this association cross-sectionally and longitudinally in a healthy sample of European helicopter emergency medical services pilots. Using a retrospective cohort design, we constructed indices of (a) physiological dysregulation state (N = 52 participants) and (b) pace of change (N = 41 participants, average follow-up 7.9 years) based on measurements of 18 health risk-associated biomarkers, as documented in the participating pilots' aeromedical examination records. The relationship of these two indices with average age (range 27.9-60.6 years) and average baseline age (29.3-57.2 years), respectively, was modeled by weighted least squares regression, adjusting for region of origin, smoking, and medication. Longitudinally, dysregulation significantly increased over the follow-up period, but the pace of change did not increase with baseline age. Cross-sectionally, we observed a significant negative quadratic effect of age on physiological dysregulation state, with maximum estimated physiological dysregulation at ages 45-50. As hypothesized, progression of dysregulation over approximately 8 years is observable in a healthy occupational sample but it does not accelerate (i.e., does not increase with baseline age). Increases in dysregulation are counteracted by what appears to be a strong healthy worker survivor effect. The study shows that the development of physiological dysregulation in high-risk occupations differs from that in the general population and that physiological dysregulation indices may be helpful in reconciling safety concerns and fair treatment of aging workers in safety-critical jobs.


Soil edaphic factors and climate seasonality explain the turnover of methanotrophic communities in riparian wetlands.
Aerobic CH4-oxidizing bacteria (methanotrophs) represent a biological model system for the removal of atmospheric CH4, which is sensitive to the dynamics of water tables. However, little attention has been given to the turnover of methanotrophic communities across wet and dry periods in riparian wetlands. Here, by sequencing the pmoA gene, we investigated the turnover of soil methanotrophic communities across wet and dry periods in typical riparian wetlands that experience intensive agricultural practices. The results demonstrated that the methanotrophic abundance and diversity were significantly higher in the wet period than in the dry period, probably owing to the climatic seasonal succession and associated variation in soil edaphic factors. The co-occurrence patterns of the interspecies association analysis demonstrated that the key ecological clusters (i.e., Mod#1, Mod#2, Mod#4, Mod#5) showed contrasting correlations with soil edaphic properties between wet and dry periods. The linear regression slope of the relationships between the relative abundance of Mod#1 and the carbon to nitrogen ratio was higher in the wet period than in the dry period, whereas the linear regression slope of the relationships between the relative abundance of Mod#2 and soil nitrogen content (i.e., dissolved organic nitrogen, nitrate, and total nitrogen) was higher in the dry period than in the wet period. Moreover, Stegen's null model combined with phylogenetic group-based assembly analysis demonstrated that the methanotrophic community exhibited a higher proportion of drift (55.0%) and a lower contribution of dispersal limitation (24.5%) in the wet period than in the dry period (43.8% and 35.7%, respectively). Overall, these findings demonstrate that the turnover of methanotrophic communities across wet and dry periods were soil edaphic factors and climate dependent.


Choosing Wisely for Birth.
In this column, the editor of The Journal of Perinatal Education describes the American Academy of Nursing's engagement in the national Choosing Wisely campaign and how it has implications for promoting normal birth. The editor also describes the contents of this issue, which offer a broad range of resources, research, and inspiration for childbirth educators in their efforts to promote, support, and protect natural, safe, and healthy birth.


Hypertrophic osteoarthropathy: what a rheumatologist should know about this uncommon condition.
This article presents an updated overview of hypertrophic osteoarthropathy and digital clubbing for the practicing rheumatologist. Discussion includes a brief historical perspective, its definition, incidence and prevalence, classification, pathology and pathophysiology, clinical manifestations, demographics, findings on physical examination, imaging techniques for its detection, differential diagnosis, and treatment modalities.


Spatial transcriptomics reveals heterogeneity of macrophages in the tumor microenvironment of granulomatous slack skin.
Granulomatous slack skin (GSS) is an extremely rare subtype of cutaneous T-cell lymphoma accompanied by an abundant number of macrophages and is clinically characterized by the development of pendulous skin folds. However, the characteristics of these macrophages in GSS remain unclear. Here, we conducted a spatial transcriptomic study on one frozen GSS sample and drew transcriptomic maps of GSS for the first time. Gene expression analysis revealed the enrichment of three clusters with macrophage transcripts, each exhibiting distinct characteristics suggesting that their primary composition consists of different subpopulations of macrophages. The CD163+ /CD206+ cluster showed a tumor-associated macrophage (TAM) M2-like phenotype and highly expressed ZFP36, CCL2, TNFAIP6, and KLF2, which are known to be involved in T-cell interaction and tumor progression. The APOC1+ /APOE+ cluster presented a non-M1 or -M2 phenotype and may be related to lipid metabolism. The CD11c+ /LYZ+ cluster exhibited an M1-like phenotype. Notably, these cells strongly expressed MMP9, MMP12, CHI3L1, CHIT1, COL1A1, TIMP1, and SPP1, which are responsible for extracellular matrix (ECM) degradation and tissue remodeling. This may partially explain the symptoms of cutaneous relaxation in GSS. Further immunohistochemistry on four GSS cases demonstrated that CD11c predominantly marked granulomas and multinucleated giant cells, whereas CD163 was mainly expressed on scattered macrophages, appearing as a mutually exclusive pattern. The expression pattern of MMP9 overlapped with that of CD11c, implying that CD11c+ macrophages may be a source of MMP9. Our data shed light on the characteristics of macrophages in the GSS microenvironment and provide a theoretical basis for the application of MMP9 inhibitors to prevent cutaneous relaxation of GSS. © 2023 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.


Headspace gas chromatographic determination of ethanol: the use of factorial design to study effects of blood storage and headspace conditions on ethanol stability and acetaldehyde formation in whole blood and plasma.
Our headspace gas chromatographic flame ionization detection (HS-GC-FID) method for ethanol determination showed slightly, but consistently, low ethanol concentrations in whole blood (blood) in proficiency testing programs (QC-samples). Ethanol and acetaldehyde were determined using HS-GC-FID with capillary columns, headspace equilibration temperature (HS-T degrees ) of 70 degrees C and 20 min equilibration time (HS-EqT). Full factorial designs were used to study the variables HS-T degrees (50 degrees -70 degrees C), HS-EqT (15-25 min), ethanol concentration (0.20-1.20 g/kg) and storage at room temperature (0-6 days) with three sample-sets; plasma, hemolyzed blood and non-hemolyzed blood. A decrease in the ethanol concentration in blood was seen as a nearly equivalent increase in the acetaldehyde concentration. This effect was not observed in plasma, indicating chemical oxidation of ethanol to acetaldehyde in the presence of red blood cells. The variables showed different magnitude of effects in hemolyzed and non-hemolyzed blood. A decrease in ethanol concentration was seen even after a few days of storage and also when changing the HS-T degrees from 50 to 70 degrees C. The formation of acetaldehyde was dependent on all the variables and combinations of these (interactions) and HS-T degrees was involved in all the significant interaction effects. Favorable instrumental conditions were found to be HS-T degrees of 50 degrees C and HS-EqT of 15-25 min. The ethanol concentrations obtained for the range 0.04-2.5 g/kg after analyzing authentic forensic blood samples with a HS-T degrees of 50 degrees C were statistically significantly higher than at 70 degrees C (+0.0154 g/kg, p < 0.0001, n = 180). In conclusion, chemical oxidation of ethanol to acetaldehyde in the presence of red blood cells has been shown to contribute to lowered ethanol concentrations in blood samples. Storage conditions before analysis and the headspace equilibration temperature during analysis were important for the determination of blood ethanol concentrations.


Case report: Patient specific combination of surgery and immunotherapy in advanced squamous cell carcinoma of the head and neck - a case series and review of literature.
Prognosis of patients with recurrent or metastatic head and neck cancer is generally poor. Adjuvant immunotherapy (IT) featuring immune checkpoint inhibition (ICI) is standard of care in advanced stage head and neck squamous cell carcinoma (HNSCC) and cutaneous squamous cell carcinoma (CSCC). ICI response rates in CSCC are described as higher than in HNSCC. IT is constantly shifting into earlier disease stages which confronts the surgeon with immunotherapeutically pre-treated patients. It is therefore becoming increasingly difficult to assess which patients with symptomatic tumor disease and a lack of curative surgical option might benefit from salvage surgery. The following 6 cases describe therapeutic decision-making regarding ICI and (salvage) surgery in patients with advanced stage HNSCC or CSCC. Cases A and B focus on neoadjuvant ICI followed by salvage surgery. In Cases C and D salvage surgery was performed after short-term stabilization with partial response to ICI. The last two cases (Cases E and F) address the surgical approach after failure of ICI. All cases are discussed in the context of the current study landscape and with focus on individual decision-making. For better understanding, a timetable of the clinical course is given for each case. ICI is rapidly expanding its frontiers into the neoadjuvant setting, frequently confronting the surgeon with heavily pretreated patients. Salvage surgery is a viable therapeutic concept despite the rise of systemic treatment options. Decision-making on surgical intervention in case of a salvage surgery remains an individual choice. For neoadjuvant ICI monitoring regarding pathological tumor response or tumor necrosis rate, we suggest correlation between the initial biopsy and the definite tumor resectate in order to increase its significance as a surrogate marker. Scheduling of neoadjuvant ICI should be further investigated, as recent studies indicate better outcomes with shorter time frames.


Associations of the 5-hydroxytryptamine (serotonin) receptor 1B gene (HTR1B) with alcohol, cocaine, and heroin abuse.
Abnormal serotonergic pathways are implicated in numerous neuropsychiatric disorders including alcohol and drug dependence (abuse). The human 5-hydroxytryptamine (serotonin) receptor 1B, encoded by the HTR1B (5-HT1B) gene, is a presynaptic serotonin autoreceptor that plays an important role in regulating serotonin synthesis and release. Although there was evidence of associations of the HTR1B gene variants in the etiologies of substance use disorders, negative findings were also reported. To clarify the roles of commonly reported single nucleotide polymorphisms (SNPs) of the HTR1B gene underlying alcohol and drug dependence (abuse), we performed a meta-analysis based on the available genotype data from individual candidate gene-based association studies. Evidence of association was found between the functional SNP -161A>T (rs130058) and alcohol, cocaine, and heroin dependence (e.g., P = 0.03 and odds ratio (OR) = 1.2 (1.02, 1.42) in the combined European, Asian, African, and Hispanic populations). SNP -261T>G (rs11568817) also showed evidence of association but with different directions in Europeans and non-Europeans (e.g., P = 0.0018 with OR = 1.42 (1.14, 1.76) and P = 0.01 with ORs = 0.5 (0.3, 0.85), respectively). This meta-analysis supports the associations of HTR1B -261T>G and -161A>T with alcohol and drug abuse and further investigations are warranted in larger samples.


Effect of mandibular bone atrophy on maxillary and mandibular bone remodeling and quality of life with an implant-retained mandibular overdenture after 3 years.
The medium-term effect of an implant-retained mandibular overdenture on bone remodeling in the maxilla and posterior mandible of edentulous patients and the effects on quality of life have not been established. The purpose of this prospective observational clinical study was to evaluate the 3-year effects of implant-retained mandibular overdentures on oral-health-related quality of life (OHRQoL) and bone remodeling in different regions of the maxilla and mandible in participants with atrophic or nonatrophic mandibles. Twenty-six edentulous participants received 2 narrow-diameter implants in the anterior mandible. Mandibular bone atrophy was categorized from presurgical panoramic radiographs according to the Cawood and Howell criteria. OHRQoL was assessed by using the OHIP-EDENT questionnaire. Participants were evaluated annually for 3 years to measure the marginal bone loss and bone area of the posterior mandible, and the anterior and posterior regions of the maxilla were assessed annually through panoramic radiographs. The data were analyzed by using a mixed-effects linear regression to estimate time-dependent trends and a mixed-effect linear regression model to verify differences between groups. The Pearson correlation coefficients between bone variables and 3-year OHIP-EDENT outcomes were calculated. In the third year, atrophic participants had a significantly lower marginal bone loss (0.02 mm) than nonatrophic participants (-0.39 mm) (P=.030). Differences were also found in the functional limitation (nonatrophic=1.82 ±1.75, atrophic participants=1.92 ±1.54; P=.018) and handicap domains (nonatrophic=0.36 ±0.54, atrophic participants=0.08 ±0.27; P=.003). For nonatrophic participants, comparisons between baseline and 3-year outcomes showed significant bone resorption as indicated by the area ratio in the anterior maxilla (P=.035), posterior maxilla (P=.022), and posterior mandible (P=.009). Conversely, the bone area of the anterior maxilla (P=.019) decreased in atrophic participants between baseline and year 1, while the bone area of the anterior maxilla and posterior mandible increased (P<.001) between years 1 and 3. Higher effect sizes were observed in the OHRQoL domains of the atrophic participants. Bone atrophy influenced both the OHRQoL profile and bone remodeling profile in different regions of the mandible and maxilla in mandibular overdenture users. In atrophic participants, bone tissue in both jaws responded positively to overdenture use, with bone apposition after the first year and bone area preservation in the anterior maxilla, posterior mandible, and peri-implant regions after 3 years of follow-up.


Influence of food density on respiration rate of two crustacean plankters, Daphnia galeata and Bosmina longirostris.
The influence of food density on respiration rate was measured for two cladoceran plankton species, Daphnia galeata and Bosmina longirostris, over the range 0 to 2.5 mg C 1-1, using the modified Winkler technique in order to examine how this affects the respiration rate and whether the functional response is the same in the two species. The respiration rate for animals of equivalent body size did not differ significantly between the two species in the absence of food, but was significantly lower in Bosmina longirostris than Daphnia galeata at high food density. Within a species, the response of respiration rate to changing food density did not differ among individuals of different body size. The respiration rate of D. galeata increased with increasing food density and reached a plateau at a high food density. A similar response curve was also found with the respiration rate of B. longirostris, although the response was less conspicuous. This response curve cannot be explained by the energy cost of known feeding behavior in cladocerans. Since the respiration rate related linearly with the assimilation rate, increase in food density seemed to increase the respiration rate by increasing the energetic cost required to process food biochemically, known as specific dynamic action.


Quality Control of Protein Complex Assembly by a Transmembrane Recognition Factor.
The inner nuclear membrane (INM) is continuous with the endoplasmic reticulum (ER) but harbors a distinctive proteome essential for nuclear functions. In yeast, the Asi1/Asi2/Asi3 ubiquitin ligase complex safeguards the INM proteome through the clearance of mislocalized ER membrane proteins. How the Asi complex selectively targets mislocalized proteins and coordinates its activity with other ER functions, such as protein biogenesis, is unclear. Here, we uncover a link between INM proteome identity and membrane protein complex assembly in the remaining ER. We show that lone proteins and complex subunits failing to assemble in the ER access the INM for Asi-mediated degradation. Substrates are recognized by direct binding of Asi2 to their transmembrane domains for subsequent ubiquitination by Asi1/Asi3 and membrane extraction. Our data suggest a model in which spatial segregation of membrane protein complex assembly and quality control improves assembly efficiency and reduces the levels of orphan subunits.


Hyphenated liquid chromatography-gas chromatography technique: recent evolution and applications.
Liquid chromatography (LC) hyphenated with gas chromatography (GC) was first presented in 1979. Since then an intensive study has been carried out to explore different types of interfaces both for coupling normal-phase (NP) and reverse-phase (RP) LC with GC. The present review focuses on the technical progress and applications presented in the last decade, and it describes the most used interfaces. In fact, more flexible interfaces have been studied to improve the use of LC-GC, in particular the use of a programmed temperature vaporizer (PTV) injector. An intensive effort has also been devoted to optimizing the coupling of reverse-phase LC for analysis of water-based samples. A brief overview of comprehensive approaches (LC×GC) is discussed along with perspective for further improvement of the technique.


Glucocorticoid induction of the rat tryptophan oxygenase gene is mediated by two widely separated glucocorticoid-responsive elements.
Transcription of the gene coding for tryptophan oxygenase (TO) in rat liver is induced 10-fold by glucocorticoids. To identify DNA elements mediating the glucocorticoid-regulated expression of the TO gene we transfected mouse L cells with a fusion gene consisting of 1.95 kb TO 5'-flanking sequences linked to the coding sequence of the gene for chloramphenicol acetyltransferase (CAT). CAT assay and RNA mapping experiments demonstrate that both transient and stable expression of the TO-CAT fusion gene are inducible by dexamethasone. Analysis of transcripts from 5'-deletion mutants identifies two glucocorticoid-responsive elements (GRE), located 450 bp and 1.2 kb upstream of the cap site. The purified rat glucocorticoid receptor binds to the sequence of each GRE as evidenced from footprinting experiments. Interestingly the protected sequence of the proximal footprint is by itself not sufficient for sequence induction, but requires sequences located immediately upstream.


Charge Transport in Twisted Organic Semiconductor Crystals of Modulated Pitch.
Many molecular crystals (approximately one third) grow as twisted, helicoidal ribbons from the melt, and this preponderance is even higher in restricted classes of materials, for instance, charge-transfer complexes. Previously, twisted crystallites of such complexes present an increase in carrier mobilities. Here, the effect of twisting on charge mobility is better analyzed for a monocomponent organic semiconductor, 2,5-bis(3-dodecyl-2-thienyl)-thiazolo[5,4-d]thiazole (BDT), that forms twisted crystals with varied helicoidal pitches and makes possible a correlation of twist strength with carrier mobility. Films are analyzed by X-ray scattering and Mueller matrix polarimetry to characterize the microscale organization of the polycrystalline ensembles. Carrier mobilities of organic field-effect transistors are five times higher when the crystals are grown with the smallest pitches (most twisted), compared to those with the largest pitches, along the fiber elongation direction. A tenfold increase is observed along the perpendicular direction. Simulation of electrical potential based on scanning electron microscopy images and density functional theory suggests that the twisting-enhanced mobility is mainly controlled by the fiber organization in the film. A greater number of tightly packed twisted fibers separated by numerous smaller gaps permit better charge transport over the film surface compared to fewer big crystallites separated by larger gaps.


Lessons learnt from the six decades of chloroquine use (1945-2005) to control malaria in Madagascar.
On the island of Madagascar, malaria was nearly eradicated in the highland areas and malaria transmission was significantly decreased in the coastal areas between the 1940s and 1960s. The success of the control programme was primarily achieved by chloroquine (CQ) use at the community level. CQ was administered to children weekly on a routine basis for malaria prevention in the period 1949-1971. Then, the Malagasy Government was unable to financially support the malaria control programme. The malarial situation worsened in the 1980s, partly due to the shortage of CQ. A malaria epidemic occurred. To deal with this epidemic, massive CQ use was urgently adopted. CQ has remained the first-line drug since 1945, but the prevalence of Plasmodium falciparum carrying the pfcrt mutation associated with CQ resistance remains low (<3%). However, late CQ treatment failure has been reported and the prevalence may be as high as 35% during 14-day follow-up since 1982. In an effort to eliminate malaria as a public health problem, a shift from CQ to artemisinin-based combination therapy has been advocated by a new policy since December 2005. A change of this kind is complex and the lessons learnt from the six decades of CQ use are of the utmost importance to achieve malaria control.


β2-Adrenoceptor agonists as novel, safe and potentially effective therapies for Amyotrophic lateral sclerosis (ALS).
Amyotrophic lateral sclerosis (ALS) is a chronic and progressive neuromuscular disease for which no cure exists and better treatment options are desperately needed. We hypothesize that currently approved β2-adrenoceptor agonists may effectively treat the symptoms and possibly slow the progression of ALS. Although β2-agonists are primarily used to treat asthma, pharmacologic data from animal models of neuromuscular diseases suggest that these agents may have pharmacologic effects of benefit in treating ALS. These include inhibiting protein degradation, stimulating protein synthesis, inducing neurotrophic factor synthesis and release, positively modulating microglial and systemic immune function, maintaining the structural and functional integrity of motor endplates, and improving energy metabolism. Moreover, stimulation of β2-adrenoceptors can activate a range of downstream signaling events in many different cell types that could account for the diverse array of effects of these agents. The evidence supporting the possible therapeutic benefits of β2-agonists is briefly reviewed, followed by a more detailed review of clinical trials testing the efficacy of β-agonists in a variety of human neuromuscular maladies. The weight of evidence of the potential benefits from treating these diseases supports the hypothesis that β2-agonists may be efficacious in ALS. Finally, ways to monitor and manage the side effects that may arise with chronic administration of β2-agonists are evaluated. In sum, effective, safe and orally-active β2-agonists may provide a novel and convenient means to reduce the symptoms of ALS and possibly delay disease progression, affording a unique opportunity to repurpose these approved drugs for treating ALS, and rapidly transforming the management of this serious, unmet medical need.


Pascal short-pulse plus subthreshold endpoint management laser therapy for diabetic macular edema: the "sandwich technique".
Diabetic macular edema (DME) is the main cause of visual loss in diabetic patients. Despite the use of anti-VEGF therapy as first-line treatment, there are many patients whose response to treatment is poor or transient at best. Sophisticated laser techniques have emerged aiming at low-intensity retinal damage, avoiding excessive heat that causes tissue necrosis and related collateral effects. To evaluate the effect of combined sublethal laser modalities from short-pulse duration (SPD) with endpoint management (EpM) subthreshold laser [named the "sandwich technique" (SWiT)] on central subfield thickness (CST) and best-corrected visual acuity (BCVA) in patients with DME. In this consecutive retrospective study, 37 patients (37 eyes) with center-involved (CI) DME were treated with SWiT laser therapy from April 2017 to June 2021. The technique consisted of a mean number of 200 (range number 50-400) SPD laser burns OCT-guided thickened area performed on the juxta- and perifoveal area 500 µm away from the foveal center, overlapping with a mean number of 1000 (range number 800-1200) EpM laser burns focused on 6 mm macular diameter area but saving 300 µm toward the foveal center. All patients underwent ophthalmological evaluations, including BCVA and CST measurement by spectral-domain optical coherence tomography (SD-OCT), before and after SWiT laser therapy. The mean follow-up time was 19.2 months (range 2-60 months). Thirty-five out of 37 cases showed an improvement in CST and BCVA following treatment. At baseline, mean CST (µm) ± standard error (SE) and mean BCVA (logMAR) ± SE was 456.95 ± 37.00 and 0.71 ± 0.29, respectively. After a mean follow-up of 19.2 months, mean CST (µm) ± SE and BCVA (logMAR) ± SE were 272.09 ± 9.10 (p < 0.0001) and 0.54 ± 0.26 (p = 0.003), respectively. A statistically significant reduction in CST and improvement in BCVA was noted after laser therapy application. The anti-VEGF injection frequency was reduced during the mean 19.2 months of the study period. The novel "sandwich" laser therapy aid reduced CST and improved BCVA in this retrospective case series. Further prospective studies are warranted.


Greedy auto-augmentation for n-shot learning using deep neural networks.
The goal of n-shot learning is the classification of input data from small datasets. This type of learning is challenging in neural networks, which typically need a high number of data during the training process. Recent advancements in data augmentation allow us to produce an infinite number of target conditions from the primary condition. This process includes two main steps for finding the best augmentations and training the data with the new augmentation techniques. Optimizing these two steps for n-shot learning is still an open problem. In this paper, we propose a new method for auto-augmentation to address both of these problems. The proposed method can potentially extract many possible types of information from a small number of available data points in n-shot learning. The results of our experiments on five prominent n-shot learning datasets show the effectiveness of the proposed method.


Cordocentesis for rapid karyotyping.
Pure fetal blood was obtained by cordocentesis in 101 fetuses of 96 patients at 15 to 38 weeks' gestation. Rapid karyotype was obtained within 2 to 4 days by fetal lymphocyte culture. Chromosomal abnormality was detected in 12 (11.9%) fetuses. Abnormal karyotype was found in 5 of 44 fetuses with structural malformations, 3 of 13 fetuses with intrauterine growth retardation or oligohydramnios, 1 of 3 fetuses with nonimmune hydrops fetalis, 2 (one monozygotic set) of 10 discordant twins, 1 of 12 isoimmunized gestations, none of 8 cases with advanced gestational-maternal age, and none of 6 immune thrombocytopenia cases. This suggests that rapid karyotype should be obtained in all cases of fetal structural malformations, intrauterine growth retardation, and nonimmune fetal hydrops, and may be obtained incidentally in isoimmunized pregnancies and discordant twins to assist in clinical management.


The CAMplexities of central ghrelin.
The gut hormone ghrelin is known to activate hypothalamic AMPK, a crucial metabolic sensor controlling energy balance. In this issue of Cell Metabolism, Anderson et al. (2008) show that CaMKK2 mediates this effect by forming a unique complex of AMPKalpha/beta with acetyl-CoA carboxylase (ACC) in a pathway distinct from the more established AMP/LKB1 pathway.


Dynamic analysis of beat-to-beat fetal heart rate variability recorded by SQUID magnetometer: quantification of sympatho-vagal balance.
Quantitative analysis of fetal heart rate variability (HRV) can be used to investigate the neural control mechanisms of fetal cardiac activity. However, conventional power spectrum methods do not reveal the full complexity of the time-varying sympatho-vagal balance in the fetus. This study was carried out to explore alternative digital signal processing methods of analysing fetal HRV in time domain (rather than frequency domain), in line with most types of physiological monitoring. The beat-to-beat fetal heart rate was obtained by Superconducting Quantum Interference Device (SQUID) magnetocardiographic recording. These data were filtered within appropriately selected frequency bands: high frequency (HF) f>0.2 Hz, low frequency (LF) 0.05<f<0.2 Hz and very low frequency (VLF) f<0.05 Hz. To quantify the dynamics of sympathetic-parasympathetic interaction, the integrated amplitude of the HF curve (within a moving 4-s time window) was compared with the LF component. Also, the interaction of the VLF component with both HF and LF components was analysed. A high degree of correlation was observed between these components for extended periods of time, although the presence of a strong correlation was found to depend on the fetal behavioural state. It was further observed that the low-frequency oscillations lag high-frequency activity by 1-3 s. We propose a numerical parameter, based on the ratio of the LF to HF components of the fetal HRV as a quantitative measure of the relative gain of the parasympathetic system. Our results show this parameter to decrease with gestational age of the fetus.


Nuclear PKG localization is regulated by G₀ alpha and is necessary in the AWB neurons to mediate avoidance in Caenorhabditis elegans.
Neural circuits interpret sensory information from their environment and use this information to influence behavioral outputs. In Caenorhabditis elegans two bilaterally symmetric cephalic amphid organs each contain the ciliated termini of twelve classes of sensory neurons. Two of these sensory neuron pairs are called the AWB and the AWC neurons. The AWC neurons confer an ability to detect attractive volatile odors such as benzaldehyde, whereas the AWB neurons endow the animal with an ability to detect repulsive volatile odors such as 2-nonanone. Previously, it has been shown that transient nuclear localization of a Protein Kinase G (PKG) called EGL-4 in the AWC neuron facilitates adaptation after sustained odor input, and here we show that constitutively nuclear EGL-4 is required in the AWB neurons for the detection of repellent odors. Furthermore, we show that the G₀ alpha subunit protein GOA-1 regulates the nuclear localization of EGL-4 in both AWB and AWC neurons. These data reveal novel insight into how the localization of an individual kinase can form a turnout switch to modify output in different neurons to drive opposite sensory behaviors.


Whole-tumor histogram analysis of apparent diffusion coefficient maps with machine learning algorithms for predicting histologic grade of sinonasal squamous cell carcinoma: a preliminary study.
Accurate histologic grade assessment is helpful for clinical decision making and prognostic assessment of sinonasal squamous cell carcinoma (SNSCC). This research aimed to explore whether whole-tumor histogram analysis of apparent diffusion coefficient (ADC) maps with machine learning algorithms can predict histologic grade of SNSCC. One hundred and forty-seven patients with pathologically diagnosed SNSCC formed this retrospective study. Sixty-six patients were low-grade (grade I/II) and eighty-one patients were high-grade (grade III). Eighteen histogram features were obtained from quantitative ADC maps. Additionally, the mean ADC value and clinical features were analyzed for comparison with histogram features. Machine learning algorithms were applied to build the best diagnostic model for predicting histological grade. The receiver operating characteristic (ROC) curve was used to evaluate the performance of each model prediction, and the area under the ROC curve (AUC) were analyzed. The histogram model based on three features (10th Percentile, Mean, and 90th Percentile) with support vector machine (SVM) classifier demonstrated excellent diagnostic performance, with an AUC of 0.947 on the testing dataset. The AUC of the histogram model was similar to that of the mean ADC value model (0.947 vs 0.957; P = 0.7029). The poor diagnostic performance of the clinical model (AUC = 0.692) was improved by the combined model incorporating histogram features or mean ADC value (P < 0.05). ADC histogram analysis improved the projection of SNSCC histologic grade, compared with clinical model. The complex histogram model had comparable but not better performance than mean ADC value model.


Bortezomib is significantly beneficial for de novo pediatric AML patients with low phosphorylation of the NF-κB subunit RelA.
The addition of the proteasome inhibitor (PI) bortezomib to standard chemotherapy (ADE: cytarabine [Ara-C], daunorubicin, and etoposide) did not improve overall outcome of pediatric AML patients in the Children's Oncology Group AAML1031 phase 3 randomized clinical trial (AAML1031) . Bortezomib prevents protein degradation, including RelA via the intracellular NF-kB pathway. In this study, we hypothesized that subgroups of pediatric AML patients benefitting from standard therapy plus bortezomib (ADEB) could be identified based on pre-treatment RelA expression and phosphorylation status. RelA-total and phosphorylation at serine 536 (RelA-pSer536 ) were measured in 483 patient samples using reverse phase protein array technology. In ADEB-treated patients, low-RelA-pSer536 was favorably prognostic when compared to high-RelA-pSer536 (3-yr overall survival (OS): 81% vs. 68%, p = 0.032; relapse risk (RR): 30% vs. 49%, p = 0.004). Among low-RelA-pSer536 patients, RR significantly decreased with ADEB compared to ADE (RR: 30% vs. 44%, p = 0.035). Correlation between RelA-pSer536 and 295 other assayed proteins identified a strong correlation with HSF1-pSer326 , another protein previously identified as modifying ADEB response. The combination of low-RelA-pSer536 and low-HSF1-pSer326 was a significant predictor of ADEB response (3-yr OS: 86% vs. 67%, p = 0.013). Bortezomib may improve clinical outcome in a subgroup of AML patients identified by low-RelA-pSer536 and low-HSF1-pSer326 .


Standard practice in the treatment of unstable pelvic ring injuries: an international survey.
Unstable pelvic ring injury can result in a life-threatening situation and lead to long-term disability. Established classification systems, recently emerged resuscitative and treatment options as well as techniques, have facilitated expansion in how these injuries can be studied and managed. This study aims to access practice variation in the management of unstable pelvic injuries around the globe. A standardized questionnaire including 15 questions was developed by experts from the SICOT trauma committee (Société Internationale de Chirurgie Orthopédique et de Traumatologie) and then distributed among members. The survey was conducted online for one month in 2022 with 358 trauma surgeons, encompassing responses from 80 countries (experience > 5 years = 79%). Topics in the questionnaire included surgical and interventional treatment strategies, classification, staging/reconstruction procedures, and preoperative imaging. Answer options for treatment strategies were ranked on a 4-point rating scale with following options: (1) always (A), (2) often (O), (3) seldom (S), and (4) never (N). Stratification was performed according to geographic regions (continents). The Young and Burgess (52%) and Tile/AO (47%) classification systems were commonly used. Preoperative three-dimensional (3D) computed tomography (CT) scans were utilized by 93% of respondents. Rescue screws (RS), C-clamps (CC), angioembolization (AE), and pelvic packing (PP) were observed to be rarely implemented in practice (A + O: RS = 24%, CC = 25%, AE = 21%, PP = 25%). External fixation was the most common method temporized fixation (A + O = 71%). Percutaneous screw fixation was the most common definitive fixation technique (A + O = 57%). In contrast, 3D navigation techniques were rarely utilized (A + O = 15%). Most standards in treatment of unstable pelvic ring injuries are implemented equally across the globe. The greatest differences were observed in augmented techniques to bleeding control, such as angioembolization and REBOA, more commonly used in Europe (both), North America (both), and Oceania (only angioembolization). The Young-Burgess and Tile/AO classifications are used approximately equally across the world. Initial non-invasive stabilization with binders and temporary external fixation are commonly utilized, while specific haemorrhage control techniques such as pelvic packing and angioembolization are rarely and REBOA almost never considered. The substantial regional differences' impact on outcomes needs to be further explored.


Degradation of lignocellulose by extracellular enzymes produced by Thermomonospora fusca BD25.
Degradation products from the addition of extracellular enzymes from Thermomonospora fusca BD25 to ball-milled wheat straw, oat spelt xylan and solubilised kraft pulps were characterised by HPLC and TLC. Overall, a high percentage hydrolysis of oat spelt xylan (28.9%) occurred after 26 h incubation. However, the rates of hydrolysis of ball-milled wheat straw and kraft pulp were approximately 4-6-fold less than xylan hydrolysis, although the total percentage hydrolysis of available substrate was similar (22.2% and 25.9% respectively). Incubation of kraft pulp and ball-milled wheat straw by crude extracellular enzymes of T. fusca BD25 resulted in the detection of aromatic compounds at concentrations of 0.6 microg ml(-1) and 8.7 microg ml(-1), respectively. Hydrolysis of oat spelt xylan by T. fusca BD25 extracellular enzymes yielded a mixture of xylose, xylotriose and putative substituted-xylotriose, while the products of ball-milled wheat straw hydrolysis were xylose, glucose and a small oligomer present in the digest. The results highlight the ability of culture supernatant from T. fusca to release both simple sugars and aromatic compounds from lignocellulosic substrates and suggest a role for this organism in the biobleaching of pulp.


The best choice for low and moderate myopia patients incapable for corneal refractive surgery: implantation of a posterior chamber phakic intraocular lens.
This study aims to evaluate the early clinical outcomes of central hole phakic posterior chamber intraocular lens(ICLV4c) implantation for low and moderate myopia correction. This retrospective clinical study included 27 patients(47 eyes)who underwent ICLV4c implantation to correct myopia with spherical equivalent (SE) between - 1.75D and -6.0D. The uncorrected visual acuity (UCVA), intraocular pressure(IOP), central anterior chamber depth (ACD), vault, and endothelial cell density (ECD) were evaluated after surgery. At one year follow-up, the postoperative UCVA of patients was higher than the preoperative CDVA and showed a stable trend. There was no significant difference in ECD (p > 0.05) one month after the surgery, the vault was 0.77 ± 0.32 mm, which decreased to 0.63 ± 0.26 mm after one year of surgery. Similarly, ACD was 3.24 ± 0.25 mm in the preoperative, which decreased significantly to 2.05 ± 0.39 mm at one month, while rose to 2.2 ± 0.39 mm at one year after surgery. There was no significant correlation between IOP and ACD and vault at one month and one year after surgery. The correlation coefficient between ACD and vault was found to be - 0.72 at one month after surgery, while the same -0.71 after one year. One patient developed visual fatigue, one with glare, and while no other complications were observed with the rest of the patients. The ICL implantation is a safe, effective and stable method to correct low and moderate myopia, and may be a good alternative for patients with low and moderate myopia who cannot undergo corneal refractive surgery.


[Chromatin RNA as a factor decreasing chromatin fragmentability under endonucleolysis].
Rat liver chromatin fragments obtained after the chromatin digestion with Ca,Mg-dependent intranuclear endonuclease were studied using electrophoresis in polyacrylamide gel concentration gradient. It is found that the content of large chromatin fragments decreases in the presence of exogenous RNAse. Possible structural role of RNA in the chramatin organization is discussed.


Interesting effects of interleukins and immune cells on acute respiratory distress syndrome.
Acute respiratory distress syndrome (ARDS) is a medical condition characterized by widespread inflammation in the lungs with consequent proportional loss of gas exchange function. ARDS is linked with severe pulmonary or systemic infection. Several factors, including secretory cytokines, immune cells, and lung epithelial and endothelial cells, play a role in the development and progression of this disease. The present study is based on Pubmed database information (1987-2022) using the words "Acute respiratory distress syndrome", "Interleukin", "Cytokines" and "Immune cells". Cytokines and immune cells play an important role in this disease, with particular emphasis on the balance between pro-inflammatory and anti-inflammatory factors. Neutrophils are one of several important mediators of Inflammation, lung tissue destruction, and malfunction during ARDS. Some immune cells, such as macrophages and eosinophils, play a dual role in releasing inflammatory mediators, recruitment inflammatory cells and the progression of ARDS, or releasing anti-inflammatory mediators, clearing the lung of inflammatory cells, and helping to improve the disease. Different interleukins play a role in the development or inhibition of ARDS by helping to activate various signaling pathways, helping to secrete other inflammatory or anti-inflammatory interleukins, and playing a role in the production and balance between immune cells involved in ARDS. As a result, immune cells and, inflammatory cytokines, especially interleukins play an important role in the pathogenesis of this disease Therefore, understanding the relevant mechanisms will help in the proper diagnosis and treatment of this disease.


Quantitative magnetization transfer MRI unbiased by on-resonance saturation and dipolar order contributions.
To demonstrate the bias in quantitative MT (qMT) measures introduced by the presence of dipolar order and on-resonance saturation (ONRS) effects using magnetization transfer (MT) spoiled gradient-recalled (SPGR) acquisitions, and propose changes to the acquisition and analysis strategies to remove these biases. The proposed framework consists of SPGR sequences prepared with simultaneous dual-offset frequency-saturation pulses to cancel out dipolar order and associated relaxation (T1D ) effects in Z-spectrum acquisitions, and a matched quantitative MT (qMT) mathematical model that includes ONRS effects of readout pulses. Variable flip angle and MT data were fitted jointly to simultaneously estimate qMT parameters (macromolecular proton fraction [MPF], T2,f , T2,b , R, and free pool T1 ). This framework is compared with standard qMT and investigated in terms of reproducibility, and then further developed to follow a joint single-point qMT methodology for combined estimation of MPF and T1 . Bland-Altman analyses demonstrated a systematic underestimation of MPF (-2.5% and -1.3%, on average, in white and gray matter, respectively) and overestimation of T1 (47.1 ms and 38.6 ms, on average, in white and gray matter, respectively) if both ONRS and dipolar order effects are ignored. Reproducibility of the proposed framework is excellent (ΔMPF = -0.03% and ΔT1 = -19.0 ms). The single-point methodology yielded consistent MPF and T1 values with respective maximum relative average bias of -0.15% and -3.5 ms found in white matter. The influence of acquisition strategy and matched mathematical model with regard to ONRS and dipolar order effects in qMT-SPGR frameworks has been investigated. The proposed framework holds promise for improved accuracy with reproducibility.


Effect of histamine on gastric acid secretion "in vitro": interference with endogenous prostaglandins.
The interference between histamine and endogenous prostaglandins (PGs) was investigated in the isolated gastric fundus from immature rats by evaluating the effect of nonsteroidal antiinflammatory agents (NSAIDs) on the acid response to histamine and the effect of histamine on PGs production by the gastric mucosa. Indomethacin (10(-5) M) and diclofenac (10(-5) M) caused a dose-dependent enhancement of the response to histamine, dimaprit and DBcAMP, but did not affect bethanechol-, isoprenaline- and forskolin-induced acid production. The enhancing effect of indomethacin was abolished in low Ca2+ medium. PGE2 and 6-keto PGF1 alpha levels in the mucosal solution were not modified by histamine, while being reduced by indomethacin. From these data it is concluded that endogenous PGs may negatively modulate the secretory response to histamine. However, a direct effect of the amine on cyclooxygenase to increase PGs synthesis seems to be excluded since histamine did not modify PGs levels in the mucosal solution.


Retinal detachment.
A retinal detachment is often traumatic and shocking to patients once they become aware of the condition. Nurses have the unique opportunity to assist these patients in accepting and in coping with this potential serious vision loss through understanding, education, rehabilitation, and ocular safety.


Health benefits and consequences of the Eastern Orthodox fasting in monks of Mount Athos: a cross-sectional study.
Greek Orthodox fasting (OF), which involves 180-200 days of fasting per year, is dictated by the Christian Orthodox religion. For the first time, this cross-sectional study examines the characteristics and the effects of OF on anthropometry, cardiometabolic markers and calcium homeostasis in Athonian monks (AMs). Daily intakes of energy, macro- and micronutrients of a day during a weekend of Nativity Fast, defined as non-restrictive day (NRD), and a weekday during Great Lent, labeled as restrictive day (RD) were recorded. The daily energy intake of 70 AM (age=38.8±9.7 years) was low during both RD and NRD (1265.9±84.5 vs 1660±81 kcal, respectively, P<0.001). Paired samples t-test showed statistically significant difference between daily intakes in RD and NRD: carbohydrates (159.6±21.8 vs 294.3±23.4 g, P<0.0001) and saturated fat (12.7±0.0 vs 16.4±0.0 g, P<0.0001) were lower, whereas protein (89.2±1.3 vs 72.35±1.3 g, P<0.001) was higher during RD. A subsample of 50 monks (age=38.7±10.6 years) formed a study cohort for cardiometabolic and calcium homeostasis assessment. Body weight (74.3±12.9 kg) and body mass index (BMI; 23.8±4.1 kg/m2) were independent of level of physical activity. Optimal profiles for lipid and glucose parameters (total cholesterol: 183.4±41.7 mg/dl, LDL: 120.6±37.6 mg/dl, triglycerides: 72.2±31.3 mg/dl, HDL: 48.5±14.2 mg/dl and homeostasis model assessment of insulin resistance (HOMA-IR) 1.02±0.40) were found. Profound hypovitaminosis D (8.8±6.2 ng/ml), high parathyroid hormone (PTH): 115.5±48.0 pg/ml with normal serum calcium levels (8.9±3.2 mg/dl) was observed. Unaffected by variation in lifestyle factors, the results of this unique study offers clear evidence for the health benefits of the strict Athonian OF through optimal lipid and glucose homeostasis.


The perception of temporal structure and auditory evoked brain activity.
The processing and perception of auditory signals depends on the temporal structure of stimulus characteristics. We studied 26 healthy subjects who participated in psychophysical experiments and in electrophysiological recordings of auditory evoked potentials from C2, C3, C4, T3 and T4. Stimuli consisted of tone series presented binaurally as tones or gaps with a base duration of 100 ms. In the psychophysical experiments, difference thresholds as indicators of temporal discrimination performance were significantly lower for tones than for gaps. In the electrophysiological recordings, gaps often failed to elicit N100 components. Tones produced shortest component latencies with largest amplitudes. In addition, brain activity was strongest at C2, and showed a symmetrical fall-off over both hemispheres. N100 components had significantly longer latencies and smaller amplitudes when they were evoked by the end of the gap (i.e. with the continuation of the tone) than by tones. Our data illustrate how the temporal structure of auditory stimuli affects neuronal responses of the brain. Similar effects were observed in psychophysical and electrophysiological experiments, and we were able to demonstrate a direct relationship between subjective sensory thresholds and auditory evoked brain activity.


Primary leiomyosarcoma of the breast: A case report and literature review.
Primary leiomyosarcoma is an uncommon form of stromal breast sarcoma. Approximately 73 cases have been documented in English-language literature to date. To our knowledge, this is the first report from Indonesia of an adolescent female with primary leiomyosarcoma of the breast. A 30-year-old Southeast Asian female presented with a tumor in her left breast. Clinical examination revealed a 12 × 8-centimeter tumor. The supraclavicular, subclavicular, and axillary lymphadenopathy were not palpable. An ultrasound revealed a Breast Imaging Reporting and Data System category 5. Abdominal ultrasonography and chest x-ray were normal, as were blood chemistry and routine blood tests. A wide excision with a surgical margin of 2 cm was performed. Pathological investigation identified the mass as a leiomyosarcoma. The pelvis, abdomen, and lung CT scan metastatic workups were negative. The patient is well 8 months post-surgery, with no signs of recurrence. Wide local excision has been the mainstay of treatment for leiomyosarcoma; however, there is no accepted standard of treatment due to the rarity of the disease. Breast leiomyosarcomas have a more favorable prognosis than other breast neoplasms; however, patients must be closely monitored for recurrence or metastases. While there are no known predictors of outcomes, the margins of the initial surgery, mitotic activity, and atypia cellularity are more indicative of malignancy.


Helper interleukins are produced by both CD4 and CD8 splenic T cells after mitogen stimulation.
We have earlier described (Cardell, S. and Sander, B., Eur. J. Immunol. 1990. 20:389) mitogen-induced production of interleukin (IL)2, IL4 and IL5 mRNA by murine spleen cells, analyzed by in situ hybridization. In the present study we have investigated the potential of CD8 T cells to produce these interleukins, normally associated with the helper function of CD4 T cells. When concanavalin A (Con A)-activated spleen cells were restimulated with Con A and phorbol 12-myristate 13-acetate (PMA), higher levels of IL2, IL4 and IL5 mRNA were induced, as detected both by increased frequencies of positive cells, and by more mRNA per cell. Four-to-six-day Con A blasts were enriched for CD4+ or CD8+ T cells, and restimulated with Con A and PMA. Both CD4 and CD8 cells were found to produce all three kinds of mRNA when restimulated. The frequencies of IL2 mRNA-containing CD8 cells were half of those found for CD4 cells (3.5% as compared to 7%). On the average 1% of the CD8 cells were induced to produce IL4 and IL5 mRNA, while 9% and 3% of the activated CD4 cells contained IL4 and IL5 mRNA, respectively. CD4 and CD8 cells displayed different sensitivities to the reagents when tested alone. Con A induced the synthesis of IL4 and IL5 in CD4 cells, but not CD8 cells, independently of PMA. PMA alone induced extensive thymidine incorporation in CD8 cells, but not in CD4 cells, in the absence of detectable lymphokine mRNA. The results suggest that some CD8 cells have the capacity to give help in immune responses, by secretion of IL2, IL4 and IL5.


Unusual properties of platelet shape in coronary and cerebral artery disease.
Platelet shape was examined in freshly fixed whole blood, using phase contrast microscopy, from 7 patients post-myocardial infarction, 9 patients undergoing elective angiography for the investigation of angina, and 17 patients with stroke. Platelet shape was abnormal in 29/33 patients and in 20 of them the distribution of platelet shape did not resemble that produced by known platelet activators, by venipuncture techniques known to produce platelet activation, nor by shearing platelet-rich plasma. In particular, the proportion of spherical platelets is too high relative to the proportion of smooth discoid shaped platelets. Addition of platelet-poor plasma from a patient post-myocardial infarction to normal platelet-rich plasma did not reproduce the abnormality in platelet shape. In contrast, for patients post-myocardial infarction platelet shape was normal in platelet-rich plasma, but the platelets were hypersensitive to ADP-induced aggregation. Incubation of citrated whole blood at 37 degrees C resulted in an incomplete disappearance of the spherical platelets suggesting that the abnormal distribution of platelet shape consists of both a reversible and a more persistent abnormality. The reversible portion of the platelet shape abnormality seen in patients with stroke and coronary artery disease may be related to venipuncture-induced platelet activation; however, the nature of the persistent abnormality in platelet shape is presently unknown.


Identification of potential diagnostic biomarkers and therapeutic targets for endometriosis based on bioinformatics and machine learning analysis.
Endometriosis (EMs) is a major gynecological condition in women. Due to the absence of definitive symptoms, its early detection is very challenging; thus, it is crucial to find biomarkers to ease its diagnosis and therapy. Here, we aimed to identify potential diagnostic and therapeutic targets for EMs by constructing a regulatory network and using machine learning approaches. Three Gene Expression Omnibus (GEO) datasets were merged, and differentially expressed genes (DEGS) were identified after preprocessing steps. Using the DEGs, a transcription factor (TF)-mRNA-miRNA regulatory network was constructed, and hub genes were detected based on four different algorithms in CytoHubba. The hub genes were used to build a GaussianNB diagnostic model and also in docking analysis that were performed using Discovery Studio and AutoDock Vina software. A total of 119 DEGs were identified between EMs and non-EMs samples. A regulatory network consisting of 52 mRNAs, 249 miRNAs, and 37 TFs was then constructed. The diagnostic model was introduced using the hub genes selected from the network (GATA6, HMOX1, HS3ST1, NFASC, and PTGIS) that its area under the curve (AUC) was 0.98 and 0.92 in the training and validation cohorts, respectively. Based on docking analysis, two chemical compounds, rofecoxib and retinoic acid, had potential therapeutic effects on EMs. In conclusion, this study identified potential diagnostic and therapeutic targets for EMs which demand more experimental confirmations.


Evidence for placental-derived iron-nitrosyls in the circulation of the fetal lamb and against a role for nitrite in mediating the cardiovascular transition at birth.
Circulating metabolites of nitric oxide, such as nitrite, iron nitrosyls (FeNO), and nitrosothiols, have vasodilatory bioactivity. In both human and sheep neonates, plasma concentrations of these NO metabolite (NOx) concentrations fall >50% within minutes after birth, raising the possibility that circulating NOx plays a role in maintaining low fetal vascular resistance and in the cardiovascular transition at birth. To test whether the fall in plasma NOx concentrations at birth is due to either ligation of the umbilical cord or oxygenation of the fetus to newborn levels, plasma NOx concentrations were measured during stepwise delivery of near-term fetal lambs. When fetal lambs were intubated and mechanically ventilated with 100% O2 to oxygenate the arterial blood while still in utero with the umbilical circulation still intact, there was no change in plasma NOx levels. In contrast, when the umbilical cord was ligated while fetal lambs were mechanically ventilated with O2 levels that maintained fetal arterial blood gases, plasma NOx levels decreased by nearly 50%. Characterization of the individual NOx species in plasma revealed that the overall fall in NOx at birth was attributable mainly to FeNO compounds. Finally, when the typical fall in NOx after birth was prevented by intravenous nitrite infusion, birth-related changes in blood pressure, heart rate, and carotid flow changes were little affected, suggesting the cardiovascular transition at birth is not dependent on a fall in plasma NOx. In conclusion, this study shows FeNO is released from the placenta and that its decline accounts for most of the measured fall in plasma NOx at birth.


Heel pad thickness following calcaneal fractures: ultrasound findings.
Ultrasound measurements of heel pad thickness in 21 patients who had suffered unilateral fractures of the calcaneum were taken. Body weight was also recorded. Contrary to previous assertions in the literature, heel pad thinning was not found to occur and in fact a significant increase in heel pad thickness on the affected side was noted.


Surgical wound bed preparation of chronic and acute wounds.
Our aging population has presented us with many new challenges. One such challenge is the need to manage an increase in wound-related problems effectively and efficiently. Over time, two parallel, yet divergent, management systems have developed. One strategy, used by medical specialists, uses a variety of dressings, topical enzymes, and local and systemic medications ultimately aimed at the promotion of healing by secondary intention or, in some cases, optimization of the wound for surgical reconstruction. In the second strategy, used by surgeons, early surgical intervention is used to prepare the wound for reconstruction in a timelier manner while promoting the healing process. This article reviews the development of these two distinct management systems and their areas of commonality and sets forth a new model to support the role of surgery in the treatment of problematic wounds.


Lifestyle, health and social class in adolescence.
The paper considers mechanisms for indirect health selection in adolescence, as part of the explanation for health inequalities between social groups. Aspects of adolescent lifestyles are identified as potentially important factors for the production of class based differences in adult health status. Survey data from a Scottish longitudinal study of adolescent socialization and lifestyles are utilized in order to locate such health lifestyle factors within the wider contexts of the individual's personal and social environment at this stage of the life cycle. Relationships and attitudes to family, school and peers in middle adolescence at 15-16 years of age are first examined, and distinctive patterns of integration within these contexts are identified. The inter-connections between these broader aspects of lifestyle, social class and individual health behaviours are then examined. Mid-adolescent patterns of social integration are found to have a clear structural basis, and most importantly, they anticipate social position in later adolescence at 17-18 years of age. It is also found that such patterns of integration into the family, peer and school contexts are linked to subsequent health related behaviours and to self assessed health in later adolescence, and that these links are independent of the young person's social class background. Thus, we conclude that behavioural--cultural lifestyle factors, when these are located within a broader social context, provide a clear and plausible mechanism for indirect health selection in adolescence.


Usefulness of intraoperative Indocyanine green video angiography to select the recipient artery for bypass surgery in arteriosclerotic occlusion of the middle cerebral artery: a technical case report.
In superficial temporal artery-to-middle cerebral artery (STA-MCA) bypass surgery, indocyanine green video angiography (ICG-VA) is usually used to verify bypass patency. Less-commonly reported is the ability to use this technique to evaluate candidate recipient vessels based on either collateral flow or identification of the distal branch of interest. An 82-year-old man presented with progressive cerebral infarction due to infarction of the M2 inferior trunk of the right middle cerebral artery. He underwent superficial temporal artery-middle cerebral artery bypass to prevent further ischemia 1 week after the initial stroke. In the surgery, M4 cortical arteries fed by the inferior trunk could not be identified as recipient arteries. Intraoperative ICG-VA showed delayed luminescence of some M4 arteries. Because the M4 arteries fed by the inferior trunk showed delayed retrograde flows from the anterior cerebral artery on preoperative digital subtraction angiography, the M4 arteries with delayed luminescence on ICG-VA were considered to be M4 arteries fed by the inferior trunk and selected as the recipient arteries. ICG-VA shows differences in flow speed as delayed luminescence. This finding may be useful for detecting target vessels.


Effect of stimulus dimension on perception and cognition.
Stimuli characteristics ha a decisive role in our perception and cognition. In the present study, we aimed to evaluate the effects of dimension of stimuli, two-dimensional (2D) versus three-dimensional (3D), on perception and working memory. In the first experiment, using the method of eye tracking, a higher blink rate, pupil size, and the number of saccade for three compared to 2D stimuli revealed a higher perceptual demand of 3D stimuli. In the second experiment, visual search task shows a higher response time for 3D stimuli and an equal performance with 2- and 3D stimuli in spatial working memory task. In the third experiment, four working memory tasks with high and low cognitive and perceptual load revealed 3D stimuli are memorized better in the both low and high load of working memory. We can conclude that 3D stimulus, compared 2D, imposes a higher load on perceptual system, but it is memorized better. It could be concluded that the phenomenon of filtering should occur in the early perceptual system for preventing overload.


Divergent Synthesis of Ultrabright and Dendritic Xanthenes for Enhanced Click-Chemistry-Based Bioimaging.
Biorthogonal labelling with fluorescent small molecules is an indispensable tool for diagnostic and biomedical applications. In dye-based 5-ethynyl-2'-deoxyuridine (EdU) cell proliferation assays, augmentation of the fluorescent signal entails an overall enhancement in the sensitivity and quality of the method. To this end, a rapid, divergent synthetic procedure that provides ready-to-click pH-insensitive rhodamine dyes exhibiting outstanding brightness was established. Compared to the shortest available synthesis of related high quantum-yielding rhodamines, two fewer synthetic steps are required. In a head-to-head imaging comparison involving copper(I)-catalyzed azide alkyne cycloaddition reactions with in vitro administered EdU, our new 3,3-difluoroazetidine rhodamine azide outperformed the popular 5-TAMRA-azide, making it among the best available choices when it comes to fluorescent imaging of DNA. In a further exploration of the fluorescence properties of these dyes, a set of bis-MPA dendrons carrying multiple fluorescein or rhodamine units was prepared by branching click chemistry. Fluorescence self-quenching of fluorescein- and rhodamine-functionalized dendrons limited the suitability of the dyes as labels in EdU-based experiments but provided new insights into these effects.


Submolecular partitioning of morphine hydrate based on its experimental charge density at 25 K.
The electron density distribution of morphine hydrate has been determined from high-resolution single-crystal X-ray diffraction measurements at 25 K. A topological analysis was applied and, in order to analyze the submolecular transferability based on an experimental electron density, a partitioning of the molecule into atomic regions was carried out, making use of Bader's zero-flux surfaces to yield atomic volumes and charges. The properties obtained were compared with the theoretical calculations of smaller fragment molecules, from which the complete morphine molecule can be reconstructed, and with theoretical studies of another opiate, Oripavine PEO, reported in the literature.


Rapid size determination of mRNAs complementary to cloned DNA sequences: plaque and colony hybrid-selection of cDNAs.
We describe a novel screening method for the rapid size determination of mRNAs. This method has been used successfully to identify the cDNA and genomic DNA clones that had been constructed using the plasmid vector pBR322 or phage vectors M13mp7 and lambda EMBL3. Poly(A) RNA is reverse-transcribed into 32P-labeled cDNA under conditions that yield a high proportion of full-length cDNA copies. This radioactive cDNA is then hybridized in situ to bacterial colonies or phage plaques harboring recombinant DNA molecules. Colonies or plaques containing sequences complementary to abundant or moderately abundant mRNAs are detected by autoradiography and excised from nitrocellulose filters. The hybridized cDNA is eluted from the filter by alkaline denaturation and sized directly by alkaline agarose gel electrophoresis. The mRNAs characterized thus far by this technique measure between 450 and 2100 nucleotides and account for 0.03% to 15% of the mass of cytoplasmic poly(A) RNA.


Effect of the relaxation time of mineral oil and monoesters on the fractal dimension and mutual information of creeping discharges propagating along a pressboard.
This paper presents a new method for analysing creeping discharges based on information theory as it applies to medical imaging. The analysis of information surface data is used to determine the impact of relaxation time on the characteristic parameters of creeping discharges. The same information is used to make a comparative study of the morphology of discharges propagating in palm kernel oil methyl ester (PKOME) and in mineral oil (MO). Other comparative methods based on fractal analysis and normality hypothesis tests associated with Anderson Darling (AD), Kolmogorov-Smirnoff (KS) and Shapiro-Wilk (SW) statistics are used. The results show that very short relaxation times increase the error on the measurement of the fractal dimension and the maximum extension of the discharges. A growth of the mutual information between 0 and 60% is observed for relaxation times varying between 60s and 420s respectively. For the same time interval, the P-value increases from 0.027 to 0.821 according to the AD statistic, from 0.01 to more than 0.150 according to KS and from 0.083 to more than 0.1 according to SW. This result indicates that the data are from a normal distribution. After 420s of relaxation, the error on the maximum extension measurement is reduced by 94% in PKOME and 92% in MO. Similarly, the error on the mean fractal dimension in MO is reduced by 86.7% for a relaxation time between 301s and 420s, and by 84.6% in PKOME for a time between 180s and 420s. These different results imply that the impact of the discharge can be predicted when it is in its initial phase during which the number of discharge occurrences is reduced. On the other hand, the physicochemical characteristics of the insulating liquid used dictate the relaxation time to be allowed for the laboratory measurements.


Gender and cystectomy for bladder cancer: A high-volume tertiary urologic care center experience.
Differences have often been reported in the outcomes of bladder cancer (BC) patients according to gender. This study aims to provide data on patients undergoing radical cystectomy (RC) in a high-volume tertiary urologic center and to assess whether gender discrepancies do exist in terms of surgical options and clinical outcomes. Consecutive BC patients treated between 2016 and 2020 at a single center (Careggi University Hospital, Florence, Italy) were included in the study. The impact of gender on disease stage at diagnosis, overall survival (OS), and type of surgery was analyzed. The study series comprised 447 patients (85 females and 362 males). At a median follow-up of 28.3 months (IQR: 33.5), OS was 52.6% and cancer-specific survival was 67.6%. Significant differences in OS emerged for age, acute myocardial infarction (AMI), Charlson Comorbidity Index (CCI), pT, and pN. OS rates were higher in patients undergoing robot-assisted surgery and in those receiving open orthotopic neobladder (ONB) (p = 0.0001). No statistically significant differences were found between male and female patients regarding surgical offer in any age group, surgical time, early postoperative complications, pathologic stage, and OS. After adjustment for pathologic tumor stage and treatment modalities, female and male patients showed similar oncologic outcomes. Further studies should be undertaken to evaluate functional results in women subjected to RC.


Comparison of the Effect of Different Types of Experimental Anisometropia on Stereopsis Measured with Titmus, Randot and TNO Stereotests.
To assess the effect of experimental anisometropia and monovision on stereopsis using the Titmus, Randot, and TNO stereoacuity tests. Sixty adult volunteers were enrolled in the present study. Four different types of anisometropia-myopia, hyperopia, and astigmatism (both 90° and 45°)-were induced by placing trial lenses over the dominant eye (from 3 to 1 D). Stereoacuity was measured using the Titmus, Randot, and TNO tests. In all the anisometropia types, stereopsis deteriorated with increase in anisometropia in the three stereoacuity tests performed (P < 0.001). The largest decrease in stereopsis was attributed to 3 D myopic anisometropia-6.51 ± 2.10, 6.59 ± 2.35, and 7.36 ± 1.89 arc seconds in Titmus circles, Randot circles, and TNO, respectively. Minimal change in stereopsis was observed in 1 D astigmatism of 45°. Any type of anisometropia may reduce stereoacuity; this reduction is most noticeable with myopic anisometropia, especially in the TNO test, probably due to the lack of monocular cues.


Alteration in protein kinase C activity and subcellular distribution in sickle erythrocytes.
In agreement with previous data, membrane protein phosphorylation was found to be altered in intact sickle cells (SS) relative to intact normal erythrocytes (AA). Similar changes were observed in their isolated membranes. The involvement of protein kinase C (PKC) in this process was investigated. The membrane PKC content in SS cells, measured by [3H]phorbol ester binding, was about 6-times higher than in AA cells. In addition, the activity of the enzyme, measured by histone phosphorylation was also found to be increased in SS cell membranes but decreased in their cytosol compared to the activity in AA cell membranes and cytosol. The increase in membrane PKC activity was observed mostly in the light fraction of SS cells, fractionated by density gradient, whereas the decrease in cytosolic activity was only observed in the dense fraction. PKC activity, measured in cells from the blood of reticulocyte-rich patients, exhibited an increase in both membranes and cytosol, thus explaining some of the effects observed in the SS cell light fraction, which is enriched in reticulocytes. The increase in PKC activity in the membranes of SS cells is partly explained by their young age but the loss of PKC activity in their cytosol, particularly in that of the dense fraction, seems to be specific to SS erythrocytes. The relative decrease in membrane PKC activity between the dense and the light fractions of SS cells might be related to oxidative inactivation of the enzyme.


T-lymphocyte sub-populations in orbito-ocular granulocytic sarcoma (OOGS) and acute myelocytic leukemia (AML): a preliminary study.
The aetiology of GS remains obscure and a little is known about the immune competence of these patients. Interestingly, all children with OOGS were from low 'socio-economic status' and showed diminished delayed hypersensitivity reactions and reduced T cell counts (E-R) in our previous observation. We present herewith a preliminary data on evaluation of T cell sub-populations determined by monoclonal antibodies (CD3, CD4, CD8 and CD16 cells) in 10 patients with OOGS and AML prior to treatment. Quantitative immunoglobulin determinations of IgA, IgM, IgG were also made. The percentage of Pan T (CD3), CD4, CD8 cells were significantly lower than those in the controls (p < 0.01). The immunoglobulin levels were slightly elevated suggesting normal B cell functions. In conclusion, these preliminary findings suggest that cellular immune deficiency may be an underlying cause.


The impact of age and gender on right ventricular diastolic function among healthy adults.
Doppler echocardiography is ideally suited for assessment of diastolic function, being widely available, non-invasive, and less expensive than other techniques. However, data regarding age- and gender-matched reference values of right ventricular diastolic function are limited. This study aims to explore the physiologic variations of right ventricle (RV) diastolic function in a large cohort of healthy adults, and to investigate clinical and echocardiographic correlates. From June 2007 to February 2014, 1168 healthy Caucasian subjects [mean age 45.1±15.6 years, range 16-92; 555 (47.5%) men] underwent comprehensive transthoracic echocardiography (TTE) following current guidelines. The following RV main diastolic measurements were measured: peak early inflow velocity (E), annular both early (e') and atrial (a') velocities, E/e' ratio. RV E/e' constantly increases with age in females, but do not change substantially in males. RV E/A constantly decreases with age in both genders. Stepwise multiple linear regression analysis underlined a close significant association of RV diastolic function with both right and left heart morphologic measurements (right atrial area, RV diameters, left atrial volume) and functional indexes (TAPSE, RV tissue Doppler peak systolic velocity, left ventricular E/Ee'), as well as with indexes of increased pulmonary resistance. Our data highlight the potential usefulness of different normal reference values according to the age and gender to correctly evaluate RV diastolic function. Differences in terms of demographic and anthropometric parameters could be useful to avoid potential misclassification of RV diastolic function when based on dichotomously suggested normal cut-off values.


Poly[(μ4-5,7-di-hydro-1H,3H-dithieno[3,4-b:3',4'-e]pyrazine-κ4 N:N':S:S')tetra-μ3-iodido-tetra-copper]: a three-dimensional copper(I) coordination polymer.
The reaction of ligand 5,7-di-hydro-1H,3H-dithieno[3,4-b:3',4'-e]pyrazine (L) with CuI lead to the formation of a three-dimensional coordination polymer, incorporating the well known [Cu x I x ] n staircase motif (x = 4). These polymer [Cu4I4] n chains are linked via the N and S atoms of the ligand to form the three-dimensional coordination polymer poly[(μ4-5,7-di-hydro-1H,3H-dithieno[3,4-b:3',4'-e]pyrazine-κ4 N:N':S:S')tetra-μ3-iodido-tetra-copper], [Cu4I4(C8H8N2S2)] n (I). The asymmetric unit is composed of half a ligand mol-ecule, with the pyrazine ring located about a center of symmetry, and two independent copper(I) atoms and two independent I- ions forming the staircase motif via centers of inversion symmetry. The framework is consolidated by C-H⋯I hydrogen bonds.


Segregation-induced ordered superstructures at general grain boundaries in a nickel-bismuth alloy.
The properties of materials change, sometimes catastrophically, as alloying elements and impurities accumulate preferentially at grain boundaries. Studies of bicrystals show that regular atomic patterns often arise as a result of this solute segregation at high-symmetry boundaries, but it is not known whether superstructures exist at general grain boundaries in polycrystals. In bismuth-doped polycrystalline nickel, we found that ordered, segregation-induced grain boundary superstructures occur at randomly selected general grain boundaries, and that these reconstructions are driven by the orientation of the terminating grain surfaces rather than by lattice matching between grains. This discovery shows that adsorbate-induced superstructures are not limited to special grain boundaries but may exist at a variety of general grain boundaries, and hence they can affect the performance of polycrystalline engineering alloys.


Association of human leucocyte-DR and DQ antigens in coeliac disease: a family study.
No family studies regarding the association of coeliac disease with the human leucocyte antigen (HLA)-DQ locus are available. Moreover, no HLA studies have been carried out in coeliac disease patients from India. The aim of this study was to study the HLA class II (DR and DQ) antigens in children with coeliac disease and in their first-degree relatives. Fifteen children with coeliac disease and their first-degree relatives (birth parents of all the coeliac disease patients and fifteen siblings) were studied. A group of 123 healthy unrelated and ethnically matched subjects were used as controls. The HLA-DR and -DQ typing was carried out by a complement-dependent microlymphocytotoxicity assay. The transmission disequilibrium test was used for analysis of results. There was no association of coeliac disease with DR phenotypes. Ninety-three per cent of patients (14/15) carried the DQ2 allele. DQ2 was transmitted in 15 of 19 informative cases (transmission probability of 79%, chi2 6.368 with 1 df, nominal P=0.012 and P value corrected for multiple test=0.035). The haplotype relative risk associated with DQ2 was 5.71 (95% confidence interval 1.71-16.28). Coeliac disease in Indian children is predominantly associated with HLA-DQ2.


Growth of human skin fibroblasts in dialyzed fetal bovine serum.
Human diploid fibroblast cultures plated at or below a density of 2 X 10(3) cells per cm2 grew very slowly or not at all in MEM supplemented with 10% fetal bovine serum that had been dialyzed for 24 hr. Adding serine (0.2 mM) or pyruvate (1.0 mM) to MEM and 10% dialyzed serum restored growth to the level observed with 10% nondialyzed serum. Serine and pyruvate also were able to overcome partially the growth arrest induced by a reduced serum concentration (1 or 2%). Human fibroblast cultures grew very well in 100% fetal bovine serum that had been dialyzed against MEM. For cells grown in dialyzed serum, the final number increased with increasing serum concentration, in contrast to the well established toxic effects of high concentrations of nondialyzed serum.


Differential modulation of phospholipase D and phosphatidate phosphohydrolase during aging in rat cerebral cortex synaptosomes.
Phosphatidylcholine (PC) hydrolysis generates two important second messengers: phosphatidic acid (PA) and diacylglycerol (DAG). Phospholipase D (PLD) and phosphatidate phosphohydrolase (PAPase) are involved in their generation and therefore are key enzymes in signal transduction. Specific isoforms of these enzymes are activated by receptor occupancy in brain. Phosphatidylinositol 4,5-bisphosphate-dependent PLD (PIP2-PLD) and N-ethylmaleimide-insensitive PAPase (PAP2) have been suggested to act in series to generate the biologically active lipids PA and DAG. In the present study we examine age-induced changes mainly in PIP2-PLD and PAP2 activities in cerebrocortical synaptosomes from adult (4 months) and aged (28 months) Wistar rats. Aging increases the activity of both enzymes. Guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) and cytosol (from cerebral cortex) stimulate PLD activity in adult and senescent synaptosomal membranes, the effect being greater in the latter. Under the same experimental conditions PAP2 activity was stimulated in aged membranes whereas in adult membranes GTPgammaS had no effect and cytosol showed a slight inhibitory effect. Diacylglycerol lipase (DGL) activity differs from that of PAP2 in aged rats and it was 21% inhibited with respect to synaptosomal membranes from adult rats. Increased sinaptosomal PLD activity in aged membranes appears to be independent of G protein regulation, whereas PAP2 activity is differentially regulated by GTPgammaS in aged membranes with respect to adult membranes. Our results suggest that under G-protein activation conditions, DAG production by the serial activation of PLD and PAP2 activities is increased in synaptosomal membranes from aged brain. The present paper demonstrates that PA generation (PLD activity) and degradation (PAPase activity) are differentially modulated during the aging process.


Leptomeningeal metastases from small cell lung carcinoma.
The current study was performed to investigate the frequency of leptomeningeal metastases (LMM) in patients with small cell lung carcinoma (SCLC) as well as the effect of LMM on survival, any correlation between the location of the LMM and survival, and a possible increased risk of LMM among patients with brain metastases (BM) located in the posterior fossa. Between 1980-2003, 458 consecutive patients with SCLC were enrolled in the current study. Patients underwent regular neurologic examination and imaging of the brain before, during, and after treatment. The diagnosis of LMM was established by either the presence of malignant cells in the cerebrospinal fluid or positive clinical symptoms and signs supported by radiologic findings on magnetic resonance imaging. The group of patients in the current study had a 2% prevalence of LMM and the 2-year cumulative incidence of LMM was found to be 10%. The median survival after the diagnosis of LMM was reported to be 1.3 months. The median survival among patients with LMM located in the spinal cord was 2.4 months. The reported LMM-free survival 2 years after the diagnosis of SCLC was 78% for patients without BM and 61% for those patients with BM. Approximately 15% of the patients with BM located in the posterior fossa developed LMM, whereas only 10% of patients with cerebral BM did. The current prospective study found a 2-year cumulative incidence of LMM of 10%, with a prevalence of 2%. Patients with LMM located in the spinal cord appeared to survive longer than patients with cranial LMM. SCLC patients with BM located in the posterior fossa may be at a higher risk of developing LMM compared with patients with cerebral BM.


The effect of forebrain lesions in the neonatal rat: survival of midbrain dopaminergic neurons and the crossed nigrostriatal projection.
Lesions were placed in the striatum and the olfactory tubercle of 1-day-old rat pups. Control and experimental animals were raised to adulthood. Efferent projections of mesencephalic neurons were examined by injecting the retrograde tracers horseradish peroxidase or Fast Blue into the undamaged striata of some experimental animals. The survival of the mesencephalic dopaminergic neurons was monitored by using immunocytochemical localization of tyrosine hydroxylase. Small lesions in the caudate-putamen had no appreciable effect on the survival of tyrosine hydroxylase-containing neurons in the mesencephalon, but the density of dopaminergic terminals adjacent to the lesion increased in the remaining caudate-putamen. Striatal lesions that involved an estimated area of more than one-third resulted in loss of dopamine neurons of the substantia nigra compacta. Rostral lesions in the striatum affected mostly rostrally positioned neurons in the substantia nigra. Dorsal lesions of the caudate-putamen resulted in disappearance of dorsal A9 neurons. Reduction of the A10 and A8 dopamine neuron groups occurred if the neonatal lesions involved the olfactory tubercle and nucleus accumbens. Some tyrosine hydroxylase-containing neurons persisted even after the largest lesions. These dopaminergic neurons formed a crossed nigrostriatal pathway which was confirmed by retrogradely transported tracers. The density of this crossed projection in the adult appeared unaffected by the neonatal lesion. We concluded that dopaminergic neurons form topographically ordered projections with their targets in the newborn rat. Rearrangement of these fibers appeared limited, but compensatory increase of axon terminal density was evident in partially lesioned target areas.


Detection of soluble type II receptor in the presence of its natural ligand IL-1 beta. Quantification by sandwich ELISA.
The type II interleukin-1 receptor (IL-1R II) is a newly described 60-68 kDa protein expressed on monocytes, neutrophils, and lymphocytes. It is hypothesized that a 45 kDa soluble form of the IL-1R II attenuates the proinflammatory effects of IL-1 by preventing its binding to the type I IL-1 receptor. However, very little information exists regarding the detection of soluble IL-1R II. Specifically, there are no reports to date characterizing IL-1R II detection by enzyme-linked immunoassay in the presence of IL-1 beta or characterizing IL-1 beta detection in the presence of IL-1R II. This study addresses the detection and quantitation of IL-1R II and IL-1 beta by a number of sandwich ELISA formats and characterizes the sensitivity of detection in the presence of competitive cytokines. We generated two distinct IL-1R II sandwich ELISAs that can detect receptor down to a level of 50 pg/ml. One, M22/R2, detects only unbound IL-1R II and the other, M2/R2, detects both bound and unbound IL-1R II. In this context, a 4:1 molar ratio of IL-1 beta to IL-1R II interferes with the IL-1R II detection by the M22/R2 but not the M2/R2 ELISA. Conversely, IL-1R II at physiologically relevant concentrations interferes with the detection of IL-1 beta by three distinct IL-1 beta ELISA formats. Taken together, these studies suggest that when measuring samples that may contain both IL-1 beta and IL-1R II, careful attention must be given to assay specificity.


Lentiviral hematopoietic cell gene therapy for X-linked adrenoleukodystrophy.
X-linked adrenoleukodystrophy (X-ALD) is a severe genetic demyelinating disease caused by a deficiency in ALD protein, an adenosine triphosphate-binding cassette transporter encoded by the ABCD1 gene. When performed at an early stage of the disease, allogeneic hematopoietic stem cell transplantation (HCT) can arrest the progression of cerebral demyelinating lesions. To overcome the limitations of allogeneic HCT, hematopoietic stem cell (HSC) gene therapy strategy aiming to perform autologous transplantation of lentivirally corrected cells was developed. We demonstrated the preclinical feasibility of HSC gene therapy for ALD based on the correction of CD34+ cells from X-ALD patients using an HIV1-derived lentiviral vector. These results prompted us to initiate an HSC gene therapy trial in two X-ALD patients who had developed progressive cerebral demyelination, were candidates for allogeneic HCT, but had no HLA-matched donors or cord blood. Autologous CD34+ cells were purified from the peripheral blood after G-CSF stimulation, genetically corrected ex vivo with a lentiviral vector encoding wild-type ABCD1 cDNA, and then reinfused into the patients after they had received full myeloablative conditioning. Over 3 years of follow-up, the hematopoiesis remained polyclonal in the two patients treated with 7-14% of granulocytes, monocytes, and T and B lymphocytes expressing the lentivirally encoded ALD protein. There was no evidence of clonal dominance or skewing based on the retrieval of lentiviral insertion repertoire in different hematopoietic lineages by deep sequencing. Cerebral demyelination was arrested 14 and 16months, respectively, in the two treated patients, without further progression up to the last follow-up, a clinical outcome that is comparable to that observed after allogeneic HCT. Longer follow-up of these two treated patients and HSC gene therapy performed in additional ALD patients are however needed to evaluate the safety and efficacy of lentiviral HSC gene therapy in cerebral forms of X-ALD.


Artificial Intelligence to Preoperatively Predict Proximal Junction Kyphosis Following Adult Spinal Deformity Surgery: Soft Tissue Imaging May Be Necessary for Accurate Models.
Retrospective cohort. In a cohort of patients undergoing adult spinal deformity (ASD) surgery, we used artificial intelligence to compare three models of preoperatively predicting radiographic proximal junction kyphosis (PJK) using: (1) traditional demographics and radiographic measurements, (2) raw preoperative scoliosis radiographs, and (3) raw preoperative thoracic magnetic resonance imaging (MRI). Despite many proposed risk factors, PJK following ASD surgery remains difficult to predict. A single-institution, retrospective cohort study was undertaken for patients undergoing ASD surgery from 2009 to 2021. PJK was defined as a sagittal Cobb angle of upper-instrumented vertebra (UIV) and UIV+2>10° and a postoperative change in UIV/UIV+2>10°. For model 1, a support vector machine was used to predict PJK within 2 years postoperatively using clinical and traditional sagittal/coronal radiographic variables and intended levels of instrumentation. Next, for model 2, a convolutional neural network (CNN) was trained on raw preoperative lateral and posterior-anterior scoliosis radiographs. Finally, for model 3, a CNN was trained on raw preoperative thoracic T1 MRIs. A total of 191 patients underwent ASD surgery with at least 2-year follow-up and 89 (46.6%) developed radiographic PJK within 2 years. Model 1: Using clinical variables and traditional radiographic measurements, the model achieved a sensitivity: 57.2% and a specificity: 56.3%. Model 2: a CNN with raw scoliosis x-rays predicted PJK with a sensitivity: 68.2% and specificity: 58.3%. Model 3: a CNN with raw thoracic MRIs predicted PJK with average sensitivity: 73.1% and specificity: 79.5%. Finally, an attention map outlined the imaging features used by model 3 elucidated that soft tissue features predominated all true positive PJK predictions. The use of raw MRIs in an artificial intelligence model improved the accuracy of PJK prediction compared with raw scoliosis radiographs and traditional clinical/radiographic measurements. The improved predictive accuracy using MRI may indicate that PJK is best predicted by soft tissue degeneration and muscle atrophy.


Impact of bronchoalveolar lavage multiplex polymerase chain reaction on microbiological yield and therapeutic decisions in severe pneumonia in intensive care unit.
The purpose of the study is to evaluate the impact of adding bronchoalveolar lavage multiplex polymerase chain reaction (M-PCR) to conventional cultures (CC) on microbiological yield and therapeutic decisions in adult intensive care unit patients with pneumonia and severe sepsis or septic shock. In this retrospective case-control study, bronchoalveolar lavage cultures were taken for control (58 patients, 58 admissions) and study arms (57 patients, 58 admissions). Bronchoalveolar lavage M-PCR was sent simultaneously for the latter. A total of 267 microorganisms were identified (M-PCR alone, 211; CC alone, 15; both, 41) in the study arm vs 64 in controls. Concordance between M-PCR and culture was complete in 32 (55.17%), partial in 4 (6.9%), and discordant in 22 (37.93%) including 17 with positive M-PCR but negative CC. Time to antibiotic therapy modification was significantly less (P < .001) in M-PCR group compared to controls (32.40 ± 14.41 vs 41.74 ± 45.61 hours). There was no significant difference in index episode resolution (48.3% vs 50%; P = 1), intensive care unit mortality (57.4% vs 51.2%; P = .67), and hospital mortality (59.6% vs 61.5%; P = 1) in study and control arms, respectively, despite more septic shock patients in the study arm (89.7% vs 75.9%; P = .05). Bronchoalveolar lavage M-PCR with culture leads to higher microbiological yield and earlier modification of antibiotics compared to conventional culture.