Monoclonal antibodies against human astrocytomas and their reactivity pattern

The establishment of hybridomas after fusion of X63-Ag8.653 mouse myeloma cells and splenocytes from mice hyperimmunized against human astrocytomas is presented. The animals were primed with 5 × 106 chemically modified uncultured or cultured glioma cells. Six weeks after the last immunization step an intrasplenal booster injection was administrated and 3 days later the spleen cells were prepared for fusion experiments. According to the specificity analysis of the generated antibodies 7 hybridoma products (MUC 7-22, MUC 8-22, MUC 10-22, MUC 11-22, MUC 14-22, MUC 15-22 and MUC 2-63) react with gliomas, neuroblastomas and melanomas as well as with embryonic and fetal cells but do not recognize non-neurogenic tumors. The selected monoclonal antibodies (McAbs) of IgG1 and IgG2a isotypes are not extensively characterized but these antibodies have been demonstrated to be reactive with a panel of glioma cell lines with varying patterns of antigen distribution. Using the McAbs described above and a series of cryosections of glioma biopsies and paraffin sections of the same material as well as glioma cultures established from these, variable antigenic profiles among glioma cell populations could be demonstrated. From these results it is evident that there is not only a distinct degree of antigenic heterogeneity among and within brain tumors, but also that the pattern of antigenic expression can change continuously. Some of the glioma associated antigens recognized by the selected antibodies persist after fixation with methanol/acetone and Karnovsky's fixative and probably are oncoembryonic/oncofetal antigen(s). The data suggest that the use of McAbs recognizing tumor associated oncofetal antigens in immunohistochemistry facilitates objective typing of intracranial malignancies and precise analysis of fine needle brain/tumor biopsies in a sensitive and reproducible manner

High-Pressure Synthesis of a Pentazolate Salt

The pentazolates, the last all-nitrogen members of the azole series, have been notoriously elusive for the last hundred years despite enormous efforts to make these compounds in either gas or condensed phases. Here we report a successful synthesis of a solid state compound consisting of isolated pentazolate anions N5-, which is achieved by compressing and laser heating cesium azide (CsN3) mixed with N2 cryogenic liquid in a diamond anvil cell. The experiment was guided by theory, which predicted the transformation of the mixture at high pressures to a new compound, cesium pentazolate salt (CsN5). Electron transfer from Cs atoms to N5 rings enables both aromaticity in the pentazolates as well as ionic bonding in the CsN5 crystal. This work provides a critical insight into the role of extreme conditions in exploring unusual bonding routes that ultimately lead to the formation of novel high nitrogen content species

Refined Approaches for Estimating the Strength of Rock Blocks

Micro-discrete fracture networks (μDFNs) have been integrated into grain-based models (GBMs) within the numerical software UDEC to assess rock block strength through a series of unconfined compressive strength (UCS) tests of progressively larger in size numerical specimens. GBMs were generated by utilizing a Voronoi tessellation scheme to capture the crack evolution processes within the intact rock material, and μDFNs were separately created and embedded into the GBMs to simulate the effect of pre-existing defects. Various μDFNs realisations were generated stochastically within the software FracMan to assess the combined impact of defect intensity, persistence, strength and specimen size. The resulting synthetic rock block models were used to assess the “flawed” material strength at block scale through a rigorous sensitivity numerical analysis. The acquired results predict a progressive strength reduction with decreasing intact rock quality and certain trends are captured when rock block strength is expressed as a function of a newly proposed “Defect Intensity× Persistence” factor. This allow us to standardise the data along specific strength reduction envelopes and to propose generic relationships that cover a wide range of defect geometrical combinations, defect strengths and sample sizes. Accordingly, an attempt is undertaken to refine two existing empirical approaches that consider the effect of scale and micro-defects explicitly for predicting the UCS of rock blocks

COL4A5 and LAMA5 variants co-inherited in familial hematuria: Digenic inheritance or genetic modifier effect?

Background: About 40-50% of patients with familial microscopic hematuria (FMH) caused by thin basement membrane nephropathy (TBMN) inherit heterozygous mutations in collagen IV genes (COL4A3, COL4A4). On long follow-up, the full phenotypic spectrum of these patients varies a lot, ranging from isolated MH or MH plus low-grade proteinuria to chronic renal failure of variable degree, including end-stage renal disease (ESRD). Methods: Here, we performed Whole Exome Sequencing (WES) in patients of six families, presenting with autosomal dominant FMH, with or without progression to proteinuria and loss of renal function, all previously found negative for severe collagen IV mutations. Hierarchical filtering of the WES data was performed, followed by mutation prediction analysis, Sanger sequencing and genetic segregation analysis. Results: In one family with four patients, we found evidence for the contribution of two co-inherited variants in two crucial genes expressed in the glomerular basement membrane (GBM); LAMA5-p.Pro1243Leu and COL4A5-p.Asp654Tyr. Mutations in COL4A5 cause classical X-linked Alport Syndrome, while rare mutations in the LAMA5 have been reported in patients with focal segmental glomerulosclerosis. The phenotypic spectrum of the patients includes hematuria, proteinuria, focal segmental glomerulosclerosis, loss of kidney function and renal cortical cysts. Conclusions: A modifier role of LAMA5 on the background of a hypomorphic Alport syndrome causing mutation is a possible explanation of our findings. Digenic inheritance is another scenario, following the concept that mutations at both loci more accurately explain the spectrum of symptoms, but further investigation is needed under this concept. This is the third report linking a LAMA5 variant with human renal disease and expanding the spectrum of genes involved in glomerular pathologies accompanied by familial hematurias. The cystic phenotype overlaps with that of a mouse model, which carried a Lama5 hypomorphic mutation that caused severely reduced Lama5 protein levels and produced kidney cysts. 2018 The Author(s).The work was supported from the Cyprus Research Promotion Foundation through the grant NEW INFRASTRUCTURE/STRATEGIC/0308/24 to CD (co-funded by the European Regional Development Fund and the Republic of Cyprus). The funding body did not contribute to the design of study, collection, analysis and interpretation of data, or in manuscript writing.Scopu

AAV9-mediated SH3TC2 gene replacement therapy targeted to Schwann cells for the treatment of CMT4C

Type 4C Charcot-Marie-Tooth (CMT4C) demyelinating neuropathy is caused by autosomal recessive SH3TC2 gene mutations. SH3TC2 is highly expressed in myelinating Schwann cells. CMT4C is a childhood-onset progressive disease without effective treatment. Here, we generated a gene therapy for CMT4C mediated by an adeno-associated viral 9 vector (AAV9) to deliver the human SH3TC2 gene in the Sh3tc2−/− mouse model of CMT4C. We used a minimal fragment of the myelin protein zero (Mpz) promoter (miniMpz), which was cloned and validated to achieve Schwann cell-targeted expression of SH3TC2. Following the demonstration of AAV9-miniMpz.SH3TC2myc vector efficacy to re-establish SH3TC2 expression in the peripheral nervous system, we performed an early as well as a delayed treatment trial in Sh3tc2−/− mice. We demonstrate both after early as well as following late treatment improvements in multiple motor performance tests and nerve conduction velocities. Moreover, treatment led to normalization of the organization of the nodes of Ranvier, which is typically deficient in CMT4C patients and Sh3tc2−/− mice, along with reduced ratios of demyelinated fibers, increased myelin thickness and reduced g-ratios at both time points of intervention. Taken together, our results provide a proof of concept for an effective and potentially translatable gene replacement therapy for CMT4C treatment

Logic shrinkage: learned connectivity sparsification for LUT-based neural networks

FPGA-specific DNN architectures using the native LUTs as independently trainable inference operators have been shown to achieve favorable area-accuracy and energy-accuracy tradeoffs. The first work in this area, LUTNet, exhibited state-of-the-art performance for standard DNN benchmarks. In this article, we propose the learned optimization of such LUT-based topologies, resulting in higher-efficiency designs than via the direct use of off-the-shelf, hand-designed networks. Existing implementations of this class of architecture require the manual specification of the number of inputs per LUT, K. Choosing appropriate K a priori is challenging, and doing so at even high granularity, e.g. per layer, is a time-consuming and error-prone process that leaves FPGAs’ spatial flexibility underexploited. Furthermore, prior works see LUT inputs connected randomly, which does not guarantee a good choice of network topology. To address these issues, we propose logic shrinkage, a fine-grained netlist pruning methodology enabling K to be automatically learned for every LUT in a neural network targeted for FPGA inference. By removing LUT inputs determined to be of low importance, our method increases the efficiency of the resultant accelerators. Our GPU-friendly solution to LUT input removal is capable of processing large topologies during their training with negligible slowdown. With logic shrinkage, we better the area and energy efficiency of the best-performing LUTNet implementation of the CNV network classifying CIFAR-10 by 1.54 × and 1.31 ×, respectively, while matching its accuracy. This implementation also reaches 2.71 × the area efficiency of an equally accurate, heavily pruned BNN. On ImageNet with the Bi-Real Net architecture, employment of logic shrinkage results in a post-synthesis area reduction of 2.67 × vs LUTNet, allowing for implementation that was previously impossible on today’s largest FPGAs. We validate the benefits of logic shrinkage in the context of real application deployment by implementing a face mask detection DNN using BNN, LUTNet and logic-shrunk layers. Our results show that logic shrinkage results in area gains versus LUTNet (up to 1.20 ×) and equally pruned BNNs (up to 1.08 ×), along with accuracy improvements

De-installation of the multi-organic dysfunction syndrome by associating the mitochondrial microcirculatory recruitment with multiple organ support therapy in extracorporeal life support organization

World Academy of Medical Sciences (WAMS), Netherland, Fundeni Institute, Bucharest, Romania, Institute of Scientific Research in the field of Mother and Child Health Care, Republic of Moldova, State University of Medicine and Pharmacy “Nicolae Testemitanu”, Republic of Moldova, State Medical University of Samara, The Russian Federation, Aretaieion University Hospital, Athens, Medical School, National, and Kapodistrian The University of Athens, Greece, Odesa National Medicine University, Ukraine, Private Hospital Medical Institution “Via - Intosana”, Republic of Moldova, Medicine University of Buenos Aires, Argentina, Al VI-lea Congres Național de Obstetrică și Ginecologie cu participare internațională, 13-15 septembrie 2018, Chișinău, Republica MoldovaIntroduction: The installation of macro-circulation centralization in MODS triggering in critical obstetric states caused by intravascular coagulation, HELLP, shock, SIRS, septicemia, CARS, embolism of the pulmonary artery, cerebral and other, – microcirculation will also be seriously damaged, as the reduction in blood flow perfusion affects the venous return to eliminate the waste of cellular metabolism, where a marker of tissue hypoxia is the increase in carbon dioxide. Objective: The mitochondrial microcirculatory recruitment with multiple organ support therapy in extracorporeal life support. Material and methods: This is a retrospective study over 35 years, in a lot of critical situations in obstetrics. Results: This disorder generates microcirculatory - mitochondrial distress syndrome, mitochondrial energy collapse, which can be recovered by microcirculation – mitochondrial recruitment to optimize systemic perfusion pressure (SPP), in turn, dependent on mean blood pressure and capillary resistance. Microcirculation - mitochondrial recruitment decentralizes macrocirculation benefits microcirculation in the capillary-cell metabolic area. In cases of manifestation respiratory-pulmonary CO2 ↑ (ARDS), confirmed ↓ PaO2/FiO2 ↓300 to Acute Respiratory Distress Syndrome (Berlin definition, 2012), thus also aggravates the microcirculatory-mitochondrial distress syndrome, mitochondrial collapse and the recruitment of the microcirculatory-mitochondrial is supplemented with multi-organ support therapy (MOST). 1. Alveolar recruitment through respiratory support in specific ventilation modes, predominantly APRV, with permissive hypercapnia at a normal pH. 2) MOST - extracorporeal with technical support. Extracorporeal Life Support Organization – ELSO. 3) Modeling of extra-vascular pulmonary fluid; 4) Th4 - Th5 thoracic epidural block. Conclusion: The absence of decreasing of the pCO2 tissue hypoxia marker at the A-V difference after microcirculatory - mitochondrial recruitment, rejects the necrosis /apoptosis, cellular hypo-(an)ergic and proves the mitochondrial eu-energetic metabolic remodeling with the elimination of the hypo (an) ergic mitochondria performed by clearance lysosomal (mitophagy), thus demonstrating eu-ergic mitochondria with the normalization of mitochondrial uniporter-Ca ++ and mitochondrial permeability pore transition, which productively inactivate the toxic forms of oxygen and nitrogen.Rezumat. Instalarea centralizării macro-circulaţiei în declanşarea MODS în stări critice de obstetrică cauzate de coagularea intravasculară, HELLP, şoc, SIRS, septicemie, CARS, embolie a arterei pulmonare, cerebrală şi altele; - microcirculaţia va fi de asemenea grav afectată, iar perfuzia fluxului sanguin afectează revenirea venoasă pentru a elimina deşeurile de metabolism celular, unde un marker al hipoxiei tisulare este creşterea dioxidului de carbon, la diferenţa A-V. Această tulburare generează sindromul detresei microcirculator – mitocondriale (MMDs), colapsul energetic mitocondrial, care poate fi de-instalat (recuperat) prin recrutarea microcirculator - mitocondrială odată cu optimizarea presiunii de perfuzie sistemică, în dependenţă de tensiunea arterială medie şi rezistenţa capilară. Recrutarea microcirculator - mitocondrială descentralizează macrocirculaţia şi ameliorează microcrculaţia în spaţiul metabolic capilar-celulă. În cazurile de manifestare a ↑CO2-dependent respirator-pulmonar, confirmat ↓ PaO2 / FiO2 ↓ 300 pentru ARDS, sindromul de detresă respiratorie acută (definiţia de la Berlin, 2012), agravează de asemenea, şi sindromul detresei microcirculator-mitocondriale, colapsul mitocondrial iar recrutarea microcirculator - mitocondrială este suplimentată cu terapia de sprijin multi-organ (MOST). 1. Recrutarea alveolară prin suport respirator în moduri de ventilaţie specifice preponderent APRV, cu hipercapnie permisivă la un pH normal. 2) MOST - extracorporal cu suport tehnic în managmentul vital prin sprijin extracorporeal - ELSO. 3) modelarea fluidului pulmonar extra-vascular; 4) Blocul epidural T4-Th5 toracic. Reducerea markerului hipoxiei tisulare pCO2 la diferenţa A-V după recuperarea microcirculator - mitocondrială, respinge necroza / apoptoza, hipo-(an)ergicul celular şi dovedeşte remodelarea metabolică eu-energetică mitocondrială prin eliminarea hipo (an) mitocondriilor ergice efectuate prin clearance-ul lizozomal (mitofagie), demonstrând astfel mitocondriile eu-ergice cu normalizarea tranziţiei porilor permeabilităţii mitocondriale şi canalului uniporter-Ca ++ , care inactivează productiv formele toxice de oxigen şi azot

Estrogen receptor alpha gene polymorphism and endometrial cancer risk – a case-control study

Background: Estrogen is an established endometrial carcinogen. One of the most important mediators of estrogenic action is the estrogen receptor alpha. We have investigated whether polymorphic variation in the estrogen receptor alpha gene (ESR1) is associated with endometrial cancer risk. Methods: In 702 cases with invasive endometrial cancer and 1563 controls, we genotyped five markers in ESR1 and used logistic regression models to estimate odds ratios (OR) and 95 percent confidence intervals (CI). Results: We found an association between rs2234670, rs2234693, as well as rs9340799, markers in strong linkage disequilibrium (LD), and endometrial cancer risk. The association with rs9340799 was the strongest, OR 0.75 (CI 0.60–0.93) for heterozygous and OR 0.53 (CI 0.37–0.77) for homozygous rare compared to those homozygous for the most common allele. Haplotype models did not fit better to the data than single marker models. Conclusion: We found that intronic variation in ESR1 was associated with endometrial cancer risk