Conditions for combining gene therapy with bone marrow transplantation in murine Krabbe disease.

Introduction: Krabbe disease (KD) is an autosomal recessive lysosomal disorder caused by mutations in the galactocerebrosidase (GALC) gene. This results in defective myelination in the peripheral and central nervous systems due to low GALC activity. Treatment at this time is limited to hematopoietic stem cell transplantation (HSCT) in pre-symptomatic individuals. While this treatment extends the lives of treated individuals, most have difficulty walking by the end of the first decade due to peripheral neuropathy. Studies in the murine model of KD, twitcher (twi) combining bone marrow transplantation (BMT) with AAVrh10-mGALC showed a great extension of life from 40 days to about 400 days, with some living a full life time. Methods: In order to find the optimum conditions for dosing and timing of this combined treatment, twi mice were injected with five doses of AAVrh10-mGALC at different times after BMT. Survival, as well as GALC expression were monitored along with studies of sciatic nerve myelination and possible liver pathology. Results: Dosing had a pronounced effect on survival and measured GALC activity. There was window of time after BMT to inject the viral vector and see similar results, however delaying both the BMT and the viral injection shortened the lifespans of the treated mice. Lowering the viral dose too much decreased the correction of the sciatic nerve myelination. There was no evidence for hepatic neoplasia. Conclusion: These studies provide the conditions optimum for successfully treating the murine model of KD. There is some flexibility in dosing and timing to obtain a satisfactory outcome. These studies are critical to the planning of a human trial combining the "standard of care", HSCT, with a single iv injection of AAVrh10-GALC

Can early treatment of twitcher mice with high dose AAVrh10-GALC eliminate the need for BMT?

Introduction: Krabbe disease (KD) is an autosomal recessive disorder caused by mutations in the galactocerebrosidase (GALC) gene resulting in neuro-inflammation and defective myelination in the central and peripheral nervous systems. Most infantile patients present with clinical features before six months of age and die before two years of age. The only treatment available for pre-symptomatic or mildly affected individuals is hematopoietic stem cell transplantation (HSCT). In the animal models, combining bone marrow transplantation (BMT) with gene therapy has shown the best results in disease outcome. In this study, we examine the outcome of gene therapy alone. Methods: Twitcher (twi) mice used in the study, have a W339X mutation in the GALC gene. Genotype identification of the mice was performed shortly after birth or post-natal day 1 (PND1), using polymerase chain reaction on the toe clips followed by restriction enzyme digestion and electrophoresis. Eight or nine-day-old affected mice were used for gene therapy treatment alone or combined with BMT. While iv injection of 4 × 1013 gc/kg of body weight of viral vector was used originally, different viral titers were also used without BMT to evaluate their outcomes. Results: When the standard viral dose was increased four- and ten-fold (4X and 10X) without BMT, the lifespans were increased significantly. Without BMT the affected mice were fertile, had the same weight and appearance as wild type mice and had normal strength and gait. The brains showed no staining for CD68, a marker for activated microglia/macrophages, and less astrogliosis than untreated twi mice. Conclusion: Our results demonstrate that, it may be possible to treat human KD patients with high dose AAVrh10 without blood stem cell transplantation which would eliminate the side effects of HSCT

A Preliminary Report on Cleft Deformities of the Face and associated Anomalies in Abha, Saudi Arabia

Cleft deformities of the face are one of the major congenital anomalies seen in our environment. There is a dearth of data from this major Southern city of Saudi Arabia on the incidence of the deformities and associated anomalies. This preliminary report aims to record the pattern of cleft lip/palate and associated anomalies at Aseer Central Hospital in Abha, Saudi Arabia. Materials and methods: A retrospective study was conducted at Aseer Central Hospital, Abha, to identify all cleft lip and palate patients that reported or were treated between 2005 and 2011. Cleft lip and/or palate records were obtained from patient’s files in Department of Maxillofacial Surgery, Pediatrics and Medical records. Gender of patient, type of cleft and any associated congenital anomalies/syndromes were recorded. Results: Of the patient records studied, 60% were patients male and 40% female. Isolated cleft palate was the most common at 60% followed by combined cleft lip palate at 24% and isolated cleft lip at 16%. Eleven percent of the patient’s studies showed associated anomalies with isolated cleft palate. Conclusion: The study concluded that isolated cleft palate cases are more common than other variants of orofacial clefts. The pattern of cleft observed in the current study was similar to reported studies for Arab populations. This preliminary report lays the foundation for large population and birth registry based studies for prevalence and frequency of orofacial anomalies in Saudi Arabia

BioImpacts: An emerging global journal

The toddling BioImpacts has now grown into a young adult with strong opinions and perspectives, to a high-quality journal, and it has not been raised but by a family of professional editors, reviewers, authors, and even readers who had fantasized about a bright future and that fantasies are now coming true one-by-one

Combined effects of hyperglycemic conditions and HIV-1 Nef: a potential model for induced HIV neuropathogenesis

Hyperglycemic conditions associated with diabetes mellitus (DM) or with the use of antiretroviral therapy may increase the risk of central nervous system (CNS) disorders in HIV-1 infected patients. In support of this hypothesis, we investigated the combined effects of hyperglycemic conditions and HIV-1 accessory protein Nef on the CNS using both in vitro and in vivo models. Astrocytes, the most abundant glial cell type required for normal synaptic transmission and other functions were selected for our in vitro study. The results show that in vitro hyperglycemic conditions enhance the expression of proinflammatory cytokines including caspase-3, complement factor 3 (C3), and the production of total nitrate and 8-iso-PGF2 α as reactive oxygen species (ROS) in human astrocytes leading to cell death in a dose-dependent manner. Delivery of purified recombinant HIV-1 Nef protein, or Nef expressed via HIV-1-based vectors in astrocytes showed similar results. The expression of Nef protein delivered via HIV-1 vectors in combination with hyperglycemia further augmented the production of ROS, C3, activation of caspase-3, modulation of filamentous protein (F-protein), depolarization of the mitochondria, and loss of astrocytes. To further verify the effects of hyperglycemia and HIV-1 Nef protein on CNS individually or in combination, in vivo studies were performed in streptozotocin (STZ) induced diabetic mice, by injecting HIV-1 Nef expressing viral particles into the sub-cortical region of the brain. Our in vivo results were similar to in vitro findings indicating an enhanced production of caspases-3, ROS (lipid oxidation and total nitrate), and C3 in the brain tissues of these animals. Interestingly, the delivery of HIV-1 Nef protein alone caused similar damage to CNS as augmented by hyperglycemia conditions. Taken together, the data suggests that HIV-1 infected individuals with hyperglycemia could potentially be at a higher risk of developing CNS related complications

Evaluasi Karakter Agro-fisiologi dan Analisis Kekerabatan 10 Aksesi Tempuyung (Sonchus arvensis L.) di Lingkungan Alami

ABSTRACTPerennial sowthistle (Sonchus arvensis L.) is known as a medicinal plant but rarely cultivated, perennial sowthistle grows wild. Characterization is needed to determine the variation and relationship of perennial sowthistle in situ. Differences environment affect plant growth and bioactive content. Phenotype characterization of plant species is the basis for selection and improvement of properties. The purpose of this study was to evaluate the variation of agrophysiological characters and to analyze the relationship of 10 accessions of sowthistle based on agro-physiological characters. The experiment was conducted in October 2015 to February 2016. Descriptive variation characteristic of agrophysiological, correlation coefficient, principal component analysis (PCA) and cluster analysis (CA) were used to evaluate the phenotypic variability. The PCA and CA generated similar results. The first five principal component axes explained 91.7% of the total variation with PC1 (43.7%) and PC2 (22.9%). The CA showed that the degree of intraspecific similarity was 52.04%. Three clusters were formed among the 10 accessions especially with the separation of accessions that were collected from similar environments.Keywords: cluster, similarity, principal component, Sonchus arvensis L.ABSTRAKTempuyung dikenal sebagai tanaman obat tradisional namun belum banyak dibudidayakan, tempuyung tumbuh liar di alam. Perbedaan lingkungan tumbuh aksesi berpengaruh terhadap pertumbuhan dan kadar bioaktif tempuyung. Karakterisasi diperlukan untuk mengetahui keragaman dan kekerabatan tempuyung in situ. Karakterisasi fenotip spesies tanaman merupakan dasar untuk seleksi dan perbaikan sifat. Penelitian ini bertujuan mengevaluasi keragaman karakter agro-fisiologi, dan menganalisis kedekatan hubungan antar 10 aksesi tempuyung in situ berdasarkan karakter agro-fisiologi. Penelitian dilaksanakan pada bulan Oktober 2015 sampai Februari 2016. Deskripsi keragaan karakter agro-fisiologi, koefisien korelasi, analisis gerombol dan analisis komponen utama digunakan untuk mengevaluasi keragaman fenotip. Aksesi Tawangmangu menunjukkan keragaan tertinggi pada karakter jumlah daun, lebar daun, diameter batang, bobot basah daun, bobot kering daun, total bobot basah, total bobot kering, tebal daun dan total flavonoid. Analisis gerombol (AG) dan analisis komponen utama (AKU) memberikan hasil yang mirip. Lima sumbu komponen utama menjelaskan 91.7% total keragaman dengan KU 1 (43.7%) dan KU 2 (22.9%). AG menunjukkan tingkat kemiripan sebesar 52.04%. Tiga kelompok terbentuk dari 10 aksesi yang dikoleksi yang diduga berdasarkan kesamaan lingkungan tumbuh.Kata kunci: gerombol, kemiripan, komponen utama, Sonchus arvensis L

Sustainable Management of Phosphorus in Agriculture for Environmental Conservation

Phosphorus (P) is an essential macronutrient for plant growth and development. Although the P-concentration in soil is 1000 folds higher than in plants, it is rarely available for plant uptake due to low diffusion and high fixation rate in soil. Hence, plants experience P-deficiency in the absence of P-fertilization, which may cause approximately a 30–40% decrease in crop yield. This highlights the importance of using a large amount of phosphate fertilizers to meet crop demands. As P-fertilizer is derived from a nonrenewable and finite source of rock phosphate, this resource is decreasing over time. In addition, farmers are applying P-fertilizers randomly without considering the soil stock, which leads to the loss of P-resources. The low P-use-efficiency (PUE) of plants in the field condition (15–20%) highlights that most of the soil-applied P remains unavailable to plants, and excess P causes ground and surface water contamination (i.e., eutrophication) through leaching and runoff, which ultimately results in environmental pollution. Therefore, it is crucial to apply P-fertilizers considering the soil test value and PUE to protect the environment from contamination and sustainable management of P-resources. This chapter mainly focuses on the sustainable management of P in agricultural fields for environmental conservation

Polypyrrole-Fe[sub]2O[sub]3 nanocomposites with high dielectric constant : in situ chemical polymerisation

Novel nanocomposites of polypyrrole (PPy) dispersed with iron oxide (Fe2O3) particles have been synthesised by in-situ chemical oxidative polymerisation of pyrrole in the presence of ammonium persulfate (APS) as an oxidising agent. The concentration of Fe2O3 was varied between 10-50wt% of PPy. The simultaneous polymerisation of pyrrole and oxide addition led to the complete synthesis of nanocomposites. A maximum dielectric constant of ~28500 was observed at 20wt% of Fe2O3. The nanocomposites were characterised by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). XRD analysis confirmed the structure and crystallinity of the nanocomposites, and a strong interaction between PPy and the Fe2O3 nanoparticles was observed by FTIR technique. SEM and TEM images showed that Fe2O3 particles had been coated with PPy by establishing a network during the polymerisation process. The values of dielectric constant were obtained from capacitance measurements. The value of dielectric constant for nanocomposites with 20wt% of Fe2O3 was observed to be almost 12 times that of the pure PPy matrix. The high value of dielectric constant indicated a high packing density of Fe2O3 particles in the PPy matrix. These nanocomposites have potential applications in electronic or biomedical devices

Environmental and anthropogenic drivers of watercress (Nasturtium officinale) communities in char-lands and water channels across the Swat River Basin: implication for conservation planning

Recent anthropogenic sources and excess usage have immensely threatened the communities and habitat ecology of this region’s medicinally and economically significant crops. Therefore, our study aims to evaluate the community structure and related environmental characteristics sustaining Nasturtium officinale communities along the river basin (RB) in Northwest Pakistan, using the clustering procedure (Ward’s method) and Redundancy analysis (RDA). From 340 phytosociological plots (34 × 10 = 340), we identified four ecologically distinct assemblages of N. officinale governed by different environmental and anthropogenic factors for the first time. The floristic structure shows the dominance of herbaceous (100%), native (77%), and annual (58.09%) species indicating relatively stable communities; however, the existence of the invasive plants (14%) is perturbing and may cause instability in the future, resulting in the replacement of herbaceous plant species. Likewise, we noticed apparent variations in the environmental factors, i.e., clay percentage (p = 3.1 × 10−5), silt and sand percentage (p< 0.05), organic matter (p< 0.001), phosphorus and potassium (p< 0.05), and heavy metals, i.e., Pb, Zn, and Cd (p< 0.05), indicating their dynamic role in maintaining the structure and composition of these ecologically distinct communities. RDA has also demonstrated the fundamental role of these factors in species–environment correlations and explained the geospatial variability and plants’ ecological amplitudes in the Swat River wetland ecosystem. We concluded from this study that N. officinale communities are relatively stable due to their rapid colonization; however, most recent high anthropogenic interventions especially overharvesting and sand mining activities, apart from natural enemies, water deficit, mega-droughts, and recent flood intensification due to climate change scenario, are robust future threats to these communities. Our research highlights the dire need for the sustainable uses and conservation of these critical communities for aesthetics, as food for aquatic macrobiota and humans, enhancing water quality, breeding habitat, fodder crop, and its most promising medicinal properties in the region

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020