Natural deep eutectic solvent-A novel green solvent for protein stabilization.
Natural deep eutectic solvents (NADESs) have been explored to provide a favorable environment for protein stabilization. In this context, NADESs were prepared with the molar ratio of trehalose to betaine ranging from 1:3 to 1:9 (NADES 1-3 to NADES 1-9). There was a strong hydrogen bond interaction between trehalose and betaine, and the interaction weakened with the reduction of trehalose. The NADES 1-7 had good thermal stability (-60-100 °C), low viscosity, and suitable pH (around 7). Trypsin had the highest relative enzyme activity in 50 % (v/v) NADES 1-7 under different temperatures, pH, and storage time. Furthermore, the changes in kinetic parameters indicated that the hydrogen bond environment of 50 % NADES 1-7 increased the contact between the substrate and the trypsin, speeding up the enzymatic reaction rate. This stabilizing effect mainly derived from the virtue of NADES 1-7 itself rather than the superposition of individual components. Additionally, spectral analysis revealed that the NADES 1-7 promoted trypsin conformational folding, effectively protecting the natural structure of trypsin. Importantly, the NADES 1-7 had good biocompatibility, further expanding its application.


Developmental lead exposure differentially alters the susceptibility to chemoconvulsants in rats.
Sprague-Dawley rats were exposed to lead (Pb) acetate (0.2% w/v) in drinking water from postnatal day (PN) 1 through PN 25 followed by discontinuing Pb exposure for 25 days. The convulsion signs produced by the i.v. infusion of convulsants were observed at PN 25 and PN 50. Pb exposure significantly decreased pentylenetetrazol (PTZ)-, picrotoxin (PIC)-, and strychnine (STRY)-induced, but increased N-methyl-D-aspartate (NMDA) and 4-aminopyridine (4-AP)-induced convulsion thresholds at PN 25. After rehabilitation, the Pb-induced changes in PTZ-, NMDA- and 4-AP- induced convulsions persisted till PN 50. In contrast, the thresholds for PIC- and STRY-induced convulsions were reversed to an increase at PN 50. Blood Pb concentrations were 46.9 +/- 6.8 and 11.4 +/- 1.1 microg/dl at PN 25 and PN 50, respectively. The results demonstrate that developmental Pb exposure does not result in a global and persistent lowering of threshold in response to convulsive stimuli and the Pb-induced changes may be due to a selective modulation of inhibitory and excitatory neurotransmission depending on age and Pb concentration in blood.


Newcastle Disease Virus-Vectored H7 and H5 Live Vaccines Protect Chickens from Challenge with H7N9 or H5N1 Avian Influenza Viruses.
Sporadic human infections by a novel H7N9 virus occurred over a large geographic region in China. In this study, we show that Newcastle disease virus (NDV)-vectored H7 (NDV-H7) and NDV-H5 vaccines are able to induce antibodies with high hemagglutination inhibition (HI) titers and completely protect chickens from challenge with the novel H7N9 or highly pathogenic H5N1 viruses, respectively. Notably, a baculovirus-expressed H7 protein failed to protect chickens from H7N9 virus infection.


Steering restoration of coal mining degraded ecosystem to achieve sustainable development goal-13 (climate action): United Nations decade of ecosystem restoration (2021-2030).
For millennium, mining sector is a source not only of mineral extraction for industrialization, economic expansion, and urban sprawling, but also of socio-environmental concern. It, therefore, has been the central attention of the business and public policy sustainable development scheme for several years. Thus, gradually, mining industries are getting involved with the concerns such as carbon emissions mitigation and carbon accounting to govern a rhetorical shift towards "sustainable mining". However, there is scarce knowledge about how the emergence of a "green and self-sustaining" forestry reclamation strategy coupled with potential carbon sequestration capacity in degraded mining areas will be an impeccable option for achieving sustainable development goal-13 (SDG-13: climate action) and ecosystem services during United Nation decade of ecosystem restoration. This paper reviews the extent to which reforestation and sustainable land management practices that employed to enhance ecosystem carbon pool and atmospheric CO2 sequestration capacity to offset CO2 emission and SOC (soil organic carbon) losses, as consequences of coal mining, to partially mitigate global climate crisis. Moreover, future research is required on mining innovation concepts and its challenges for designing an SDG impact framework, so that it not only synergies amongst SDGs, but also trade-offs between each individual "politically legitimized post-2015 development agenda" (i.e. UNSDGs) could be depicted in a systematic way. In a developing country like India, it is also an utmost need to assess the environmental impact and economic performance of such technological innovation and its possible synergistic effect.


Li2RbSO4Cl with a Short Ultraviolet Absorption Edge and an Acentric Structure through Assembling Heteroleptic [LiO3Cl] Tetrahedra.
Currently, short-wavelength nonlinear optical materials are urgently needed. Through substituting homoleptic [LiO4] in centrosymmetric LiRbSO4 with heteroleptic [LiO3Cl] tetrahedra, an acentric sulfate chloride, Li2RbSO4Cl, was designed and synthesized by the high-temperature melting method. Li2RbSO4Cl shows a relatively short ultraviolet absorption edge (<200 nm) among newly reported sulfate chlorides. Millimeter-sized crystals were grown due to the congruent melting behavior and high thermal stability of the compound.


Exploring the risk factors for early-life sugar consumption: A birth cohort study.
Sugar consumption in early childhood is the primary cause of negative health outcomes, including early childhood caries. To investigate risk factors associated with early-life sugar consumption. Explanatory variables were collected at baseline of a birth cohort in Porto Alegre, Southern Brazil. At six months of age, data were collected on child feeding practices, including the number of foods and beverages containing sugar. Multivariate Poisson regression analysis with robust variance was performed. Virtually all children (98.3%) had consumed sugar by the age of 6 months. Multivariable analysis showed that the number of sweet items was significantly larger in children whose mothers were less than 20 years of age (MR = 1.19; 95% CI: 1.05-1.36), those from non-nuclear families (MR = 1.12; 95% CI: 1.04-1.20), those whose mothers had less than eight years of schooling (MR = 1.34; 95% CI: 1.20-1.50) and those whose mothers smoked (MR = 1.23; 95% CI: 1.13-1.35). Moreover, the number of sweet items was significantly lower among children who breastfed in the first hour of life (MR = 0.85; 95% CI: 0.76-0.95). Sugar consumption begins very early, especially in children with no access to breastfeeding in the first hours of life and those from younger, less educated, and smoking mothers.


Effect of hyperglycemia on the growth hormone response to L-dopa in diabetic subjects.
Plasma growth hormone (GH) levels were measured before and after oral administration of L-dopa in 8 insulin-dependent juvenile diabetic patients, and in 6 non insulin-dependent maturity-onset diabetics, while they were hyperglycemic, and values were compared with those of normal children and adult subjects. Maturity-onset diabetics failed to show an increase in serum GH levels after L-dopa administration, while serum peak GH values of juvenile diabetic patients were not significantly different from those of normal adults and children. Hyperglycemia thus failed to prevent L-dopa induced growth hormone response in insulin-dependent juvenile diabetic patients.


Epidermal Basement Membrane in Health and Disease.
Skin, as the organ protecting the individual from environmental aggressions, constantly meets external insults and is dependent on mechanical toughness for its preserved function. Accordingly, the epidermal basement membrane (BM) zone has adapted to enforce tissue integrity. It harbors anchoring structures created through unique organization of common BM components and expression of proteins exclusive to the epidermal BM zone. Evidence for the importance of its correct assembly and the nonredundancy of its components for skin integrity is apparent from the multiple skin blistering disorders caused by mutations in genes coding for proteins associated with the epidermal BM and from autoimmune disorders in which autoantibodies target these molecules. However, it has become clear that these proteins not only provide mechanical support but are also critically involved in tissue homeostasis, repair, and regeneration. In this chapter, we provide an overview of the unique organization and components of the epidermal BM. A special focus will be given to its function during regeneration, and in inherited and acquired diseases.


Role of single disulfide linkages in the folding and activity of scyllatoxin-based BH3 domain mimetics.
Anti-apoptotic Bcl-2 proteins are implicated in pathogenic cell survival and have attracted considerable interest as therapeutic targets. We recently developed a class of synthetic peptide based on scyllatoxin (ScTx) designed to mimic the helical BH3 interaction domain of the pro-apoptotic Bcl-2 protein Bax. In this communication, the contribution of single disulfides in the folding and function of ScTx-Bax peptides was investigated. We synthesized five ScTx-Bax variants, each presenting a different combination of native disulfide linkage and evaluated their ability to directly bind Bcl-2 in vitro. It was determined that the position of the disulfide linkage had significant implications on the structure and function of ScTx-Bax peptides. This study underscores the importance of structural dynamics in BH3:Bcl-2 interactions and further validates ScTx-based ligands as potential modulators of anti-apoptotic Bcl-2 function. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.


Development of a versatile LCM-Seq method for spatial transcriptomics of fluorescently tagged cholinergic neuron populations.
Single-cell transcriptomics are powerful tools to define neuronal cell types based on co-expressed gene clusters. Limited RNA input in these technologies necessarily compromises transcriptome coverage and accuracy of differential expression analysis. We propose that bulk RNA-Seq of neuronal pools defined by spatial position offers an alternative strategy to overcome these technical limitations. We report a laser-capture microdissection (LCM)-Seq method which allows deep transcriptome profiling of fluorescently tagged neuron populations isolated with LCM from histological sections of transgenic mice. Mild formaldehyde fixation of ZsGreen marker protein, LCM sampling of ∼300 pooled neurons, followed by RNA isolation, library preparation and RNA-Seq with methods optimized for nanogram amounts of moderately degraded RNA enabled us to detect ∼15,000 different transcripts in fluorescently labeled cholinergic neuron populations. The LCM-Seq approach showed excellent accuracy in quantitative studies, allowing us to detect 2891 transcripts expressed differentially between the spatially defined and clinically relevant cholinergic neuron populations of the dorsal caudate-putamen and medial septum. In summary, the LCM-Seq method we report in this study is a versatile, sensitive, and accurate bulk sequencing approach to study the transcriptome profile and differential gene expression of fluorescently tagged neuronal populations isolated from transgenic mice with high spatial precision.


Improving Outcomes through a Neonatal Abstinence Syndrome Collaborative in Maryland.
A statewide Maryland Perinatal Neonatal Quality Collaborative, facilitated by the Maryland Patient Safety Center (MPSC), identified the three specific, measurable, attainable, relevant, and time-limited (SMART) aims to improve outcomes of neonatal abstinence syndrome (NAS) care as follows: (1) to reduce hospital length of stay (LOS), (2) to reduce interhospital transfers, and (3) to reduce 30-day readmission rates of infants with NAS. The Maryland collaborative developed a bundle of best practices for care of infants with NAS. MPSC partnered with Vermont Oxford Network (VON) to utilize the VON NAS toolkit and provided its standardized NAS educational curriculum to address the three objectives for participating birthing hospitals. Efforts began in quarter 4 (Q4) of 2016 and continued for 2 years. Thirty-one of Maryland's 32 delivery hospitals (97%) participated in the 2-year collaborative. Additionally, one specialty pediatric hospital with an NAS unit participated in the group learnings. Participating facilities implemented components of the MPSC NAS bundle and provided their staff caring for infants with NAS and their mothers access to the VON standardized educational curriculum. MPSC partnered with VON to conduct two audits of implementation of policies and procedures in Q1 of 2016 and Q3 of 2018. The Maryland Department of Health supplied quarterly aggregate hospital information on LOS, interhospital transfers, and 30-day readmissions of infants with a discharge diagnosis of the International Classification of Disease, 10th Revision (ICD-10), P96.1. Among term infants with NAS with total hospital stay greater than 5 days, we observed a nonsignificant reduction in both mean and median LOS of 1.5 days. In this same group, the rate of interhospital transfers fell significantly from 20.1% in 2016 to 13.8 and 11.0% in 2017 and 2018, respectively. The best practice bundle created by the Maryland collaborative was associated with a reduction in the percentage of infants with NAS who required interhospital transfer, thereby reducing family disruption. · A state NAS collaborative engaged 97% of delivery hospitals in education and standardization of care.. · The collaborative witnessed a 1.5-day decrease in length of stay, similar to that observed in other state collaboratives.. · The unique outcome of our collaborative was a 50% decrease in the rate of interhospital transfer..


Iron regulation of ferritin gene expression.
Ferritin is a ubiquitous iron-storage protein found in the cells of animals, plants, molds, and bacteria which it protects from toxic intracellular levels of iron. Ferritin stores iron within a hollow protein shell formed by subunits of two types, H and L. The 5' untranslated regions of the two subunit mRNAs contain an almost identical 28-nucleotide sequence which regulates translation by binding to a specific cell sap protein. When cell iron level is low, this repressor protein obstructs translation of stored ferritin mRNAs, whereas increased iron levels release this protein, thus permitting extensive ferritin subunit synthesis to respond rapidly. Similar motifs in the 3' untranslated region of transferrin receptor mRNA interact with this protein to regulate breakdown of the mRNA and thus change the receptor population. Finally, transcription of the H and L genes can be independently increased by iron and other factors. In the case of iron, synthesis of the L-mRNA is increased preferentially since ferritin shells with a preponderance of L-subunits store iron more efficiently. Thus regulation of ferritin synthesis at the translational and transcriptional levels and by transferrin receptor mRNA abundance at the level of breakdown provide a coordinated mechanism for protecting cells against the effects of excess iron.


The mammalian homolog of the Drosophila discs large tumor suppressor protein up-regulates expression of the ELR+ CXC chemokine Scyb5.
The mammalian homolog of the Drosophila discs large tumor suppressor protein Dlg functions as a scaffolding protein that facilitates the transmission of diverse signals. In the present study, we attempted to identify the downstream target genes of Dlg, and found that Dlg up-regulates expression of the ELR+ CXC chemokine Scyb5, which has been implicated in the immune system. Our finding suggests that Scyb5 may play an important role in the tumor suppressor function of Dlg.


Ipsilateral radial-head dislocation and distal fractures of both forearm bones in a child.
Monteggia's fracture-dislocation is a common fracture pattern seen in children. In this case report, an unusual injury pattern, consisting of a dislocation of the radial head with ipsilateral fractures of the distal radius and ulna, is described. This fracture-dislocation may represent a variation of Monteggia's lesion.


SFPQ and Tau: critical factors contributing to rapid progression of Alzheimer's disease.
Dysfunctional RNA-binding proteins (RBPs) have been implicated in several neurodegenerative disorders. Recently, this paradigm of RBPs has been extended to pathophysiology of Alzheimer's disease (AD). Here, we identified disease subtype specific variations in the RNA-binding proteome (RBPome) of sporadic AD (spAD), rapidly progressive AD (rpAD), and sporadic Creutzfeldt Jakob disease (sCJD), as well as control cases using RNA pull-down assay in combination with proteomics. We show that one of these identified proteins, splicing factor proline and glutamine rich (SFPQ), is downregulated in the post-mortem brains of rapidly progressive AD patients, sCJD patients and 3xTg mice brain at terminal stage of the disease. In contrast, the expression of SFPQ was elevated at early stage of the disease in the 3xTg mice, and in vitro after oxidative stress stimuli. Strikingly, in rpAD patients' brains SFPQ showed a significant dislocation from the nucleus and cytoplasmic colocalization with TIA-1. Furthermore, in rpAD brain lesions, SFPQ and p-tau showed extranuclear colocalization. Of note, association between SFPQ and tau-oligomers in rpAD brains suggests a possible role of SFPQ in oligomerization and subsequent misfolding of tau protein. In line with the findings from the human brain, our in vitro study showed that SFPQ is recruited into TIA-1-positive stress granules (SGs) after oxidative stress induction, and colocalizes with tau/p-tau in these granules, providing a possible mechanism of SFPQ dislocation through pathological SGs. Furthermore, the expression of human tau in vitro induced significant downregulation of SFPQ, suggesting a causal role of tau in the downregulation of SFPQ. The findings from the current study indicate that the dysregulation and dislocation of SFPQ, the subsequent DNA-related anomalies and aberrant dynamics of SGs in association with pathological tau represents a critical pathway which contributes to rapid progression of AD.


Family understanding of psychiatric illness.
Family members of patients with a diagnosis of schizophrenia were surveyed to ascertain their understanding of their relative's illness. This paper reports on deficits in knowledge of family members as they struggle to absorb the repercussions of chronic illness. It was useful for the development of a psycho-education treatment curriculum while dramatically demonstrating the need for it.


CRISPR/Cas technology promotes the various application of Dunaliella salina system.
Dunaliella salina (D. salina) has been widely applied in various fields because of its inherent advantages, such as the study of halotolerant mechanism, wastewater treatment, recombinant proteins expression, biofuel production, preparation of natural materials, and others. However, owing to the existence of low yield or in the laboratory exploration stage, D. salina system has been greatly restricted for practical production of various components. In past decade, significant progresses have been achieved for research of D. salina in these fields. Among them, D. salina as a novel expression system demonstrated a bright prospect, especially for large-scale production of foreign proteins, like the vaccines, antibodies, and other therapeutic proteins. Due to the low efficiency, application of traditional regulation tools is also greatly limited for exploration of D. salina system. The emergence of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system offers a precise editing tool to overcome the obstacles of D. salina system. This review not only comprehensively summarizes the recent progresses of D. salina in domain of gene engineering but also gives a deep analysis of problems and deficiencies in different fields of D. salina. Moreover, further prospects of CRISPR/Cas system and its significant challenges have been discussed in various aspects of D. salina. It provides a great referencing value for speeding up the maturity of D. salina system, and also supplies practical guiding significance to expand the new application fields for D. salina. KEY POINTS: • The review provides recent research progresses of various applications of D. salina. • The problems and deficiencies in different fields of D. salina were deeply analyzed. • The further prospects of CRISPR/Cas technology in D. salina system were predicted. • CRISPR/Cas system will promote the new application fields and maturity for D. salina.


Poorly controlled ulcerative colitis treated by colectomy during remission induced by extracorporeal leukocyte removal therapy.
Both monocyte-granulocytapheresis (M-GCAP) and leukocytapheresis (LCAP) are categorized as extracorporeal leukocyte removal therapies (ECCTs). These therapies have been recognized as efficient adjuncts for patients of steroid-resistant ulcerative colitis (UC). This study aimed to consider the adaptation and the limitation of these new therapies from the clinical standpoint based on a case of UC showing strong resistance to high-dose continuous steroid injection therapy. The patient successfully underwent a scheduled colectomy while maintaining remission after applying M-GCAP and LCAP independently. Surgical therapy was chosen because of a deep ulcer in the patient's sigmoid colon, which was assumed to constitute a future risk for perforation. This case suggests that combining ECCT with steroid therapy can maintain such poorly controlled and high-risk UC patients safely for the scheduled colectomy while improving the prognosis by reducing the dosage of steroid efficiently prior to operation.


SATB2 and NGR1: potential upstream regulatory factors in uterine leiomyomas.
We attempted to identify the genes involved in the pathogenesis of uterine leiomyomas, under a hypothesis that the aberrant expression of upstream regulatory genes caused by aberrant DNA methylation is involved in the onset and development of uterine leiomyomas. To find such genes, we compared genome-wide mRNA expression and DNA methylation in uterine leiomyomas and adjacent normal myometrium. Analysis of the data by Ingenuity Pathway Analysis software identified SATB2 which is known to be an epigenetic regulator, and NRG1 as candidate upstream regulatory genes. To infer the functions of these genes, human uterine smooth muscle cell lines overexpressing SATB2 or NRG1 genes were established (SATB2 or NRG1 lines), and their transcriptomes and pathways were analyzed. SATB2 and NRG1 were confirmed to be hypermethylated and upregulated in most uterine leiomyoma specimens (nine to 11 of the 11 cases). Among the established cell lines, morphological changes from spindle-like forms to fibroblast-like forms with elongated protrusions were observed in only the SATB2 line. Pathway analysis revealed that WNT/β-catenin and TGF-β signaling pathways which are related to the pathogenesis of uterine leiomyomas were activated in both SATB2 and NRG1 lines. In addition, signaling of growth factors including VEGF, PDGF, and IGF1, and retinoic acid signaling were activated in the SATB2 and NRG1 lines, respectively. These results indicate that SATB2 and NRG1 overexpression induced many of the signaling pathways that are considered to be involved in the pathogenesis of uterine leiomyomas, suggesting that these genes have roles as upstream regulatory factors.


Prevalence, variation, and transmission patterns of human respiratory syncytial virus from pediatric patients in Hubei, China during 2020-2021.
Human respiratory syncytial virus (RSV) is a severe threat to children and a main cause of acute lower respiratory tract infections. Nevertheless, the intra-host evolution and inter-regional diffusion of RSV are little known. In this study, we performed a systematic surveillance in hospitalized children in Hubei during 2020-2021, in which 106 RSV-positive samples were detected both clinically and by metagenomic next generation sequencing (mNGS). RSV-A and RSV-B groups co-circulated during surveillance with RSV-B being predominant. About 46 high-quality genomes were used for further analyses. A total of 163 intra-host nucleotide variation (iSNV) sites distributed in 34 samples were detected, and glycoprotein (G) gene was the most enriched gene for iSNVs, with non-synonymous substitutions more than synonymous substitutions. Evolutionary dynamic analysis showed that the evolutionary rates of G and NS2 genes were higher, and the population size of RSV groups changed over time. We also found evidences of inter-regional diffusion from Europe and Oceania to Hubei for RSV-A and RSV-B, respectively. This study highlighted the intra-host and inter-host evolution of RSV, and provided some evidences for understanding the evolution of RSV.


miR-138 suppresses cell proliferation and invasion by inhibiting SOX9 in hepatocellular carcinoma [Retraction].
[This retracts the article on p. 2159 in vol. 8, PMID: 27347323.].


How does prestrain in the tympanic membrane affect middle-ear function? A finite-element model study in rabbit.
Experiments have shown that prestrain exists in the rabbit tympanic membrane (TM), also in the absence of external loads. To date, it is unclear how prestrain influences the vibration response of the middle ear (ME). In this study, a detailed 3D finite-element model of the rabbit ME was constructed based on experimentally validated material properties. The model incorporates different degrees of prestrain in the TM and simulates the ME vibration response to sound as a linear harmonic perturbation around the prestressed reference state. To account for finite deformations associated with large prestrains, a framework was developed that iteratively updates the initial unstrained geometry until the prestrained geometry is in agreement with the given reference geometry. After validating the model using quasi-static and acoustic measurement data, it was shown that small levels of prestrain already have a substantial impact on the normal umbo and footplate response due to a phenomenon known as prestress stiffening. Although the approach is not preferable, it was possible to replicate the effect of prestrain in the normal ME by appropriately scaling the elastic moduli and damping factors in the base model. To evaluate the effect of possible changes in TM prestrain when the normal state of the ear is altered due to pathological modifications in the ME structure, we created a model with a perforation in the TM. It was shown that the change in vibration response after perforation is affected at low frequencies by a release of TM prestrain. In future studies, it may be necessary to incorporate prestrain in ME models to better understand the function of the diseased or reconstructed ME, which may be relevant for the development of reconstructive tissue grafts in the middle ear.


Live-cell imaging of slow axonal transport in cultured neurons.
Cytoskeletal polymers and other cytosolic protein complexes are transported along axons in the slow components of axonal transport. Studies on the movement of neurofilaments and microtubules in the axons of cultured neurons indicate that these polymers actually move at fast rates and that the movements are also infrequent and highly asynchronous. These observations indicate that the slow rate of slow axonal transport is due to rapid movements interrupted by prolonged pauses which presents special challenges for studies on the mechanism of movement. This chapter describes the procedures that the author's laboratory has used to observe and analyze the movement of neurofilaments and microtubules in axons of cultured neurons from the superior cervical ganglia of neonatal rats. In particular, the author describes how to culture these neurons, how to transfect them by nuclear injection, and how to detect the rapid and infrequent movement of cytoskeletal polymers using time-lapse fluorescence imaging.


Quantitative evaluation of intercellular local deformation of human oocytes during Piezo-assisted intracytoplasmic sperm injection using video-based motion analysis.
To evaluate the relationship between the quantitative properties of local deformation of the ooplasm during Piezo-assisted intracytoplasmic sperm injection (Piezo-ICSI) and oocyte developmental fate after Piezo-ICSI. Video-based motion analysis and case-control study. Private fertility center. A total of 99 couples assigned to the standard Piezo-ICSI program. Video recordings of the sperm injection process during Piezo-ICSI. Quantitative deformation properties consisting of the orientation of deformation perpendicular (Perp) or parallel (Para) to the oolemma and the mechanical response of the ooplasm (elongation, contraction, and total): Perp-elongation, Perp-contraction, Perp-total, Para-elongation, Perp-contraction, and Para-total. The deformation parameters were compared among different angular ranges of 30°-60°, 60°-90°, 90°-120°, and 120°-150° (defined as 0° at the 3 o'clock puncture site). The Perp-total value at 30°-60° was significantly greater than that at the other angles (130.2 vs. 57.7-85.5, respectively) primarily due to substantially greater Perp-elongation values. The Para-total value at 30°-60° was significantly greater than that at the other angles (20.1 vs. 6.7-10.3, respectively) primarily due to substantially smaller Para-contraction values. The deformation parameters of the ooplasm adjacent to the first polar body were also compared among different developmental fates (fertilization failure, cleavage arrest, and blastocyst development). The Perp-total value of oocytes derived from blastocyst formation was significantly smaller than those derived from other developmental fates (58.1 vs. 70.0-87.3, respectively). The Piezo-ICSI process induced deformation with various mechanical behaviors in different regions within a single oocyte. Furthermore, the exposure of the ooplasm adjacent to the first polar body to excessive deformation was found to be a possible trigger for adverse fertilization and embryo development after Piezo-ICSI.


Refusing to Treat Sexual Dysfunction in Sex Offenders.
This article examines one kind of conscientious refusal: the refusal of healthcare professionals to treat sexual dysfunction in individuals with a history of sexual offending. According to what I call the orthodoxy, such refusal is invariably impermissible, whereas at least one other kind of conscientious refusal-refusal to offer abortion services-is not. I seek to put pressure on the orthodoxy by (1) motivating the view that either both kinds of conscientious refusal are permissible or neither is, and (2) critiquing two attempts to buttress it.


Babesia in North America: An Update.
Canine babesiosis results from infection of 1 of 5 identified protozoal species in the United States (Babesia conradae, Babesia sp. "coco," Babesia gibsoni, Babesia vogeli, and Babesia vulpes). They are part of the Apicomplexa family of protozoa and are obligate intraerythrocytic parasites. Domestic and wild canids are suspected of being intermediate hosts. This updated article aims to provide practical guidance about the clinical manifestations of disease, treatment options, and outcomes. In addition, the authors hope to provide some clarity about the taxonomy and nomenclature of these organisms, as they have undergone multiple changes since their initial discovery.


The diabetic patient and dental treatment: an update.
This paper has been written to both refresh and update clinicians' knowledge of diabetes. Treatment for patients with diabetes continues to develop with the majority of Type 1 diabetics now using multiple daily injections and an increasing minority using insulin pumps. Blood glucose monitoring and patient education programmes have resulted in more patient involvement in controlling this condition. Type 2 diabetics have had improvement in care provision through the development of shorter acting sulphonylureas and the potential for GLP1 injections. The impact of diabetes on both oral health and quality of life is discussed. Practical suggestions are made regarding the dental treatment of diabetic patients using both local anaesthetic and under sedation. Diabetes continues to be a fickle master for those affected by this condition. The paper is written from the perspective of the 'expert patient'. It is hoped that a greater understanding of this chronic condition will improve both access to, and safety of, dental care for those patients with diabetes.


The single miss ng tooth: conventional fixed and removable solution capabilities are expanded with implant supported restorations.
Implants are a major emerging field in dental technology. And along with this emergence are different procedures required from dental laboratories. This article begins a five part series focusing on "Implant Supported Restorations."


Contributions of private landowners to the conservation of the gopher tortoise (Gopherus polyphemus).
Private landowners play a pivotal role in determining whether or not rare species persist in regions where privately owned land is extensive. The range of the gopher tortoise (Gopherus polyphemus) is confined to the Southeastern U.S., a region predominantly under private ownership, and thus the status of this species is largely dependent upon land management decisions made by private landowners. We sent an anonymous mail survey to 2,584 individuals to examine factors affecting gopher tortoise occurrence on private lands in Mississippi (adjusted response rate of 23%). Few respondents (19%) reported currently having tortoises on their property, although many had them in the past (30%). Tortoises were persisting primarily on larger properties with longleaf pine that were not managed chiefly for timber production. In general, respondents were largely unaware of habitat requirements of tortoises or effects of various land management practices on them, and few reported using management techniques that benefit tortoises, such as prescribed burning. Most respondents (57%) knew of wildlife incentive programs, but were hesitant to enroll because they did not want to commit to managing their property in a particular manner (34%). We suggest actions that could improve the likelihood of tortoise persistence in this region, as well as changes that could be made to incentive programs to increase landowner participation. These suggestions should be relevant to the conservation of other rare species on private lands in other regions.


PCLO gene: its role in vulnerability to major depressive disorder.
A recent genome-wide association study on Major Depressive Disorder (MDD) identified a specific association with a non-synonymous polymorphism (rs2522833) of a gene encoding the presynaptic protein piccolo (PCLO). A high percentage of patients who develop MDD have particular temperamental traits, such as passivity, pessimism, indecisiveness, and low self-esteem, which are related to the subsequent development of depression. The aims of this study were to perform a replicate case-control study and to conduct the first association study between the rs2522833 polymorphism and depression-related personality traits using the Temperament and Character Inventory (TCI) in a healthy subject sample. A total of 522 MDD patients and 375 healthy volunteers were enrolled in the study. Two hundred and forty-six controls agreed to fill out the TCI. The results showed that rs2522833 CC homozygotes were more frequent among the depressed patients than in the controls (p<0.01). The C allele distribution showed a trend in the same direction (p=0.08). Among controls, we found that the C allele carriers were associated with personality traits increasing vulnerability to depression, including higher Harm Avoidance (HA) and lower in Novelty Seeking (NS). In particular, C allele carriers were more fearful (HA2) and fatigable (HA4), and less impulsive/more deliberate (NS2) and less extravagant/more frugal (NS3). The absence of possible epistatic interaction effect. These results provide further support for the involvement of the PCLO gene in MDD and show that this effect may be mediated by influencing personality traits that increase the risk of major depression.


Community changes following shrub invasion of grassland.
We studied the development of the shrub Baccharis pilularis ssp consanguinea and its effects on herbs of the annual grassland in Northern California. A series of shands of Baccharis was sampled of ages ranging from 1 yr to>9 yr, representing most of the life cycle of the shrub. In each stand we examined shrub biomass, structure and litterfall. We also determined cover and biomass of all herbaceous species and estimated seed production, seed rain and storage of seed in the soil. Abundances of all herbaceous species declined greatly after Baccharis formed a closed canopy at 2-3 yr, and little seed of herbaceous species was either dispersed into shrub stands or stored in the soil. Exclosures suggested that herbivory by small mammals in the closed shrub stands may be important in reducing the abundance of herbaceous species following shrub invasion of grassland.


Usefulness of an angioplasty balloon as selective bronchial blockade device after transbronchial cryobiopsy.
Transbronchial cryobiopsy (TBCB) is a technique in which frozen samples of lung are obtained using a probe inserted through a bronchoscope. We performed a retrospective study to assess the performance of the TBCB procedure complemented by segmental bronchial blockade using an angioplasty balloon, in terms of diagnostic yield and safety in diffuse parenchymal lung disease (DPLD). Data from 100 patients with suspected DPLD (clinical and radiological findings), who underwent TBCB in our institution to establish a definitive diagnosis, were reviewed. In our institution, TBCB is monitored with fluoroscopy and performed under general anaesthesia by a multidisciplinary team (an anaesthesiologist, a pulmonologist and an interventional radiologist). In each patient, four samples were collected using a 2.4-mm distal diameter cryoprobe. To control bleeding, the biopsied segmental bronchus was blocked with a 6-mm diameter angioplasty balloon, inserted over a 0.035-inch angled hydrophilic guidewire. After the cryoextraction, the balloon was inflated for 3 min intervals until bleeding stopped. Overall, 98% of samples had diagnostic value. In 85% of cases, DPLD was confirmed, while in 7%, cancer was diagnosed. Complications were observed in 16% of the patients: 13 patients developed moderate haemorrhage, and 3 developed pneumothorax. Transbronchial cryobiopsy had a high diagnostic yield for DPLD. Performing the procedure under fluoroscopy guidance and using angioplasty balloon for selective bronchial blockade achieved a low rate of iatrogenic complications directly associated with the technique.


The neutralization mechanism of Vipera palaestinae neurotoxin by a purified factor from homologous serum.
The serum of Vipera palaestinae contains two separate factors which neutralize the hemorrhagic and the neurotoxic activities of its venom. The purified antineurotoxic factor shows one major band in disc electrophoresis and an 800-fold increase in specific activity. Its molecular weight is approximately 56 000 and the isoelectric point is 4.0. These data, together with the failure to form precipitin lines in immuno-diffusion tests, suggest that the antineurotoxic factor of Vipera palaestinae serum is probably an albumin-like or alpha-globulin fraction rather than an immunoglobulin fraction. In vivo experiments show that the neutralizing protein does not protect or block the neurotoxin target sites. When the neurotoxic fraction is injected into mice intravenously at different intervals following injection of the snake serum, the neutralizing activity decreases with time. Experiments carried out in vitro show that the antineurotoxic factor is inactivated upon heating at 95 degrees C for 10 min but resists boiling when mixed with the neurotoxic component, suggesting the formation of a thermostable complex between the two components. This assumed complex dissociates at pH 2.5 and the neurotoxic activity reappears. Further investigation of the reaction with 131I-labeled neurotoxin and neurotoxin components shows that a stable complex is formed between the neutralizing protein and one of the synergistic components of the neurotoxic fraction.


Anti-virulence activity of novel (1-heteroaryloxy-2-hydroxypropyl)- phenylpiperazine derivatives against both wild-type and clinical drug-resistant Candida albicans strains.
Candida albicans is an important human fungal pathogen. Our previous study disclosed that aryloxy-phenylpiperazine skeleton was a promising molecule to suppress C. albicans virulence by inhibiting hypha formation and biofilm formation. In order to deeply understand the efficacy and mechanism of action of phenylpiperazine compounds, and obtain new derivatives with excellent activity against C. albicans, hence, we synthesized three series of (1-heteroaryloxy-2-hydroxypropyl)-phenylpiperazines and evaluated their inhibitory activity against C. albicans both in vitro and in vivo in this study. Compared with previously reported aryloxy-phenylpiperazines, part of these heteroaryloxy derivatives improved their activities by strongly suppressing hypha formation and biofilm formation in C. albicans SC5314. Especially, (9H-carbazol-4-yl)oxy derivatives 25, 26, 27 and 28 exhibited strong activity in reducing C. albicans virulence in both human cell lines in vitro and mouse infection models in vivo. The compound 27 attenuated the virulence of various clinical C. albicans strains, including clinical drug-resistant C. albicans strains. Moreover, additive effects of the compound 27 with antifungal drugs against drug-resistant C. albicans strains were also discussed. Furthermore, the compound 27 significantly improved the composition and richness of the faecal microbiota in mice infected by C. albicans. These findings indicate that these piperazine compounds have great potential to be developed as new therapeutic drugs against C. albicans infection.


Using hyperrealistic simulations to improve digital informed consent for endourological procedures.
To show that digital informed consent (DIC) improves the subjective understanding of information and, therefore, informed consent. A nonblinded randomized controlled trial was performed in 84 patients who had undergone transurethral resection of bladder, transurethral resection of prostate, or ureterorenoscopy between July 2017 and March 2018. The DIC group watched a hyperrealistic simulation on a tablet device before surgery. After surgery and again 30 days later, both groups completed a validated questionnaire that measured subjective understanding, anxiety, and utility of and need for information. The mean ± SD age of the participants was 68.7 ± 11.1 years. Nine of 84 patients (10.7%) did not complete the questionnaire. A total of 42 patients were allocated to the DIC group and 42 to the control group. The mean ± SD score for immediate subjective understanding in the DIC group was 14.5% higher than in the control group (72% ± 17.5% vs 57.5% ± 23.5%, respectively; P = 0.006). There was no statistical difference in anxiety, utility of and need for information relative to delayed subjective understanding. In subgroup analysis, subjective understanding scores were higher, but not significantly so, among patients with low and higher education levels in the DIC group than in the control group (68% ± 18.1% vs 54% ± 22.5% [P = 0.06] and 76% ± 18.3% vs and 66% ± 21.9%, respectively [P = 0.89]). Hyperrealistic simulations improved subjective understanding of information and, therefore, informed consent for endourological procedures.


Genetic studies of low-abundance human plasma proteins. VII. Heterogeneity of the C1S subcomponent of the first complement component.
Charge-based structural variation has been observed in the C1s subcomponent of the first complement component C1 after isoelectric focusing and immunoblotting. One common and two uncommon autosomal co-dominantly expressed alleles, designated C1S*1, C1S*2 and C1S*3, have been recognized at the C1S structural locus. The frequency of these alleles was 0.979, 0.016 and 0.005, respectively, in a U.S. white population. No variation at the C1S locus was observed in a U.S. black sample (n = 95).


Measurement of elastic properties of prostate cancer cells using AFM.
This communication reports that three prostate cancer cells of differing metastatic potential were discriminated based on their Young's moduli (LNCaP - 287 +/- 52 N m(-2), PC-3 - 1401 +/- 162 N m(-2) and BPH - 2797 +/- 491 N m(-2)) which were determined using AFM and the Hertz model.


Models of primary care service delivery in Ontario: why such diversity?
A surprisingly large and ever-growing number of alternative models of primary care service delivery have been developed in Ontario. The models are relatively poorly understood, and it is unclear why there are so many of them. What needs of providers and of government as payer are they attempting to address? Through a literature review and interviews with key informants, we sought to explain why there are so many models.


Abnormal whole-brain functional connection in amnestic mild cognitive impairment patients.
Amnestic mild cognitive impairment (aMCI) patients are thought to be particularly vulnerable to convert to clinical AD where functional disconnection is a major feature of the cortical neuropathology. However, the presence and extent of whole-brain connectivity disturbances is largely unknown in aMCI patients. Twenty-six aMCI patients and eighteen matched healthy subjects were evaluated at baseline and at mean 20 months follow up. Temporal correlations between spatially distinct regions were evaluated by using longitudinal resting-state fMRI. Compared to normal aging controls, patterns of abnormal interregional correlations in widely dispersed brain areas were identified in the patients, which also changed with disease progression. These disturbances were found particularly in subcortical regions and frontal cortex. Importantly, significantly decreased negative functional connection may be specifically associated with the development of aMCI patients. This suggests a compensatory mechanism is underway where local processing deficits are offset by recruitment of more dispersed cortical regions. In addition, the presence of this increased connectivity is seen to eventually weaken with disease progression. The results suggest that patterns of whole-brain functional connection may be a useful risk marker for conversion to AD in aMCI patients.


Bioactivity studies on TiO₂-bearing Na₂O-CaO-SiO₂-B₂O₃ glasses.
Soda lime silica borate glasses mixed with different concentrations of TiO2 are synthesized by the melt-quenching technique. As a part of study on bioactivity of these glasses, the samples were immersed in simulated body fluid (SBF) solution for prolonged times (~21 days) during which weight loss along with pH measurements is carried out at specific intervals of time. The XRD and SEM analyses of post-immersed samples confirm the formation of crystalline hydroxyapatite layer (HA) on the surface of the samples. To assess the role of TiO2 on the formation of HA layer and degradability of the samples the spectroscopic studies viz. optical absorption and IR spectral studies on post- and pre-immersed samples have been carried out. The analysis of the results of degradability together with spectroscopic studies as a function of TiO2 concentration indicated that about 6.0 mol% of TiO2 is the optimal concentration for achieving better bioactivity of these glasses. The presence of the maximal concentration octahedral titanium ions in this glass that facilitates the formation of HA layer is found to be the reason for such a higher bioactivity.


Socioeconomic Inequalities in Elective and Nonelective Hospitalizations in Older Men.
Among older adults, there is limited and inconsistent evidence on the association between socioeconomic position (SEP) and elective and nonelective hospitalization. To evaluate the association between SEP and all-cause and cause-specific elective and nonelective hospitalization and hospital length of stay among older men. This population-based, prospective cohort study used data from the Concord Health and Aging in Men Project (CHAMP). CHAMP recruited 1705 men aged 70 years or older between January 28, 2005, and June 4, 2007, in Sydney, Australia. Data were analyzed from February 1 to September 30, 2021. Indicators of SEP, including education (university degree certificate, diploma or no postschool qualifications), occupation (professionals and managers; small employers and self-employed; or lower clerical, service, sales workers, skilled, and unskilled workers), and source of income (other sources of income than government pension, reliance on government pensions and other sources of income, or reliant solely on a government pension), and a cumulative SEP score (tertiles) as SEP indicators; 3-level variables present high, intermediate, and low SEP. All-cause and cause-specific elective and nonelective hospitalizations, number of hospitalizations, and length of stay were the study outcomes, ascertained through data linkage. Associations were quantified using competing-risks survival regression and negative binomial regression. A total of 1566 men (mean [SD] age, 76.8 [5.4] years) were included. During a mean (SD) 9.07 (3.53) years of follow-up, 1067 men had at least 1 elective hospitalization, and 1255 men had at least 1 nonelective hospitalization. No associations were found between SEP and elective hospitalizations. Being in the lowest tertile for educational level (subhazard ratio [SHR], 1.32; 95% CI, 1.11-1.58), occupational position (SHR, 1.30; 95% CI, 1.12-1.50), sources of income (SHR, 1.33; 95% CI, 1.17-1.52), and cumulative SEP tertile groups (SHR, 1.45; 95% CI, 1.24-1.68) were all associated with having at least 1 nonelective hospitalization compared with those in the highest tertiles. Significant associations were found between being in the lowest SEP groups and increased numbers and longer length of stay of nonelective hospitalizations. In this prospective cohort study, low SEP was inversely associated with nonelective hospitalizations but not elective hospitalization in older men in Australia. These findings point to the existence of socioeconomic inequalities in health care use, indicative of a need to take action to reduce these inequalities.


[Epidemiological features of patients hospitalized in coronary care units at the military hospital, Tunisia, for 1994-1998: preliminary results].
The aim of this study was to evaluate the frequency of cardiovascular disease and risk factors associated in patients hospitalised in coronary care units at Military Hospital, Tunis, over the period 1994-1998. The clinical features of 3513 patients (2389 men and 1124 women) on hospital admission were analysed. 47.3% of patients were hospitalised for coronary disease, 12.5% for valvular heart disease, 5.2% for cardiomyopathy, 16.2% for arrhythmia and conduction disturbance, 6.4% for hypertension and 12.2% for other pathologies. With this risk factor profile Tunisia has to implement a national strategy of primary prevention and heart heath promotion in addition to the efforts recently made in secondary prevention of some chronic disease such as hypertension and diabetes.


Bumping up kinase activity with an ATP-derived neo-substrate.
Pharmacologic agents capable of increasing kinase function would be useful for treating diseases associated with reduced kinase activity, such as inherited forms of Parkinson's disease. In this issue, Hertz et al. report an innovative approach for activating the Parkinson's-associated kinase PINK1 in cells with an ATP-derived neo-substrate.


Paradigm shift in antibiotic-resistome of petroleum hydrocarbon contaminated soil.
The increasing prevalence of antibiotic-resistant microorganisms in both clinical and environmental samples is of great concern for public health. In the present study, environmental samples from seven different sites, heavily contaminated with petroleum hydrocarbons has been examined for the antimicrobial resistome through metagenomic approach. The soil samples were found to be contaminated with high concentration of total petroleum hydrocarbons (average 45 g/kg), polyaromatic hydrocarbons (average ∑16PAH = 280 mg/kg), and heavy metals, which shapes the microbial community and their function. Proteobacteria was found to be predominant phylum in the contaminated habitat with the highest diversity (55.91%) followed by Actinobacteria (9.86%). Although the taxonomical abundance of the non-contaminated sample was not significantly different from contaminated samples, the functional abundance of genes related to antibiotic resistance was found to be higher up to 2 fold in contaminated samples. The comparative metagenomic analysis revealed a higher abundance of different antibiotic resistance genes, especially genes for fluoroquinolones was found to be higher up to 10 fold in contaminated samples. Moreover, the study has shown a significant difference in total functional diversity and abundance, mainly genes for aromatic compound metabolism and genes for phages, mobile genetic elements. These higher abundances of well recognized antibiotic resistance genes, multidrug efflux pumps, and integrons, suggest that the petroleum hydrocarbon contaminated sites can act as reservoirs for development of antibiotic resistance genes (ARGs). From this study, a significant link between the presence of petroleum hydrocarbon and the development of antibiotic resistance in the microbiome of contaminated habitat has been established.


Adverse events associated with fluoroscopically guided sacroiliac joint injections.
To describe the type, incidence, and factors that contribute to adverse events associated with fluoroscopically guided intra-articular sacroiliac joint injections (IASIJ). A retrospective cohort study. Tertiary, academic, outpatient physical medicine and rehabilitation interventional spine clinic. English-speaking adults aged 18-90 years who underwent fluoroscopically guided IASIJ injections between March 8, 2004, and April 19, 2007. After IASIJ injections, 3 senior researchers recorded the presence and types of adverse events. The relationship of adverse events with age, gender, fluoroscopy time, vital signs, and trainee presence was analyzed with the Fisher exact or the Wilcoxon rank sum 2-sided tests. The frequency of immediate (during or immediately after the procedure) or delayed (within 24-72 hours after the procedure) adverse events. A total of 162 patients (133 women) underwent 191 procedures. The range of subject age was from 20 to 90 years (15.8 years, standard deviation [SD]). The range (SD) of the preprocedure 11-point Likert Pain Scale was from 1.0 to 10.0 (2.0) and for the postprocedure 11-point Likert Pain Scale was from 0.0 to 9.0 (2.5). Trainees were involved in 57% of the procedures. Reported immediate adverse events were vasovagal reaction (2.1% [n = 4]) and steroid-clogged needle (0.5% [n = 1]). Follow-up data were available for 132 of 191 procedures (69%). There were 32 adverse events reported at a mean follow-up interval of 2 days, of which, the most frequent adverse events were injection-site soreness (12.9% [n = 17]), pain exacerbation (5.3% [n = 7]), and facial flushing and/or sweating (2.3% [n = 3]). Delayed adverse events decreased with older age (P = .0029). The patients who underwent bilateral procedures experienced more delayed adverse events than the patients who underwent unilateral procedures (P = .024). Fluoroscopically guided IASIJ injection is associated with minimal adverse effects. The most common immediate adverse event was vasovagal reaction, and the most common delayed adverse event was injection-site soreness. Younger age is significantly related to reported delayed adverse events.


Unilateral reduction of the early and late blink reflex component in hemiparkinson syndrome.
Electrically evoked blink reflexes were recorded in 10 patients with unilateral tremor and/or rigor, mostly diagnosed as hemiparkinson syndrome. Five of the patients could be investigated before and after stereotaxic thalamo-subthalamotomy. The EMG activity of the early and late components was quantified by means of averaging and integrations techniques. In addition, the latencies of the reflex components were determined. The major finding was a unilateral decrease of the EMG activity of both the early and late components confined to the clinically affected side. This pattern points to a lowered state of excitability of neurons in or close to the facial nucleus, probably due to a dysfunction of contralateral EPMS structures. A loss of facilitatory influences from EPMS centers, e.g. the nigro-striatal system, on brain stem neurons in the area of the facial nucleus appears most probable. The latencies of the early and late components were mostly normal. After stereotaxic surgery, the BR activity was decreased bilaterally. The difference between the affected and unaffected sides, however, was nearly unchanged. At present an unspecific postoperative effect cannot be excluded.


Cloning and structural analysis of a gene encoding a mouse mastocytoma proteoglycan core protein; analysis of its evolutionary relation to three cross hybridizing regions in the mouse genome.
Serglycin (SGC) is a Ser-Gly-repeat-containing protein, used as a proteoglycan core protein in the parietal yolk sac and in mast cells, where glycosaminoglycan side chains are attached to the serine residues of the repeat region. In this article, the structure of the gene SGC encoding mouse SGC is reported. The gene is divided into three exons, which are all contained within a region of approximately 13 kb. Nucleotide (nt) sequence analysis was carried out on a region of 1.2 kb upstream from the first exon. The region containing the two promoters (active in parietal yolk sac and in mast cells, respectively) was analyzed for the presence of recognition sites for known DNA-binding proteins. A number of sequences closely related to known recognition sites were found in both promoters, and one consensus octamer-binding site could be identified in the putative yolk-sac promoter. Multiple regions in the mouse genome hybridizing with DNA fragments covering the Ser-Gly repeat region have previously been described, and it has been suggested that these loci may represent other proteoglycan core proteins. Analysis of nt sequence was carried out on three out of the more than 15 of these regions present in the mouse genome. However, none of the clones analyzed was found to have any open reading frame in the region of cross-hybridization which possibly could code for a SGC protein. Instead, one of the clones was found to contain an exon encoding a highly basic protein, unrelated to SGC. Hence, no evidence was found for a multigene family of Ser-Gly-repeat-containing proteoglycan-encoding genes.


Spatial-temporal variations of proline and related amino acids reveal distinct nitrogenous utilization strategies in rice during detoxification of exogenous cyanide.
Cyanide (CN-) pollution in agricultural systems impairs amino acid metabolism in rice plants, hence decreasing their quality and yield. Meanwhile, little is known about the effects of CN- assimilation on the innate pool of proline (Pro) and its synthesis-related amino acids (Pro-AAs) in rice plants. In this study, a hydroponic experiment was carried out to investigate the effect of exogenous KCN on indigenous levels of Pro-AAs, i.e., Pro, glutamate (Glu), arginine (Arg), and ornithine (Orn) in rice seedlings fertilized with either nitrate (NO3-) or ammonium (NH4+) through the biochemical and RT-qPCR analysis. At the same KCN treatment concentration, the relative growth rate of NH4+-fed rice seedlings was considerably higher than that of NO3--fed rice seedlings, but the residual concentration of CN- in NH4+-fed rice tissues was lower than that of NO3--fed rice tissues. Based on the UPLC and stoichiometry molar ratio calculations, it is evident that the Glu pathway contributed significantly to Pro synthesis in rice under KCN + NO3- treatments; whereas the Orn pathway governed the synthesis of Pro in rice under KCN + NH4+ treatments. Moreover, transcriptional and bioinformatics analysis revealed that NH4+ fertilization resulted in spatial-temporal differences in the genetic response in rice tissue during detoxification of CN- compared with KCN + NO3- treatments. These findings suggested that the innate level of Pro serves as "a fishing float" to balance the flux between Pro and Pro-AAs in exogenous KCN-treated rice plants under different nitrogenous nutritional conditions.


LC-MS-Based Metabolomic Investigation of Chemopreventive Phytochemical-Elicited Metabolic Events.
Phytochemicals are under intensive investigation for their potential use as chemopreventive agents in blocking or suppressing carcinogenesis. Metabolic interactions between phytochemical and biological system play an important role in determining the efficacy and toxicity of chemopreventive phytochemicals. However, complexities of phytochemical biotransformation and intermediary metabolism pose challenges for studying phytochemical-elicited metabolic events. Metabolomics has become a highly effective technical platform to detect subtle changes in a complex metabolic system. Here, using green tea polyphenols as an example, we describe a workflow of LC-MS-based metabolomics study, covering the procedures and techniques in sample collection, preparation, LC-MS analysis, data analysis, and interpretation.


The GAPDH gene system of the red alga Chondrus crispus: promoter structures, intron/exon organization, genomic complexity and differential expression of genes.
Our previous phylogenetic analysis based on cDNA sequences of chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenases (GAPDH; genes GapA and GapC, respectively) of the red alga Chondrus crispus suggested that rhodophytes and green plants are sister groups with respect to plastids and mitochondria and diverged at about the same time or somewhat later than animals and fungi. Here we characterize the genomic sequences of genes GapC and GapA of C. crispus with respect to promotor structures, intron/exon organization, genomic complexity, G + C content, CpG suppression and their transcript levels in gametophytes and protoplasts, respectively. To our knowledge this is the first report on nuclear protein genes of red algae. The GapC gene is G + C-rich, contains no introns and displays a number of classic sequence motifs within its promotor region, such as TATA, CAAT, GC boxes and several elements resembling the plant-specific G-box palindrome. The GapA gene has a moderate G+C content, a single CAAT box motif in its promotor region and a single intron of 115 bp near its 5' end. This intron occupies a conserved position corresponding to that of intron 1 in the transit peptide region of chloroplast GAPDH genes (GapA and GapB) of higher plants. It has consensus sequences similar to those of yeast introns and folds into a conspicuous secondary structure of -61.3 kJ. CpG profiles of genes GapC and GapA and their flanking sequences show no significant CpG depletion suggesting that these genomic sequences are not methylated. Genomic Southern blots hybridized with generic and gene specific probes indicate that both genes are encoded by single loci composed of multiple polymorphic alleles. Northern hybridizations demonstrate that both genes are expressed in gametophytes but not in protoplasts where appreciable amounts of transcripts can only be detected for GapC.


Perceptions of caregivers and adolescents of the use of telemedicine for the child sexual abuse examination.
Childhood sexual abuse is a common cause of morbidity and mortality. All victims should receive a timely comprehensive medical exam. Currently there is a critical shortage of child abuse pediatricians who can complete the comprehensive child sexual abuse examination. Telemedicine has emerged as an innovative way to provide subspecialty care to this population. Despite the growing popularity of telemedicine, no literature exists describing patient and caregiver perceptions of telemedicine for this sensitive exam. To explore caregiver and adolescent perspectives of the use of telemedicine for the child sexual abuse examination and discover factors that drive satisfaction with the technology. Caregivers and adolescents who presented for a child sexual abuse medical evaluation at our county's child advocacy center. We completed semi structured interviews of 17 caregivers and 10 adolescents. Guided by the Technology Acceptance Model interviews assessed perceptions about: general feelings with the exam, prior use of technology, feelings about telemedicine, and role of the medical team. Interviews were audio-recorded, transcribed, coded and analyzed using content analysis with constant comparative coding. Recruitment ended when thematic saturation was reached. There was an overwhelming positive response to telemedicine. Participants reported having a good experience with telemedicine regardless of severity of sexual abuse or prior experience with technology. Behaviors that helped patients and caregivers feel comfortable included a clear explanation from the medical team and professionalism demonstrated by those using the telemedicine system. Telemedicine was widely accepted by adolescents and caregivers when used for the child sexual abuse examination.


Socio-economic inequality in prevalence of type 2 diabetes among adults in north-west of Iran: a Blinder-Oaxaca decomposition.
The aim of this study was to estimate the socio-economic inequality in prevalence of type 2 diabetes among adults in north-west of Iran. A cross-sectional study was conducted in Ardabil with data from the PERSIAN Cohort Study. Diabetes has been measured by combining self-reported and clinical records. Based on the socio-economic status score, households divided into five quintiles. A multiple logistic regression model was used to examine the association between having diabetes and independent variables and the Blinder-Oaxaca (BO) method was used to decompose the socioeconomic inequality, respectively. The Overall age-adjusted prevalence of diabetes among 20,419 Ardabil's adults was 14.3% (95% CI: 13.6 to 14.9). The prevalence of type 2 diabetes for the poorest and richest groups was 16.07% and 7.60%, the gap between the poorest and richest groups was 8.47%. The prevalence type 2 diabetes was significantly increasing with increasing in age (OR = 4.05, 95% CI = 3.27-5.02), BMI (OR: 3.10, 95%CI = 1.25-7.68), blood pressure (OR: 2.61, 95% CI = 2.37-2.88), and decreases with higher education level (OR = 0.78, 95% CI = 0.63-0.97). The richest-economic group has lower prevalence of diabetes (OR = 0.73, 95% CI = 0.60-0.88). The decomposition showed that most important factors affecting the difference between poorest and richest group in the prevalence of type 2 diabetes were age (86.1%), years of schooling (46.9%) and having chronic diseases such as hypertension (26.9%). The present study showed that the prevalence of type 2 diabetes was significantly higher among the elderly, women, uneducated, obese, and poor populations. Policies that address people poverty such as increasing job opportunities, increasing the minimum income etc. could reduce diabetes risk for poor people.


Successful therapy of the prostaglandin defect "Wien-Hietzing" (lack of platelet high-affinity binding sites for prostaglandin I2).
Some years ago we detected a lack of platelet high-affinity PGI2 binding sites in a 10 year-old girl who presented to the outpatient unit with clinical symptoms and signs of acute popliteal artery occlusion. We named this new defect in the prostaglandin system "Wien-Hietzing". Acute surgery was successful. On the basis of earlier findings that aspirin is able to sensitize platelets to the action of PGI2 and produce beneficial changes in platelet sensitivity, we decided to treat this girl with a daily dosage of 20 mg aspirin orally. Repeated control examinations during the total follow-up period of about 6 years revealed normalized platelet sensitivity and normalized receptor behaviour. The girl is symptom-free to date. It is concluded that this prostaglandin defect may be successfully treated with long-term, low-dose aspirin administration.


Prediction and verification of survival in patients with non-small-cell lung cancer based on an integrated radiomics nomogram.
To develop and validate a nomogram to predict 1-, 2-, and 5-year survival in patients with non-small-cell lung cancer (NSCLC) by combining optimised radiomics features, clinicopathological factors, and conventional image features extracted from three-dimensional (3D) computed tomography (CT) images. A total of 172 patients with NSCLC were selected to construct the model, and 74 and 72 patients were selected for internal validation and external testing, respectively. A total of 828 radiomics features were extracted from each patient's 3D CT images. Univariable Cox regression and least absolute shrinkage and selection operator (LASSO) regression were used to select features and generate a radiomics signature (radscore). The performance of the nomogram was evaluated by calibration curves, clinical practicability, and the c-index. Kaplan-Meier (KM) analysis was used to compare the overall survival (OS) between the two subgroups. The radiomics features of the NSCLC patients correlated significantly with survival time. The c-indexes of the nomogram in the training cohort, internal validation cohort, and external test cohort were 0.670, 0.658, and 0.660, respectively. The calibration curves showed that the predicted survival time was close to the actual survival time. Decision curve analysis shows that the nomogram could be useful in the clinic. According to KM analysis, the 1-, 2- and 5-year survival rates of the low-risk group were higher than those of the high-risk group. The nomogram, combining the radscore, clinicopathological factors, and conventional CT parameters, can improve the accuracy of survival prediction in patients with NSCLC.


Head-space gas-liquid chromatography for the rapid laboratory diagnosis of urinary tract infections caused by enterobacteria.
Urine specimens were analysed in parallel in a hospital laboratory by routine methods which were regarded as the standard of correct diagnosis and by the rapid test already developed in a research laboratory. Technical modifications made to the rapid test ensured that its results agreed with those from routine methods and increased its rapidity so that a result was possible 4 h after receipt of the specimen. When 382 urine specimens were analysed by the modified test which is described in detail, there were neither false negatives nor false positives for infections with Escherichia, Klebsiella, Citrobacter or Proteus species.


Embryonic development of choline acetyltransferase and nitric oxide synthase in the spinal cord of pigeons and chickens with special reference to the superficial dorsal horn.
The superficial dorsal horn of birds as well as mammals contains both cholinergic and nitrergic neuronal structures as evident from the presence of the synthesizing enzymes such as choline acetyltransferase and nitric oxide synthase, which is an NADPH diaphorase. In the rat, both systems develop only postnatally. Rats are altricial at birth whereas pigeons and chickens are semiprecocial or precocial, respectively, at the time of hatching. Immunocytochemical studies of choline acetyltransferase and nitric oxide synthase in the developing avian spinal cord (starting with embryonic day 12 of 18 in the pigeon and 14 of 21 in the chicken) showed that both systems are well developed in the superficial dorsal horn at the time of hatching in both avian species. In the pigeon, choline acetyltransferase-positive superficial dorsal horn neurons appear only on the day of hatching (E18), whereas nitric oxide synthase-positive neurons can be first detected at stage E14. In the chicken, nitric oxide synthase-positive neurons are present already at stage E14, whereas choline acetyltransferase-positive neurons appear at stage E20. Autonomic and somatic motor neurons show adult-like choline acetyltransferase-immunoreactivity and/or nitric oxide synthase-immunoreactivity at the earliest stages investigated. It is concluded that the stage of maturation at birth or hatching plays an important role in the development of superficial dorsal horn cholinergic and nitrergic systems.


Larva lights: a decade of photoaffinity labeling with juvenile hormone analogues.
The introduction of photoaffinity labeling into the mode of action of insect hormones and pheromones started 12 yr ago with the photoaffinity labeling of juvenile hormone binding proteins (JHBPs) from cockroaches in the laboratory of the late John K. Koeppe. Applying this technique to Manduca sexta led ultimately to a three-laboratory collaborative project that has begun to dissect the molecular basis for JH transport, metabolism, and nuclear binding and gene activation in Lepidoptera. This review provides (1) a history of the first experiments; (2) an idea of the breadth of the technique in the arthropod classes Insecta, Crustacea, and Arachnida; and (3) evidence for the depth of the technique in unearthing key details about three different types of the molecular action of JH in M. sexta.


Synthesis and biological evaluation of a new family of anti-benzylanilinosulfonamides as CA IX inhibitors.
We report the synthesis and the pharmacological evaluation of a new class of human carbonic anhydrase (hCA) inhibitors prepared regio- and stereoselectively by reacting sulfanilamide with ethyl trans-phenylglycidate in the presence of cobalt(II) chloride. Various derivatizations of the ester moiety in the parent compound led to a small library of derivatives (2R,3R and 2S,3S) which displayed interesting inhibitory activities towards the human tumor-associated isoform CA IX. One of the new compounds shows high selectivity in inhibiting hCA IX compared to the two physiologically relevant, cytosolic isozymes hCA I and hCA II. A molecular modeling study was conducted in order to simulate the binding mode of this new family of enzyme inhibitors within the active sites of hCA IX and hCA II.


Columnar cell variant of papillary thyroid carcinoma: A diagnostic dilemma in fine-needle aspiration cytology.
Columnar cell variant of papillary thyroid carcinoma (PTC) is an uncommon variant with an aggressive course as compared to classic papillary carcinoma. Cytologic diagnosis of these tumors is difficult due to absence of characteristic nuclear features of classic pattern of papillary carcinoma. We present a case of columnar cell variant in a young female misdiagnosed on aspiration cytology. A 21-year-old female presented with solitary nodule in the left aspect of thyroid. A diagnosis of medullary thyroid carcinoma was rendered. The resected thyoroidectomy specimen revealed a columnar cell variant of PTC which was further supported by immunohistochemical staining. Diagn. Cytopathol. 2016;44:816-819. © 2016 Wiley Periodicals, Inc.


Feeding Characteristics of Deep-Sea Acorn Worm (Hemichordata, Enteropneusta, Torquaratoridae) from the Bering Sea.
Deep-sea hemichordates Torquaratoridae gen. sp. reach high abundance up to 12 spec. m-2 at the depths of 1830-2130 m on the slope of the Volcanologists Massif in the south-western part of the Bering Sea, dominating in the benthic community at these depths. Their abundance exceeds by two orders the values recorded earlier. In order to clarify this phenomenon, we examined the gut contents of Torquaratoridae gen. sp. The detritus particles and frustules of planktonic diatoms Thalassiosira, Coscinodiscus, Actinocyclus, Chaetoceros, Neodenticula, and Grammatophora were the most common in the gut, as well as the remains of skeletons of benthic invertebrates with little admixture of mineral particles. According to obtained data, Torquaratoridae gen. sp. are mobile deposit feeders with high selectivity to fresh phytodetritus, able to compete with holothurians occupying similar trophic niche. Unusually high abundance of acorn worms is apparently related to high organic matter flux to the seafloor as a result of spring phytoplankton bloom in the surface water layer of the Bering Sea.


Optimal setting of image bounding box can improve registration accuracy of diffusion tensor tractography.
When we register diffusion tensor tractography (DTT) to anatomical images such as fast imaging employing steady-state acquisition (FIESTA), we register the B0 image to FIESTA. Precise registration of the DTT B0 image to FIESTA is possible with non-rigid registration compared to rigid registration, although the non-rigid methods lack convenience. We report the effect of image data bounding box settings on registration accuracy using a normalized mutual information (NMI) method METHODS: MRI scans of 10 patients were used in this study. Registration was performed without modification of the bounding box in the control group, and the results were compared with groups re-registered using multiple bounding boxes limited to the region of interest (ROI). The distance of misalignment after registration at 3 anatomical characteristic points that are common to both FIESTA and B0 images was used as an index of accuracy. Mean ([Formula: see text]SD) misalignment at the 3 anatomical points decreased significantly from [Formula: see text] to [Formula: see text] mm, [Formula: see text]), [Formula: see text] to [Formula: see text] mm, ([Formula: see text], and [Formula: see text] to [Formula: see text] mm, ([Formula: see text], each showing improvement compared to the control group CONCLUSION: Narrowing the image data bounding box to the ROI improves the accuracy of registering B0 images to FIESTA by NMI method. With our proposed methodology, accuracy can be improved in extremely easy steps, and this methodology may prove useful for DTT registration to anatomical image.


B Cell Disorders in Children: Part II.
B cell disorders result in decreased levels or function of immunoglobulins in an individual. Genetic mutations have been reported in a variety of B cell disorders. This review, in follow-up to a previous review, describes some rare B cell disorders as well as their known underlying genetic etiologies. Genetic studies identify and permit precise classification of an increasing number of B cell disorders, leading to a greater understanding of B cell development and function. The B cell disorders are rare diseases. While clinicians are most familiar with X-linked agammaglobulinemia and so-called common variable immunodeficiency (CVID), there are many causes of hypogammaglobulinemia. Genetic testing provides a specific diagnosis, offers useful information for genetic counseling, and can identify previously unrecognized B cell disorders.


Etiological agents of bovine mastitis.
A total of 137 microbial species, subspecies and serovars have been isolated from the bovine mammary gland. Nucleic acid hybridization studies have restructured the classification of many mastitis pathogens. Availability of defined species descriptions has permitted greater insight into the distribution and pathogenicity of many previously unrecognized microorganisms associated with bovine mastitis. Precise epidemiological studies are needed to better delineate the role of some microorganisms in bovine mastitis and to aid development of improved control methods.


Functional disomy for Xq22-q23 in a girl with complex rearrangements of chromosomes 3 and X.
A 5-year-old girl with developmental and growth retardation is reported with complex chromosome rearrangements consisting of a partial Xq deletion and an abnormal chromosome 3 with multiple breakpoints. GTG-banding, and multiplex and conventional FISH studies showed that a 6.6-Mb Xq22-q23 segment was inserted into 3q, in addition to three intrachromosomal insertions in chromosome 3. Her karyotype was thus interpreted as 46,X,der(X)(Xpter-->Xq22::Xq23-->Xqter),der(3)(3pter-->3p26::3p12-->3q25.3::3p12-->3p26::Xq22-->Xq23::3q25.3-->3qter). Replication R-banding study showed that the der(X) was inactivated in all blood lymphocytes analyzed. Methylation-specific PCR at the androgen receptor gene (HUMARA) locus at Xq11-q12 showed a skewed inactivation pattern with the active/inactive X chromosome ratio of 92/8. These data indicated the presence, in the majority of cells, of a functioning Xq22-q23 segment in both the normal X and the der(3) chromosomes. Her growth retardation, developmental delay, and other minor anomalies were most likely caused by dosage effects of the genes in the functionally disomic Xq22-q23 region. Despite the presence of two active copies of the proteolipid protein 1 gene (PLP1), she did not show the symptoms of Pelizaeus-Merzbacher disease, a subset of which has been known to be caused by the duplication of PLP1.


Antitumour activity and side effects of combined treatment with chitosan and cisplatin in sarcoma 180-bearing mice.
We examined the possible modulation by chitosan of the antitumour effects and side effects of cisplatin (cis-diaminedichloroplatinum, CDDP). The study showed that CDDP had potent antitumour activity when administered orally as well as intraperitoneally. We also compared the antitumour activity and side effects of orally administered CDDP plus orally administered chitosan versus intraperitoneally administered CDDP plus orally administered chitosan in sarcoma 180-bearing mice. When CDDP (1.25 mgkg(-1) x 2 day(-1)) was intraperitoneally administered to sarcoma 180-bearing mice, myelotoxicity (the reduction of leucocyte and platelet numbers), nephrotoxicity (the increase of blood nitrogen urea level), immunotoxicity (the reduction of spleen and thymus weight) and a reduction in body weight resulted. These intraperitoneally administered CDDP-induced side effects were not prevented by oral administration of chitosan (150 mgkg(-1) x 2 day(-1) and 750 mgkg(-1) x 2 day(-1)) for 14 consecutive days. On the other hand, the side effects such as the reductions of body and spleen weights induced by orally administered CDDP (1.25 mgkg(-1) x 2 day(-1)) were prevented by the oral administration of chitosan (150 mgkg(-1) x 2 day(-1) and 750 mg kg(-1) x 2 day(-1)). From these results, we conclude that the orally administered chitosan plus CDDP might be useful for the prevention of body weight reduction and immunotoxicity (the reduction of spleen weight) induced by the orally administered CDDP without diminishing antitumour activity.


Transcriptome analysis of GVHD reveals aurora kinase A as a targetable pathway for disease prevention.
Graft-versus-host disease (GVHD) is the most common complication of hematopoietic stem cell transplant (HCT). However, our understanding of the molecular pathways that cause this disease remains incomplete, leading to inadequate treatment strategies. To address this, we measured the gene expression profile of nonhuman primate (NHP) T cells during acute GVHD. Utilizing microarray technology, we measured the expression profiles of CD3(+) T cells from five cohorts: allogeneic transplant recipients receiving (i) no immunoprophylaxis (No Rx), (ii) sirolimus monotherapy (Siro), (iii) tacrolimus-methotrexate (Tac-Mtx), as well as (iv) autologous transplant recipients (Auto) and (v) healthy controls (HC). This comparison allowed us to identify transcriptomic signatures specific for alloreactive T cells and determine the impact of both mTOR (mechanistic target of rapamycin) and calcineurin inhibition on GVHD. We found that the transcriptional profile of unprophylaxed GVHD was characterized by significant perturbation of pathways regulating T cell proliferation, effector function, and cytokine synthesis. Within these pathways, we discovered potentially druggable targets not previously implicated in GVHD, prominently including aurora kinase A (AURKA). Utilizing a murine GVHD model, we demonstrated that pharmacologic inhibition of AURKA could improve survival. Moreover, we found enrichment of AURKA transcripts both in allo-proliferating T cells and in sorted T cells from patients with clinical GVHD. These data provide a comprehensive elucidation of the T cell transcriptome in primate acute GVHD and suggest that AURKA should be considered a target for preventing GVHD, which, given the many available AURKA inhibitors in clinical development, could be quickly deployed for the prevention of GVHD.


Cross-linking of immune complexes by human mononuclear phagocytes.
Incubation of immune complexes (IC) bound to plastic surfaces with human blood monocytes for 48 hours resulted in the cross-linking of a proportion of antibody molecules. This process was largely inhibited by the addition of sodium azide to the cultures. Cross-linking was defined as the inability of strong chaotropic solutions (3 M MgCl2 or 5 M guanidine) or acid pH (0.1 N HCl) to solubilize 125I-labeled rabbit anti-human serum albumin attached to plastic-bound antigen. Addition to the cultures of a suitable hydrogen donor such as catechol (0.5 mM) resulted in a large increase in cross-linking of IC. This process was shown to depend on the presence of viable phagocytic cells because incubation with dead monocytes or with viable T lymphocytes failed to induce cross-linking. Quantitation of rabbit immunoglobulin remaining in the wells by enzyme-linked immunoassay techniques excluded the possibility that the increase in 125I bound was merely due to a transiodination reaction. Experiments using various oxygen metabolite inhibitors and scavengers indicated that catechol-dependent protein cross-linking depended on the action of hydrogen peroxide and enzyme systems inhibitable by sodium azide, probably monocyte-peroxidase. Superoxide dismutase, 1O2, and OH X radical scavengers failed to inhibit cross-linking, whereas addition of catalase resulted in almost complete abolition of the process. These observations suggest that catechol-dependent cross-linking of IC may be due to oxidation of catechol to orthoquinone and that this strong oxidant is responsible for nonenzymic chemical action on proteins leading to intermolecular covalent bond formation. Cell-mediated protein cross-linking by oxidative mechanisms may be a prominent feature of drug-related reactions and of acute and chronic inflammatory processes in general. The possible mechanisms involved in catechol-dependent and -independent cross-linking of IC by human mononuclear phagocytes are discussed.


Long-term Behavioral Consequences of the COVID-19 Pandemic for Autistic Individuals and Their Mothers.
Long-term effects of the COVID-19 pandemic on autistic individuals and their families are recently emerging. This study investigated these effects in 40 mother-child dyads by measuring the behavioral problems of autistic individuals (via Aberrant Behavior Checklist) and their mothers' anxiety levels (via Beck Anxiety Inventory) during the pre-pandemic period, one month after, and one year after its onset. One year into the pandemic, aberrant behaviors of autistic individuals worsened only for those whose mothers had high anxiety levels. The continued negative impact of the COVID-19 pandemic on the behavior of autistic individuals is related to their mother's anxiety level, highlighting the need to support maternal mental health in families with individuals with autism.


Results of a community-based screening programme for chronic kidney disease and associated risk factors, (obesity, diabetes and hypertension) in a Samoan cohort.
In 2019, under the World Kidney Day theme of 'Kidney health for everyone everywhere', the National Kidney Foundation of Samoa undertook an extensive community screening campaign to detect the estimated prevalence of chronic kidney disease (CKD) and its associated risk factors in the community. Fifteen screening sites, with 11 urban and rural sites on the main island of Upolu, and 4 in different rural areas on the island of Savaii. All participants were self-referrals to the various screening sites. In total, 1163 Samoans were screened, with similar numbers from both urban and rural areas and similar numbers of female and male. All participants were screened for CKD using point of care serum creatinine determinations, with calculation of estimated glomerular filtration rate using the CKD-EPI formula and dipstix urinalysis. A standardised screening survey was used to capture demographic and medical history with associated risk factors of obesity, diabetes, using point of care determination of HbA1c and hypertension. Logistic regression was used to investigate the association of CKD with risk factors. In total, 1163 people were screened for CKD within the month of March 2019. The prevalance of CKD (grades 1-5) was 44.5% (95% CI 41.6% to 47.4) with individual grade prevalence CKD 1: 3.7%, CKD 2: 6.1%, CKD 3: 30.7%, CKD 4: 2.9% and CKD 5: 1.0%. The prevalence of obesity (body mass index ≥32), diabetes and hypertension was 66.3%, 30.8% and 54.3%, respectively. This is the first paper to report the estimated prevalence of CKD in Samoa or any other Pacific Island nation. It reveals an urgent need for further studies on the epidemiology of CKD in Samoa, to develop country-specific prevention strategies to mitigate this growing burden and prevent subsequent CKD associated complications including development of kidney failure and premature death.


Youth depression in the family context: familial risk factors and models of treatment.
Research on parent risk factors, family environment, and familial involvement in the treatment of depression in children and adolescents is integrated, providing an update to prior reviews on the topic. First, the psychosocial parent and family factors associated with youth depression are examined. The literature indicates that a broad array of parent and family factors is associated with youth risk for depression, ranging from parental pathology to parental cognitive style to family emotional climate. Next, treatment approaches for youth depression that have been empirically tested are described and then summarized in terms of their level of parent inclusion, including cognitive-behavioral therapy, interpersonal therapy, and family systems approaches. Families have mostly not been incorporated into clinical treatment research with depressed adolescents, with only 32% of treatments including parents in treatment in any capacity. Nonetheless, the overall effectiveness of treatments that involve children and adolescents exclusively is very similar to that of treatments that include parents as agents or facilitators of change. The article concludes with a discussion of the implications of these findings and directions for further research.


Chemometric quality control of chromatographic purity.
It is common practice in chromatographic purity analysis of pharmaceutical manufacturing processes to assess the quality of peak integration combined by visual investigation of the chromatogram. This traditional method of visual chromatographic comparison is simple, but is very subjective, laborious and seldom very quantitative. For high-purity drugs it would be particularly difficult to detect the occurrence of an unknown impurity co-eluting with the target compound, which is present in excess compared to any impurity. We hypothesize that this can be achieved through Multivariate Statistical Process Control (MSPC) based on principal component analysis (PCA) modeling. In order to obtain the lowest detection limit, different chromatographic data preprocessing methods such as time alignment, baseline correction and scaling are applied. Historical high performance liquid chromatography (HPLC) chromatograms from a biopharmaceutical in-process analysis are used to build a normal operation condition (NOC) PCA model. Chromatograms added simulated 0.1% impurities with varied resolutions are exposed to the NOC model and monitored with MSPC charts. This study demonstrates that MSPC based on PCA applied on chromatographic purity analysis is a powerful tool for monitoring subtle changes in the chromatographic pattern, providing clear diagnostics of subtly deviating chromatograms. The procedure described in this study can be implemented and operated as the HPLC analysis runs according to the process analytical technology (PAT) concept aiming for real-time release.


Ticlopidine induces embryonic development toxicity and hepatotoxicity in zebrafish by upregulating the oxidative stress signaling pathway.
Ticlopidine exerts its anti-platelet effects mainly by antagonizing platelet p2y12 receptors. Previously, a few studies have shown that ticlopidine can induce liver injury, but the exact mechanism of hepatotoxicity remains unclear. Oxidative stress, metabolic disorders, hepatocyte apoptosis, lipid peroxidation, and inflammatory responses can all lead to hepatic liver damage, which can cause hepatotoxicity. In this study, in order to deeply explore the potential molecular mechanisms of ticlopidine -induced hepatotoxicity, we used zebrafish as a model organism to comprehensively evaluate the hepatotoxicity of ticlopidine and its associated mechanism. Three days post-fertilization, zebrafish larvae were exposed to varying concentrations (1.5, 1.75 and 2 μg/mL) of ticlopidine for 72 h, in contrast, adult zebrafish were exposed exposure to 4 μg/mL of ticlopidine for 28 days. Ticlopidine-exposed zebrafish larvae showed changes in liver morphology, shortened body length, and delayed development of the swim bladder development. Liver tissues of ticlopidine-exposed zebrafish larvae and adults stained with Hematoxylin & Eosin revealed vacuolization and increased cellular interstitial spaces in liver tissues. Furthermore, using Oil Red O and periodic acid-Schiff staining methods and evaluating different metabolic enzymes of ticlopidine-exposed zebrafish larvae and adults suggested abnormal liver metabolism and liver injury in both ticlopidine-exposed zebrafish larvae and adults. Ticlopidine also significantly elevated inflammation and oxidative stress and reduced hepatocyte proliferation. During the rescue intervention using N-acetylcysteine, we observed significant improvement in ticlopidine-induced morphological changes in the liver, shortened body length, delayed swim bladder development, and proliferation of liver tissues showed significant improvement. In conclusion, ticlopidine might inhibit normal development and liver proliferation in zebrafish by upregulation of oxidative stress levels, thus leading to embryonic developmental toxicity and hepatotoxicity. In this study, we used zebrafish as a model organism to elucidate the developmental toxicity and hepatotoxicity induced by ticlopidine upregulation of oxidative stress signaling pathway in zebrafish, providing a theoretical basis for clinical application.


MECONIUM ASPIRATION SYNDROME : A CLINICAL STUDY.
Forty two babies with Meconium aspiration syndrome managed in neonatal intensive care unit of a service hospital were analysed. Incidence was 1.7 per cent. Twelve babies were delivered by caserean section and 30 babies by vaginal route. Seventeen deliveries were conducted by team of gynaecologists, pediatrician and nursing staff whereas 25 deliveries were conducted by nursing staff alone. Out of total babies with meconium aspiration syndrome, thin meconium detected in 9 and thick meconium detected in 33 babies during delivery. Five babies were preterm and none were less than 35 weeks of gestation. Pneumonia detected on right side in 24 patients, on left side in 6, and both sides in 12 patients. Twenty seven babies had asphyxia at birth, 13 babies had seizure and 6 babies had air leak syndrome and overall mortality was 13 (31%).


Activity of Sulfa Drugs and Their Combinations against Stationary Phase B. burgdorferi In Vitro.
Lyme disease is a most common vector-borne disease in the US. Although the majority of Lyme patients can be cured with the standard two- to four-week antibiotic treatment, at least 10%-20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS). While the cause for this is unclear, one possibility is that persisting organisms are not killed by current Lyme antibiotics. In our previous studies, we screened an FDA drug library and an NCI compound library on B. burgdorferi and found some drug hits including sulfa drugs as having good activity against B. burgdorferi stationary phase cells. In this study, we evaluated the relative activity of three commonly used sulfa drugs, sulfamethoxazole (Smx), dapsone (Dps), sulfachlorpyridazine (Scp), and also trimethoprim (Tmp), and assessed their combinations with the commonly prescribed Lyme antibiotics for activities against B. burgdorferi stationary phase cells. Using the same molarity concentration, dapsone, sulfachlorpyridazine and trimethoprim showed very similar activity against stationary phase B. burgdorferi enriched in persisters; however, sulfamethoxazole was the least active drug among the three sulfa drugs tested. Interestingly, contrary to other bacterial systems, Tmp did not show synergy in drug combinations with the three sulfa drugs at their clinically relevant serum concentrations against B. burgdorferi. We found that sulfa drugs combined with other antibiotics were more active than their respective single drugs and that four-drug combinations were more active than three-drug combinations. Four-drug combinations dapsone + minocycline + cefuroxime + azithromycin and dapsone + minocycline + cefuroxime + rifampin showed the best activity against stationary phase B. burgdorferi in these sulfa drug combinations. However, these four-sulfa-drug-containing combinations still had considerably less activity against B. burgdorferi stationary phase cells than the Daptomycin + cefuroxime + doxycycline used as a positive control which completely eradicated B. burgdorferi stationary phase cells. Future studies are needed to evaluate and optimize the sulfa drug combinations in vitro and also in animal models.


Erratum to clinical presentation and imaging characteristics of clear cell sarcoma-like tumour of the gastrointestinal tract with liver metastasis: a case description.
[This corrects the article DOI: 10.21037/qims-21-186.].