Deprotonation of 3(2H)-furanone and 3(2H)-thiophenone by carbanions in the gas phase: disproportionately high aromaticity of the transition state: an ab initio study.
The reversible deprotonation of 3(2H)-furanone (3H-O) and 3(2H)-thiophenone (3H-S) by a series of delocalized carbanions and by CN(-), and the identity proton transfer of 3H-O to its conjugate base (3(-)-O) and of 3H-S to 3(-)-S have been studied at the MP2//6-31+G** level. The main objective has been to examine to what extent the aromaticity of 3(-)-O and 3(-)-S is expressed at the transition state of these reactions and how the intrinsic barriers are affected by the transition state aromaticity. Aromaticity parameters such as NICS values, HOMA and Bird Indices indicate a disproportionately high degree of aromatic stabilization of the transition state. This stabilization results in a reduction of the intrinsic barriers which is most clearly manifested in the identity reactions. However, these reductions are relatively modest compared to those reported previously for the identity proton transfers from the benzenium ion to benzene and of cyclopentadiene to its conjugate base, reflecting the smaller aromatic stabilization of 3(-)-O and 3(-)-S compared to those of benzene and cyclopentadienyl anion.


Development of Water-in-Oil Emulsions as Delivery Vehicles and Testing with a Natural Antimicrobial Extract.
Water-in-oil (W/O) emulsions have high potential for several industrial areas as delivery systems of hydrophilic compounds. In general, they are less studied than oil-in-water (O/W) systems, namely in what concerns the so-called fluid systems, partly due to problems of instability. In this context, this work aimed to produce stable W/O emulsions from a natural oil, sweet almond oil, to be further tested as vehicles of natural hydrophilic extracts, here exemplified with an aqueous cinnamon extract. Firstly, a base W/O emulsion using a high-water content (40/60, v/v) was developed by testing different mixtures of emulsifiers, namely Tween 80 combined with Span 80 or Span 85 at different contents. Among the tested systems, the one using a 54/46 (v/v) Span 80/Tween 80 mixture, and subjected to 12 high-pressure homogenizer (HPH) cycles, revealed to be stable up to 6 months, being chosen for the subsequent functionalization tests with cinnamon extract (1.25-5%; w/v; water-basis). The presence of cinnamon extract leaded to changes in the microstructure as well as in the stability. The antimicrobial and antioxidant analysis were evidenced, and a sustained behavior compatible with an extract distribution within the two phases, oil and water, in particular for the higher extract concentration, was observed.


An organotypic in vitro model system for studying pulmonary surfactant production by type II alveolar pneumonocytes.
This report describes an in vitro model system in which monodisperse fetal rat lung cells reorganize to form alveolar-like structures when cultured on a gelatin sponge matrix. The alveolar-like structures are composed of cells that have morphologic characteristics like those of the type II alveolar pneumonocytes of intact lung. These morphologic characteristics include lightly stained nuclei, microvilli on the apical surface, and osmiophilic lamellar bodies in the cytoplasm. The presence of osmiophilic lamellar bodies and tubular myelin in the lumen of the alveolar-like structures suggests that the cells in these structures are producing pulmonary surfactant. The formation and long-term maintenance of these alveolar-like structures provide a unique in vitro model system for studies of the synthesis, storage, and secretion of pulmonary surfactant.


Genetically modified keratinocytes transplanted to wounds reconstitute the epidermis.
Normal and retrovirally transfected keratinocyte suspensions expressing either the beta-galactosidase gene or the human growth hormone (hGH) gene were transplanted into chamber-enclosed skin full-thickness wounds of Yorkshire pigs. Immunostaining of sequential skin biopsies obtained for 4 weeks after transplantation showed survival of the transplanted keratinocytes as well as expression of beta-galactosidase. Transfected keratinocytes were first seen in the neodermal portions of the wounds, then in the regenerating basal epidermal layer, and finally in the terminally differentiating cells of the stratum spinosum. When keratinocytes transfected with the hGH gene were transplanted into similar wounds, hGH was detected for 10 days in wound fluid. In contrast, hGH was detected in vitro for 47 days. Wounds transplanted with either transfected or normal keratinocytes restored the epithelial barrier function significantly faster than nontransplanted controls (P < 0.05). The study confirms the successful transplantation of keratinocyte suspensions, their reconstitution of the epidermis, and their acceleration of repair. Further, this apparently normal incorporation of genetically engineered transplanted keratinocytes expressing either beta-galactosidase or hGH suggests the possibility of introducing other genes expressing therapeutic proteins into wounds to favorably affect healing. Wound fluid detection of the expressed peptide provided early demonstration of successful transfer of the hGH gene.


Treatment of condylar fractures in children and youths: the clinical value of the occlusal plane orientation and correlation with facial development (case reports).
The relative position of the plane of occlusion to the cranial base determines the direction of the forces generated in the cranium during occlusal function. When the plane of occlusion is level and when the neuromuscular system is in harmony, the vectors of forces created by the closing muscles are directed to the central area of the cranium in a symmetrically balanced way. Unfortunately, TMJ fractures may alter completely this balance with loss of the support to the mandible against the temporal component and loss of the functional effect of the lateral pterygoid muscle on the mandible. Changes in orientation of the occlusal plane may result in facial alteration and asymmetries. In our experience, the restoration of a plan of occlusion orthogonally aligned to the forces of occlusion for a correct transfer of forces through the maxilla to the rest of the cranial bones is essential to allow proper face development. Two, quite similar cases of unilateral, dislocated condylar fracture treated in a different way, will be reported to demonstrate how this can occur. Available clinical data will be illustrated.


Enhanced sensitivity of skin fibroblasts from neurofibromatosis patients to transformation by the Kirsten murine sarcoma virus. A potential laboratory assay for individuals at risk of cancer.
The present work describes a laboratory assay for individuals predisposed to cancer within NF pedigrees. The assay is based on the association between the increased sensitivity of human skin fibroblasts to transformation by the Kirsten murine sarcoma virus and predisposition to cancer in clinically affected patients and in otherwise apparently healthy individuals within NF pedigrees. The more sensitive the cells are to transformation by KiMSV, the greater the probability that a person from whom such cells have been derived will develop cancer. The results show a strong correlation with the NF trait. Together with the clinical data this laboratory assay could, therefore, be used to ascertain the NF genotype.


Ethical and practical problems in blood sampling for research purposes during pre-hospital emergencies.
Research in the pre-hospital phase of emergencies involves certain ethical and practical challenges. Severely ill or injured patients are not able to give informed consent in the immediate stressful situation. The aim of this pilot study was to find an ethically acceptable method to carry out research in acutely ill or injured patients before any treatment was given in order to be able to study the acute and unmodified systemic inflammatory response to trauma. Younger physicians were assigned to the mobile emergency care unit (MECU) on a volunteer basis. They drew blood samples 'at the scene' from patients exposed to accidental injury or from patients with acute medical emergencies. The ethical committee accepted that informed consent to blood analyses could be postponed until later or given by relatives. Pro- and anti-inflammatory cytokines, catecholamines and cortisol were measured. During 6 months, the study group accompanied the MECU on approximately 500 call-outs. Blood samples were drawn from 42 patients. Consent to the analysis of blood samples was obtained in 30 cases. In 20 cases, it was not possible to draw the blood sample before medication. This pilot study showed that it was possible to conduct blood sampling from acutely ill patients 'at the scene'. However, the present legislation on informed consent makes this type of research very time consuming. When patients die in hospital and no relatives can be found, consent cannot be obtained, and information from these severely affected patients is lost.


Growth hormone treatment for non-GHD disorders: Excitement tempered by biology.
The success of growth hormone (GH) replacement in children with classical GH deficiency led to excitement that other causes of short stature may benefit similarly. However, clinical experience has shown less consistent and generally less dramatic effects on adult height, perhaps not surprising in light of increased understanding of GH and growth plate biology. Nonetheless, clinical demand for GH treatment continues to grow. Upon the 20th anniversary of the U.S. FDA's approval of GH treatment for idiopathic short stature, this review will consider the factors underlying the expansion of GH treatment, the biological mechanisms of GH action, the non-GH deficient uses of GH as a height-promoting agent, biological constraints to GH action, and future directions.


Herpes simplex virus 2 causes apoptotic infection in monocytoid cells.
Increasing evidence indicates that apoptosis can be associated with several viral infections. Here we demonstrate, that infection of monocytoid cells by Herpes simplex virus 2 (HSV-2) resulted, in time- and dose-dependent induction of apoptosis as an exclusive cytopathic effect. The phenomenon was confirmed using four different techniques. Conversely, apoptosis was not observed in the Vero cell line. Virus yield in monocytoid cells was delayed and reduced, although well detectable, in comparison with that observed in Vero cells. Nevertheless, released virions exhibited full infecting capability. Apoptosis induced by HSV-2 was not inhibited by cycloheximide and only partially by an UV-treatment which completely abrogated infectivity. Virus-induced apoptosis was partly inhibited by indomethacin and was associated with a down-regulation of Bcl-2. A similar, but less pronounced, apoptosis-inducing effect in monocytoid cells was also observed with HSV-1 infection. Depending on the target cells, therefore, HSV could complete a cycle of infection which is characterized by apoptosis of infected cells.


Association of ERK2 mitogen-activated protein kinase with human immunodeficiency virus particles.
Here we report the presence of a protein kinase activity associated with human immunodeficiency virus type 1 (HIV-1) particles. We observed phosphorylation of five major proteins by the endogenous protein kinase activity. Phosphoamino acid analysis revealed phosphorylated serine and threonine residues. In addition, we observed autophosphorylation of two proteins in the presence of gamma-ATP in an in-gel phosphorylation assay. These two proteins are not linked by a disulfide bond, suggesting that two different protein kinases are associated with HIV-1 virions. Our results indicate the presence of ERK2 mitogen-activated protein kinase and of a 53,000-molecular-weight protein kinase associated with virions. Moreover, the use of different HIV strains derived from T cells and promonocytic cells, as well as the use of human T-cell leukemia virus type 1 particles, demonstrates that ERK2 is strongly associated with retrovirus particles in a cell-independent manner. Exogenous substrates, such as histone proteins, and a viral substrate, such as Gag protein, are phosphorylated by virus-associated protein kinases.


Development of a lumbar EMG-based coactivation index for the assessment of complex dynamic tasks.
The objective of this study was to develop and test an EMG-based coactivation index and compare it to a coactivation index defined by a biologically assisted lumbar spine model to differentiate between tasks. The purpose was to provide a universal approach to assess coactivation of a multi-muscle system when a computational model is not accessible. The EMG-based index developed utilised anthropometric-defined muscle characteristics driven by torso kinematics and EMG. Muscles were classified as agonists/antagonists based upon 'simulated' moments of the muscles relative to the total 'simulated' moment. Different tasks were used to test the range of the index including lifting, pushing and Valsalva. Results showed that the EMG-based index was comparable to the index defined by a biologically assisted model (r2 = 0.78). Overall, the EMG-based index provides a universal, usable method to assess the neuromuscular effort associated with coactivation for complex dynamic tasks when the benefit of a biomechanical model is not available. Practitioner Summary: A universal coactivation index for the lumbar spine was developed to assess complex dynamic tasks. This method was validated relative to a model-based index for use when a high-end computational model is not available. Its simplicity allows for fewer inputs and usability for assessment of task ergonomics and rehabilitation.


Epigenetic discrimination by mouse metaphase II oocytes mediates asymmetric chromatin remodeling independently of meiotic exit.
In mammalian fertilization, paternal chromatin is exhaustively remodeled, yet the maternal contribution to this process is unknown. To address this, we prevented the induction of meiotic exit by spermatozoa and examined sperm chromatin remodeling in metaphase II (mII) oocytes. Methylation of paternal H3-K4 and H3-K9 remained low, unlike maternal H3, although paternal H3-K4 methylation increased in zygotes. Thus, mII cytoplasm can sustain epigenetic asymmetry in a cell-cycle dependent manner. Paternal genomic DNA underwent oocyte-mediated cytosine demethylation and acquired maternally-derived K12-acetylated H4 (AcH4-K12) independently of microtubule assembly and maternal chromatin. AcH4-K12 persisted without typical maturation-associated deacetylation, irrespective of paternal pan-genomic cytosine methylation. Contrastingly, somatic cell nuclei underwent rapid H4 deacetylation; sperm and somatic chromatin exhibited asymmetric AcH4-K12 dynamics simultaneously within the same mII oocyte. Inhibition of somatic histone deacetylation revealed endogenous histone acetyl transferase activity. Oocytes thus specify the histone acetylation status of given nuclei by differentially targeting histone deacetylase and acetyl transferase activities. Asymmetric H4 acetylation during and immediately after fertilization was dispensable for development when both parental chromatin sets were hyperacetylated. These studies delineate non-zygotic chromatin remodeling and suggest a powerful model with which to study de novo genomic reprogramming.


[Causes of stillbirths according to the period of pregnancy].
Stillbirths represent 50% of all perinatal mortality and remain significant and unanswered problem. To investigate the causes of stillbirths in term and preterm fetuses and place of intrauterine growth retardation in the pathogenesis of antenatal death. The study was retrospective for the period 2008 to 2013, covers 217 cases of stillbirths in University Hospital "Maichin dom" Sofia. Gestational age was from 23 to 41 weeks. Inclusion criteria for the study were: antenatal occurred dead and singleton pregnancy. Exclusion criteria: fetal malformations and multiple pregnancies. Thus number of enrolled cases was 153. The causes of stillbirths are different in different groups according gestational age. The cases of unknown cause, abruption of placenta and preeclampsia prevail in preterm group. The term stillborns headed with pathology of the umbilical cord, followed by cases of unknown cause, diabetes and infection. Overall, preterm group stillborn with mass below 10 percentile for the gestational age - 53 % predominate, compared with those born at term - 39%. The incidence of stillbirths is higher in preterm than term born fetuses. The causes of antenatal death in preterm is mainly abruption of placenta and preeclampsia, whereas in term - pathology of the umbilical cord. In premature stillborn intrauterine growth retardation is more common in comparison with those on term. Unexplained antenatal fetal death is more common in preterm fetuses.


Computational insights into pH-dependence of structure and dynamics of pyrazinamidase: A comparison of wild type and mutants.
The mycobacterial enzyme pyrazinamidase (PZase) is the target of key tuberculosis drug, pyrazinamide. Mutations in PZase cause drug resistance. Herein, three point mutations, W68G, L85P, and V155G, were investigated through over 8 µs of molecular dynamics simulations coupled with essential dynamics and binding pocket analysis at neutral (pH = 7) and acidic (pH = 4) ambient conditions. The 51-71 flap region exhibited drastic displacement leading to enlargement of binding cavity, especially at the lower pH. Accessibility of solvent to the active site of the mutant enzymes was also reduced. The protonation of key surface residues at low pH results in more contribution of these residues to structural stability and integrity of the enzyme and reduced interactions with solvent molecules, which acts as a cage, keeping the enzyme together. The observed results suggest a pattern of structural alterations due to point mutations in PZase, which is consistent with other experimental and theoretical investigations and, can be harnessed for drug design purposes.


Alternate paths of evolution for the photosynthetic gene rbcL in four nonphotosynthetic species of Orobanche.
We have determined the nucleotide sequence for the Rubisco large subunit from four holoparasitic species of Orobanche. Intact open reading frames are present in two species (O. corymbosa and O. fasciculata), whereas the remaining species (O. cernua and O. ramosa) have rbcL pseudogenes. Sequences for rbcL 5'-UTRs from species of Orobanche have few changes in the promoter and ribosome binding sites compared to photosynthetic higher plants. Comparison of rbcL 3'-UTR sequences for Nicotiana, Ipomoea, Cuscuta, and Orobanche reveal that nucleotide sequences from parasitic plants have regions capable of forming stem-loop structures, but 56-69 nt are deleted upstream of the stem-loop in the parasitic plants compared to their photosynthetic relatives. Although rbcL pseudogenes of O. cernua and O. ramosa have many large and small deletions, few indels are shared in common, implying that their common ancestor probably had an intact rbcL reading frame. Intact rbcL reading frames in O. corymbosa and O. fasciculata retain a bias of synonymous over nonsynonymous substitutions and deduced protein sequences are consistent with potentially functional Rubisco large subunit proteins. A conservative model of random substitution processes in pseudogene sequences estimates that the probability is low (P < 0.028) that these sequences would retain an open reading frame by chance. Species of Orobanche have either had recent photosynthetic ancestors, implying multiple independent losses of photosynthesis in this genus, or the rbcL gene may serve an unknown function in some nonphotosynthetic plants.


Focal macular electroretinograms before and after removal of choroidal neovascular lesions.
To evaluate the changes in focal macular electroretinograms (fmERGs) after surgical removal of choroidal neovascular (CNV) lesions. Fourteen patients (14 eyes) with subfoveal or juxtafoveal CNV associated with age-related macular degeneration and 1 patient with idiopathic CNV underwent vitrectomy and removal of the lesions. fmERGs elicited by a 15 degree stimulus were recorded before and 3 months after surgery. Optical coherence tomography (OCT) was performed to measure the foveal and parafoveal thickness before and 3 months after surgery. Preoperative fmERGs were markedly reduced in all eyes. The mean amplitude of the b-wave in 15 eyes recorded 3 months after surgery increased significantly (P = 0.0022, Wilcoxon signed rank test). In all eyes except two with nearly nonrecordable a- and b- waves, the mean b-wave-to-a-wave ratio after surgery increased significantly in all eyes (P = 0.0330, Wilcoxon signed rank test). The percentage increase in the b-wave amplitude correlated significantly with the percentage decrease in the mean parafoveal retinal thickness (r = 0.688, P = 0.0076). The decreased macular ERGs were partially recoverable in the early postoperative period. The decreased retinal edema after surgery may have contributed to this recovery.


How the principles of geriatric assessment are shaping managed care.
In traditional geriatric medicine, comprehensive assessment is considered crucial to the care of frail older patients. The principles of geriatric assessment--identifying high-risk patients and targeting them for preventive interventions--are also practiced by managed care organizations (MCOs). Self-reported health surveys and administrative data are two methods used by MCOs to identify members at high risk for adverse health outcomes and functional decline who may benefit from geriatric case management. For a successful partnership with primary care physicians, it is very important that geriatric care managers should be knowledgeable in the principles of geriatric medicine.


Succinic acid exacerbates experimental autoimmune uveitis by stimulating neutrophil extracellular traps formation via SUCNR1 receptor.
To investigate the effect of succinic acid on the development of experimental autoimmune uveitis (EAU) and the underlying mechanism. Succinic acid was administrated intraperitoneally to evaluate its effects on immune response and EAU in mice. Intraocular inflammation was evaluated by histopathological scoring. Frequencies of Th1/Th17 cells were measured by flow cytometry. Concentrations of IFN-γ/IL-17A, neutrophil elastase (NE) and myeloperoxidase (MPO) were determined by enzyme-linked immunosorbent test. Infiltration of neutrophils and generation of neutrophil extracellular traps (NETs) within the eye were assessed by immumofluorescence. NETs formation in extracellular matrix was visualised by laser scanning confocal microscopy. Succinate receptor (SUCNR1) antagonist was used to investigate its effect on the generation of NETs. Intraperitoneal injection of succinic acid exacerbated EAU severity as evidenced by severe histological changes in association with elevated frequencies of splenic Th1/Th17 cells, and upregulated levels of IFN-γ/IL-17A and NETs in plasma. In vitro experiments showed that succinic acid could promote the generation of NETs by neutrophils as shown by increased expression of NE and MPO.NETs could increase the frequencies of Th1/Th17 cells in CD4+ T cells and their expression of IFN-γ/IL-17A. In the experiment of receptor antagonism, the upregulatory effect of succinic acid on NETs could be significantly blocked by SUCNR1 antagonist. Succinic acid could worsen EAU induced by IRBP in mice. This effect was possibly mediated by its upregulation on NETs generation and frequencies of Th1/Th17 cells in affiliation with increased production of IFN-γ/IL-17A through succinic acid-SUCNR1 axis.


Assessment of replication rates of human keratinocytes in engineered skin substitutes grafted to athymic mice.
Stable closure of skin wounds with engineered skin substitutes (ESS) requires indefinite mitotic capacity to generate the epidermis. To evaluate whether keratinocytes in ESS exhibit the stem cell phenotype of label retention, ESS (n = 6-9/group) were pulsed with 5-bromo-2'-deoxyuridine (BrdU) in vitro, and after grafting to athymic mice (n = 3-6/group). Pulse and immediate chase in vitro labeled virtually all basal keratinocytes at day 8, with label uptake decreasing until day 22. Label retention in serial chase decreased more rapidly from day 8 to day 22, with a reorganization of BrdU-positive cells into clusters. Similarly, serial chase of labeled basal keratinocytes in vivo decreased sharply from day 20 to day 48 after grafting. Label uptake was assessed by immediate chases of basal keratinocytes, and decreased gradually to day 126, while total labeled cells remained relatively unchanged. These results demonstrate differential rates of label uptake and retention in basal keratinocytes of ESS in vitro and in vivo, and a proliferative phenotype with potential for long-term replication in the absence of hair follicles. Regulation of a proliferative phenotype in keratinocytes of ESS may improve the biological homology of tissue-engineered skin to natural skin, and contribute to more rapid and stable wound healing.


Antioxidant Effects, Antiproliferative Effects, and Molecular Docking of Clinacanthus nutans Leaf Extracts.
Clinacanthus nutans is a well-known herb that has been used as an alternative and therapeutic medicine, however more selective C. nutans extracts are needed. In this study, leaves were extracted with 80% methanol and further fractionated with n-hexane, dichloromethane, chloroform, n-butanol, and aqueous residue. Subsequently, the total phenolic content (TPC), total flavonoid content (TFC), antioxidant scavenging activity, and antiproliferative effects on breast cancer (Michigan Cancer Foundation-7 [MCF7]) and normal breast (Michigan Cancer Foundation-10A [MCF 10A]) cells of the extracts were measured. Additionally, molecular docking simulation of the major compounds from C. nutans extracts was conducted. The aqueous residue had the highest TPC and TFC, whereas the crude extract had the highest scavenging activity. Among the extracts, dichloromethane extract (CN-Dcm) was selected as it had the highest selectivity index (SI) (1.48). Then, the chosen extract (CN-Dcm) was proceed for further analysis. The compounds from CN-Dcm were identified using gas chromatography-mass spectrometry (GC-MS). The major compounds from CN-Dcm were further investigated through molecular docking studies. Palmitic acid and linolenyl alcohol were the compounds found in the CN-Dcm extract that exhibited the highest binding affinities with p53-binding protein Mdm-2. These results highlight the potential of C. nutans as a source of anticancer activities.


Recurrent Trigeminal Neuralgia: A Case Series and a Review of the Literature.
Trigeminal neuralgia is a peripheral neuropathy characterized by intermittent episodes of severe facial pain originating in the sensory nucleus of the trigeminal nerve. The most commonly involved area is the mandibular division with a higher prevalence on the right side. Advances in the field of MRI have played an important role in its diagnosis, especially in presurgical assessment, to probe into any secondary causes of nerve compression and/or neurovascular conflict. The condition is primarily managed medically, although many patients may require surgical or radiotherapeutic intervention. A recurrence rate ranging from 6 to 41% has been postulated. Reasons for recurrence are mainly attributed to improper operative techniques, dislocation of the Teflon implant, or granuloma formation. MRI serves as a powerful tool in the segmental evaluation of the trigeminal nerve. A proper diagnosis with a structured treatment protocol is critical for managing such cases of trigeminal neuralgia. In this report, we present a series of two cases of recurrent trigeminal neuralgia.


The development of the Systems and Individual Responsibility for Poverty (SIRP) Scale.
Poverty is a widespread social problem that affects a substantial number of Americans each year. Attitudes can affect a range of judgments, behavioral intentions, and actions related to addressing this problem. However, existing tools that measure attitudes toward those in poverty do not fully capture the deficit ideology that is a critical component of beliefs about social class. We developed the Systems and Individual Responsibility for Poverty (SIRP) Scale to address this gap. This article describes the development of an initial pool of 20 items, and then the refinement and validation of the final 17-item measure. Exploratory factor analysis (N = 260) and confirmatory factor analysis (N = 280) yielded 2 factors: Individual Responsibility and Systems Responsibility. The Individual Responsibility factor comprises two subscales, differentiating between reasons for getting into and not getting out of poverty. The SIRP is a new tool for evaluating community psychologists' efforts to shift deficit-oriented thinking about poverty, and is likely to be particularly useful in education contexts. In future work, community psychologists might adapt the measure to assess deficit oriented thinking about other problems, and to explore how such attitudes are related to behavior.


Ketoconazole-evoked [Ca2+]i rises and non-Ca2+-triggered cell death in rabbit corneal epithelial cells (SIRC).
The effect of ketoconazole on cytosolic free Ca2+ concentrations ([Ca2+]i) and proliferation has not been explored in corneal cells. This study examined whether ketoconazole alters Ca2+ levels and causes cell death in SIRC rabbit corneal epithelial cells. [Ca2+]i and cell viability were measured by using the fluorescent dyes fura-2 and WST-1, respectively. Ketoconazole at concentrations of 5 microM and above increased [Ca2+]i in a concentration-dependent manner. The Ca2+ signal was reduced partly by removing extracellular Ca2+. The ketoconazole-induced Ca2+ influx was insensitive to L-type Ca2+ channel blockers and protein kinase C modulators. In Ca2+-free medium, after pretreatment with 50 microM ketoconazole, thapsigargin-(1 microM)-induced [Ca2+]i rises were abolished; conversely, thapsigargin pretreatment nearly abolished ketoconazole-induced [Ca2+]i rises. Inhibition of phospholipase C with 2 microM U73122 did not change ketoconazole-induced [Ca2+]i rises. At concentrations between 5 and 100 microM, ketoconazole killed cells in a concentration-dependent manner. The cytotoxic effect of 50 microM ketoconazole was not reversed by prechelating cytosolic Ca2+ with BAPTA. In summary, in corneal cells, ketoconazole-induced [Ca2+]i rises by causing Ca2+ release from the endoplasmic reticulum and Ca2+ influx from unknown pathways. Furthermore, the cytotoxicity induced by ketoconazole was not caused via a preceding [Ca2+]i rise.


Homogeneous time-resolved fluoroimmunoassay of thyroxin in serum.
We describe a rapid, simple nonseparation fluoroimmunoassay for determination of thyroxin in serum. The assay is based on the labeling of thyroxin directly with a fluorescent europium chelate, the fluorescence of which is quenched on binding to an antithyroxin antibody. With the assay buffer we used, maximum quenching is 90%. The rapid achievement of equilibrium in the assay solution, regardless of the sequence of reagent additions, allows fast measurement of thyroxin. Precision was good (CV less than 5%) within the clinical range for total thyroxin (50-300 nmol/L), and results correlated well with those by a commercial radioimmunoassay.


Renal angiomyolipoma, a diagnostic dilemma: a case report.
Angiomyolipomas are the most common benign mesenchymal tumours of the kidney. Most of the angiomyolipomas in adults are asymptomatic and discovered incidentally. We report a case in a symptomatic 55 yrs old male who had a huge retroperitoneal mass abdomen and in whom diagnosis and organ of origin remained unclear until operation and histopathological study was performed. Radiological and fine needle aspiration cytology (FNAC) report of a malignant retroperitoneal tumour was given, but histological examination of the respective tumorous mass showed a benign renal tumour - angiomyolipoma. Our emphasis by reporting this case is to make cytopathologists and radiologists aware of such kind of diagnostic pitfall encountered while reporting retroperitoneal masses.


Percutaneous sacroplasty for sacral metastatic tumors under fluoroscopic guidance only.
Percutaneous sacroplasty is a safe and effective procedure for sacral insufficient fractures under CT or fluoroscopic guidance; although, few reports exist about sacral metastatic tumors. We designed a pilot study to treat intractable pain caused by a sacral metastatic tumor with sacroplasty. A 62-year-old man and a 38-year-old woman with medically intractable pain due to metastatic tumors of S1 from lymphoma and lung cancer, respectively, underwent percutaneous sacroplasty. Over the course of the follow-up period, the two patients experienced substantial and immediate pain relief that persisted over a 3-month and beyond. The woman had deposition of PMMA (polymethyl methacrylate) in the needle track, but did not experience significant symptoms. No other peri-procedural complications were observed for either patient.


Perioperative experience of pelvic organ prolapse repair with the Prolift and Elevate vaginal mesh procedures.
We compared the operative and immediate postoperative experience of the trocar-based Prolift and non-trocar-based Elevate techniques used to repair vaginal prolapse. A retrospective review of Prolift and Elevate repairs was performed. Baseline characteristics and operative and postoperative variables evaluated included compartment(s) repaired, adjacent organ injury, operative time (OT), change in hemoglobin (ΔH), pain score, narcotic use, length of stay (LOS), and short-term complications. Categorical variables were assessed as counts and percent frequency. Data were compared using chi-squared analysis and paired t test. Prolift (n = 143) and Elevate (n = 77) patients were similar in age (p = 0.19). Concurrent hysterectomy was done in 22 (15.4 %) and 24 (31.2 %), respectively, and concurrent midurethral sling placed in 100 (70 %) and 50 (65 %), respectively. LOS (median, 25th,75th) after anterior/apical compartment repairs was shorter with Elevate, whether with (1.0; 1.0,1.5 vs. 2.0 days;1.0, 2.0; p = 0.003) or without (2.0; 1.0, 2.0 vs. 2.0 days; 2.0, 3.0; p = 0.024) hysterectomy, but no differences in OT, ΔH, pain score, or narcotic use occurred. Posterior compartment mean pain scores were lower with Prolift (3.6 ± 2.2 vs. 1.7 ± 1.5, p = 0.035), and three-compartment-repair pain scores were lower with Elevate (0.6 ± 1.3 vs 2.5 ± 1.9; p = 0.013). Three bladder injuries occurred with Prolift but none with Elevate. Operative and postoperative experiences were similar between groups; however, Elevate anterior/apical repairs had shorter LOS, which might reflect more aggressive discharge planning. There were no bowel or major vascular injuries, and the Prolift trocar bladder injuries did not alter the surgical procedure.


One- and Two-Coordinate Detectors in BINP.
One- and two-coordinate detectors with proportional chambers developed at the Budker Institute of Nuclear Physics (BINP) are presented. The parallax-free 10 MHz one-coordinate OD-3.1 and OD-3.2 detectors are used in synchrotron radiation powder diffraction and SAXS experiments. The two-coordinate DED-3 detector with a multiwire proportional chamber (MWPC) is used in Laue diffraction. The latest modification of this detector, DED-5, with a working area of 384 x 384 mm, is briefly described. The micro-strip detector prototype MSGC-100 has passed test synchrotron radiation experiments and the next modification (MSGC-500), with 500 channels for high energy, is under construction. The one-coordinate MWPC OD-160 detector, with an angle aperture of 160 degrees and a count rate of 3.3 GHz, is under construction. It will be used for high-resolution powder diffraction. Two types of gas chamber will be used: L for low energies (5-30 keV) and H for high energies (30-70 keV). A 16 degrees section with an H-chamber has been produced and tested on the synchrotron radiation beamline.


Synthesis of Benzodihydrofurans by Asymmetric C-H Insertion Reactions of Donor/Donor Rhodium Carbenes.
Metal carbenes appended with two electron-donating groups, known as "donor/donor" carbenes, undergo diastereo- and enantioselective rhodium-catalyzed C-H insertion reactions with ether substrates to form benzodihydrofurans. Unlike the reactions of metal carbenes with electron-withdrawing groups attached, the attenuated electrophilicity enables these reactions to be conducted in Lewis basic solvents (e.g., acetonitrile) and in the presence of water. The diazo precursors for these species are prepared in situ from hydrazone using a mild and chemoselective oxidant (MnO2 ). Although this sequence often can be performed in one-pot, control experiments have elucidated why a "two-pot" process is often more efficient. A thorough screening of achiral catalysts demonstrated that sterically encumbered catalysts are optimal for diastereoselective reactions. Although efficient insertion into allylic and propargylic C-H bonds is observed, competing dipolar cycloaddition processes are noted for some substrates. The full substrate scope of this useful method of benzodihydrofuran synthesis, mechanisms of side reactions, and computational support for the origins of stereoselectivity are described.


4-Phenyl-4-[1H-imidazol-2-yl]-piperidine derivatives, a novel class of selective delta-opioid agonists.
A novel series of 4-phenyl-4-[1H-imidazol-2-yl]-piperidine derivatives has been prepared and their synthesis described herein. In vitro affinities for delta-, mu-, and kappa-opioid receptors, as well as the functional activity in the [(35)S]GTPgammaS assay are reported. The most potent and selective delta-opioid agonist 18a exhibited a K(i) of 18 nM, and was >258-fold and 28-fold selective over mu- and kappa-receptors, respectively; the compound is a full agonist with an EC(50) value of 14 nM.


Measurement of pain and analgesia in cancer patients.
Measurement of pain in cancer patients requires all the procedural safeguards essential for the measurement of subjective responses, including the employment of active and inactive controls, double-blind techniques, randomization, and statistical verification of results. Pain is traditionally measured in analgesic studies by employing verbal descriptors of intensity, but more recently visual analogues of pain intensity have been used and generally provide more sensitive measures of pain intensity. Patients with chronic pain tend to rate the categories representing more intense pain as lower in the visual analogue scale than do patients with postoperative pain. This may well reflect differences in the prior pain experiences of the two groups. Patients with chronic cancer pain have greater positive mood effects after the narcotic, morphine, than after the non-steroidal antiinflammatory analgesic, zomepirac, and this appears to be independent of analgesic activity. It is possible to design crossover analgesic studies in cancer patients so as to minimize carry-over effects, and such studies are more efficient than parallel group assays. Crossover studies also provide the ability to measure carry-over when it occurs.


Stacking and T-shape competition in aromatic-aromatic amino acid interactions.
The potential of mean force of interacting aromatic amino acids is calculated using molecular dynamics simulations. The free energy surface is determined in order to study stacking and T-shape competition for phenylalanine-phenylalanine (Phe-Phe), phenylalanine-tyrosine (Phe-Tyr), and tyrosine-tyrosine (Tyr-Tyr) complexes in vacuo, water, carbon tetrachloride, and methanol. Stacked structures are favored in all solvents with the exception of the Tyr-Tyr complex in carbon tetrachloride, where T-shaped structures are also important. The effect of anchoring the two alpha-carbons (C(alpha)) at selected distances is investigated. We find that short and large C(alpha)-C(alpha) distances favor stacked and T-shaped structures, respectively. We analyze a set of 2396 protein structures resolved experimentally. Comparison of theoretical free energies for the complexes to the experimental analogue shows that Tyr-Tyr interaction occurs mainly at the protein surface, while Phe-Tyr and Phe-Phe interactions are more frequent in the hydrophobic protein core. This is confirmed by the Voronoi polyhedron analysis on the database protein structures. As found from the free energy calculation, analysis of the protein database has shown that proximal and distal interacting aromatic residues are predominantly stacked and T-shaped, respectively.


Rapid Genomic Evolution Drives the Diversification of Male Reproductive Genes in Dung Beetles.
The molecular basis for the evolution of novel phenotypes is a central question in evolutionary biology. In recent years, dung beetles have emerged as models for novel trait evolution as they possess distinct precopulatory traits such as sexually dimorphic horns on their head and thorax. Here, we use functional and evolutionary genomics to investigate the origins and the evolution of postcopulatory reproductive traits in male dung beetles. Male ejaculates that underlie postcopulatory sexual selection are excellent candidates to study novel trait evolution as they are complex, fast evolving, and often highly divergent in insects. We assemble de novo transcriptomes of male accessory glands and testes of a widespread dung beetle, Catharsius molossus, and we perform an evolutionary analysis of closely and distantly related insect genomes. Our results show there is rapid innovation at the genomic level even among closely related dung beetles. Genomic expansion and contraction drive the divergence of male reproductive traits and their functions. The birth of scores of completely novel reproductive genes is reinforced by the recruitment of these genes for high expression in male reproductive tissues, especially in the accessory glands. We find that male accessory glands of C. molossus are specialized for secretory function and express female, egg, and embryo-related genes as well as serine protease inhibitors, whilst the testes are specialized for spermatogenesis and sperm function. Finally, we touch upon putative functions of these evolutionary novelties using structure-function analysis as these proteins bear no homology to any other known proteins.


Clinical endpoints in allergen immunotherapy: State of the art 2022.
110 years after the classical study by Noon, numerous studies have confirmed the efficacy of allergen immunotherapy. A variety of clinical endpoints have been used in these trials. This review gives an overview of clinical endpoints for randomized clinical trials on allergen immunotherapy (AIT) in rhinitis and asthma. In addition, real-life studies have been carried out with the same kind of endpoints. In general, AIT studies are characterized by a lack of standardized and validated outcome measures. For allergic rhinoconjunctivitis, digital tools have been developed to monitor patients. Such tools are particularly useful to obtain real-world evidence for AIT. Finally, well-accepted outcome measures are available for cost-effectiveness studies.


Comprehensive review on intravertebral intraspinal, intrajoint, and intradiscal vacuum phenomenon: From anatomy and physiology to pathology.
The term 'vacuum phenomenon' (VP), is characterized by gas-like density areas due to a rapid increase in the joint space volume ('acute VP') or represent a chronic gas collection. It can occur within a collapsed vertebral body, the spinal canal, joints but mainly the intervertebral disc. Studies support that VP is originated by a dynamic process involving the balance between tissues' liquid and gaseous components, influenced by the duration and the depth of mechanical and metabolic alterations, by the nature of neighboring tissues and the variability in both pressure and permeability of disc or vertebral or joint structures. Prevalence of VP in the general population is about 2%, reaching 20% in the elderly with disc degeneration. Although it's often a random finding in asymptomatic patients, VP is an eventually painful expression of disc degeneration, or disc or vertebral fracture, or bone lesions. In sporadic cases, intradiscal gas can be expelled (all-in-one or gradually), resulting in a gaseous cyst, causing pain and neurological symptoms. Considering that spontaneous resolution and recurrence after surgery are both possible, most of the authors recommend conservative treatment in patients with intradiscal and intravertebral VP; occasionally percutaneous CT(computed tomography) -guided aspiration or vertebral stabilization.


Early Life Pb Exposure and its Effect on Later Life Retinal Degeneration.
The study presents the longitudinal effect of early life lead exposure on retinal ischemia. Swiss albino mice were exposed to lead acetate at two different timepoints viz. postnatal day 1-20 and at 7th week of age. These mice were then followed till 10 week of age and subjected to retinal ischemia. Retinal ischemia was induced using pterygopalatine artery ligation. The effect of prior lead exposure on ischemic insult was determined using various histological and molecular parameters. Although toxic effects of Pb are well reported, but the results in this study showed not much significant effect of early life Pb exposure on later life retinal degeneration. While retinal thickness was decreased in surgery group and 7th week Pb exposed group, PND Pb exposed mice showed retinal thickness comparable to normal control. There was no difference in TUNEL positive cells in Pb exposed group when compared to surgery group. The molecular studies revealed overexpression of BDNF and GFAP in PND Pb exposed mice reflecting more injury and inflammation. The combined results revealed that Pb exposure have mild effect on overall susceptibility to retinal damage in later life. The Pb may exert its toxic effects at longer duration and thus the toxic effects may be mapped at longer timepoints. The study provides a new dimension to the already well known toxic effects of lead which needs to be further explored. J. Cell. Biochem. 118: 3213-3224, 2017. © 2017 Wiley Periodicals, Inc.


Lattice Boltzmann simulations of heat transfer in fully developed periodic incompressible flows.
Flow and heat transfer in periodic structures are of great interest for many applications. In this paper, we carefully examine the periodic features of fully developed periodic incompressible thermal flows, and incorporate them in the lattice Boltzmann method (LBM) for flow and heat transfer simulations. Two numerical approaches, the distribution modification (DM) approach and the source term (ST) approach, are proposed; and they can both be used for periodic thermal flows with constant wall temperature (CWT) and surface heat flux boundary conditions. However, the DM approach might be more efficient, especially for CWT systems since the ST approach requires calculations of the streamwise temperature gradient at all lattice nodes. Several example simulations are conducted, including flows through flat and wavy channels and flows through a square array with circular cylinders. Results are compared to analytical solutions, previous studies, and our own LBM calculations using different simulation techniques (i.e., the one-module simulation vs. the two-module simulation, and the DM approach vs. the ST approach) with good agreement. These simple, however, representative simulations demonstrate the accuracy and usefulness of our proposed LBM methods for future thermal periodic flow simulations.


Association between anlotinib trough plasma concentration and treatment outcomes in advanced non-small-cell lung cancer.
Efficacy and toxicities of anlotinib (ANL) show large inter-patient variation, which may partly be explained by differences in ANL exposure. Exposure-response/toxicities relationship have not been investigated for ANL. Therefore, the aim of the present study was to explore the association between the trough plasma concentration (Ctrough) of ANL and treatment outcomes in Chinese patients with advanced non-small cell lung cancer (NSCLC). Patients with advanced NSCLC who started third-line or further ANL alone therapy between January 2021 and October 2022. This study examined the ANL Ctrough and clinical response evaluation at day 43 after initiation of ANL treatment. We evaluated the association between the ANL Ctrough and clinical efficacy and toxicities. Additionally, this study defined patients with complete response (CR), partial response (PR) and stable disease (SD) as responder. The receiver-operating characteristic (ROC) curve combined with Youden index was identify the potential threshold value of ANL Ctrough for the responder. 52 patients were evaluated for analyses. The median ANL Ctrough was 11.45ng/ml (range, 3.69-26.36 ng/ml). The ANL Ctrough values in the PR group (n=6, 15.51 ng/ml (range, 8.19-17.37 ng/ml)) was significantly higher than in the PD group (n=8, 7.44 ng/ml (range, 5.41-14.69 ng/ml), p=0.001). The area under the ROC curve (AUCROC) was 0.76 (95% confidence interval (CI), 0.58-0.93; p=0.022) and threshold value of ANL Ctrough predicting responder was 10.29 ng/ml (sensitivity 65.9% and specificity 87.5%, the best Youden index was 0.53). The disease control rate (DCR) was 84.6%, and DCR was significantly higher in the high-exposure group (≥10.29ng/ml) than low-exposure group (<10.29ng/ml) (96.67% vs 68.18%, p=0.005). Although there was no significant difference in ANL Ctrough between grade ≥ 3 and grade ≤2 toxicities, the incidence of any grade hand-foot syndrome (70.0% vs 36.36%, p=0.016) and thyroid-stimulating hormone elevation (53.33% vs 22.73%, p =0.026) was significantly higher in the high-exposure group compared with the low-exposure group. Considering these results, we propose that maintaining ANL Ctrough ≥ 10.29ng/ml was important for achieving the response in advanced NSCLC patients treated with ANL.


Molecular dynamics and metadynamics simulations of electrosprayed water nanodroplets including sodium bis(2-ethylhexyl)sulfosuccinate micelles.
The behavior of aqueous solutions of sodium bis(2-ethylhexyl)sulfosuccinate (AOTNa) under conditions of electrospray ionization (ESI) has been investigated by molecular dynamics (MD) and well-tempered metadynamics (WTM) simulations at 300 K and 400 K. We have examined water droplets with initial fixed numbers of water molecules (1000) and AOT- anions (100), and with sodium cations in the range of 70-130. At 300 K, all charged droplets show the water evaporation rate increasing with the absolute value of the initial droplet charge state (Z), accompanied by ejection of an increasing number of solvated sodium ions or by expulsion of AOT- anions depending on the sign of Z and by fragmentation in the case of high |Z|. At 400 K, the water evaporation becomes more rapid and the fission process more extensive. In all cases, the AOTNa molecules, arranged as a direct micelle inside the aqueous system, undergo a rapid inversion in vacuo so that the hydrophilic heads and sodium ions surrounded by water molecules move toward the droplet interior. At the end of the 100-ns MD simulations, some water molecules remain within the aggregates at both temperatures. The subsequent metadynamics simulations accelerate the droplet evolution and show that all systems become anhydrous, in agreement with the experimental results of ESI mass spectrometry. This complete water loss is accompanied by sodium counterion emission for positively charged aggregates at 300 K. The analysis shows how the temperature and droplet charge state affect the populations of the generated surfactant aggregates, providing information potentially useful in designing future ESI experimental conditions.


BTB and TAZ domain scaffold proteins perform a crucial function in Arabidopsis development.
In Arabidopsis, bric-a-brac, tramtrack and broad (BTB) domain scaffold proteins form a family of 80 proteins that have involvement in various signaling pathways. The five members of the subfamily of BTB AND TAZ DOMAIN proteins (BT1-BT5) have a typical domain structure that is only observed in land plants. Here, we present a functional analysis of the BT family, of which at least four members are encoded by auxin-responsive genes. BT1 is a short-lived protein that is characteristically targeted for degradation by the 26S proteasome. Expression pattern, gene structure and sequence analyses indicate that BT1 and BT2 are closely related. They both localize to the nucleus and the cytosol, whereas the remaining BT proteins were determined as cytosolic proteins. Detailed molecular and phenotypic analysis of plants segregating for null mutations in the BT family revealed substantial redundancy among the BT members, and highlighted that BT proteins perform crucial roles in both male and female gametophyte development. BT2 seems to be the predominant gene in this process, in which it is functionally replaced by BT3 and BT1 through reciprocal transcription regulation. Compensational expression alters the steady-state mRNA levels among the remaining BT family members when other BT members are lost, and this contributes towards functional redundancy. Our data provide a surprising example of functional redundancy among genes required during gametophyte development, something that could not be detected in the current screens for gametophyte mutants.


Multiparametric in vitro genotoxicity assessment of different variants of amorphous silica nanomaterials in rat alveolar epithelial cells.
The hazard posed to human health by inhaled amorphous silica nanomaterials (aSiO2 NM) remains uncertain. Herein, we assessed the cyto- and genotoxicity of aSiO2 NM variants covering different sizes (7, 15, and 40 nm) and surface modifications (unmodified, phosphonate-, amino- and trimethylsilyl-modified) on rat alveolar epithelial (RLE-6TN) cells. Cytotoxicity was evaluated at 24 h after exposure to the aSiO2 NM variants by the lactate dehydrogenase (LDH) release and WST-1 reduction assays, while genotoxicity was assessed using different endpoints: DNA damage (single- and double-strand breaks [SSB and DSB]) by the comet assay for all aSiO2 NM variants; cell cycle progression and γ-H2AX levels (DSB) by flow cytometry for those variants that presented higher cytotoxic and DNA damaging potential. The variants with higher surface area demonstrated a higher cytotoxic potential (SiO2_7, SiO2_15_Unmod, SiO2_15_Amino, and SiO2_15_Phospho). SiO2_40 was the only variant that induced significant DNA damage on RLE-6TN cells. On the other hand, all tested variants (SiO2_7, SiO2_15_Unmod, SiO2_15_Amino, and SiO2_40) significantly increased total γ-H2AX levels. At high concentrations (28 µg/cm2), a decrease in G0/G1 subpopulation was accompanied by a significant increase in S and G2/M sub-populations after exposure to all tested materials except for SiO2_40 which did not affect cell cycle progression. Based on the obtained data, the tested variants can be ranked for its genotoxic DNA damage potential as follows: SiO2_7 = SiO2_40 = SiO2_15_Unmod > SiO2_15_Amino. Our study supports the usefulness of multiparametric approaches to improve the understanding on NM mechanisms of action and hazard prediction.


Unraveling trauma and stress, coping resources, and mental well-being among older adults in prison: empirical evidence linking theory and practice.
A theoretical integration of the life course perspective, cumulative advantage, disadvantage or inequality, and stress processing theories provide an important integrated lens to study the relationship between accumulated interpersonal, social-structural, and historical trauma and stressful experiences on mental well-being mental well-being in later life. This study builds upon the extant literature by examining the mediating role of coping resources on the relationship between trauma and stressful life experiences, post traumatic stress symptoms, and mental well-being among a sample of 677 adults aged 50 and older in prison. The majority (70%) reported experiencing one or more traumatic or stressful life experiences during their life span. Participants also reported on average 11 occurrences of multilevel trauma and stressful life events and lingering subjective distress related to these events. Results of a structural equation model revealed that internal and external coping resources (e.g., cognitive, emotional, physical, spiritual, and social) had a significant and inverse effect on the relationship between trauma and stressful life experiences and mental well-being. As prisons are forced to deal with an aging population, research in this area can take the preliminary steps to enhance understanding of risk and resilience among older adults in prison. This understanding will aid in the development and improvement of integrated theory-based interventions seeking to increase human rights, health, and well-being among older adults in prison.


Sorbinil: a member of the novel class of spirohydantoin aldose reductase inhibitors.
A decade ago, we discovered that spirohydantoins are a novel class of aldose reductase inhibitors characterized by very potent in vivo activity. This important discovery resulted from a systematic screening effort for in vitro activity against aldose reductase isolated from bovine lens and subsequent testing of active compounds for in vivo activity in a streptozotocin-diabetic rat model, measuring inhibition of sorbitol formation in sciatic nerve. In this in vivo model, spirohydantoins were clearly more potent than all known carboxylic acid-type aldose reductase inhibitors. Structure-activity studies in the spirohydantoin class demonstrated that potent in vitro and in vivo activity were obtained when the spiro junction was adjacent to an aromatic ring and when this junction was part of a 5- or 6-membered ring system fused to the aromatic ring. Excellent in vitro and in vivo activity was achieved in 6-halogenated chromane derivatives with the spirohydantoin group attached in the 4-position. In this series, biologic activity is highly stereospecific and associated with the S configuration. Sorbinil, the S isomer of the 6-fluoro derivative in this series, is one such example: it is 30 times more potent than its R isomer in vitro (IC50 0.15 v 4.4 mumol/L) and 100 times more potent in vivo (ED50 0.25 mg/kg po v 25 mg/kg po).


Limits of Surgical Resection for Bile Duct Cancer.
Perihilar cholangiocarcinoma is the most frequent cholangiocarcinoma and poses difficulties in preoperative evaluation. For its therapy, often major hepatic resections as well as resection and reconstruction of the hepatic artery or the portal vein are necessary. In the last decades, great advances were made in both the surgical procedures and the perioperative anesthetic management. In this article, we describe from our point of view which facts represent the limits for curative (R0) resection in perihilar cholangiocarcinoma. Retrospective data of a 6-year period (2008-2014) was collected in an SPSS 22 database and further analyzed with focus on the surgical approach and the postoperative as well as histological results. Out of 96 patients in total we were able to intend a curative resection in 73 patients (76%). In 58/73 (79.5%) resections an R0 situation could be reached (R1 n = 14; R2 n = 1). 23 patients were irresectable because of peritoneal carcinosis (n = 8), broad infiltration of major blood vessels (n = 8), bilateral advanced tumor growth to the intrahepatic bile ducts (n = 3), infiltration of the complete liver hilum (n = 2), infiltration of the gallbladder (n = 1), and liver cirrhosis (n = 1). Patients with a T4 stadium were treated with curative intention twice, and in each case an R1 resection was achieved. Most patients with irresectable tumors can be suspected to have a T4 stadium as well. In a T3 situation (n = 6) we could establish five R0 resections and one R1 resection. The limit of surgical resection for bile duct cancer is the advanced tumor stage (T stadium). While in a T3 stadium an R0 resection is possible in most cases, we were not able to perform an R0 resection in a T4 stadium. From our point of view, early T stadium cannot usually be estimated through expanded diagnostics but only through surgical exploration.


A multinational review of recent trends and reports in dementia caregiver burden.
This systematic review of the literature focuses on the influence of ethnic, cultural, and geographic factors on the caregivers of patients with dementia. In particular, we explore the impact of cultural expectations on five important questions: 1) Do the characteristics of dementia affect caregiver burden? 2) Do characteristics of the caregiver independently predict burden? 3) Does the caregiver affect patient outcomes? 4) Does support or intervention for caregiver result in reduced caregiver burden or improved patient outcomes? 5) Finally, do patient interventions result in reduced caregiver burden or improved patient outcomes? Our findings suggest that noncognitive, behavioral disturbances of patients with dementia result in increased caregiver burden and that female caregivers bear a particularly heavy burden across cultures, particularly in Asian societies. Caregiver burden influences time to medical presentation of patients with dementia, patient condition at presentation, and patient institutionalization. Moreover, interventions designed to reduce caregiver burden have been largely, although not universally, unsuccessful. Pharmacological treatments for symptoms of dementia were found to be beneficial in reducing caregiver burden. The consistency of findings across studies, geographic regions, cultural differences, and heathcare delivery systems is striking. Yet, there are critical differences in cultural expectations and social resources. Future interventions to reduce caregiver burden must consider these differences, identify patients and caregivers at greatest risk, and develop targeted programs that combine aspects of a number of interventional strategies.


Metabolomic profiling of cardiac allografts after controlled circulatory death.
Assessment of myocardial viability during ex situ heart perfusion (ESHP) is based on the measurement of lactate concentrations. As this provides with limited information, we sought to investigate the metabolic signature associated with donation after circulatory death (DCD) and the impact of ESHP on the myocardial metabolome. Porcine hearts were retrieved either after warm ischemia (DCD group, N = 6); after brain-stem death (BSD group, N = 6); or without DCD nor BSD (Control group, N = 6). Hearts were perfused using normothermic oxygenated blood for 240 minutes. Plasma and myocardial samples were collected respectively every 30 and 60 minutes, and analyzed by an untargeted metabolomic approach using liquid chromatography coupled to high-resolution mass spectrometry. Median duration of warm ischemia was 23 minutes [19-29] in DCD animals. Lactate level within myocardial biopsies was not significantly different between groups at T0 (p = 0.281), and remained stable over the 4-hour period of ESHP. More than 300 metabolites were detected in plasma and heart biopsy samples. Compared to BSD animals, metabolomics changes involving energy and nucleotide metabolisms were observed in plasma samples of DCD animals before initiation of ESHP, whereas 2 metabolites (inosine monophosphate and methylbutyrate) exhibited concentration changes in biopsy samples. Normalization of DCD metabolic profile was remarkable after 4 hours of ESHP. A specific metabolic profile was observed in DCD hearts, mainly characterized by an increased nucleotide catabolism. DCD and BSD metabolomes proved normalized during ESHP. Complementary investigations are needed to correlate these findings to cardiac performances.


Asymptotically stable compensation of the soliton self-frequency shift.
We report on the stable cancellation of the soliton self-frequency shift (SSFS) in nonlinear optical fibers. A soliton, which scatters a group velocity matched pump wave in a so-called optical event horizon regime, may experience a blueshift in frequency, which counteracts the SSFS induced by Raman scattering. The SSFS can easily be compensated by a suitably prepared pump wave, but usually the compensation is unstable and is destroyed after a certain propagation length. We study this kind of soliton-pump wave interaction by an adiabatic approach and quantify the parameter range in which the stable SSFS compensation is possible. The theory enjoys agreement with numerical simulations of soliton propagation.


Internal thoracic artery blood flow is not affected by major side branch.
We report a case of a 73-year-old man in which coronary angiography demonstrated a large side branch of the internal thoracic artery (ITA). The side branch was identified intraoperatively. As assessed intraoperatively by transit time Doppler flow measurement, temporary occlusion of the side branch showed no effect on ITA flow, which remained unchanged at 49 mL/min after anastomosis to left anterior descending coronary artery.


High frequency of rotavirus viremia in children with acute gastroenteritis: discordance of strains detected in stool and sera.
Recently, rotavirus antigenemia and viremia have been identified in patients with acute gastroenteritis. This study examined rotavirus viremia in children hospitalized for acute gastroenteritis in order to establish its association with fecal shedding of rotavirus, infecting genotypes and antibody marker of acute infection. Thirty-one pairs of stool-serum specimens were collected from November 2004 to February 2005 together with clinical information. All paired specimens were screened for rotavirus RNA by RT-PCR using the VP6 gene primers. All stool and serum specimens were tested for rotavirus antigen and anti-rotavirus IgM respectively by ELISA. Sixteen of 31 stool-serum pairs showed the presence of rotavirus RNA. Nine stool and two serum specimens were positive only by RT-PCR. The total positivity in rotavirus RNA was significantly higher in both stools (80.6%) and sera (58.1%) than that of stool antigen (38.7%) and anti-rotavirus IgM (25.8%) (P < 0.01). All PCR positive paired specimens were typed for the VP7 (G) and VP4 (P) genes. Five of sixteen pairs could be typed for both genes. Three of the five pairs showed concordance (G2P[4]/G2P[4]) while two showed discordance (G12P[8]/G2P[4], G8P[4]/G2P[4]) in the genotypes detected in stool and serum specimens respectively. The study documents a high frequency of rotavirus viremia in patients with acute diarrhea. The discordance of rotavirus strains at the genotypic level in the serum and stool of individual patients with diarrhea suggests the susceptibility of extra-intestinal sites for rotavirus infection and the possibility of differential dissemination of rotavirus strains from the intestine.


Peromyscus populations and their Cuterebra parasites display congruent phylogeographical structure.
The relationships between populations of the Deer Mouse (Peromyscus maniculatus) and the White-footed Mouse (P. leucopus) and their respective Cuterebra parasites were examined. Population genetic structure of hosts and parasites was inferred using cytochrome oxidase mitochondrial sequences of specimens from 7 populations. Genetic analyses revealed that isolation-by-distance applies for P. maniculatus and its associated parasite (C. grisea). A significant correlation was also observed between the genetic distances of these host and parasite species. Furthermore, populations of P. maniculatus and C. grisea from the North and South shores of the St Lawrence River were found to be significantly different. This structure may be explained by the St Lawrence River being a dispersal barrier for both species. A robust analysis of the other species pair (P. leucopus and C. fontinella) could not be performed because of limited sample sizes.


Serum extracellular vesicle depletion processes affect release and infectivity of HIV-1 in culture.
Extracellular vesicles (EVs) are involved in intercellular communication and affect processes including immune and antiviral responses. Blood serum, a common cell culture medium component, is replete with EVs and must be depleted prior to EV-related experiments. The extent to which depletion processes deplete non-EV particles is incompletely understood, but depleted serum is associated with reduced viability and growth in cell culture. Here, we examined whether serum depleted by two methods affected HIV-1 replication. In cell lines, including HIV-1 latency models, increased HIV-1 production was observed, along with changes in cell behavior and viability. Add-back of ultracentrifuge pellets (enriched in EVs but possibly other particles) rescued baseline HIV-1 production. Primary cells were less sensitive to serum depletion processes. Virus produced under processed serum conditions was more infectious. Finally, changes in cellular metabolism, surface markers, and gene expression, but not miRNA profiles, were associated with depleted serum culture. In conclusion, depleted serum conditions have a substantial effect on HIV-1 production and infectivity. Dependence of cell cultures on "whole serum" must be examined carefully along with other experimental variables, keeping in mind that the effects of EVs may be accompanied by or confused with those of closely associated or physically similar particles.


Functional affinity of antibody to the Haemophilus influenzae type b polysaccharide.
The functional affinity, or avidity, of antibody to the capsular polysaccharide of Haemophilus influenzae (Hib-PS) was measured in serum samples from 12 adult male subjects before and after immunization with Hib-PS vaccine. Mean avidity increased from 0.74 nM-1 to 1.5 nM-1 after immunization (P less than .05, paired Student's t test). The Bureau of Biologics reference antiserum had an avidity constant of 12 nM-1, which was the highest of all 25 serum samples studied.


[New, innovative prognosis calculator for patients with metastatic spinal tumors].
The aim of our research was to create a scoring system that predicts prognosis and recommends therapeutic options for patients with metastatic spine tumor. Increasing oncological treatment opportunities and prolonged survival have led to a growing need to address clinical symptoms caused by meta-stases of the primary tumor. Spinal metastases can cause a significant reduction in quality of life due to the caused neurological deficits. A scoring system that predicts prognosis with sufficient accuracy could help us to achieve personalised treatment options. Methods - We performed a retrospective clinical research of data from patients over 18 years of age who underwent surgery due to symptomatic spinal metastasis at the National Institute of Mental Disorders, Neurology and Neurosurgery between 2008 and 2018. Data from 454 patients were analysed. Survival analysis (Kaplan-Meier, log-rank, Cox model) was performed, network science-based correlation analysis was used to select the proper prognostic factors of our scoring system, such that its C value (predictive ability index) was maximized. Multivariate Cox analysis resulted in the identification of 5 independent prognostic factors (primary tumour type, age, ambulatory status, internal organ metastases, serum protein level). Our system predicted with an average accuracy of 70.6% over the 10-year study period. Our large case series of surgical dataset of patients with symptomatic spinal metastasis was used to create a risk calculator system that can help in the choice of therapy. Our risk calculator is also available online at https://emk.semmelweis.hu/gerincmet.


Indonesian caretaker experiences in providing child care to Taiwanese children.
The purpose of this study was to learn about the experiences of Indonesian nannies in caring for Taiwanese children in Taiwan. An ethnography method was adopted, using in-depth interviews with 52 respondents, 20 of whom were Indonesians employed as caretakers for Taiwanese children and 32 of whom were general informants. Twenty-six of the general informants were Indonesian child caretakers, and six were employers. Audio-taping of interview sessions was permitted by all respondents. Transcriptions of the interviews were written up verbatim. Leininger's (1991) Phases of Analysis for qualitative data was adopted to analyze data. Four themes were extracted from the analysis, including "Learning diverse information from the beginning--the employer's family," "Complying with the advice of the child's mother," "Maintaining the child's daily activities and health," and "Awareness of external context influences on child care." The findings indicated that Indonesian child caretakers carry with them the values, beliefs, and lifestyles of their own culture and have their own ingrained ideas regarding how best to provide care for their Taiwanese charges. Health care providers, by better understanding the general experiences of Indonesian caretakers in Taiwan, can apply competent intervention to help caretakers overcome the challenges of working within a different cultural environment. It is hoped that the results of this study can promote cultural competence in childcare within the context of internationalism.


Why too soon? Predictors of time to diabetic peripheral neuropathy among newly diagnosed diabetes mellitus patients: a multicenter follow-up study at health-care setting of Ethiopia.
Due to the rising number of diabetic patients, the burden of diabetic peripheral neuropathy (DPN) is clearly posing a major challenge to the long-term viability of the health-care system. Despite this, most DPN epidemiological research in eastern Africa, including Ethiopia, has so far been limited to survey studies. Thus, we determined the incidence of DPN and its predictors among diabetic patients in tertiary health-care setting of southwest Ethiopia. A multicenter retrospective follow-up study was carried out on 567 randomly selected diabetic patients. Data were entered using Epi-Data v4.6 and analyzed using R v4.0.4. The survival curves were estimated using the Kaplan-Meier, and compared using Log-rank test between groups of categorical variables. The PHA were evaluated using the Schoenfeld residuals test. Multivariable Gompertz proportional hazard model was used to examine the predictors of DPN at 5% level of significance. Overall, of 567 DM patients 119 developed DPN with an incidence rate of 3.75, 95%CI [3.13, 4.49] per 100 PY. About 15.13% and 69% of DPN cases occurred within 2 and 5 years of DM diagnosis, respectively. In the multivariable Gompertz PH model, being female [AHR = 1.47; 95% CI (1.01, 2.15)], T2DM [AHR = 3.49 95% CI (1.82, 6.71)], having diabetic retinopathy [AHR = 1.9 95% CI (1.25, 2.91)], positive proteinuria [AHR = 2.22 95% CI (1.35, 3.65)], being obese [AHR = 3.94 95% CI (1.2, 12.89)] and overweight [AHR = 3.34 95% CI (1.09, 10.25)] significantly predicts the future risk of DPN. Nearly, 7 in 10 of DPN cases occurred within short period of time (5 year) of DM diagnosis. Being female, T2DM, DR, positive proteinuria, obese and overweight significantly predicts the risk of DPN. Therefore, we recommend screening and early diagnosis of diabetes with its complication. While doing so, attention should be given for DM patients with DR and positive proteinuria at baseline.


Pharmacotherapy of alcohol use disorders and concurrent psychiatric disorders: a review.
Alcohol use disorders (AUDs) are among the most prevalent psychiatric disorders. Epidemiologic studies have shown a high prevalence of concurrent psychiatric disorders among people with AUDs as well as a higher prevalence of AUDs in people with psychiatric disorders than in the general population. Though psychiatric patients with concurrent AUDs are at increased risk for morbidity and mortality, they are commonly undertreated for their alcohol-related disorders. The efficacy of pharmacotherapy for AUDs is well documented. Our paper reviews the common pharmacotherapies available for AUDs and focuses on the available research regarding treatment of AUDs among psychiatric populations with mood, anxiety, and psychotic disorders. Despite the high prevalence of concurrent AUDs and psychiatric disorders, very limited information has been collected using a randomized controlled trial design targeting those concurrent conditions. Several prevalent psychiatric disorders have not been studied when co-occurring with AUDs. Further research of pharmacological treatments for concurrent AUDs and psychiatric diagnoses is urgently needed.


Phenolic acid glucosides from the seeds of Entada phaseoloides Merill.
Phytochemical investigation of the defatted seeds of Entada phaseoloides Merill. (Mimosaceae) led to the isolation of three new phenolic acid glucosides, which were characterized as 2-hydroxy-5-methylbenzoyl-β-L-glucopyranoside (p-cresotyl glucoside, 1), 2-hydroxy-5-methylbenzoyl-β-L-glucopyranosyl (2 → 1)-β-L-glucopyranosyl (2 → 1)-β-L-glucopyranoside (p-cresotyl triglucoside, 2), and 2-hydroxybenzoyl-β-L-glucopyranosyl (2 → 1)-β-L-glucopyranosyl (2 → 1)-β-L-glucopyranosyl (2 → 1)-β-L-glucopyranoside (salicylic acid tetraglucoside, 5), along with sucrose and triglucoside. The structures of these phytoconstituents have been established on the basis of spectral data analysis and chemical reactions.


Thermal inactivation of avian influenza and Newcastle disease viruses in chicken meat.
Avian influenza viruses (AIV) and Newcastle disease viruses (NDV) of high pathogenicity cause severe systemic disease with high mortality in chickens and can be isolated from the meat of infected chickens. Although AIV and NDV strains of low pathogenicity are typically not present in chicken meat, virus particles in respiratory secretions or feces are possible sources of carcass contamination. Because spread of AIV and NDV is associated with movement of infected birds or their products, the presence of these viruses in chicken meat is cause for concern. This study presents thermal inactivation data for two viruses of high pathogenicity in chickens (AIV strain A/chicken/Pennsylvania/1370/1983 and NDV strain APMV-1/ chicken/California/S0212676/2002) and two viruses of low pathogenicity in chickens (AIV strain A/chicken/Texas/298313/ 2004 and NDV strain APMV-1/chicken/Northern Ireland/Ulster/1967). Under the conditions of the assay, high-pathogenicity AIV was inactivated more slowly in meat from naturally infected chickens than in artificially infected chicken meat with a similar virus titer. In contrast, high-pathogenicity NDV was inactivated similarly in naturally and artificially infected meat. Linear regression models predicted that the current U.S. Department of Agriculture-Food Safety and Inspection Service time-temperature guidelines for cooking chicken meat to achieve a 7-log reduction of Salmonella also would effectively inactivate the AIV and NDV strains tested. Experimentally, the AIV and NDV strains used in this study (and the previously studied H5N1 high-pathogenicity AIV strain A/chicken/Korea/ES/2003) were effectively inactivated in chicken meat held at 70 or 73.9 degrees C for less than 1 s.


Critical behavior and scaling functions of the three-dimensional O(6) model.
We numerically investigate the three-dimensional O(6) model on 12(3) to 120(3) lattices within the critical region at zero magnetic field, as well as at finite magnetic field on the critical isotherm and for several fixed couplings in the broken and the symmetric phase. We obtain from the Binder cumulant at vanishing magnetic field the critical coupling J(c)=1.42865(3). The universal value of the Binder cumulant at this point is g(r)(J(c))=-1.94456(10). At the critical coupling, the critical exponents gamma=1.604(6), beta=0.425(2), and nu=0.818(5) are determined from a finite-size-scaling analysis. Furthermore, we verify predicted effects induced by massless Goldstone modes in the broken phase. The results are well described by the perturbative form of the model's equation of state. Our O(6) result is compared to the corresponding Ising, O(2) and O(4) scaling functions. Finally, we study the finite-size-scaling behavior of the magnetization on the pseudocritical line.


The relationship of body image with symptoms of depression and anxiety in patients with anorexia nervosa during outpatient psychotherapy: Results of the ANTOP study.
Body image disturbance represents a central characteristic of anorexia nervosa (AN). Depression and anxiety are the most common mental comorbidities in patients with AN. This study aims to investigate the relationship of body image with symptoms of depression and anxiety during outpatient psychotherapy in AN. Analyses were conducted using the data set of the Anorexia Nervosa Treatment Outpatient Study (ANTOP) randomized controlled trial. The ANTOP study included N = 242 females with AN between 18 and 56 years of age. The trial was designed to compare enhanced cognitive behavioral therapy (CBT-E) and focal psychodynamic therapy (FPT) with optimized treatment as usual (TAU-O) for patients with AN. The analyses on body image dimensions were conducted using measures of correlations and multiple linear regression analyses to assess the relationship and longitudinal prediction of symptoms of depression and anxiety by body image dimensions. Results showed that body image perceptions were significantly associated with symptoms of depression and anxiety in patients with AN at all treatment stages. In addition, body image dimensions at early treatment stages predict depression and anxiety in follow-up measurements. The correlation of symptoms of depression and anxiety by body image perceptions increased along treatment course. The persistence of body image disturbance, while body mass index increases under treatment (persistency effect), may constitute a relevant factor contributing to the course of the most common affective comorbidities of depression and anxiety in patients with AN. Body image disturbances in patients with AN should therefore be explicitly targeted within the specialized psychotherapy of affected patients. (PsycINFO Database Record


A profile of Australian family caregivers: diversity of roles and circumstances.
Research on family caregiving has been based largely on small samples, often drawn from support organisations or services, and has tended to focus on particular disability groups. Our study was population-based and included all ages and disabilities. As the first stage in a longitudinal research and health promotion program for informal caregivers, a statewide random survey of over 26,000 households was conducted by telephone: 78 per cent of self-identified carers (N = 976) agreed to participate in a one-hour interview. This paper presents a sociodemographic profile of Australian caregivers. Four types of relationship between carers and care recipients (adult offspring, spouses, parents and other relatives and friends) provide the framework for results. Group differences were observed on most characteristics: for example, age, living arrangements, work status and duration of care. Care recipient characteristics, including difficult behaviours and need for assistance, are also reported, as well as use of and need for community services. What the findings reflected most was the heterogeneity of both caregivers and care recipients and the diversity of caregiving roles and circumstances.


Recent developments in dye-sensitized solar cells.
The knowledge of dye-sensitized solar cells (DSCs) has expanded considerably in recent years. They are multiparameter and complex systems that work only if various parameters are tuned simultaneously. This makes it difficult to target to a single parameter to improve the efficiency. There is a wealth of knowledge concerning different DSC structures and characteristics. In this review, the present knowledge and recent achievements are surveyed with emphasis on the more promising cell materials and designs.


Helminths and lipid peroxidation in Astyanax aeneus (Pisces: Characidae) from a river in the humid subtropics of southeastern Mexico.
Xenobiotic mixtures in aquatic ecosystems may compromise fish health by inducing oxidative stress, decreasing resistance, and fostering susceptibility to pathogens such as helminths. Lipid peroxidation (LPO) is one of the most commonly used methods to evaluate oxidative stress. The present study examines the potential correlations between LPO in liver, muscle, and gill and the presence of helminths in Astyanax aeneus from a subtropical river in Mexico. Five helminth species were identified: the trematodes Uvulifer ambloplitis, a subdermal larval parasite, and Clinostomum complanatum, which infected the base of the gills; the nematodes Contracaecum sp. and Spiroxys sp.; and the acanthocephalan Polymorphus sp. The latter 3 species occurred in the posterior half of the intestine. Total helminth abundance was highest in April and July. Helminth species richness increased along a gradient from the headwaters to the middle reaches of the river. An integrated biomarker response for LPO (IBR-LPO) was assessed in the 3 tissues analyzed. IBR-LPO peaked in July. A direct correlation was found between gill and muscle LPO and mean abundance of Clinostomum complanatum, Contracaecum sp., and orthophosphate values, as well as between liver LPO and mean abundance of Polymorphus sp., whereas no relationship was found between LPO and U. ambloplitis or Spiroxys sp. This research revealed an association of both orthophosphate values and the presence or abundance of parasites with oxidative stress levels in A. aeneus, without any causal association, but rather as a result of synergistic processes associated with these variables.


Evaluation of pharmacokinetic, pharmacodynamic, efficacy, and safety data of low-dose ticagrelor versus standard dose in East Asians: a systematic review.
East Asians are prone to higher systemic exposure and increased risk of bleeding compared to other races after administration of antiplatelet agent(s). The aim of this systematic review was to compare and evaluate the appropriateness of a lower dose versus standard dose of ticagrelor in East Asians. A systematic search was conducted in PubMed, EMBASE, and Cochrane databases for studies comparing low versus standard doses of ticagrelor in East Asian populations; a total of seven studies were included in the review. Pharmacokinetic (PK), pharmacodynamic (PD), clinical efficacy, and safety data were collected. PK data demonstrated that the Cmax and area under the curve (AUC) of ticagrelor increased dose-proportionally. PD data indicated that while the low dose of ticagrelor resulted in less antiplatelet activity in three of the five PD studies, in the other studies, there were no differences between low and standard doses. There were two clinical studies included in this review, which showed that the efficacy was similar between the low and standard doses, but both studies were limited by the number of patients included. While there were generally greater incidence of adverse events observed in the standard ticagrelor dose, the magnitude or actual difference between the two doses is difficult to determine due to lack of consistent data. In East Asians, the appropriateness of the low-dose ticagrelor cannot be determined based on the currently available evidence. Additional large-scale and longer duration studies are warranted.


Clinical profile of keratitis treated within 3 months of acute COVID-19 illness at a tertiary care eye centre.
To report the spectrum of keratitis treated within 3 months of acute COVID-19 infection. Retrospective, descriptive case series study of 19 eyes of 16 patients who presented at tertiary eye care centre in Southern India. Median age of the patients was 43(IQR 35-55.5) years. Majority (15/16, 93.75%) were males. Unilateral affliction was predominant (13/16, 81.25% patients). Nine had a history of hospitalization, five had received oxygen supplementation and five had been treated with steroids during COVID-19 illness. The median duration between COVID-19 diagnosis and the ocular symptoms in the eye was 29 (IQR 22-57) days. Microbiological diagnosis consisted of microsporidia in nine eyes of seven patients, fungus in six patients, Pythium in one patient, and herpes zoster ophthalmicus in one patient. One patient had neurotrophic keratitis. Therapeutic penetrating keratoplasty was performed in five patients, glue application in two patients and three were managed with tarsorrhaphy with/without amniotic membrane grafting or tenonplasty. There was medical and surgical cure in all patients. Microsporidia was the commonest cause of keratitis, followed by fungal infection. Majority of the microsporidia infections were keratoconjunctivitis. The fungal isolates identified were Aspergillus and Mucor species. All patients responded to conventional management guidelines with favourable outcomes.


Umbilical cord blood: The promise and the uncertainty.
Unfortunately, many patients referred for hematopoietic cell transplant will not have a fully matched related donor, and finding matched unrelated donors through the registry may be difficult, especially if the recipient is not of Northern European descent [N Engl J Med 2014;371:339-348]. Umbilical cord blood (UCB) has been an available graft source for hematopoietic cell transplant for more than 30 years, since the first UCB transplant was performed in the late 1980s [N Engl J Med 1989;321:1174-1178]. UCB is readily available, has low immunogenicity, and does not require as strict of human leukocyte antigen (HLA) matching compared to other graft sources [N Engl J Med 2004;351:2265-2275]. According to data from the Center for International Blood and Marrow Transplant Research (CIBMTR), an estimated 500 patients in the US will have received a UCB transplant in 2018. Since 2014, haploidentical transplants have surpassed UCB transplants performed in the United States (CIBMTR Summary Slides, 2018, available at https://www.cibmtr.org). Increased use of haploidentical transplants has brought to light concerns about UCB transplants, including delayed engraftment and graft failure, increased nonrelapse mortality, increased infection risk, and UCB acquisition costs [Lancet Oncol 2010;11:653-660; Biol Blood Marrow Transplant 2019;1456-1464]. These concerns will need to be addressed for UCB to remain a viable option as a graft source for hematopoietic cell transplant. Other promising therapeutic benefits for UCB, in addition to hematopoietic cell transplant, is its use in regenerative medicine and immune modulation, which is currently being evaluated in ongoing clinical trials.


Analyses of Gingival Adhesion Molecules in Periodontitis: Theoretical In Silico, Comparative In Vivo, and Explanatory In Vitro Models.
A deeper understanding of periodontitis pathophysiology is central to future development of novel biomarkers and therapeutics. The following is reported here: 1) an in silico network model of interactions among cell adhesion molecules and a network-focused microarray analysis of the corresponding genes in periodontitis; 2) analysis of secretions of adhesion molecules in gingival tissue samples from patients with periodontitis and healthy controls; and 3) effect of the human neutrophilic peptide-1 (HNP-1) on epithelial adhesion molecules. The network model identified 85 nodes in relation to the interactions of adhesion molecules. Subsequently, the relative gene expression was overlaid on the network model. Differential gene expression was analyzed, and false discovery rate control was performed for statistical assessment of the microarray data. Both tissue and cell culture samples were immunostained for desmocollin (DSC)2, occludin (OCLN), desmoglein (DSG)1, tight junction protein 2, and gap junction protein α. The differential gene expression analysis revealed that the epithelial adhesion molecules were significantly lower in abundance in individuals with periodontitis than controls. In contrast, the genes for leukocyte adhesion molecules showed a significant upregulation. Immunostainings revealed elevated secretions of both DSG1 and OCLN in periodontitis. An in vitro model suggested reduced DSC2 and OCLN secretions in the presence of HNP-1. Gene expression of gingival adhesion molecules in periodontitis is regulated by leukocyte transmigration, whereas the neutrophilic antimicrobial peptide HNP-1 is noted as a putative regulator of epithelial adhesion molecules. These observations contribute to the key mechanisms by which future biomarkers might be developed for periodontitis.


Modulation of the enzymic activity of chicken pepsin by the covalent modification of its single -- SH group.
The single cysteinyl residue of chicken pepsin was modified with a wide spectrum of reagents to produce mixed disulfides or alkylated derivatives. All these derivatives showed enhanced catalytic activity towards the synthetic peptide Z-Ala-Ala-Phe-OPrP, where OPrP is the 3-(4-pyridyl)-propyl-1-oxy group. The overall catalytic constant kcat/Km for these derivatives was 4 -- 25-fold larger than that of the native enzyme. The activity of the enzyme towards denatured hemoglobin was slightly decreased (10--45%) by these modifications. When the mixed disulfide derivatives were treated with excess mercaptan, the sulfhydryl group was regenerated and activity reverted to that of the native enzyme. The --SH group of chicken pepsin reacted preferentially with reagents containing an aromatic group. The reaction was found to depend on the ionization of a single group, presumably the --SH itself, with a pKa = 7.5. The rate of reaction of the fully deprotonated species with various disulfides was 100--1000-fold smaller than that of the --SH group of glutathione. It is suggested that the groups attached covalently to the sulfhydryl also interact with other amino acid side chains in the protein thereby affecting the active center of chicken pepsin.


4-Aminopyridine-induced ultrastructural alterations of pinched-off nerve terminals from rat cerebral cortex.
Pinched-off nerve terminals (synaptosomes) from rat cerebral cortex were depolarized with 60 mM KCl and treated with 20 mM 4-aminopyridine in order to evaluate ultrastructural alterations. The empty presynaptic terminals were counted and their number was given as a percentage of the normal terminals. The proportion of empty terminals increased from 10.47 +/- 1.56% to 32.45 +/- 1.88% (P less than 0.001) following treatment with 20 mM 4-aminopyridine. This effect of 4-aminopyridine depended on the presence of Ca++ in the incubation medium. The results are discussed in terms of facilitation by 4-aminopyridine of exocytotic transmitter release. We think that the increase of the empty synaptosomes was due to the exhaustion or inhibition of the synaptic vesicle recycling mechanism.


[Relationship between regulation effect of salvia miltiorrhiza on AQP2 in kidney and promoting blood circulation and diuresis].
Partial nature of "promoting blood circulation and dieresis" of Salvia Miltiorrhizain was initially demonstrated by investigating the regulation effect of AQP2 expression in kidney of trauma blood stasis model rats with the Salvia Miltiorrhizain so as to provide guidance for its clinical deployment of administration. Random allocation was taken to averagely divide 30 SD rats into two groups: 10 rats in normal group and 20 rats in blood stasis syndrome group. Trauma blood stasis rat model was established by quantitatively beating. Then the rat model group was divided into model group and salvia group. After 7 days of treatment, the rat kidney AQP2 expression was detected, the content of urine AQP2 was compared and the damaged local muscle and kidney pathological changes were observed by immunohistochemical method and western blot method. Compared with that of the normal group, rats in model group had inflammatory cells infiltration, blood stasis and edema of the injured local muscles and up-regulated AQP2 expression, decreasing urinary output, and kidney tissues blood stasis and edema (P < 0.05). On the other hand, compared with that of the model group, those parameters of rats in salvia group were all decreasing except urine output (P < 0.05). Such result indicated that Salvia Miltiorrhiza can reduce trauma blood stasis rat content of urine AQP2 and down-regulated AQP2 expression in kidney tissue, so as to reduce the reabsorption of water by renal tubular and increase urine output. The promoting blood circulation effect of Salvia Miltiorrhizain can alleviate the degree of the damaged tissue edema and encourage urine drainage. This therapy is closely related to the effect of regulating AQP2 in kidney by salvia, so the purpose of this study by verifying "promoting blood circulation and diuresis" as the mechanism for the regulation effect of the salvia on AQP2 expression.


Antibody responses after dose-sparing intradermal influenza vaccination.
Reduced-dose intradermal (ID) influenza vaccination is an attractive approach to increase availability of vaccine supply in an event of vaccine shortage. We conducted a randomized open-label study, in which 500 subjects were randomly assigned to receive an ID injection of 0.1 ml dose of inactivated split-virion influenza vaccine or an IM injection of 0.5 ml dose. The subjects who had hemagglutination inhibition (HI) antibody titer of at least 1:40 at day 28 post-vaccination in ID and IM groups were 93.3% versus 98.0% for influenza A(H1N1) virus, 86.3% versus 95.0% for A(H3N2) virus, and 43.5% versus 57.0% for influenza B virus. Subjects in the ID group had an increase in geometric mean titer by a factor of 16 for the H1N1 strain, 8 for the H3N2 strain, and 2 for the B strain on day 28, as compared with respective increase in the IM group of 31, 20, and 3. Local reactions were significantly more frequent among subjects in the ID group than those in the IM group, but the reactions were mild and transient. In this study, ID administration of one-fifth dose of influenza vaccine elicited significantly lower levels of antibody response as compared to full-dose IM injection. However, the antibody responses elicited by the ID vaccination were still sufficiently high to meet the requirement guidelines of the European Committee for Proprietary Medicinal Products (CPMP) for the annual relicensure of influenza vaccines.