Multicomponent Assembly of Bicyclic Aza-peroxides Catalyzed by Samarium Complexes and Their Cytotoxic Activity.
An original strategy toward bridged tetraoxazaspirobicycloalkanes was developed. The synthesis is based on a three-component condensation-cyclization reaction of primary arylamines with 1,1'-peroxybis (1-hydroperoxycycloalkanes) and pentane-1,5-dial catalyzed by Sm(NO3)3·6H2O. The structures and conformations of the products were determined by X-ray diffraction analysis and 1H and 13C NMR spectroscopy. High cytotoxic activity and biological potential toward ferroptosis induction were found for the synthesized bicyclic aza-peroxides.


Activity-guided isolation and identification of anti-staphylococcal components from Senecio tenuifolius Burm. F. leaf extracts.
To investigate activity-guided isolation and identification of anti-Staphylococcus aures components from Senecio tenuifolius Burm. F. (S. tenuifolius). Hexane, chloroform, ethyl acetate, methanol and aqueous extracts of S. tenuifolius were prepared by soxilation for antimicrobial activity against one registered Staphylococcus aureus (S. aureus) (ATCC No: 25923) and two clinical isolates, methicillin resistant and methicillin sensitive S. aureus. NCCL standard methods were followed for antibacterial activity. GC-MS was performed to identify the chemical composition of bio active fraction. Among all solvent extracts, methanol extract significantly reduced the growth of S. aureus (ATCC No: 25923), methicillin resistant and methicillin sensitive S. aureus with the best zone of inhibition at 16.23, 14.06 and 15.23 mm and minimum inhibition concentration (MIC) values at 426.16, 683.22 and 512.12 µg/mL, respectively. In order to detect the active component in methanol extract, it was further purified by column chromatography, which yielded four fractions (St1, St2, St3, and St4). Among these four fractions, St3 was effective against the tested strains of S. aures, with the best zone of inhibition at 15.09, 13.25 and 14.12 mm and with best MIC values at 88.16, 128.11 and 116.12 µg/mL, respectively. Effective fraction partially purified from S. tenuifolius (St3) yielded MIC's that were at least 20 fold less when compared to crude extract. GC-MS analysis of St3 revealed the presence of 3-[methyl-6,7-dihydro benzofuran-4 (5H)-one], 1,2-benzenedicarboxylic acid, hydroquinone, methyl ester and 3 unknown compounds. The study provides scientific evidence for traditional and folklore medicinal use of S. tenuifolius in skin infections treatment.


Immunotherapy in Patients with Recurrent and Metastatic Squamous Cell Carcinoma of the Head and Neck.
Head and neck squamous cell carcinoma (HNSCC) is the most common malignant cancer occurring in the head and neck area, approximately 90% of the cases. Even in the cases of primary radical treatment (surgical, concomitant chemoradiotherapy or radiotherapy alone), subsequent local recurrence or distant metastases are often observed. In patients with recurrent disease who are unable to receive radical treatment, the results of palliative chemotherapy are not satisfactory. In this review, we summarized the standard treatment options, current development of new drugs and future perspectives in the treatment of patients with recurrent locally advanced and/or metastatic HNSCC. PubMed databases with words 'head and neck cancer treatment', 'immunotherapy in head and neck cancer treatment' were searched and yielded 186512 and 2249 papers respectively. We selected the most cited articles and reports presenting new immunotherapy agents and drug combinations in HNSCC. Recently, two new agents been approved in the treatment of recurrent locally advanced and/or metastatic HNSCC. These are immune-checkpoint inhibitors targeting PD1 (nivolumab and pembrolizumab) which are the most active drugs in the second line treatment of advanced HNSCC. Still, the first line 'golden standard' is the chemotherapy regimen (cisplatin, 5-fluorouracyl) combined with cetuximab. Many phase 3 studies are currently ongoing, evaluating the efficacy of combinational treatment-anti-CTLA4 with anti-PD1 or anti-PDL1. Very encouraging results have been shown in early phase studies evaluating the combination of immunecheckpoint inhibitors with tumor microenvironment immunosuppressive inhibitors. Despite the huge progress in the systemic treatment of patients with recurrent locally advanced and/or metastatic HNSCC, the disease at this stage remains incurable. Undoubtedly, further research in the field of biomarkers for effective immunotherapy is needed in order to select a group of patients whose will benefit from this therapy, as the treatment is still ineffective in most patients.


[Strategies for reducing the CT radiation dose].
The rapid technical advances in computed tomography have led to an increased number of clinical indications. Unfortunately, at the same time the radiation exposure to the population has also increased due to the increased total number of CT examinations. In the last few years various publications have demonstrated the feasibility of radiation dose reduction for CT examinations with no compromise in image quality and loss in interpretation accuracy. The majority of the proposed methods for dose optimization are easy to apply and are independent of the detector array configuration. This article reviews indication-dependent principles (e.g. application of reduced tube voltage for CT angiography, selection of the collimation and the pitch, reducing the total number of imaging series, lowering the tube voltage and tube current for non-contrast CT scans), manufacturer-dependent principles (e.g. accurate application of automatic modulation of tube current, use of adaptive image noise filter and use of iterative image reconstruction) and general principles (e.g. appropriate patient-centering in the gantry, avoiding over-ranging of the CT scan, lowering the tube voltage and tube current for survey CT scans) which lead to radiation dose reduction.


The methyltransferase domain of DNMT1 is an essential domain in acute myeloid leukemia independent of DNMT3A mutation.
Aberrant DNA methylation patterns are a prominent feature of cancer. Methylation of DNA is mediated by the DNA methyltransferase (DNMT) protein family, which regulates de novo (DNMT3A and DNMT3B) and maintenance (DNMT1) methylation. Mutations in DNMT3A are observed in approximately 22% of acute myeloid leukemia (AML). We hypothesized that DNMT1 or DNMT3B could function as a synthetic lethal therapeutic strategy for DNMT3A-mutant AML. CRISPR-Cas9 tiling screens were performed to identify functional domains within DNMT1/DNMT3B that exhibited greater dependencies in DNMT3A mutant versus wild-type cell lines. Although increased sensitivity to DNMT1 mutation was observed in some DNMT3A mutant cellular models tested, the subtlety of these results prevents us from basing any conclusions on a synthetic lethal relationship between DNMT1 and DNMT3A. Our data suggests that a therapeutic window for DNMT1 methyltransferase inhibition in DNMT3A-driven AML may exist, but validation in more biologically relevant models is required.


The importance of the urine flow rate in semi-quantitative assessment of the standard renogram.
Renograms were recorded at different rate of urine flow in 30 healthy Nigerian children to establish semi-quantitative criteria to assess the standard renogram. The results indicate that knowledge of the urine flow rate during renography is necessary for meaningful interpretation of the study. They further showed that for semi-quantitative assessment of the renogram the time to peak and time to peak half values are easily identified indexes that can be measured accurately with negligible observer variation. Regression equations for the relationship between urine flow rate on the one hand, and time to peak and time to peak half values on the other have been obtained and used to compute nomograms, which should be useful for future work in this area.


Origin of mesenchymal tissue in the septum primum: a structural and ultrastructural study.
A structural, ultrastructural and histochemical study in chick embryos indicates that the septum primum mesenchymal tissue originate between 3 and 5 days of development and that their origin may be related to an activation of endocardial cells that cover the septum primum. By day 3, endocardial cells display migratory appendages, cell hypertrophy and an increase in secretory and mitotic activity. In later stages (day 4) hypertrophic endocardial cells undergoing division seem to delaminate and translocate toward the subendocardial space to give rise to free mesenchymal-type cells. These results suggest that the endocardium makes up the bulk of the septum primum mesenchymal tissue as has been demonstrated during mesenchymal tissue formation in the atrioventricular canal and outflow tract. Before and during mesenchymal tissue formation an accumulation of extracellular matrix components like proteoglycans can be visualized using tannic acid. These extracellular components might be related to the promotion of cellular events described during endocardial activation. The fusion of the septum primum with the atrioventricular (AV) endocardial cushions which would obliterate the foramen primum, occurs between mesenchymal tissues. Therefore, any alteration in the normal development of these mesenchymal tissues could be related to pathological cases of persistent atrial communications. Light microscopy preliminary observations of embryonic mouse heart indicate that septum primum mesenchymal tissue formation occurs similarly between mouse and chick embryos.


A Surgical Technique to Preserve the Subvalvular Apparatus in Patients Undergoing Mitral Valve Replacement for Severe Ischemic Regurgitation.
Severe functional mitral valve regurgitation should be treated in patients undergoing myocardial revascularization. When replacement is considered the best therapeutic option, preservation of the mitral subvalvular apparatus is crucial, especially in the emergency setting, because of its primary role in preserving geometry and function of left and right ventricles. Here we present a simple and quick technique, where subvalvular apparatus is preserved in toto in patients undergoing mitral valve replacement with a bioprosthesis.


The use of cytotoxic T cell lines to detect the segregation of a human minor alloantigen within families.
A cytotoxic T cell (CTL) line, which detected a minor alloantigen provisionally called W was generated in vitro with lymphocytes from a multiply transfused individual, S1. Lymphocytes from S1 were first stimulated with cells from an unrelated known from previous studies to express the minor antigen. The primary CTL were then restimulated with cells from a W +/ve HLA identical sib, S2, in the presence of IL-2. As in previous work, recognition of the W antigen by these CTL was restricted by HLA-B7. Antigen assignments of W + W -, based upon cold target inhibition studies, confirmed previous assignments which had depended upon the ability of lymphocytes either to stimulate the generation of or to be killed by anti-W CTL effectors. Testing of lymphocyte targets from members of several unrelated families in which HLA-B7 segregated showed that the CTL lines could detect the expression of W on cells of individuals in the general population. In 3 of 5 cases, members of an HLA identical sib pair differed for W. These results open up the possibility of designing studies using CTL lines to determine whether differences for minor alloantigens play a role in clinical transplantation.


Effect of nicotine on exocytotic pancreatic secretory response: role of calcium signaling.
Nicotine is a risk factor for pancreatitis resulting in loss of pancreatic enzyme secretion. The aim of this study was to evaluate the mechanisms of nicotine-induced secretory response measured in primary pancreatic acinar cells isolated from Male Sprague Dawley rats. The study examines the role of calcium signaling in the mechanism of the enhanced secretory response observed with nicotine exposure. Isolated and purified pancreatic acinar cells were subjected to a nicotine exposure at a dose of 100 μM for 6 minutes and then stimulated with cholecystokinin (CCK) for 30 min. The cell's secretory response was measured by the percent of amylase released from the cells in the incubation medium Calcium receptor antagonists, inositol trisphosphate (IP3) receptor blockers, mitogen activated protein kinase inhibitors and specific nicotinic receptor antagonists were used to confirm the involvement of calcium in this process. Nicotine exposure induced enhanced secretory response in primary cells. These responses remained unaffected by mitogen activated protein kinases (MAPK's) inhibitors. The effects, however, have been completely abolished by nicotinic receptor antagonist, calcium channel receptor antagonists and inositol trisphosphate (IP3) receptor blockers. The data suggest that calcium activated events regulating the exocytotic secretion are affected by nicotine as shown by enhanced functional response which is inhibited by specific antagonists… The results implicate the role of nicotine in the mobilization of both intra- and extracellular calcium in the regulation of stimulus-secretory response of enzyme secretion in this cell system. We conclude that nicotine plays an important role in promoting enhanced calcium levels inside the acinar cell.


[Dynamics of the activity of rabbit visual cortex neurons during elaboration of elementary conditioned reflexes to time and association].
Elaboration of a reflex to time (with omissions of stimuli) and of association; light--light + electrocutaneous stimulation of the paw was studied on neurones of the rabbit visual cortex. It was shown that by similarity of patterns of the discharge to a conditioned stimulus and to reinforcement, 83% of neurones reveal the formation of a reflex to time, and 57%--elaboration of association. Three ways of coding plastic reorganizations of the neuronal discharges are suggested; by the total number of spikes in the response, by the total sum of inhibitory periods, and by the pattern of the response. It has been found that in the reflex to time the early components of the response to the stimulus are reproduced during omissions; in elaboration of the association the late components of the discharge reaction to reinforcement are mainly repeated. Most cells with plastic reorganizations possessed polysensority and revealed discharge sensitization during habituation.


Mycobacteriosis in the rabbit and rodent.
Spontaneous mycobacteriosis is rare in rabbits and rodents with the exception of the pygmy rabbit, and there are only a handful of reported cases involving other rodents. Mycobacterium avium complex was the most commonly identified organism in reports of spontaneous mycobacteriosis involving rabbits and rodents. The resistance of rabbits and rodents to mycobacterial disease has been useful in understanding the disease in humans and other animals. Preventing or controlling Mycobacterium sp transmission from wildlife to domestic animals will require collaboration between agriculture, wildlife, environmental, and political entities. Understanding the ecology and epidemiology of mycobacteria is needed for better worldwide management of tuberculosis.


Accountability in protection of children against violence: Monitoring and measurement.
Violence against children (VAC) is a pervasive and persistent injustice around the world. Thirty years after the adoption of the United Nations Convention on the Rights of the Child (CRC), the right of children to be protected against all forms of violence is still routinely violated. It is only within the last twenty years that the scope of this problem has been studied; as many point out, understanding the scope of the problem through comprehensive data collection is necessary to improve child protection. Data collection and data quality have improved drastically over this time. The Office of the High Commissioner on Human Rights recommends using comprehensive indicator systems that collect data on States Parties' structure- and process-related commitments to fulfil human rights, and on the outcomes of these commitments. However, most existing indicator sets on VAC fall short of this comprehensive approach, often focusing only on structural or outcome indicators. An international team of child rights experts is now developing GlobalChild, which will be an online platform consisting of comprehensive sets of structure-, process-, and outcome-related indicators for all 41 substantive rights of the CRC. This system will advance data collection, facilitate the monitoring function of the Committee on the Rights of the Child, and enable research on many rights of children, including their right to protection against violence. VAC is a complex problem, and protection against VAC is a right that is indivisible from other rights under the CRC. Therefore, a comprehensive and systematic approach is required to improve accountability to children around the world.


What does degeneration at the cervicothoracic junction tell us? A kinematic MRI study of 93 individuals.
Current decision-making in multilevel cervical fusion weighs the potential to protect adjacent levels and reduce reoperation risk by crossing the cervicothoracic junction (C7/T1) against increased operative time and risk of complication. Careful planning is required, and the planned distal and adjacent levels should be assessed for degenerative disc disease (DDD). This study assessed whether DDD at the cervicothoracic junction was associated with DDD, disc height, translational motion, or angular variation in the adjacent superior (C6/C7) or inferior (T1/T2) levels. This study retrospectively analyzed 93 cases with kinematic MRI. Cases were randomly selected from a database with inclusion criteria being no prior spine surgery and images having sufficient quality for analysis. DDD was assessed using Pfirrmann classification. Vertebral body bone marrow lesions were assessed using Modic changes. Disc height was measured at the mid-disc in neutral and extension. Translational motion and angular variation were calculated by assessing translational or angular motion segment integrity respectively in flexion and extension. Statistical associations were assessed with scatterplots and Kendall's tau. DDD at C7/T1 was positively associated with DDD at C6/C7 (tau = 0.53, p < 0.01) and T1/T2 (tau = 0.58, p < 0.01), with greater disc height in neutral position at T1/T2 (tau = 0.22, p < 0.01), and with greater disc height in extended position at C7/T1 (tau = 0.17, p = 0.04) and at T1/T2 (tau = 0.21, p < 0.01). DDD at C7/T1 was negatively associated with angular variation at C6/C7 (tau = - 0.23, p < 0.01). No association was appreciated between DDD at C7/T1 and translational motion. The association of DDD at the cervicothoracic junction with DDD at the adjacent levels emphasizes the necessity for careful selection of the distal level in multilevel fusion in the distal cervical spine.


Managed death in a PACE: pathways in present and advance directives.
To test the effect of an innovative method of documenting present and advance health care wishes on the rates of completion and the qualitative choices of health care wishes. Interventional prospective cohort (pre- and post-). Program for All-inclusive Care of the Elderly (PACE) site in St. Louis, MO. Enrolled PACE participants. A documentation tool that captures both present and advance directives in a framework of "pathways," blending goals of care with typical procedure-oriented directives. Data from medical records to calculate rates of health care wishes (HCW) completion, proportions of qualitative choices, and compliance with wishes at death. Baseline prevalences of present directives (PD) and advance directives (AD) were 77% and 36%, respectively, while Do Not Resuscitate (DNR) wishes were documented in 48% of PD and 26% of AD. After implementation of the Pathways Tool, completion rates increased to 99% for both PD and AD. Documented DNR wishes decreased to 38% of PD and increased to 66% of AD. Qualitative choices for care (Longevity vs Function vs Palliation) changed toward a palliation pathway for AD (from 9% to 53%). The rate of dying at home increased from 24% to 65%. Compliance with end-of-life wishes increased from 72% to 96%. These are statistically significant. Introduction of a novel pathways method of documenting HCW in a PACE site was associated with increased completion, preferences toward less invasive levels of care at life's end, and increased compliance with participants' wishes and deaths at home. Future research to validate the methodology employed in this intervention should be conducted in other long-term care settings.


Ocular manifestations of trichothiodystrophy.
Trichothiodystrophy (TTD) is a rare, autosomal recessive disorder characterized by sulfur-deficient brittle hair and multisystem abnormalities. Many TTD patients have a defect in known DNA repair genes. This report systematically evaluates the ocular manifestations of the largest-to-date cohort of TTD patients and xeroderma pigmentosum (XP)/TTD patients. Case series. Thirty-two participants, ages 1 to 30 years, referred to the National Eye Institute for examination from 2001 to 2010; 25 had TTD and 7 had XP/TTD. Complete, age- and developmental stage-appropriate ophthalmic examination. Visual acuity (VA), best-corrected VA, ocular motility, state of the ocular surface and corneal endothelial cell density, corneal diameter, and lens assessment. Developmental abnormalities included microcornea (44% TTD), microphthalmia (8% TTD, 14% XP/TTD), nystagmus (40% TTD), and infantile cataracts (56% TTD, 86% XP/TTD). Corrective lenses were required by 65% of the participants, and decreased best-corrected VA was present in 28% of TTD patients and 71% of XP/TTD patients. Degenerative changes included dry eye (32% TTD, 57% XP/TTD) and ocular surface disease identified by ocular surface staining with fluorescein (32% TTD) that usually are exhibited by much older patients in the general population. The 2 oldest TTD patients exhibited clinical signs of retinal/macular degeneration. Four XP/TTD patients presented with corneal neovascularization. These TTD and XP/TTD study participants had a wide variety of ocular findings including refractive error, infantile cataracts, microcornea, nystagmus, and dry eye/ocular surface disease. Although many of these can be ascribed to abnormal development--likely owing to abnormalities in basal transcription of critical genes--patients may also have a degenerative course. Proprietary or commercial disclosures may be found after the references.


Role of TSC-mTOR pathway in diabetic nephropathy.
TSC-mTOR signaling plays a crucial role in the regulation of cell growth and survival control. Mammalian target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine kinase that forms two distinct functional complexes, termed TOR complex 1 (TORC1) and TORC2, respectively. TORC1 is a rapamycin-sensitive complex and regulates a wide array of cellular processes including translation, transcription, and autophagy. Tuberous sclerosis complex (TSC) gene products, TSC1 and TSC2 are tumor suppressors and specifically suppress TORC1 activity. Mutation of either TSC1 or TSC2 causes TSC disease, which is characterized by formation of hamartomas in multiple organs. Although the role of TSC-mTOR pathway in tumor and cancer development has been extensively studied, more recent studies have indicated a role for mTOR function in appetite, memory, aging, and energy metabolism. Dysregulation of the TSC-mTOR pathway may cause not only tumor development but also metabolic disorders such as diabetes and its complications.


Mechanism of histone deacetylase HDAC2 in FOXO3-mediated trophoblast pyroptosis in preeclampsia.
Histone deacetylase 2 (HDAC2) has been demonstrated to regulate trophoblast behaviors. However, its role in trophoblast pyroptosis remains unknown. This study sought to analyze the molecular mechanism of HDAC2 in trophoblast pyroptosis in PE. Expression levels of HDAC2, forkhead box O3 (FOXO3), and protein kinase R-like endoplasmic reticulum kinase (PERK) in placenta tissues and HTR8/SVneo cells and H3K27ac levels in cells were determined. Levels of IL-1β and IL-18 in placenta tissues were determined, and their correlation with HDAC2 was analyzed. Cell proliferation, migration, and invasion were evaluated, and levels of pyroptosis-associated proteins and cytokines were determined. The enrichments of H3K27 acetylation (H3K27ac) and FOXO3 in the FOXO3/PERK promoter region were determined. HDAC2 was downregulated, and FOXO3, PERK, IL-1β, and IL-18 levels were elevated in PE placenta tissues. In HTR8/SVneo cells, HDAC2 downregulation suppressed cell proliferation, migration, and invasion and increased pyroptosis. HDAC2 erased H3K27ac in the FOXO3 promoter region and repressed FOXO3, and FOXO3 bound to the PERK promoter and increased PERK transcription. Functional rescue experiments revealed that silencing FOXO3 or PERK counteracted HDAC2 downregulation-induced cell pyroptosis. Overall, HDAC2 downregulation enhanced H3K27ac to activate FOXO3 and PERK, leading to the occurrence of trophoblast pyroptosis in PE.


[Investigation of the problem of safety of computer models in automatic analysis of nystagmus (author's transl)].
The aim of the investigations was to demonstrate by means of statistical methods, that there was no signifikant difference between manual calculation and automatical computer analysis of ENG.


Microscopic colitis: clinical findings, topography and persistence of histopathological subgroups.
Uncertainty remains on topography and persistence of histological subgroups of microscopic colitis (MC). To assess longitudinal clinical, endoscopic, histological, and therapeutic description of MC subgroups including patients with incomplete findings of MC (MCi). Retrospective review of a consecutive cohort with MC and histological reassessment of MCi. Clinical characteristics of 168 patients with lymphocytic colitis (LC), 270 with collagenous colitis (CC) and 101 with MCi were similar. At colonoscopy 95% (95% CI: 91-98%) of CC and 98% (93-100%) of LC cases had diagnostic histopathology of MC in both left and right colon. Eight and three patients had characteristics of MC only in the left and right colon, respectively. Histology findings resembling coexistence of the other MC subtype was present in 48% (40-55%) with CC and 24% (18-31%) with LC. A first diagnosis of MC was made in 49 (30%) of 164 patients only at repeat endoscopy. Another 34 of 115 (30%) with MC in the first endoscopy did not fulfil the MC criteria at repeat endoscopy. Only seven cases had a primary endoscopy without histopathological abnormalities. Fifteen percentage of MCi were reclassified as MC. Ileal inflammation was present in 33 of 81 patients. Budesonide was efficacious in all MC subgroups irrespective of bile acid malabsorption. Clinical characteristics of microscopic colitis subgroups are indistinguishable. Biopsies from the left colon suffice to exclude microscopic colitis, and the histological diagnosis of microscopic colitis is inconsistent over time. Ileal inflammation is common. The term microscopic colitis should perhaps be considered one clinical entity and include lymphocytic colitis, collagenous colitis, and incomplete findings of microscopic colitis.


Using weak supervision to generate training datasets from social media data: a proof of concept to identify drug mentions.
Twitter has been a remarkable resource for research in pharmacovigilance in the last decade. Traditionally, rule- or lexicon-based methods have been utilized for automatically extracting drug tweets for human annotation. The process of human annotation to create labeled sets for machine learning models is laborious, time consuming and not scalable. In this work, we demonstrate the feasibility of applying weak supervision (noisy labeling) to select drug data, and build machine learning models using large amounts of noisy labeled data instead of limited gold standard labelled sets. Our results demonstrate the models built with large amounts of noisy data achieve similar performance than models trained on limited gold standard datasets, hence demonstrating that weak supervision helps reduce the need to rely on manual annotation, allowing more data to be easily labeled and useful for downstream machine learning applications, in this case drug mention identification.


Galactose inhibits lactase expression by mouse jejunal enterocytes.
Present work uses a combination of quantitative cytochemistry and measurements of cell migration rates to describe galactose effects on lactase expression by mouse enterocytes. Mice fed galactose were found to eat less, weigh less and drink more than mice maintained on a low-carbohydrate isocalorific diet. The enterocyte migration rate in these mice was also only one third of that determined in low-carbohydrate-fed animals. The rate at which lactase activity increased in the brush border membrane of migrating enterocytes was 3-times greater in low-carbohydrate- compared with galactose-fed mice. The time during which this increase persisted was, however, 3-times less in low-carbohydrate-fed animals. The maximum rate of sucrase-maltase appearance, measured as control in these experiments, remained unaffected by galactose feeding. Galactose effects on lactase expression might in part result from mice being unable to metabolise this substrate. Previously it has been stated that galactose increases lactase biosynthesis in rat intestine (Koldovsky, O., Bustamonte, S. and Yamada (1981) In Mechanisms of intestinal adaptation (Robinson, J.W.L., Dowling, R.H. and Ricken, E.O., eds.), pp. 153-156, MTP Press, Lancaster). This result is discussed in relation to the opposite finding reported in the present work for mouse jejunal enterocytes. The need to relate enzyme appearance to age and developmental state of enterocytes in this type of study is also emphasized.


[Cross reticular bone grass used in slotted acetabular augmentation].
To investigate the technique of cross reticular bone grass used in slotted acetabular augmentation ( SAA), and enrich the surgical procedures in the treatment of the DDH. Thirty seven cases were treated with cross reticular bone grass used in SAA, including developmental dysplasia of the hip (DDH) and coaxial plana caused by Legg-Perthes disease. SAA provided an easy way to shape an adept acetabulum with cancellous bone graft. After the operation a stable hip was obtained. Thirty cases were good, 2 remained limping, 1 had a limitation of abduction of the hip. SAA is a safe and effective way to provide an adequate coverage of the femoral head. It is a developed shelf procedure with good stability and for a surgical frame of the hip. SAA method is safe and technically simple.


[Fixed femoral implant on the lateral side - Long term results in hip arthritis in young patients].
The total Hip Arthroplasty of Sir John Charnley was a revolutionary step in the treatment of Hip diseases. Currently, the long-term follow-up of such a prosthesis can be estimated from fifteen to twenty years. The question is: is that delay sufficient to treat young patients with severe hip disease?In a recent publication, Rik Huiskes [3] claims that since, Charnley, no so-called innovation has either scientific proof in real efficacity nor in terms of longer longevity. This should not be a surprise for all total hip prostheses are placed in the medullary canal, either with or without cement. In fact, the so-called innovations are minimal modifications in the design of the stem. It is obvious that if these modifications are not efficient, one should not follow line of research. It seems that almost every thing has been written on total hip arthroplasty failure. We must therefore work in another direction, with another criteria.Let us suppose that it is bone, with all its specifications, which is the possible reason for long-term failure. Is the "kuntscher" imperial road, the only highway to deal with the diaphysis of a long bone? Our point of view is that the bio-mechanical aspect of a stem hip prosthesis is no longer of interest. It seems to us that it is in working on bone physiology that a new concept, a new philosophy of a hip stem prosthesis becomes apparent. If the stem can be placed inside the medullary canal, it can be fixed to on the external part of the diaphyseal cortex, below the periosteal layer.This is the purpose of the following article.


The time course of glycogen depletion in single fibers of chronically stimulated rabbit fast-twitch muscle.
A time course study was conducted to investigate the possibility of a relationship between fiber degeneration and glycogen depletion in chronically nerve-stimulated extensor digitorum longus muscle of the rabbit. Muscles were stimulated 12 h daily at 10 Hz using alternating one-hour periods of stimulation and rest. When measured for the first time after 3 h (1 h stimulation, 1 h rest, 1 h stimulation), microphotometry revealed complete glycogen depletion of all fiber types (fast glycolytic, FG; fast oxidative glycolytic, FOG; slow oxidative, SO). Different responses were noted beginning at day 4. At this time point, all FOG and SO fibers recovered their glycogen stores with some of the FOG population attaining levels higher than the FOG fibers in the unstimulated, contralateral muscle. Approximately 28% of the FG fibers recovered to normal glycogen values, whereas 58% remained depleted and 14% displayed "overshooting glycogen" levels. Fifteen percent of all fibers were glycogen-depleted after 12 days of stimulation. At this time, classic fiber types could no longer be distinguished. Fiber degeneration, which was recognized by the invasion of nonmuscle cells, began after 6 days and was restricted to the glycogen-depleted fibers. By this time, there was also a significant increase in DNA content. Exhaustion of glycogen, the main fuel of the FG fibers, is believed to cause a collapse of energy-supply and ATP-driven ionic pumps. The latter could be the initial step of fiber deterioration.


Review of positive direct antiglobulin tests found on cord blood sampling.
Until recently, all babies born in Wellington had umbilical cord blood sampling for direct antiglobulin test (DAT). It is considered to be an important test in identifying babies who are at risk of haemolytic disease of the newborn (HDN). The aim of this review was to examine the utility of positive DAT results and ascertain: -- How many cases required phototherapy? -- Were any babies readmitted for phototherapy? -- Did the positive DAT influence the detection and treatment of HDN? The clinical records of all newborn babies found to have positive DATs by Wellington Hospital Blood Bank, over a 6-month period (January 2001-June 2001) were reviewed. Blood group serological results of all babies that received phototherapy during this period were also reviewed. Ninety-four babies had a positive DAT, of which 22 (23%) received phototherapy. The incidence of a positive cord blood DAT was found to be 5.5%. In total, 1724 cord blood samples were analysed by Blood Bank over the first 6 months in 2001. Overall 145 babies received phototherapy, 117 were DAT-negative and six were not tested. Six of the 22 (27%) DAT-positive babies that received phototherapy were alerted by a positive DAT, leading to measurement of serum bilirubin (SBR). Twelve of the 22 (55%) were initially alerted by clinical jaundice, leading to measurement of SBR. Two DAT-positive cases were diagnosed antenatally, both were due to anti-D. Overall 10 babies were readmitted for phototherapy, two had a positive DAT. One baby received an exchange transfusion in addition to phototherapy. Two babies that received phototherapy had SBRs in the exchange transfusion range. Eighty-six per cent of the DAT-positive cases treated with phototherapy were due to anti-A. There were four cases of DAT-negative ABO HDN. The positive predictive value of a positive DAT for HDN is 23%. The sensitivity was estimated to be 86%. Ten babies required readmission for phototherapy, two of these were DAT-positive. Jaundice, rather than the positive DAT, was the first alert in the majority of cases of HDN requiring phototherapy. Recommendations for testing are discussed but remain controversial in practice. Assessment for hyperbilirubinaemia in all infants early in life is fundamental.


Reduced volume of orbitofrontal cortex in major depression.
Functional neuroimaging studies have implicated dysfunction of orbitofrontal cortex in the symptoms of depression, and a recent postmortem study of depressed patients found reduced density of neurons and glia in this area. The purpose of this study was to measure volume of orbitofrontal cortex and other frontal cortical subregions in patients with major depression. Magnetic resonance imaging was used to measure volume of the orbitofrontal cortex and other frontal cortical regions in patients with major depression in remission (n = 15) and comparison subjects (n = 20). Patients with depression had a statistically significant 32% smaller medial orbitofrontal (gyrus rectus) cortical volume, without smaller volumes of other frontal regions including anterior cingulate Brodmann's area 24 (subgenual gyrus), anterior cingulate Brodmann's area 32, subcallosal gyrus (Brodmann's area 25), or whole brain volume. The findings were significant after statistically controlling for brain size. These findings are consistent with smaller orbitofrontal cortical volume in depression.


The phosphodiesterase inhibitor, pentoxifylline, alters rat intestinal epithelial cell proliferation via changes in the expression of transforming growth factors.
Phosphodiesterase (PDE) inhibitors, among which pentoxifylline (PTX), are candidate molecules for the treatment of TNF-alpha-dependent inflammatory diseases. Based on the controversial effects of PTX observed in experimentally-induced colitis, the aim of this work was to analyse its influence on intestinal epithelial cell proliferation and growth factor expression using the well-established IEC18 cell line. The effects of PTX, and of an activation (addition of dibutyryl-cAMP, db-cAMP) or inhibition (by a specific cAMP-protein kinase inhibitor, PKI) of the cAMP pathway, were examined after 3 days of culture. The IEC18 cell proliferation and [3H] thymidine incorporation, as well as the expression of TGF-alpha, TGF-beta1 and -beta2 mRNAs, were analysed in basal culture conditions and in the presence of the pro-inflammatory cytokine, TNF-alpha. PTX, like exogenous db-cAMP, inhibited in a dose-dependent manner the basal and TNF-alpha-modulated IEC18 cell proliferation; this effect was partly prevented by PKI. We confirmed that PTX induced a dose-related increase in intracellular cAMP. Concomitantly, the expression of TGF-alpha mRNA dropped and that of TGF-beta2 increased. Addition of db-cAMP instead of PTX also decreased TGF-alpha mRNA, but did not change TGF-beta2 transcripts. The decrease in the expression of TGF-alpha mRNA caused by PTX and db-cAMP was completely abolished by PKI; in contrast, TGF-beta2 remained unaltered. Yet, anti-TGF-beta2 antibodies partially restored the PTX-inhibited cell proliferation. The phosphodiesterase inhibitor, PTX, inhibits IEC18 cell proliferation via a differential modulation of TGF-alpha and TGF-beta2 expression. The drop in TGF-alpha mRNA is related to increasing intracellular cAMP, whereas the effect upon TGF-beta2 appears cAMP-independent.


Anterior prefrontal involvement in episodic retrieval reflects contextual interference.
Different patterns of prefrontal activation are commonly found in studies of episodic and source memory (typically anterior and lateral) compared to those found in studies of autobiographical memory (typically ventromedial). We investigated a proposal that the former pattern reflects contextual interference when retrieving events that occurred in similar contexts. We used virtual reality to simulate contextually varied life-like events, in which subjects received distinct objects from a number of people in a number of locations. We compared fMRI data from two experiments in which the number of events per context varied. The first experiment (Burgess, N., Maguire, E.A., Spiers, H.J., and O'Keefe, J. 2001. A temporoparietal and prefrontal network for retrieving the spatial context of lifelike events. Neuroimage 14, 439-453) involved 16 objects received from one of two people in one of two locations. The second experiment involved 20 objects, each received from a different person in a different location. The first experiment showed extensive bilateral activation of anterior and lateral prefrontal cortex, as well as a medial temporal and parietal network characteristic of both autobiographical and episodic memory. In the second study, the prefrontal activations were largely absent, while the medial parietal and temporal activations remained, and a ventromedial prefrontal area was additionally activated. Direct comparisons revealed large areas of significantly reduced activation in BA10, with lesser reductions in lateral prefrontal regions. We suggest that involvement of these prefrontal regains in episodic and source memory reflects the use of paradigms involving many events and few sources rather than any fundamental processing requirement of contextual retrieval in the absence of interference.


Unraveling the sources of ground level ozone in the Intermountain Western United States using Pb isotopes.
Ozone as an atmospheric pollutant is largely produced by anthropogenic precursors and can significantly impact human and ecosystem health, and climate. The U.S. Environmental Protection Agency has recently proposed lowering the ozone standard from 75 ppbv (MDA8 = Maximum Daily 8-Hour Average) to between 65 and 70 ppbv. This will result in remote areas of the Intermountain West that includes many U.S. National Parks being out of compliance, despite a lack of significant local sources. We used Pb isotope fingerprinting and back-trajectory analysis to distinguish sources of imported ozone to Great Basin National Park in eastern Nevada. During discrete Chinese Pb events (> 1.1 ng/m(3) & > 80% Asian Pb) trans-Pacific transported ozone was 5 ± 5.5 ppbv above 19 year averages for those dates. In contrast, concentrations during regional transport from the Los Angeles and Las Vegas areas were 15 ± 2 ppbv above the long-term averages, and those characterized by high-altitude transport 3 days prior to sampling were 19 ± 4ppbv above. However, over the study period the contribution of trans-Pacific transported ozone increased at a rate of 0.8 ± 0.3 ppbv/year, suggesting that Asian inputs will exceed regional and high altitude sources by 2015-2020. All of these sources will impact regulatory compliance with a new ozone standard, given increasing global background.


Guanine based purines inhibit [(3)H]glutamate and [(3)H]AMPA binding at postsynaptic densities from cerebral cortex of rats.
Extracellular guanine-based purines (GBPs) have been implicated in neuroprotective effects against glutamate toxicity by modulating the glutamatergic system through mechanisms without the involvement of G proteins. Accordingly, GBPs have been shown to inhibit the binding of glutamate and its analogs in different brain membrane preparations. However, brain membrane preparations used for these studies are comprised of both post- and pre-neuronal and glial synaptic components. In this study we investigated the ability of GBPs to displaced glutamate and AMPA binding at postsynaptic densities (PSDs). PSDs are markedly prominent in glutamatergic synapses and retains the native apposition of membrane components and post synaptic receptors. The PSD fraction was prepared from cerebral cortex of Wistar rats and it was characterized as PSDs by electron microscopy and by an enrichment of PSD-95, a protein marker of PSDs (90% of immunodetection). Moreover, we detected an enrichment of glutamate receptors subunits that including NR1 subunit of NMDA receptors and GluR1 subunit of AMPA receptors. GppNp (poor hydrolyzable GTP analog) and GMP displaced 40 and 36% of glutamate binding, respectively, and guanosine only 23%. AMPA binding was not affected by guanosine and was inhibited 21 and 25% by GppNp and GMP, respectively. Hence, this study demonstrates that guanine based purines inhibited glutamate and AMPA binding at postsynaptic membrane preparations, contributing for a better understanding of the mechanisms by which GBPs antagonize glutamatergic neurotoxicicity, e.g. the possible involvement of glutamatergic postsynaptic receptors in their neuroprotective roles.


Comparison of electron microscopy, enzyme-linked immunosorbent assay, solid-phase radioimmunoassay, and indirect immunofluorescence for detection of human rotavirus antigen in faeces.
Four techniques were compared for their practicability, speed, and sensitivity for the detection of human rotavirus. Radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were found to be the most sensitive means of identifying rotavirus, and, once processed, up to 40 specimens could be examined daily. Electron microscopy, although less sensitive than these techniques, had the advantage of being able to detect other viral agents present in faecal extracts. Indirect immunofluorescence failed to detect rotavirus as often as the other three methods. In laboratories where routine examination of faecal specimens from patients with gastroenteritis is required, ELISA and RIA are useful alternatives to electron microscopy.


Endocytosis of Bacteriophages.
Endocytosis is used by eukaryotic cells for ingesting external objects. Importantly, endocytosis is a major process that determines phage pharmacokinetics in vivo. Either dissemination of phages throughout the system or phage clearance engages cellular uptake of phage particles. Here we discuss phage uptake by mammalian cells, focusing on mechanisms and pathways involved. Of note, cellular uptake of phage virions was first observed in professional phagocytes, such as macrophages or granulocytes. For this reason, it was historically referred to as 'phagocytosis'. The modern definition of phagocytosis, however, identifies this process as a type of endocytosis within a larger repertoire of endocytic pathways, such as macropinocytosis, clathrin-mediated endocytosis, and caveolar endocytosis, which have all been included in the scope of this review.


Modulating hierarchically microporous biochar via molten alkali treatment for efficient adsorption removal of perfluorinated carboxylic acids from wastewater.
This work presented a three-dimensional (3D) hierarchically microporous biochar (HMB) via molten alkali treatment that achieved efficient adsorption of perfluorinated carboxylic acids (PFCAs), which was a significant environment concern due to the global distribution and potential health risks. The systematic optimization of fabrication process rendered the HMB large surface area and uniform microporous structure, leading to a high adsorption capacity and adsorption rate of 1269 mg/g and 197 mg/(g·min), respectively, when perfluorooctanoic acid (PFOA) was as a representative. The adsorption mechanisms were explored via controlling the interaction between PFCAs and the HMB900-2.4. Specifically, hydrophobic effect was verified by the enhanced adsorption performance with the increase of the PFCAs homologues hydrophobicity. The observed highly pH-dependent adsorption capacity additionally suggested the dominant contribution of electrostatic interaction. For long-chain PFCAs (CnF2n+1COOH, n > 5), the HMB900-2.4 presented a high removal efficiency (> 90%) within 30 min. Even for short-chain PFCAs (CnF2n+1COOH, n = 4-5), the removal efficiency reached to over 60%. The synthesized HMB900-2.4 exhibited high stability during recycling experiments and superior performance over commercial adsorbents, suggested a promise of utilizing it to remove PFCAs from wastewater.


Pancytopenia in Zimbabwe.
There has been little systematic study of the clinical spectrum of pancytopenia, and the optimal diagnostic approach to pancytopenia remains undefined. The authors studied 134 hospitalized pancytopenic patients in Zimbabwe in both consecutive and nonconsecutive fashion. The most common cause of pancytopenia was megaloblastic anemia, followed by aplastic anemia, acute leukemia, acquired immunodeficiency syndrome (AIDS), and hypersplenism. Severe pancytopenia was usually due to aplastic anemia. Patients with aplastic anemia and acute leukemia were usually children, whereas those with megaloblastic anemia were adults. Moderate to severe anemia was noted throughout the series, but was most striking in patients with megaloblastic anemia, aplastic anemia, and acute leukemia. The mean corpuscular volume (MCV) was elevated in most patients with megaloblastic hematopoiesis, aplastic anemia, and acute nonlymphocytic leukemia. Normal or low MCV values were noted in almost one third of patients with megaloblastic anemia. Anisocytosis, poikilocytosis, macroovalocytosis, microcytosis, fragmentation, and teardrop erythrocytes were more prominent on the blood films of patients with megaloblastic anemia. Megaloblastic anemia, aplastic anemia, and AIDS are the most common causes of pancytopenia in Zimbabwe. Aplasia is the most frequent cause of severe pancytopenia. The authors have formulated tentative guidelines for the evaluation of pancytopenic patients in this setting.


Stability of phenobarbital N-glucosides: identification of hydrolysis products and kinetics of decomposition.
The two diastereomers of 1-(1-beta-D-glucopyranosyl)phenobarbital, (1A) and (1B), decompose to 1-(1-beta-D-glucopyranosyl)-3-(2-ethyl-2-phenylmalonyl)urea (2A or 2B) followed by decarboxylation to 1-(1-beta-D-glucopyranosyl)-3-(2-phenylbutyryl)urea (3A and 3B) under physiological conditions of temperature and pH. The sigmoidal pH-rate profile and the Arrhenius parameters indicate that degradation takes place by hydroxide ion attack on the undissociated and monoanion forms of 1A and 1B. The rates of hydrolysis of the nonionized species of 1A and 1B are more than two orders of magnitude faster than those of common 5,5-disubstituted or 1,5,5-trisubstituted barbiturates. Molecular modeling studies suggest that rate enhancement is due to intramolecular hydrogen bonding in the transition state of the C2' hydroxyl with the tetrahedral hydrated C6 carbonyl as well as hindered rotation around the N1-C1' of phenobarbital and glucose. Based on these studies it is recommended that any data related to the quantitation of 1A and 1B be reevaluated depending on how the samples were collected, stored, and analyzed.


Analysis of paramagnetic centers in X-ray-irradiated enamel, bone, and carbonate-containing hydroxyapatite by electron spin resonance spectroscopy.
Carbonate-containing hydroxyapatite, enamel, and bone were irradiated by an X-ray and investigated between 77 degrees and 350 degrees K by means of electron spin resonance (ESR) spectroscopy. The ESR spectrum of enamel irradiated at 77 degrees K in vacuum and observed at the same temperature was almost the same as that of the carbonate-containing hydroxyapatite. The temperature dependence of signal intensities confirms a spin-energy exchange between the mineral and organic constituents in bone, but in enamel no or very little spin-energy exchange between the mineral and organic constituents. Considerable similarity among the ESR spectra of enamel, bone, and carbonate-containing apatite was obtained after X-ray irradiation in air at 300 degrees K with both an X-band and a Q-band ESR spectrometer. The Q-band spectrum can be interpreted in terms of two paramagnetic species. One is identified as a CO3(3-) anion radical which has an axial symmetry with g factors of 2.0029 and 1.9972. The other species is found to be centered at g = 2.0019.


Retropharyngeal and mediastinal abscess following adenoidectomy.
Adenoidectomy is one of the most common surgical procedures in children. On rare occasions it can be complicated by a retropharyngeal abscess (RA). We report the case of a 9-year-old girl with an RA extending to the mediastinum following adenoidectomy. The mediastinal collection was successfully treated with computed tomography-guided percutaneous drainage to avoid extensive surgical procedure. Surgeons should be aware of these dramatic complications and should avoid overzealous surgery in the removal of the adenoid tissue.


Shape- and Material-Dependent Self-Propulsion of Photocatalytic Active Colloids, Interfacial Effects, and Dynamic Interparticle Interactions.
Active colloids powered by self-generated, local chemical concentration gradients exhibit dynamics that are a function of the particles' morphology and material properties. These characteristics also govern how the active colloids interact with surfaces, including other particles and nearby walls. Thus, by targeted design, the dynamic behavior, on average, can be engineered, despite a lack of "external" control such as an applied magnetic field. This allows for the development of new applications and the investigation of novel effects that arise when self-propelled active colloids have complex shapes and material composition. Here, we explore some of our recent work on this topic including the dynamics and interactions of photoactivated, self-propelled colloids with such multifaceted properties. We also delve into some special cases, such as a new variety of active particle-particle interaction that we recently developed, in which direct contact between the active colloids is forbidden, and the direction of propulsion for pairs of particles is correlated. The unifying theme of the work highlighted herein is the relationship between the physical, chemical, and material properties of active colloids and their motive behavior, the understanding of which opens up a wide range of new possibilities as we move toward the ultimate goal of realizing functional, man-made micro- and nanomachinery.


Effect of the P-glycoprotein inhibitor, R(+)-verapamil on the drug susceptibility of a triclabendazole-resistant isolate of Fasciola hepatica.
A study has been carried out to investigate whether the action of triclabendazole (TCBZ) against Fasciola hepatica is altered by the inhibition of P-glycoprotein (Pgp)-linked drug efflux pumps. The Sligo TCBZ-resistant and Cullompton TCBZ-susceptible fluke isolates were used for these experiments and the Pgp inhibitor selected was R(+)-verapamil [R-VPL]. In the first experiment, flukes were initially incubated for 2 h in R-VPL (100 μM), then incubated for a further 22 h in R-VPL+triclabendazole sulphoxide (TCBZ.SO) (50 μg/ml, or 0.1327 μM). For controls, flukes were incubated for 24 h in R-VPL and TCBZ.SO on their own. In a second experiment, flukes were removed from the incubation media following cessation of movement. In the third experiment, Sligo flukes were incubated in lower concentrations of R-VPL (10 μM) and TCBZ.SO (15 μg/ml, or 0.0398 μM). Morphological changes resulting from drug treatment and following Pgp inhibition were assessed by means of scanning electron microscopy. Incubation in R-VPL alone had minimal effect on either isolate. After treatment with TCBZ.SO alone, there was greater surface disruption to the Cullompton than Sligo isolate. However, combined treatment of R-VPL+TCBZ.SO led to more severe surface changes to the Sligo isolate than with TCBZ.SO on its own; this potentiation of drug activity was not seen with the Cullompton isolate. The phenomenon was evident at both concentrations of TCBZ.SO. Inclusion of R-VPL in the incubation medium also reduced the time taken for the flukes to become inactive; again, this effect was more distinct with the Sligo isolate. The results of this study support the concept of altered drug efflux in TCBZ-resistant flukes and indicate that drug transporters may play a role in the development of drug resistance.


Inferior Vena Cava Draining Into the Left Atrial Cavity Due to Atrial Septal Defect: Two Atypical Presentations.
The inferior vena cava draining to the left atrium is a rare congenital anomaly. Patients usually present with hypoxia and dyspnoea. This condition is usually diagnosed by echocardiography and sometimes by CT scan. Here we report two cases that presented with normal saturation and their surgical management.


Enzymatically produced cellulose nanocrystals as reinforcement for waterborne polyurethane and its applications.
Waterborne polyurethanes (WBPUs) have been proposed as ecofriendly elastomers with several applications in coatings and adhesives. WBPU's physicochemical properties can be enhanced by the addition of cellulose nanocrystals (CNCs). The way CNCs are isolated has a strong effect on their properties and can determine their role as reinforcement. In this work, CNCs produced using ancestral endoglucanase (EnCNCs) were used as reinforcement for WBPU and compared with CNC produced by sulfuric acid hydrolysis (AcCNC). The enzymatic method produced highly thermostable and crystalline CNCs. The addition of small contents of EnCNCs improved the thermomechanical stability and mechanical properties of WBPUs, even better than commercial AcCNCs. Besides, WBPU reinforced by adding EnCNCs was studied as a coating for paper materials, increasing its abrasion resistance and as electrospun nanocomposite mats where EnCNCs helped maintaining the morphology of the fibers.


Gemella morbillorum in primary and secondary/persistent endodontic infections.
To investigate the presence of Gemella morbillorum by culture or nested PCR in primary and secondary/persistent endodontic infections. Microbial samples were taken from 50 cases with primary and 50 cases with secondary/persistent endodontic infections. Microbiologic techniques were used for culture and identification. The DNA extracted from the samples was analyzed for the presence of the target species using species-specific primers. Culture and polymerase chain reaction (PCR) identified the species in 23 and 77, respectively, of 100 root canals. Culture yielded the test organism in 19 of 50 (38%) of root canal samples from primary and in 4 of 50 (8%) from secondary/persistent infections. PCR yielded the test organisms in 41 of 50 (82%) and 36 of 50 (72%) of the, respectively, primary and secondary/persistent root canal infections studied. Gemella morbillorum was identified more frequently in primary endodontic infections than in secondary/persistent ones. A higher frequency of the target species was detected by PCR than by culture.


Intestinal epithelial stem cell transplants as a novel therapy for cerebrovascular stroke.
Almost 2/3rds of stroke survivors exhibit vascular cognitive impairment and a third of stroke patients will develop dementia 1-3 years after stroke. These dire consequences underscore the need for effective stroke therapies. In addition to its damaging effects on the brain, stroke rapidly dysregulates the intestinal epithelium, resulting in elevated blood levels of inflammatory cytokines and toxic gut metabolites due to a 'leaky' gut. We tested whether repairing the gut via intestinal epithelial stem cell (IESC) transplants would also improve stroke recovery. Organoids containing IESCs derived from young rats transplanted into older rats after stroke were incorporated into the gut, restored stroke-induced gut dysmorphology and decreased gut permeability, and reduced circulating levels of endotoxin LPS and the inflammatory cytokine IL-17A. Remarkably, IESC transplants also improved stroke-induced acute (4d) sensory-motor disability and chronic (30d) cognitive-affective function. Moreover, IESCs from older animals displayed senescent features and were not therapeutic for stroke. These data underscore the gut as a critical therapeutic target for stroke and demonstrate the effectiveness of gut stem cell therapy.


[Free protein S (PS) in normal pregnancy: a comparison between two analytical methods].
Pregnancy is a physiological hypercoagulable state with an increased incidence of thromboembolic phenomena. There is an increase in the concentrations of most clotting factors, a decrease in concentration of some of the natural anticoagulants and reduced fibrinolytic activity. Changes in PS levels have also been reported. To establish referral range values of functional PS and free PS antigen, during the second (2nd T) and third trimester (3rd T) of normal gestation. Forty one normal pregnant women were included in our study, 20 during the 2nd T (22-24 weeks) and 21 during the 3rd T (29-38 weeks). Functional PS was measured by a clot based test and free PS antigen by ELISA. Free PS Antigen was 65.8+/-18.3% during the 2nd T and 62.3+/-16.5% during the 3rd T. The figures for normal controls were 106+/-6.5%. Functional PS was 43.8+/-13.3 and 25.9+/-14.6% during the 2nd T and 3rd T, respectively. The figures for normal controls were 97+/-24% (p <0.001 compared with pregnant women). Free PS antigen did not change from the 2nd to the 3rd T (p=NS), however functional PS fell significantly from the 2nd to the 3rd T (p <0.001) and was significantly lower than free PS antigen in both trimesters (p <0.001). Pregnancy is associated to a decrease in PS. This abnormality is more pronounced for functional PS than free PS antigen and functional PS falls progressively during pregnancy. These assays should not be used to screen for PS deficiency during pregnancy because they could lead to a misdiagnosis.


Quantitative Assessment of Orbital Decompression Surgery Using Photogrammetric Stereoimaging.
To evaluate periorbital volume changes, using quantitative photogrammetric stereoimaging, after bony orbital decompression for thyroid eye disease (TED) and compare this to changes in Hertel exophthalmometry. Tissue volumes of the upper and lower eyelids were also assessed independently. Prospective, nonrandomized, nonconsecutive series of patients. Adults with inactive TED who had elected to undergo bony orbital decompression at Moorfields Eye Hospital between 2015 and 2017. With their eyes gently closed, patients with inactive TED underwent imaging using the VECTRA M3 system (Canfields Imaging, Fairfield, NJ), both before and at least 3 months after orbital decompression. Proptosis was assessed by Hertel exophthalmometry. Using the manufacturer's software, changes in periorbital volumes between the preoperative and postoperative images were calculated for the upper and lower eyelids. The Pearson product-moment correlation coefficient was used to assess the linear relationship between changes in periorbital volume and exophthalmometry. Change in stereoimaging volumes of the upper and lower eyelids, and clinical proptosis as measured with Hertel exophthalmometry. Thirty-three patients (11 males; 33%) underwent lateral decompression (39 orbits in 26 patients), lateral decompression with complete ethmoidectomy (3 orbits in 3 patients), combined decompression of lateral wall, medial wall, and medial half of floor (10 orbits in 6 patients), or bilateral lateral, medial, and complete floor decompression (1 patient). The corresponding average volumetric changes were 1.74 ml (median 1.64; range 0.39-3.73 ml), 3.38 ml (median 3.38, range 1.89-4.88 ml), 4.05 ml (median 3.53, range 1.72-6.43 ml), and 4.52 ml (range 4.36-4.68 ml), respectively. Similarly, the average reduction in proptosis was 3.6 mm (median 3; range 1.5-7 mm), 5.3 mm (median 5; range 5-6 mm), 7.4 mm (7.5; range 6-9 mm), and 9 mm (range 8-10 mm). Periorbital volume changes were related to the reduction in exophthalmometry (r = 0.713, p < 0.0001), and reduction of lower eyelid volume did not increase further with more than two-wall decompression. Graded orbital decompression reduces both proptosis and the upper and lower eyelid tissue volumes, the eyelid changes being likely to influence decisions about future restorative eyelid surgery. The significant changes in eyelid profiles underlines the established principle of thyroid periocular rehabilitation, namely decompression, then strabismus surgery, and finally eyelid surgery.


Orally available tubulin inhibitor VERU-111 enhances antitumor efficacy in paclitaxel-resistant lung cancer.
Lung cancer is the most common cause of cancer associated mortality. Chemotherapeutic agents, such as paclitaxel, are important treatment options but drug resistance often develops upon prolonged use. We report here the preclinical evaluation of a new orally available tubulin inhibitor, VERU-111, which can overcome several ABC-transporters mediated multi-drug resistance associated with taxane treatment. In vitro, VERU-111 prevents cell proliferation, invasion, migration and colony formation in both paclitaxel-sensitive and paclitaxel-resistant A549 lung cancer cells. VERU-111 effectively inhibits tubulin polymerization, arrests cells in G2/M phase, and induces cancer cell apoptosis. Further evaluation of various apoptotic proteins revealed that treatment of VERU-111 increases the expression of cleaved-PARP, cleaved-caspase-3 and p-histone H3 proteins. In vivo, orally administered VERU-111 in a paclitaxel-sensitive A549 xenograft model strongly inhibits tumor growth in a dose-dependent manner and is equally potent with paclitaxel. When tested in a highly paclitaxel-resistant A549/TxR tumor model, VERU-111 is as effective as the parental A549 model in significantly reducing the tumor volume, whereas paclitaxel is essentially ineffective. Collectively, this study showed that VERU-111 is a promising new generation of anti-tubulin agent for the treatment of taxane-resistant lung cancer.


Chief Resident Election of Emergency Department (CREED) - An innovative approach to fair and bias-free chief resident selection in a residency program.
Emergency medicine has transitioned from developing to a developed specialty in Pakistan. It is gaining recognition in the national and international arena. Residency in emergency medicine is a tough pathway in which a resident learns and is assessed at multiple levels. The attributes that are needed for an empathetic emergency medicine physician are multifaceted. Chief resident selection has been an important step in postgraduate residency. The selection process was traditionally based on seniority and academic achievements with no consideration for soft skills. In the current write-up, we are proposing an evidence-base sequential chief resident selection process called Chief Resident Election of Emergency Department (CREED). The program was developed keeping in consideration the traditional method of election and interviews with the incorporation of reflexive, leadership, communication, and collaboration skills.


Optimization of Injection Molding Parameters for HDPE/TiO₂ Nanocomposites Fabrication with Multiple Performance Characteristics Using the Taguchi Method and Grey Relational Analysis.
The current study presents an investigation on the optimization of injection molding parameters of HDPE/TiO₂ nanocomposites using grey relational analysis with the Taguchi method. Four control factors, including filler concentration (i.e., TiO₂), barrel temperature, residence time and holding time, were chosen at three different levels of each. Mechanical properties, such as yield strength, Young's modulus and elongation, were selected as the performance targets. Nine experimental runs were carried out based on the Taguchi L₉ orthogonal array, and the data were processed according to the grey relational steps. The optimal process parameters were found based on the average responses of the grey relational grades, and the ideal operating conditions were found to be a filler concentration of 5 wt % TiO₂, a barrel temperature of 225 °C, a residence time of 30 min and a holding time of 20 s. Moreover, analysis of variance (ANOVA) has also been applied to identify the most significant factor, and the percentage of TiO₂ nanoparticles was found to have the most significant effect on the properties of the HDPE/TiO₂ nanocomposites fabricated through the injection molding process.


Spectroscopical study of bacteriopurpurinimide-naphthalimide conjugates for fluorescent diagnostics and photodynamic therapy.
Two novel bis(chromophoric) dyads ABPI-NI1 and ABPI-NI2 containing 1,8-naphthalimide and bacteriopurpurinimide units linked by p-phenylene-methylene (ABPI-NI1) and pentamethylene (ABPI-NI2) spacers were prepared to test their ability to be used in the design of effective agents for both photodynamic therapy (PDT) and fluorescent tumor imaging. Photophysical studies revealed that the emission from the naphthalimide chromophore in both conjugates was partially quenched due to resonance energy transfer between the photoactive components. Compound ABPI-NI2 with more sterically flexible oligomethylene group demonstrated higher fluorescence intensity as compared with that for ABPI-NI1.


[Traumatic thoracic aorta rupture: preclinical assessment, diagnosis and treatment options].
Traumatic aortic rupture is a life-threatening injury which is frequently associated with blunt thoracic trauma or found coincidentally in heavily traumatized patients. Depending on the degree of disruption of the damaged aortic wall, vascular injury is associated with a high primary mortality rate and a significant risk of secondary aortic rupture. Early clinical signs which may indicate a ruptured thoracic aorta are left sided thoracic pain, reduced ventilation, tachycardia and dyspnoe as well as hypotension in the lower extremities. The primary aim for emergency treatment is to maintain vital organ function and to hemodynamically stabilize the patient. Surgical treatment was previously performed by either direct aortic suture or segmental alloplastic graft interposition using the clamp and sew technique with or without extra-anatomic shunts or extracorporeal circulation. However, endovascular stent graft implantation has now become another treatment option for traumatic aortic rupture. According to the reported data and our own experience there is increasing evidence that endovascular aortic repair might become the treatment of choice for patients with traumatic aortic rupture, with the option of an early, less invasive intervention thus avoiding thoracotomy. Regular follow-up is necessary to detect possible stent graft migration or leakage which could require additional endovascular or open surgical re-interventions.


Parallel Syntheses of Peptides on Teflon-Patterned Paper Arrays (SyntArrays).
Screening of peptides to find the ligands that bind to specific targets is an important step in drug discovery. These high-throughput screens require large number of structural variants of peptides to be synthesized and tested. This chapter describes the generation of arrays of peptides on Teflon-patterned sheets of paper. First, the protocol describes the patterning of paper with a Teflon solution to produce arrays with solvophobic barriers that are able to confine organic solvents. Next, we describe the parallel syntheses of 96 peptides on Teflon-patterned arrays using the SPOT synthesis method.


Apolipoprotein E ε4 Allele was Associated With Nonlesional Mesial Temporal Lobe Epilepsy in Han Chinese Population.
Apolipoprotein E (APOE) gene has been implicated as one of the genes susceptible to temporal lobe epilepsy (TLE), but the association is inconsistent. We carried out a study to investigate the association of APOEε4 allele with a subtype of TLE-nonlesional mesial temporal lobe epilepsy (NLMTLE) in Han Chinese people.T he study consisted of total 308 NLMTLE patients and 302 controls in Han Chinese. The APOE polymorphisms were genotyped using polymerase chain reaction (PCR) DNA sequencing. We compared the frequency of APOEε4 allele and carrying status between NLMTLE patients and control subjects to test for the association of APOEε4 allele with NLMTLE clinical status. Carrying status of APOEε4 allele was significantly associated with the risk of NLMTLE. No effect of APOEε4 allele was found on the age of onset, duration of epilepsy, or frequency of seizure. Moreover, there was no association between APOEε4 allele and hippocampal sclerosis (HS) or febrile convulsion (FC) history.O ur study provided an evidence that APOEε4 allele was a possible risk factor for NLMTLE, and further study with a larger sample is needed to warrant this finding.


Needle core biopsy for the assessment of unilateral breast masses in men.
Although unilateral male breast swelling is relatively common, a histological diagnosis is infrequently obtained. From 1998 to 2003 we routinely performed needle core biopsy on all men presenting with unilateral breast swelling in whom there was diagnostic uncertainty. Of 113 patients, 93% had gynaecomastia, two patients had primary breast cancer and one had metastatic lymphoma. One patient had chronic mastitis. Gamolenic acid treatment produced a 73% response rate amongst patients presenting with pain. Core biopsy is a safe and effective method of diagnosing unilateral male breast swelling, which allows either confident reassurance or definitive treatment of those with cancer or pain.


A case-based review of troubleshooting deep brain stimulator issues in movement and neuropsychiatric disorders.
To review the spectrum of problems that can occur in the DBS patient and to suggest potential troubleshooting tips for identification and management of DBS related issues. Deep brain stimulation (DBS) has become commonplace for the treatment of medication-refractory neurological disorders. There remains no consensus on the best practices for screening, surgical techniques, and post-operative care. There are few experienced DBS programmers and scarce resources available describing approaches for troubleshooting DBS problems. We present a case-based review that offers practical tips for the management and troubleshooting of difficult to manage DBS cases. We present 10 cases to demonstrate common issues encountered in DBS management. There are many important difficulties that may be encountered with DBS devices, and practitioners should be aware of these potential problems, as well as rational management solutions. The following areas should be emphasized as potential causes of difficulties: a non-ideal initial DBS candidate, inadequate multidisciplinary team care, failure of perceived expectations, DBS procedural complication, hardware complication, suboptimal lead placement, programming, access to care, disease progression, and tolerance/habituation. Neurologists seeing DBS patients should become familiar with issues involved in difficult to manage DBS cases. Many "DBS failures" are currently treatable by appropriate medicine, programming, and surgical approaches.


Theoretical and Experimental Investigation of Thermodynamics and Kinetics of Thiol-Michael Addition Reactions: A Case Study of Reversible Fluorescent Probes for Glutathione Imaging in Single Cells.
Density functional theory (DFT) was applied to study the thermodynamics and kinetics of reversible thiol-Michael addition reactions. M06-2X/6-31G(d) with the SMD solvation model can reliably predict the Gibbs free energy changes (ΔG) of thiol-Michael addition reactions with an error of less than 1 kcal·mol(-1) compared with the experimental benchmarks. Taking advantage of this computational model, the first reversible reaction-based fluorescent probe was developed that can monitor the changes in glutathione levels in single living cells.


The gut microbiome and cancer immunotherapeutics: A review of emerging data and implications for future gynecologic cancer research.
Investigation of the gynecologic tract microbial milieu has revealed potential new biomarkers. Simultaneously, immunotherapeutics are establishing their place in the treatment of gynecologic malignancies. The interplay between the microbiome, the tumor micro-environment and response to therapy is a burgeoning area of interest. There is evidence to support that microbes, through their genetic make-up, gene products, and metabolites affect human physiology, metabolism, immunity, disease susceptibility, response to pharmacotherapy, and the severity of disease-related side effects. Specifically, the richness and diversity of the gut microbiome appears to affect carcinogenesis, response to immunotherapy, and modulate severity of immune-mediated adverse effects. These effects have best been described in other tumor types and these have shown compelling results. This review summarizes the current understanding and scope of the interplay between the human microbiome, host factors, cancer, and response to treatments. These findings support further exploring whether these associations exist for gynecologic malignancies.


Effectiveness of quality of care for patients with type 2 diabetes in China: findings from the Shanghai Integration Model (SIM).
This cross-sectional study aimed to investigate the quality of care of diabetes in Shanghai, China. A total of 173 235 patients with type 2 diabetes in 2017 were included in the analysis. Profiles of risk factors and intermediate outcomes were determined. The patients had a mean age of 66.43 ± 8.12 (standard deviation (SD)) years and a mean diabetes duration of 7.95 ± 5.53 (SD) years. The percentage of patients who achieved the target level for HbA1c (< 7.0%) was 48.6%. Patients who achieved the target levels for blood pressure (BP) < 130/80 mmHg and low-density lipoprotein-cholesterol (LDL-c) < 2.6 mmol/L reached 17.5% and 34.0%, respectively. A total of 3.8% achieved all three target levels, and the value increased to 6.8% with an adaptation of the BP target level (< 140/90 mmHg) for those over 65 years. Multivariable analysis identified the factors associated with a great likelihood of achieving all three target levels: male, young age, short diabetes duration, low body mass index, macrovascular complications, no microvascular complications, prescribed with lipid-lowering medication, and no prescription of antihypertensive medication. In conclusion, nearly 50% and one-third of the patients with diabetes met the target levels for HbA1c and LDL-c, respectively, with a low percentage achieving the BP target level. The percentage of patients who achieved all three target levels needs significant improvement.


How ought we allocate unanticipated doses of COVID-19 vaccines? A proposal based on experience in the United States, 2020-2022.
With vaccination against COVID-19 well underway, providers in the United States (U.S.) found that vials of the Pfizer/BioNTech and Moderna vaccines contained more doses than listed. Because of limited vaccine shelf life, vaccine-providing institutions have thrown extra doses away or quickly distributed them using ad-hoc allocation schemes (drawing names from waiting lists or administering doses to whomever happened to be nearby). This Viewpoint argues that these practices are ethically inadequate and proposes an alternate distribution scheme based on the system in the U.S. to allocate donated organs. The proposal allows for the allocation of unanticipated doses in a way that maximizes societal benefits, is fair, and considerate of the sensitive time constraints for preserving vaccine effectiveness.


A 3-year survey of Italian honey bee-collected pollen reveals widespread contamination by agricultural pesticides.
Honey bee (Apis mellifera L.) health is compromised by complex interactions between multiple stressors, among which pesticides play a major role. To better understand the extent of honey bee colonies' exposure to pesticides in time and space, we conducted a survey by collecting corbicular pollen from returning honey bee foragers in 53 Italian apiaries during the active beekeeping season of 3 subsequent years (2012-2014). Of 554 pollen samples analysed for pesticide residues, 62% contained at least one pesticide. The overall rate of multiresidual samples (38%) was higher than the rate of single pesticide samples (24%), reaching a maximum of 7 pesticides per sample (1%). Over 3years, 18 different pesticides were detected (10 fungicides and 8 insecticides) out of 66 analysed. Pesticide concentrations reached the level of concern for bee health (Hazard Quotient (HQ) higher than 1000) at least once in 13% of the apiaries and exceeded the thresholds of safety for human dietary intake (Acute Reference Dose (ARfD), the Acceptable Daily Intake (ADI), and the Maximum Residue Limit (MRL)) in 39% of the analysis. The pesticide which was most frequently detected was the insecticide chlorpyrifos (30% of the samples overall, exceeding ARfD, ADI, or MRL in 99% of the positive ones), followed by the fungicides mandipropamid (19%), metalaxyl (16%), spiroxamine (15%), and the neonicotinoid insecticide imidacloprid (12%). Imidacloprid had also the highest HQ level (5054, with 12% of its positive samples with HQ higher than 1000). This 3year survey provides further insights on the contamination caused by agricultural pesticide use on honey bee colonies. Bee-collected pollen is shown to be a valuable tool for environmental monitoring, and for the detection of illegal uses of pesticides.


A controlled, longitudinal study of home visits compared to telephone contacts to prevent early childhood caries.
Home visits (HV) provide excellent opportunities for health promotion. This longitudinal study compared the effects of HV and telephone contacts (TC) in preventing early childhood caries (ECC) and colonisation of mutans streptococci (MS) and lactobacilli (LB) from 0 to 24 months. A total of 325 children were recruited from community health centres at mean age of 42 days, and randomly assigned to receive either HV or TC. A total of 188 children completed three, 6 monthly HV, and another 58 had three, 6 monthly TC. An additional 40 age-matched children from childcare facilities served as reference controls (RC). At 24 months, all groups were examined at a community dental clinic. At 24 months, three HV children of 188 (1.5%) had caries, compared to four TC of 58 (6.8%) and nine RC of 40 (22.5%) (P < 0.001 for HV versus RC; P = 0.05 for HV versus TC and P = 0.03 for TC versus RC). There were also more children with MS in the TC (47%) and RC (35%) compared to HV (28%) group (P = 0.01 and P = 0.02). Home visits and telephone contacts conducted 6 monthly from birth are effective in reducing ECC prevalence by 24 months.


Genetic suppression of IKK2/NF-κB in astrocytes inhibits neuroinflammation and reduces neuronal loss in the MPTP-Probenecid model of Parkinson's disease.
Neuroinflammatory activation of glia is considered a pathological hallmark of Parkinson's disease (PD) and is seen in both human PD patients and in animal models of PD; however, the relative contributions of these cell types, especially astrocytes, to the progression of disease is not fully understood. The transcription factor, nuclear factor kappa B (NFκB), is an important regulator of inflammatory gene expression in glia and is activated by multiple cellular stress signals through the kinase complex, IKK2. We sought to determine the role of NFκB in modulating inflammatory activation of astrocytes in a model of PD by generating a conditional knockout mouse (hGfapcre/Ikbk2F/F) in which IKK2 is specifically deleted in astrocytes. Measurements of IKK2 revealed a 70% deletion rate of IKK2 within astrocytes, as compared to littermate controls (Ikbk2F/F). Use of this mouse in a subacute, progressive model of PD through exposure to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine and probenecid (MPTPp) revealed significant protection in exposed mice to direct and progressive loss of dopaminergic neurons in the substantia nigra (SN). hGfapcre/Ikbk2F/F mice were also protected against MPTPp-induced loss in motor activity, loss of striatal proteins, and genomic alterations in nigral NFκB gene expression, but were not protected from loss of striatal catecholamines. Neuroprotection in hGfapcre/Ikbk2F/F mice was associated with inhibition of MPTPp-induced astrocytic expression of inflammatory genes and protection against nitrosative stress and apoptosis in neurons. These data indicate that deletion of IKK2 within astrocytes is neuroprotective in the MPTPp model of PD and suggests that reactive astrocytes directly contribute the potentiation of dopaminergic pathology.


Ex situ resection techniques and liver autotransplantation: last resource for otherwise unresectable malignancy.
Ex vivo liver resection and hepatic autotransplantation allow treatment of otherwise unresectable hepatobiliary malignancy and expand the role of liver surgery. Few series are reported in the English literature, and they are mainly case reports focused on the technical aspects of the procedure more than on the long-term results. Postoperative mortality rate is still high, as well as early tumor recurrence; recently, improvement in outcomes in selected cases is being reported. The aim of this study is to describe our experience with two cases where we applied the combination of ex situ resection with hepatic autotransplantation for intended cure of two otherwise untreatable malignancies. These two cases are the paradigm of the possibility of performing extreme salvage procedures after the failure of more conventional treatments, allowing improvement in survival and quality of life in a controversial clinical scenario. This should be performed only AT referral centers for liver resection surgery and liver transplantation in light of the technical refinements reached in the era of living-related liver transplantation.


A CRISPR knockout negative screen reveals synergy between CDKs inhibitor and metformin in the treatment of human cancer in vitro and in vivo.
Laboratory research and pharmacoepidemiology provide support for metformin as a potential antitumor agent. However, the lack of a clear understanding of the indications of metformin limits its efficacy. Here, we performed a genome-wide CRISPR knockout negative screen to identify potential targets that might synergize with metformin. Next-generation sequencing of pooled genomic DNAs isolated from surviving cells after 18 days of metformin treatment (T18) compared to those of the untreated cells at day 0 (T0) yielded candidate genes. Knockdown of a group of cyclin-dependent kinases (CDKs), including CDK1, CDK4, and CDK6, confirmed the results of the screen. Combination treatment of the CDKs inhibitor abemaciclib with metformin profoundly inhibited tumor viability in vitro and in vivo. Although cell cycle parameters were not further altered under the combination treatment, investigation of the metabolome revealed significant changes in cell metabolism, especially with regard to fatty acid oxidation, the tricarboxylic acid cycle and aspartate metabolism. Such changes appeared to be mediated through inhibition of the mTOR pathway. Collectively, our study suggests that the combination of CDKs inhibitor with metformin could be recognized as a potential therapy in future clinical applications.


Primary ovarian Burkitt lymphoma.
Primary ovarian Burkitt lymphoma (BL) is a rare neoplasm in adults. We report a 30-year-old woman diagnosed with primary bilateral ovarian BL. She presented features of a twisted ovarian cyst and underwent bilateral salpingo-oophorectomy. The histopathologic evaluation yielded the diagnosis of BL and subsequently she received chemoimmunotherapy with CODOX-M-IVAC plus rituximab (anti-CD20 monoclonal antibody).


Measurement of cardiac output during pregnancy: validation of Doppler technique and clinical observations in preeclampsia.
In 12 patients requiring pulmonary artery catheterization, cardiac output was measured using Doppler and thermodilution techniques. The Doppler technique accurately predicted measurements made by thermodilution (r = 0.91; P less than .001). Eighteen normal patients in the third trimester and 36 preeclamptics who had not been treated with medications other than magnesium sulfate were evaluated with Doppler alone. Of note was the heterogeneity among preeclamptics. Although their mean systemic vascular resistance was elevated, it ranged from 2256-648 dyne X sec X cm-5. Cardiac output ranged from 13.2-3.9 L/minute.


A global analysis of conservative and non-conservative mutations in SARS-CoV-2 detected in the first year of the COVID-19 world-wide diffusion.
The ability of SARS-CoV-2 to rapidly mutate represents a remarkable complicancy. Quantitative evaluations of the effects that these mutations have on the virus structure/function is of great relevance and the availability of a large number of SARS-CoV-2 sequences since the early phases of the pandemic represents a unique opportunity to follow the adaptation of the virus to humans. Here, we evaluated the SARS-CoV-2 amino acid mutations and their progression by analyzing publicly available viral genomes at three stages of the pandemic (2020 March 15th and October 7th, 2021 February 7th). Mutations were classified in conservative and non-conservative based on the probability to be accepted during the evolution according to the Point Accepted Mutation substitution matrices and on the similarity of the encoding codons. We found that the most frequent substitutions are T > I, L > F, and A > V and we observe accumulation of hydrophobic residues. These findings are consistent among the three stages analyzed. We also found that non-conservative mutations are less frequent than conservative ones. This finding may be ascribed to a progressive adaptation of the virus to the host. In conclusion, the present study provides indications of the early evolution of the virus and tools for the global and genome-specific evaluation of the possible impact of mutations on the structure/function of SARS-CoV-2 variants.


Spermatozoa nucleus takes up lead during the epididymal maturation altering chromatin condensation.
Lead (Pb) alters sperm chromatin condensation (CC) and the mechanisms are investigated. During spermatogenesis, protamines replace histones and disulfide bonds formation during epididymal maturation condense the chromatin. We evaluated sperm Pb uptake in testis and epididymis and the effects on CC in mice (0.06% Pb(2+)/16 weeks/drinking water). Spermatozoa from caput epididymis (CP) and cauda epididymis-vas deferens (CE-VD) were obtained and CC was measured by SCSA. Lead levels in spermatozoa from CP were lower than those from CE-VD, and correlated with a decreased CC, while Pb in CE-VD spermatozoa correlated with an increased CC. Lead accumulation into the nucleus was observed and Pb binding to nuclear sulfhydryl groups decreased chromatin decondensation in vitro. Our results suggest that spermatozoa take up Pb during testicular development and epididymal transport and alter CC, depending of the timing of Pb incorporation into the sperm nucleus, which finally may interfere with the chromatin decondensation process after fertilization.


Structural characterization and proliferation activity of chondroitin sulfate from the sturgeon, Acipenser schrenckii.
The cartilages of marine fish, such as sharks and sturgeon, are important resources of the bioactive chondroitin sulfate (CS). To explore glycosaminoglycans from marine fish, polysaccharides from the cartilage of the sturgeon, Acipenser schrenckii, were extracted. Using enzyme-assisted extraction and anion-exchange chromatography, an uronic acid-containing polysaccharide, YG-1, was isolated. YG-1 is composed of GlcN, GlcUA, GalN, and Gal, in the ratio of 1.4: 3.4: 3.7: 1.0, and its molecular weight was determined to be 3.0 × 105 Da. YG-1 was confirmed to be chondroitin 4-sulfate (CS) composed of →4GlcAβ1→3GalNAc4Sβ1→ and minor →4GlcAβ1→3GalNAcβ1→, which was confirmed using IR spectroscopy, disaccharide composition analysis, and NMR. Bioactivity studies, including MTT assay and scratch-wound assays revealed that CS from Acipenser schrenckii had significant proliferation activity. The proliferation activity of the polysaccharide, YG-1, was related to Fibroblast growth factor 2 (FGF2). GalNAc 4S of YG-1 could be the binding sites of FGF2 and FGFR.


Identification and characterization of an equine herpesvirus 1 late gene encoding a potential zinc finger.
In this report, we present the DNA sequence and transcriptional characterization of a gene (IR5) that maps within each of the inverted repeat (IR) segments of the equine herpesvirus type 1 (EHV-1) genome. The IR5 open reading frame (ORF) is located within both IR sequences (nucleotides 9932-10,642 of the IR). DNA sequence analyses of the IR5 gene region revealed an ORF of 236 amino acids (24,793 Da) that showed significant homology to ORF64 of varicella-zoster virus and ORF3 of EHV-4 both of which map within the inverted repeats and to the US10 ORF of herpes simplex virus type 1 (HSV-1) which maps within the unique short segment. Additional analyses of the nucleotide sequence failed to reveal any overlapping ORFs that would correspond to US11 or US12 of HSV-1. Interestingly, the IR5 ORF of EHV-1 possesses a sequence of 13 amino acids (CAYWCCLGHAFAC) that is a perfect match to the consensus zinc finger motif (C-X2-4-C-X2-15-C/H-X2-4-C/H). Putative cis-acting elements flanking the IR5 ORF include a TATA box (nucleotides 9864-9870), a CAAT box (nucleotides 9709-9714), and a polyadenylation signal (nucleotides 10,645-10,650). Northern blot and S1 nuclease analyses identified a single 0.9-kb mRNA species that first appears at 2 hr postinfection, and whose synthesis is reduced in the presence of phosphonoacetic acid, an inhibitor of EHV-1 DNA synthesis. Thus, the IR5 gene of EHV-1 exhibits characteristics representative of a late gene of the gamma-1 class. The characterization of the IR5 gene at the DNA and RNA levels will facilitate ongoing studies to identify and characterize the IR5 polypeptide.


Hypoxia down-regulates DNA double strand break repair gene expression in prostate cancer cells.
Intratumoral hypoxia has been correlated with poor clinical outcome in prostate cancer. Prostate cancer cells can be genetically unstable and have altered DNA repair. We, therefore, hypothesized that the expression of DNA double-strand break (DNA-dsb) repair genes in normal and malignant prostate cultures can be altered under hypoxic conditions. The expression of homologous recombination (HR) and non-homologous recombination (NHEJ) genes following gas hypoxia (0.2%) or exposure to HIF1alpha-inducing agent, CoCl2 (100 microM), was determined for normal diploid fibroblasts (GM05757) and the pre-malignant and malignant prostate cell lines, BPH-1, 22RV-1, DU145 and PC3. RNA and protein levels were determined using RT-PCR and Western blotting. Additionally, p53 genotype and function, the level of hypoxia-induced apoptosis, and cell cycle distribution, were determined to correlate to changes in DNA-dsb gene expression. Induction of hypoxia was confirmed using HIF1alpha and VEGF expression in gas- and CoCl2-treated cultures. Hypoxia (48-72 h of 0.2% O2) decreased RNA expression of a number of HR-related genes (e.g. Rad51, Rad52, Rad54, BRCA1, BRCA2) in both normal and malignant cultures. Similar decreases in RNA pertaining to the NHEJ-related genes (e.g. Ku70, DNA-PKcs, DNA Ligase IV, Xrcc4) were observed. In selected cases, hypoxia-mediated decreases in RNA expression led to decreased DNA-dsb protein expression. CoCl2-treated cultures did not show decreased DNA-dsb protein expression. The ability of hypoxia to down-regulate Rad51 and other HR-associated genes under hypoxia was not correlated to c-Abl or c-Myc gene expression, p53 genotype or function, propensity for hypoxia-mediated apoptosis, or specific changes in cell cycle distribution. Hypoxia can down-regulate expression of DNA-dsb repair genes in both normal and cancer cells. If associated with a functional decrease in DNA-dsb repair, this observation could provide a potential basis for the observed genetic instability within tumor cells exposed to hypoxia.


Feather mites (Acariformes, Astigmata) from marine birds of the Barton Peninsula (King George Island, Antarctica), with descriptions of two new species.
We report on the first investigation of feather mites associated with birds living on the Barton Peninsula (King George Island, Antarctica). We found seven feather mite species of the superfamily Analgoidea from four host species. Two new species are described from two charadriiform hosts: Alloptes (Sternalloptes) antarcticussp. nov. (Alloptidae) from Stercorariusmaccormicki Saunders (Stercorariidae), and Ingrassiachionis sp. nov. (Xolalgidae) from Chionisalbus (Gmelin) (Chionidae). Additionally, we provide partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI), which was utilized as a DNA barcode, for all seven feather mite species.


Hematopoietic prostaglandin D2 synthase in the chicken Harderian gland.
The Harderian gland (HG), a sero-mucous secreting organ in the eye orbit, has long been recognized as immunologically important in chickens. During experimentation to characterize immune components of the gland, proteomics analysis revealed the presence of hematopoietic prostaglandin D synthase (H-PGDS). Extraction of total RNA followed by RT-PCR produced cDNA of 597 base pairs. DNA sequencing revealed nucleic acid and predicted amino acid sequences that were 99% aligned with the one published sequence for chicken H-PGDS of the spleen. Alignment with murine, rat, and human H-PGDS were 69, 69, and 66%, respectively. Ocular vaccination of chickens with a Newcastle Disease/Infectious Bronchitis vaccine (Mass.-Ark. Strain) induced an increase in H-PGDS expression determined by real-time PCR. Furthermore, immunohistochemistry of frozen HG sections showed positive stained cells for both H-PGDS and mast cell tryptase in the sub-epithelial cell layers of the HG ducts. Based on the potent vasoactive role of PGD(2), it appears that the chicken HG is a site of active mucosal immunity partially mediated by PGD(2) synthesized by H-PGDS in the gland.


Antimicrobial potential of endophytic fungi from Astragalus chinensis.
The purpose of the present study was to discover antimicrobial endophytic fungi from Astragalus chinensis. Three fungal endophytes with antibacterial activity were isolated and determined as Chaetomium sp. HQ-1, Fusarium sp. HQ-7 and Fusarium sp. HQ-9 based on the neighbor-joining phylogenetic tree. Chaetomium sp. HQ-1 showed the best antibiotic potential and was thus selected for large-scale fermentation. Bioactivity-directed separation of ME fermentation of strain HQ-1 led to the discovery of three compounds, which were identified as differanisole A (1), 2,6-dichloro-4-propylphenol (2) and 4,5-dimethylresorcinol (3), from the HR-ESI-MS and NMR data analysis. All three compounds exhibited moderate antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and methicillin-resistant S. aureus, with MIC values ranging from 16 to 128 μg/mL. Compounds 1 and 3 also displayed promising antifungal activity against Selerotium rolfsii with IC50 values of less than 16 and 32 μg/mL, respectively, which were comparable to that of actidione (8 μg/mL). The findings of the present study suggest that the endophytic fungi from A. chinensis have the potential to be used as bactericides and fungicides.