Aldosterone signaling defect in young infants: single-center report and review.
Aldosterone (Ald) is a crucial factor in maintaining electrolyte and water homeostasis. Defect in either its synthesis or function causes salt wasting (SW) manifestation. This disease group is rare, while most reported cases are sporadic. This study aimed to obtain an overview of the etiology and clinical picture of patients with the above condition and report our rare cases. A combination of retrospective review and case studies was conducted at the Pediatric Endocrine unit of The First Affiliated Hospital Sun Yat Sen University from September 1989 to June 2020. A total of 187 patients with SW were enrolled, of which 90.4% (n = 169) were diagnosed with congenital adrenal hyperplasia (CAH). SW type 21-hydroxylase deficiency accounted for 98.8% (n = 167) of CAH diagnosis, while 1.2% (n = 2) was of lipoid CAH. Non-CAH comprised 9.6% (n = 18) of the total patients whose etiologies included SF-1 gene mutation (n = 1), X-linked adrenal hypoplasia congenita (n = 9), aldosterone synthase deficiency (ASD, n = 4), and pseudo-hypoaldosteronism type 1 (PHA1, n = 1). Etiologies were not identified in three patients. All of patients with ASD and PHA1 exhibited SW syndrome in their early neonatal period. DNA sequencing showed mutations of CYP11B2 for P1-P4 and NR3C2 for P5. P1 and P2 were sibling brothers affected by compound heterozygous mutations of c.1121G > A (p.R374Q) and c.1486delC p.(L496fs); likewise, P4 was identified with compound heterozygous mutations of c.1200 + 1G > A and c.240-1 G > T; meanwhile P3 demonstrated c.1303G > A p.(G435S) homozygous mutation in CYP11B2 gene. Lastly, P5 showed c.1768 C > T p.(R590*) heterozygous mutation in the NR3C2 gene. Etiology of infant with aldosterone defect was mostly congenital. Renal and adrenal imaging are recommended to exclude renal causes. If clinical picture is suggestive, normal plasma Ald in early infancy cannot rule out aldosterone insufficiency.


Biodegradable polymeric vectors for gene delivery to human endothelial cells.
Endothelial cells are an important cell type to both cardiovascular disease and cancer, as they play critical roles in vascular function and angiogenesis. However, effective and safe gene delivery to primary endothelial cells in the presence of serum proteins is known to be particularly challenging. A library of biodegradable poly(beta-amino esters) was synthesized for use as potential vectors. Promising vectors were optimized for high efficacy and low cytotoxicity to human umbilical vein endothelial cells (HUVECs) in serum. Vector parameters including polymer type, polymer weight, and DNA loading were varied, and biophysical properties including particle size, zeta potential, and particle stability over time were studied. While many of the poly(beta-amino ester) vectors have similar biophysical properties in the presence of buffer, their biophysical properties changed differentially in the presence of serum proteins, and the properties of these serum-interacting particles correlated to transfection efficacy. Leading poly(beta-amino ester) vectors were found to transfect HUVECs in the presence of serum significantly higher (47 +/- 9% positive, n = 10) than the best commercially available transfection reagents including jetPEI (p < 0.001) and Lipofectamine 2000 (p < 0.01). These results demonstrate the potential of a new class of biomaterials, poly(beta-amino esters), for effective human endothelial cell gene therapy.


Evaluation of Hepatoprotective Effect of Leaves of Cassia sophera Linn.
In the present study, the hepatoprotective activity of ethanolic extracts of Cassia sophera Linn. leaves was evaluated against carbon-tetrachloride- (CCl(4)-) induced hepatic damage in rats. The extracts at doses of 200 and 400 mg/kg were administered orally once daily. The hepatoprotection was assessed in terms of reduction in histological damage, changes in serum enzymes, serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP), total bilirubin, and total protein levels. The substantially elevated serum enzymatic levels of AST, ALT, ALP, and total bilirubin were restored towards the normalization significantly by the extracts. The decreased serum total protein level was significantly normalized. Silymarin was used as standard reference and exhibited significant hepatoprotective activity against carbon tetrachloride-induced hepatotoxicity in rats. The biochemical observations were supplemented with histopathological examination of rat liver sections. The results of this study strongly indicate that Cassia sophera leaves have potent hepatoprotective action against carbon tetrachloride-induced hepatic damage in rats. This study suggests that possible activity may be due to the presence of flavonoids in the extracts.


Construction of a tumor cell-targeting non-viral gene delivery vector with polyethylenimine modified with RGD sequence-containing peptide.
The objective of the present study was to construct a novel type of non-viral gene delivery vector with high delivery efficiency and specific tumor cell-targeting ability. The CP9 peptide (CYGGRGDTP) containing Arg-Gly-Asp sequence was employed to be conjugated onto polyethylenimine (PEI) to act as the role of the targeting moiety. The chemical linker, N-succinimidyl-3-(2-pyridyldithio) propionate, was applied during the synthesis of the vector (CP9-PEI). The physicochemical characteristics of the vector were evaluated by the methods of 1H-nuclear magnetic resonance, Fourier transform infrared spectroscopy, gel retardation assay, electron microscope observation and particle size detection. HepG2 cells were used to verify the gene delivery efficiency and targeting ability by gene delivery procedure and free CP9 peptide inhibition tests. The results showed that the successful synthesis of CP9-PEI and the synthesized vector may efficiently condense plasmid DNA into round particles with diameters of ~200 nm at a polymer/pDNA ratio of 10. CP9-PEI may deliver the reporter gene into HepG2 cells with higher efficiency and the efficiency may be inhibited by the free CP9 peptide. The present study suggested that the modification of PEI with the CP9 peptide is an effective method to construct a novel tumor cell-targeting non-viral vector, and that the novel vector exhibits great prospect in the field of cancer gene therapy.


Clinical epidemiology of occupational neurotoxic disease.
Occupational exposures to neurotoxic chemicals have produced large outbreaks of illness in chemical and pesticide workers worldwide. Outbreaks of occupational neurologic disease in the United States have included (1) the Kepone episode in Hopewell, Virginia, in which 76 workers at a pesticide plant producing the chlorinated hydrocarbon insecticide, Kepone, developed a previously unrecognized syndrome of nervousness, tremor, ataxia, weight loss, opsoclonus, pleuritic and joint pain, and oligospermia; (2) an outbreak of 104 cases of autonomic neuropathy in polyurethane foam workers in Marblehead, Massachusetts, manifest principally by urinary bladder dysfunction, which followed exposure to a new catalyst, dimethylaminopropionitrile (DMAPN); and (3) an outbreak of acute mixed motor and sensory neuropathy in 48 plastic fabric workers in Columbus, Ohio, exposed to the solvent methyl butyl ketone (MBK). These outbreaks underscore the vulnerability of chemical workers to neurotoxins. In addition, occurrence of these large, easily detectable epidemics suggests that many more smaller clusters and single cases of neurologic disease of undetermined origin, particularly in younger adults, may be caused by exposure to occupational or to other toxic chemicals. Detection of the etiology of chemically induced neurologic illness requires a high index of suspicion and careful ascertainment of occupational history.


Pastoral Care and Counseling with Military Families.
The complex human experience of military service and the stress suffered by millions of military families each time a loved one deploys present unique challenges and opportunities in providing pastoral care and counseling. War and military service impact many facets of our society, as well as generational and interpersonal relationships. This article speaks to both academic and practitioner communities, and provides a vision for effective pastoral care and counseling with military families drawing on resources from family systems theory.


Critically ill patients with COVID-19 with ECMO and artificial liver plasma exchange: A retrospective study.
COVID-19 is an emerging infectious disease capable of causing severe pneumonia. We aimed to characterize a group of critically ill patients in a single-center study.This was a retrospective case series of 23 patients with confirmed COVID-19-related critical illness in the intensive care unit (ICU) of a hospital in Hangzhou Zhejiang Province between January 22 and March 20, 2020.Of the 23 critically ill patients, the median age was 66 years (interquartile range [IQR] 59-80 years). The median time from disease onset to ICU admission was 10 days (IQR 6-11 days), to mechanical ventilation (MV) was 11 days (IQR 7.75-13 days), to artificial liver plasma exchange was 12 days (IQR 9.75-14.75 days), and to extracorporeal membrane oxygenation (ECMO) was 22 days (IQR 17.5-30 days). Nine patients required high flow oxygen. Fourteen patients received MV. Six required ECMO. Nine received artificial liver plasma exchange. Mortality was 0 at day 28.Mortality was 0 at day 28 in our single-center study. Extracorporeal membrane oxygenation reduced the requirements for ventilator support. Artificial liver plasma exchange significantly reduced inflammatory cytokine levels. These supportive therapies helped to extend the patients' survival times and increase the chance of follow-up treatments.


Sepsis in Canadian children: a national analysis using administrative data.
Severe infection resulting in sepsis is recognized as a leading cause of morbidity and mortality worldwide. The purpose of this study is to use longitudinal, population-based data to report national-level hospital metrics, providing a current assessment of the status of sepsis hospitalizations in Canadian children. We performed an analysis of previously abstracted data from the Canadian Institute for Health Information (CIHI) Discharge Abstract Database (DAD). Children aged 0-17 years at the time of hospital admission were identified from a cohort of patients with sepsis or severe sepsis using the International Classification of Diseases and Related Health Problems, 10th Revision (ICD-10-CA) and the Canadian Classification of Health Interventions (CCI). Descriptive population-based statistics are reported. Hospitalization data for 20,130 children admitted over 5 years were reviewed. The majority of children were young, with neonates (56.3%) and infants under 2 months (18.8%) representing the majority of cases. A decline in age-adjusted hospitalization rates was demonstrated in both overall and non-severe sepsis across the study period; however, no change was demonstrated for severe sepsis. While overall in-hospital crude mortality rates did not change significantly across the study period (range 5.1%-5.4%), a significant decrease was found in children aged 3-23 months and adolescents. Multi-organ failure was reported in more than one-quarter of children with severe sepsis. Odds of mortality increased significantly with number of organs failed. Sepsis remains an important cause of morbidity and mortality in Canadian children, posing a significant burden on health care resources. Age continues to be associated with the incidence and severity of illness. Overall hospitalization rates have declined over time, as has mortality in severe sepsis. This report provides baseline metrics for future outcome-based research in Canada targeting prevention strategies and early diagnosis, as well as therapies preventing and managing organ failure.


Sudden infant death syndrome, child sexual abuse, and child development.
Since the introduction of the Back to Sleep Campaigns, there has been a dramatic reduction in sudden infant death syndrome in this country. Steven Blatt and Victoria Meguid review the literature surrounding sleep position. Investigators have continued efforts to find other modifiable risk factors of sudden infant death syndrome. A prospective study of more than 33,000 neonates found a link between a prolonged QT electrocardiogram interval and sudden infant death syndrome. Also discussed are investigations seeking to explain the relationship between smoking and sudden infant death syndrome. Ann Botash, Florence Jean-Louis and Mongkae Ploy Siripornsawan review the latest thinking on genital warts and their relation to specific viral etiologies and child sexual abuse. Other symptoms and signs of sexual abuse are the focus of a number of articles that can help the practitioner care for these unfortunate children. Catherine Church reviews medication options for children diagnosed with pervasive developmental disorders or autism spectrum disorders. Finally, in this article, risperidone, fluoxetine and naltrexone are reviewed.


A CONTROLLED TRIAL OF DIGOXIN IN THE PREVENTION OF THE RESPIRATORY DISTRESS SYNDROME.
The possibility that cardiac failure may be an important contributory or additive factor has led to the sporadic use of digitalis in the treatment of the respiratory distress syndrome in newborn infants. To assess the value of such medication a double-blind controlled study was conducted on 196 newborn infants, using digoxin and a placebo. As a result of the findings in this study the routine use of digoxin for the prevention of the respiratory distress syndrome is not recommended. The toxic effects of digitalis are outlined.


Length-biased semi-competing risks models for cross-sectional data: an application to current duration of pregnancy attempt data.
Cross-sectional length-biased data arise from questions on the at-risk time for an event of interest from those who are at-risk but have yet to experience the event. For example, in the National Survey on Family Growth (NSFG), women who were currently attempting to become pregnant were asked how long they had been attempting pregnancy. Cross-sectional survival analysis methods use the observed at-risk times to make inference on the distribution of the unobserved time-to-failure. However, methodological gaps in these methods remain such as how to handle semi-competing risks. For example, if the women attempting pregnancy had undergone fertility treatment during their current pregnancy attempt. In this paper, we develop statistical methods that extend cross-sectional survival analysis methods to incorporate semi-competing risks. They can be used to estimate the distribution of the length of natural pregnancy attempts (i.e., without fertility treatment) while correctly accounting for women that sought fertility treatment prior to being sampled using cross-sectional data. We demonstrate our approach based on simulated data and an analysis of data from the NSFG. The proposed method results in separate survival curves for: time-to-natural-pregnancy, time-to-fertility treatment, and time-to-pregnancy after fertility treatment.


Introduction of Li into Ag-Based Noncentrosymmetric Sulfides for High-Performance Infrared Nonlinear Optical Materials.
The understanding of the structure-performance relationship is beneficial to establish the balance between the second-harmonic generation (SHG) response and laser-induced damage threshold (LIDT) in exploring novel nonlinear optical (NLO) materials. In this work, a facile strategy of partial Li co-occupation with Ag is employed to investigate the structures and NLO performances of the four Ag-based noncentrosymmetric chalcogenides Ag2In2SiS6 (1), (Li0.79Ag1.21)In2SiS6 (2), (Li1.12Ag0.88)In2SiS6 (3), and (Li1.44Ag0.56)In2SiS6 (4). Both the SHG intensity and LIDT are improved as the Li element is introduced, among which 4 exhibits the optimal phase-matchable SHG response of ∼0.8 × AgGaS2 (AGS) and a high LIDT of 4 × AGS. These NLO activities are explained by theoretical calculation and the dipole moment analysis.


On-line preconcentration and determination of six sulfonylurea herbicides in cereals by MEKC with large-volume sample stacking and polarity switching.
A new MEKC method with large-volume sample stacking and polarity switching was developed for on-line preconcentration and detection of sulfonylurea herbicide (SUH) residues in cereals, including nicosulfuron (NS), thifensulfuon (methyl) (TFM), tribenuron-methly (TBM), sulfometuron-methyl (SMM), pyrazosulfuron-ethyl (PSE), and chlorimuron-ethyl (CME). In order to achieve a high resolution and enrichment factor, several parameters were optimized, such as the pH of the running buffer, the concentration of the BGE and the SDS, the separate voltage, the sample size, the pH, and the electrolyte concentration of the sample. The optimal running buffer was composed of 30 mM borate and 80 mM SDS at pH 7.0. The borate concentration in the sample was 30 mM and the pH value of the sample was the same as that of the running buffer. The concentrating voltage and the separating voltage were -15 kV and 15 kV, respectively. The sample size was 1.455 kPa × 780 s (33.11 cm). Under the optimum conditions, for NS, TFM, TBM, SMM, PSE, and CME, the enrichment factors were 613, 642, 835, 570, 709, and 599; the LODs were 0.29-0.50 ng/g, 0.22-0.36 ng/g, 0.60-0.89 ng/g, 0.39-0.72 ng/g, 0.28-0.56 ng/g, and 0.31-0.57 ng/g; the LOQs of six SUHs were all 5 ng/g; the average recoveries of the spiked sample were 86.68-92.99%, 80.73-93.65%, 81.49-94.40%, 82.97-95.1%, 82.96-98.84%, and 80.41-92.94%, respectively.


Considerations on the effects of tidal regimes in the movement of floating litter in an estuarine environment: Case study of the estuarine system of Santos-São Vicente, Brazil.
Floating litter in the estuarine system of Santos-São Vicente is common and is part of day-to-day activities of various users of the area. The objective of the present study was to carry out a quali-quantitative evaluation of the occurrence of floating litter, to infer their sources, and to identify environmental factors that are likely to control occurrence and distribution, with particular emphasis on the effects of tidal regimes. Six sampling stations were selected along the aforementioned estuary and visited monthly between July 2010 and January 2012. Floating litter was counted from a fixed sampling station. Plastics prevailed (89.64%) and their main source was domestic activities (55.41%). More litter was found during ebb spring tides, with higher concentrations obeying confluence patterns of the estuary's channels. Results indicated that occurrence can be attributed to the deficiency in basic sewage system in the area and the deliberate disposal into the estuary by the local population.


Immediate loading with fixed implant-supported restorations in an edentulous patient with an HIV infection: a case report.
Oral rehabilitation of edentulous patients with implants that are immediately loaded is a viable treatment alternative today; however, the immediate loading protocol has not been extensively documented in HIV-seropositive patients. This clinical case report presents a surgical and prosthetic implant-supported immediate loading protocol in an edentulous HIV-seropositive patient (CD4 >200 and T4/T8: 0.99). Eight implants were placed in each jaw, and no ridge augmentation procedures were required. All the implants were connected with abutments, and a temporary fixed restoration was placed immediately after surgery. The final fixed reconstruction was cemented after 10 weeks of loading. All implants showed no clinical signs of mobility or infection. Periotest values were evaluated during healing and showed a progressive reduction. Radiological findings showed stable bone levels after an observation period of 4 years. This case report presents an alternative treatment for the oral rehabilitation of an HIV-positive (but disease asymptomatic) edentulous patient. The concept of immediate loading of implants offers advantages when compared with a traditional treatment modality even in immunocompromised patients who are well controlled.


Real-Time Visualization of Embryonic Apoptosis Using a Near-Infrared Fluorogenic Probe for Embryo Development Evaluation.
Developing an accurate and reliable detection technique for early embryonic apoptosis is of great significance for real-time monitoring and evaluation of embryonic development in living systems. Herein, we have rationally designed and synthesized a novel near-infrared (NIR) fluorogenic probe CGK(QSY21)DEVD-Cy5.5 for real-time imaging of embryonic apoptosis. This probe is constructed with a NIR dye Cy5.5, a fluorescence quencher QSY21, and a peptide substrate Asp-Glu-Val-Asp (DEVD) of the caspase-3 enzyme that is a key executor of cell apoptosis. The probe was initially nonfluorescent in aqueous solution but emitted strong NIR fluorescence upon specific cleavage by activated caspase-3 in a concentration-dependent manner. Taking advantage of this unique feature, this fluorogenic probe was for the first time used for real-time imaging of caspase-3 activity in apoptotic embryos. More notably, significant fluorescence enhancement was solely determined from the apoptotic embryos with the treatment of the probe both in vitro and in vivo, highly suggesting that this probe has great potential to monitor the apoptosis of embryos. We thus envision that this probe would provide a very useful means for real-time visualization and accurate evaluation of embryonic development in the future.


The injured child.
Pre-hospital immediate care for seriously injured children is rarely required, but when it is, the response must be prompt and effective. The key to an effective and confident approach to injured children lies in understanding the age related anatomical and physiological differences between adults and children. These differences are most exaggerated in the first few years of life and excellent training courses and materials are available to help practitioners develop their confidence and skills in this age group. An easy to use length based aide memoire and a set of equipment packed according to size are essential to ensure safe management in the pre-hospital environment. Care of the seriously ill child, emergency childbirth and neonatal resuscitation are beyond the scope of this article. Nonetheless, pre-hospital practitioners should develop an understanding of assessment of ill children and normal delivery and be prepared to assist with emergency childbirth and neonatal resuscitation. Excellent educational material, courses and web based resources are available to achieve this (54). This is the sixth article in the add on series in pre-hospital care. The series is edited by Maj R Mackenzie. Maj RC Sutcliffe is a general practitioner and regimental medical officer. Maj R Mackenzie is a Specialist Registrar in Accident and Emergency Medicine and an immediate care doctor. While every effort has been made to ensure correct drug dosages are quoted, readers should always check these before use.


Separation and detection of lipoproteins in human serum by use of size-exclusion liquid chromatography: a preliminary report.
Components of human serum can be separated on the basis of differences in relative molecular mass by using size-exclusion "high-performance" liquid chromatography. Lipoproteins in fractions of the eluate can be quantitated by conventional chemical and enzymatic methods. Alternatively, if lipoproteins in the serum are selectively prestained with diformazan dye, the column effluent can be monitored spectrophotometrically at 580 nm, so that only the lipoprotein components of serum are detected. Samples of purified low-density lipoproteins, so stained and analyzed, provide peak-area values that are proportional to their concentration as evaluated by chemical methods. With this technique, the various lipoprotein classes can be quickly separated and their concentration estimated. These techniques should prove useful in clinical and research laboratories.


Discriminative stimulus effect of phenylephrine.
Rats were trained to discriminate phenylephrine in a two-lever, food-motivated operant task by increasing the i.p. administered training dose from 0.8 to 2 mg/kg. Stable discrimination to 2 mg/kg phenylephrine was established and testing of 0.5-2.5 mg/kg was shown to be dose-responsive and allowed for a calculated ED50 value of 0.87 mg/kg. Administration of methoxamine (0.5-6 mg/kg), another alpha 1-adrenoceptor agonist, produced a dose-responsive generalization, whereas only the lowest (0.04 mg/kg) and highest (0.12 mg/kg) doses of the alpha 2-adrenoceptor agonist clonidine produced generalization in the phenylephrine-trained rats. Likewise, yohimbine (2.5-4.5 mg/kg) generalized, whereas pretreatment with prazosin (0.1-0.4 mg/kg), the alpha 1-adrenoceptor antagonist, dose-responsively decreased phenylephrine discrimination. The results would indicate that phenylephrine is capable of controlling differential responding in a drug discrimination paradigm and would suggest that this effect is centrally mediated. The possibility that peripheral administration of phenylephrine (Neosynephrine) affects the brain in chronic users of nasal decongestants is discussed.


Flexible, charged biopolymers in monovalent and mixed-valence salt: Regimes of anomalous electrostatic stiffening and of salt insensitivity.
The conformations of biological polyelectrolytes (PEs), such as polysaccharides, proteins, and nucleic acids, affect how they behave and interact with other biomolecules. Relative to neutral polymers, PEs in solution are more locally rigid due to intrachain electrostatic repulsion, the magnitude of which depends on the concentration of added salt. This is typically quantified using the Odijk-Skolnick-Fixman (OSF) electrostatic-stiffening model, in which salt-dependent Debye-Hückel (DH) screening modulates intrachain repulsion. However, the applicability of this approach to flexible PEs has long been questioned. To investigate this, we use high-precision single-molecule elasticity measurements to infer the scaling with salt of the local stiffness of three flexible biopolymers (hyaluronic acid, single-stranded RNA, and single-stranded DNA) in both monovalent and mixed-valence salt solutions. In monovalent salt, we collapse the data across all three polymers by accounting for charge spacing, and find a common power-law scaling of the electrostatic persistence length with ionic strength with an exponent of 0.66±0.02. This result rules out simple OSF pictures of electrostatic stiffening. It is roughly compatible with a modified OSF picture developed by Netz and Orland; alternatively, we posit the exponent can be explained if the relevant electrostatic screening length is the interion spacing rather than the DH length. In mixed salt solutions, we find a regime where adding monovalent salt, in the presence of multivalent salt, does not affect PE stiffness. Using coarse-grained simulations, and a three-state model of condensed, chain-proximate, and bulk ions, we attribute this regime to a "jacket" of ions surrounding the PE that regulates the chain's effective charge density as ionic strength varies. The size of this jacket in simulations is again consistent with a screening length controlled by interion spacing rather than the DH length. Taken together, our results describe a unified picture of the electrostatic stiffness of polyelectrolytes in the mixed-valence salt conditions of direct relevance to cellular and intercellular biological systems.


Biogenesis, turnover, and mode of action of plant microRNAs.
MicroRNAs (miRNAs) are small RNAs that control gene expression through silencing of target mRNAs. Mature miRNAs are processed from primary miRNA transcripts by the endonuclease activity of the DICER-LIKE1 (DCL1) protein complex. Mechanisms exist that allow the DCL1 complex to precisely excise the miRNA from its precursor. Our understanding of miRNA biogenesis, particularly its intersection with transcription and other aspects of RNA metabolism such as splicing, is still evolving. Mature miRNAs are incorporated into an ARGONAUTE (AGO) effector complex competent for target gene silencing but are also subjected to turnover through a degradation mechanism that is beginning to be understood. The mechanisms of miRNA target silencing in plants are no longer limited to AGO-catalyzed slicing, and the contribution of translational inhibition is increasingly appreciated. Here, we review the mechanisms underlying the biogenesis, turnover, and activities of plant miRNAs.


Supramolecular Enhancement of Electrochemical Nitrate Reduction Catalyzed by Cobalt Porphyrin Organic Cages for Ammonia Electrosynthesis in Water.
The electrochemical nitrate (NO3 - ) reduction reaction (NO3 RR) to ammonia (NH3 ) represents a sustainable approach for denitrification to balance global nitrogen cycles and an alternative to traditional thermal Haber-Bosch processes. Here, we present a supramolecular strategy for promoting NH3 production in water from NO3 RR by integrating two-dimensional (2D) molecular cobalt porphyrin (CoTPP) units into a three-dimensional (3D) porous organic cage architecture. The porphyrin box CoPB-C8 enhances electrochemical active site exposure, facilitates substrate-catalyst interactions, and improves catalyst stability, leading to turnover numbers and frequencies for NH3 production exceeding 200,000 and 56 s-1 , respectively. These values represent a 15-fold increase in NO3 RR activity and 200-mV improvement in overpotential for the 3D CoPB-C8 box structure compared to its 2D CoTPP counterpart. Synthetic tuning of peripheral alkyl substituents highlights the importance of supramolecular porosity and cavity size on electrochemical NO3 RR activity. These findings establish the incorporation of 2D molecular units into 3D confined space microenvironments as an effective supramolecular design strategy for enhancing electrocatalysis.


Computer graphics with computerized tomography for functional neurosurgery.
A computer graphics technique for computer-assisted stereotactic surgery is presented. The program is designed to aid the surgeon by presenting an on-line graphics display of stereotactic probes and electrodes superimposed on cross sections of the human brain stem. This technique simulates an otherwise blind surgical procedure on a graphics screen for use during surgery. An earlier system based around the DEC MINC-11 BA computer system has been used by the authors for the performance of stereotactic surgery with conventional ventriculography. This system has been upgraded and is now configured about an even more compact microprocessor-based hardware system with expanded graphics capabilities, which also allows its use with computerized tomography.


Response of microorganisms to a simulated Martian environment.
A study was undertaken to determine whether terrestrial microorganisms can survive in a simulated Martian environment. The ultimate objective is to establish whether earth organisms can contaminate Mars. In addition, any demonstration of survival and growth in a simulated Martian environment will provide information relating to the biology of Mars. In the experimental design, exhaustive consideration was given to the duplication of the known and the theoretical environmental parameters of Mars. These included composition of the soil and the atmosphere, barometric pressure, moisture content, solar radiation, and diurnal temperature extremes. Based upon these considerations, a simulated Martian summer environment was defined and used in the experiments. One group of microorganisms was selected from culture collections on the basis of their known characteristics. The other group was made of microorganisms isolated from soils. The soil samples were obtained from the Antarctic, from New Mexico, and California deserts, and from the Colorado tundra. The studies showed that a number of microorganisms can survive the simulated Martian environment. However, no substantial growth under such conditions could be demonstrated. The ability of microorganisms to form spores as a mechanism for survival will be discussed. Also, experiments utilizing augmented environments to establish minimum environmental conditions which will permit growth will be described.


Tissue-specific regulation of human alpha 1-antitrypsin gene expression in transgenic mice.
The 5'-flanking sequence of the human alpha 1-antitrypsin (AAT) gene contains multiple cis-regulatory elements, including a distal enhancer and proximal sequences essential for its transcription in cultured hepatoma cells. To understand better the promoter specificity of the AAT gene in vivo, transgenic mice harboring the AAT-SV40 hybrid promoter or the natural AAT promoter fused to a reporter gene (CAT) were generated. Examination of CAT activity in various tissues indicated that the CAT gene was expressed primarily in the liver and also, to a lesser extent, in tissues known to express the AAT gene. In addition, the cis-acting elements of the human AAT gene were utilized to drive the transcription of the SV40 T antigen gene in transgenic mice. Hepatocellular malignancy was found in all founder animals examined, while sporadic occurrence of malignancy was also observed in stomach, pancreas, and kidney. These results verify that the 5'-flanking region of the human AAT gene contains cis-regulatory elements sufficient to confer tissue specificity in vivo.


Binding of cationic porphyrin to isolated and encapsidated viral DNA analyzed by comprehensive spectroscopic methods.
The complexation of tetrakis(4-N-methylpyridyl)porphyrin (TMPyP) with free and encapsidated DNA of T7 bacteriophage was investigated. To identify binding modes and relative concentrations of bound TMPyP forms, the porphyrin absorption spectra at various base pair/porphyrin ratios were analyzed. Spectral decomposition, fluorescent lifetime, and circular dichroism measurements proved the presence of two main binding types of TMPyP, e.g., external binding and intercalation both in free and in encapsidated DNA. Optical melting studies revealed that TMPyP increases the strand separation temperature of both free and native phage DNA and does not change the phase transition temperature of phage capsid proteins. From these findings we concluded that TMPyP binding does not influence the protein structure and/or the protein-DNA interaction. A combined analysis of absorption spectra and fluorescence decay curves made possible the determination of concentrations of free, externally bound, and intercalated porphyrin. As a perspective, our results facilitate a qualitative analysis of the TMPyP binding process at various experimental conditions.


Quantitative phosphoproteome analysis of Bacillus subtilis reveals novel substrates of the kinase PrkC and phosphatase PrpC.
Reversible protein phosphorylation on serine, threonine, and tyrosine (Ser/Thr/Tyr) residues plays a critical role in regulation of vital processes in the cell. Despite of considerable progress in our understanding of the role of this modification in bacterial physiology, the dynamics of protein phosphorylation during bacterial growth has rarely been systematically addressed. In addition, little is known about in vivo substrates of bacterial Ser/Thr/Tyr kinases and phosphatases. An excellent candidate to study these questions is the Gram-positive bacterium Bacillus subtilis, one of the most intensively investigated bacterial model organism with both research and industrial applications. Here we employed gel-free phosphoproteomics combined with SILAC labeling and high resolution mass spectrometry to study the proteome and phosphoproteome dynamics during the batch growth of B. subtilis. We measured the dynamics of 1666 proteins and 64 phosphorylation sites in five distinct phases of growth. Enzymes of the central carbon metabolism and components of the translation machinery appear to be highly phosphorylated in the stationary phase, coinciding with stronger expression of Ser/Thr kinases. We further used the SILAC workflow to identify novel putative substrates of the Ser/Thr kinase PrkC and the phosphatase PrpC during stationary phase. The overall number of putative substrates was low, pointing to a high kinase and phosphatase specificity. One of the phosphorylation sites affected by both, PrkC and PrpC, was the Ser281 on the oxidoreductase YkwC. We showed that PrkC phosphorylates and PrpC dephosphorylates YkwC in vitro and that phosphorylation at Ser281 abolishes the oxidoreductase activity of YkwC in vitro and in vivo. Our results present the most detailed phosphoproteomic analysis of B. subtilis growth to date and provide the first global in vivo screen of PrkC and PrpC substrates.


Predictors of disease severity and outcomes in pediatric patients with croup and COVID-19 in the pediatric emergency department.
Croup caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging disease, and data on the risk factors associated with disease severity are still limited. The Westley croup score (WS) is widely used to assess croup severity. The current study aimed to analyze biomarkers associated with the WS and clinical outcomes in patients with croup and coronavirus disease 2019 in the pediatric emergency department (PED). Patients diagnosed with croup caused by SARS-CoV-2 were admitted at two PEDs. Clinical data including age, WS, length of hospital stay, initial laboratory data, and treatment were analyzed. Clinical parameters were evaluated via multivariate logistic regression analysis. The best cutoff values for predicting croup severity and outcomes were identified using the receiver operating characteristic curve. In total, 250 patients were assessed. Moreover, 128 (51.2%) patients were discharged from the PED, and 122 (48.8%) were admitted to the hospital. Mild, moderate, and severe croup accounted for 63.6% (n = 159), 32% (n = 80), and 4.4% (n = 11) of all cases, respectively. A high mean age (years), neutrophil count (%), neutrophil-to-lymphocyte ratio (NLR), ALT (U/L), procalcitonin (ng/mL), and hemoglobin (g/dL) level, and length of hospital stay (days), and a low lymphocyte count (%) and blood pH were associated with croup severity and need for intensive care. Based on the multivariate logistic regression model, the NLR remained independent factors associated with croup severity and prognosis. Further, NLR was significantly correlated with WS. The area under the receiver operating characteristic curve of NLR for predicting a WS of ≥3 was 0.895 (0.842-0.948, p < 0.001), and that for predicting ICU admission was 0.795 (0.711-0.879, p < 0.001). The best cutoff values for a WS of ≥3 and ICU admission were 1.65 and 2.06, respectively. NLR is correlated with WS and is a reliable, easy-to-use, and cheap biomarker for the early screening and prognosis of croup severity in the PED. A higher NLR may indicate severe croup and the need for further treatment. And the WS score remains reliable for estimating the severity of croup caused by SARS-CoV-2 and the risk of intensive care.


Bioactive Peptides: Synthesis, Properties, and Applications in the Packaging and Preservation of Food.
Bioactive peptides are protein fragments which have a positive impact on the functions and conditions of living beings. Peptides have shown several useful properties for human health, including antimicrobial, antifungal, antiviral, and antitumor activities. These compounds are produced by almost all species of life. However, they are produced in limited quantities in nature. As a result, researchers have tried to synthesize bioactive peptides to study their properties and applications in various areas. Among their applications in food preservation, peptides have been incorporated into packaging materials. This review begins with a brief description of the methods used for the synthesis, purification, and characterization of peptides. Also, the main bioproperties and mechanisms of action of peptides are discussed. Finally, some applications of peptides are presented, especially their use in active packaging, their effects on the polymeric matrix, and peptide migration.


Adequate prescribing of medication does not necessarily translate into good control of diabetes mellitus.
Patients with diabetes mellitus in Lithuania have access to almost all the latest blood glucose-lowering drugs available in the rest of the world. This study evaluated the effects of prescribing of treatment (oral medications, insulin, or both) in Lithuanian patients with type 1 or 2 diabetes and poor blood glucose control. The relevant information was obtained from specialized questionnaires completed by 26 consulting endocrinologists in Lithuania between October 1, 2008 and December 31, 2008. The study cohort comprised 865 randomly selected patients with diabetes mellitus and a glycosylated (HbA1c) level ≥7%. In total, there were 95 patients with type 1 diabetes and 770 with type 2 diabetes. Linear regression for patients with type 1 diabetes revealed a weak trend towards higher doses of insulin reflecting lower HbA1c values. The mean dose of insulin in patients with type 1 diabetes before an endocrinology consultation was 57.1 ± 15.7 U/day (0.8 ± 0.2 U/kg), which increased significantly to 63.3 ± 16.5 U/day (0.9 ± 0.2 U/kg) after an endocrinology consultation (P < 0.05). Treatment prescribed for patients with type 2 diabetes depended on the duration of disease. Earlier treatment recommended for 68% of patients with type 2 diabetes was subsequently changed by the endocrinologist. Linear regression showed that the insulin dose prescribed before a specialist consultation as well as that recommended by an endocrinologist was significantly correlated with body mass index. Appropriate prescribing of blood glucose-lowering drugs does not always translate into good metabolic control of diabetes mellitus. The mean HbA1c was 8.5% ± 1.3% in patients with type 2 diabetes treated with oral drugs alone versus 9.0% ± 1.3% in those treated with insulin alone.


Activated and resting regulatory T cell exhaustion concurs with high levels of interleukin-22 expression in systemic sclerosis lesions.
Transforming growth factor-β is considered to play a key role in the process of fibrosis in systemic sclerosis (SSc) and in the development of regulatory T cells (Treg) and pro-inflammatory Th17 T cells producing interleukin 17 (IL-17) and IL-22. The authors therefore postulated that SSc could be characterised by a marked Treg/Th17 imbalance. Previous works did not distinguish between the different subsets of Treg and the non-regulatory FoxP3(+) cells leading to inconsistent results. Combined phenotypic and functional analysis of Th17 cells and FoxP3(+)CD4 T cells, discriminating activated Tregs and resting Tregs from non-regulatory FoxP3(+) T cells, in blood and skin of SSc patients. In early disease stages, there is a decreased proportion of activated Tregs. A concomitant resting Treg deficit becomes more apparent with disease progression. Active and diffuse forms of the disease are characterised by a relatively higher proportion of all FoxP3(+) subsets, including non-regulatory T cells. No peripheral or local IL-17 amplification was observed. However, the authors found significantly increased IL-22 transcription levels in SSc lesional skin, as compared with healthy skin. Cytofluorometry confirmed the existence in SSc patients and controls of a distinct subset of T cells producing IL-22 in the absence of IL-17. SSc pathogenesis does not appear to be linked to IL-17-, but rather to IL-22-producing cells with skin-homing potential and a concomitant quantitative Treg defect. Active and diffuse forms of the disease are associated with a FoxP3 signature. Altogether, our data depict a status of regulatory/pro-inflammatory T cell imbalance in SSc.


Adipose-derived mesenchymal stem cell-loaded β-chitin nanofiber hydrogel activates the AldoA/HIF-1α pathway to promote diabetic wound healing.
To identify the effect of adipose-derived mesenchymal stem cell-loaded β-chitin nanofiber (ADSC-loaded β-ChNF) hydrogel on diabetic wound healing and clarify its mechanism of action. We prepared the ADSC-loaded β-ChNF hydrogel to repair wounds of db/db diabetic mice. Wound healing rate, histopathology, enzyme-linked immunosorbent assay, and western blot were used to confirm its role and mechanism in promoting diabetic wound healing. The ADSC-loaded β-ChNF hydrogel accelerated wound healing in db/db diabetic mice, as indicated by increased cell proliferation, epithelization, and tissue granulation in the skin. Moreover, expression of vascular endothelial growth factor (VEGF) and its receptor (VEGFR), matrix metalloproteinase 9 (MMP9), and TIMP metallopeptidase inhibitor 1 (TIMP1) were upregulated. These results demonstrate the beneficial effects of this ADSC-loaded β-ChNF hydrogel on diabetic wound healing. Furthermore, we show that the ADSC-loaded β-ChNF hydrogel activated aldolase A (AldoA)/hypoxia-inducible factor 1α (HIF-1α) signaling. An inhibitor of HIF-1α markedly decreased the promotive effects of the ADSC-loaded β-ChNF hydrogel on wound healing and reduced expression of VEGF, VEGFR, MMP9, and TIMP1. Our findings suggest that the ADSC-loaded β-ChNF hydrogel activated the HIF-1α/MMP9 axis through AldoA feedback to promote diabetic wound healing.


Seasonal trends in the prevalence of hypospadias: Aetiological implications.
The aim of the present study was to examine the seasonality of hypospadias in Greece in an attempt to elucidate the aetiology. All boys born between 1991-1998, who underwent hypospadias repair at 'Aghia Sophia' Children's Hospital, Athens (n=542) were analysed. All Greek live-born males during the same period (population at risk; m=421,175) served as the controls. Seasonality by month of birth was evaluated with specific statistical tools. Meteorological parameters were also analysed. All tests yielded significant results, suggesting a simple harmonic prevalence pattern (highest/lowest: autumn, peak in October/spring, trough in April). Therefore, the first trimester of hypospadiac gestations coincides more frequently with winter. Meteorological parameters varied seasonally (maximal sunlight; air temperature in summer/minimal in winter, maximal rainfall in winter/minimal in summer) and were strongly associated pairwise. Hypospadiac birth prevalence follows a simple harmonic seasonal pattern and is associated with that of cryptorchidism in Greece. The coincidence of the first or third trimester of a potentially genetically influenced gestation with winter could lead to the phenotypic expression of hypospadias or cryptorchidism, respectively. The potential role of a cyclic-varied androgen-production stimulator, such as human chorionic gonadotrophin may be speculated. The seasonality of a common environmental factor acting directly/indirectly may contribute to these patterns, and possibly to the common pathogenesis of these congenital malformations.


Experiencing hypnotizability scale motor items by an amputee: a brief report.
The following brief report describes the experiences of a hand and arm amputee following the administration of the Harvard Group Scale of Hypnotic Susceptibility, Form A (HGSHS:A) of Shor and Orne, 1962. The participant passed two of the three motor items involving his missing limb. This report discusses the results of a postsession interview regarding our participant's experiences during hypnosis and briefly discusses phantom limb sensations in general.


Outcome of shunting in idiopathic normal-pressure hydrocephalus and the value of outcome assessment in shunted patients.
To develop guidelines for assessing shunt outcome in patients with idiopathic normal-pressure hydrocephalus (INPH). To date, the literature available on this topic has been marked by disparate definitions of clinical improvement, varying postoperative follow-up protocols and periods, and substantial differences in the postoperative management. Because specific criteria for defining clinical improvement are seldom reported, conclusions drawn about shunt outcome may be subjective. A MEDLINE search back to 1966 was undertaken using the query NPH, normal-pressure hydrocephalus, shunting, shunt treatment, shunt response, outcome, and clinical outcome. The criteria for selection were studies that included INPH from 1966 to the present in which the outcome of INPH was reported in patient groups of 20 or more. To date, there is no standard for outcome assessment of shunt treatment in INPH. The variable improvement rates reported are not only because of different criteria for selection of patients but also because of different postoperative assessment procedures and follow-up intervals. Studies that have established fixed protocols for follow-up have shown that short- and long-term periods after shunting are determined by many factors. Whereas short-term results were more likely to be influenced by shunt-associated risks, long-term results were independent of factors inherent to the shunt procedure and shunt complications, i.e., death and morbidity related to concomitant cerebrovascular and vascular diseases. Studies have shown that beyond 1 year after surgery, these factors definitely influence the clinical effect of shunting, making the 1-year postshunt period a potential determinant of the shunt outcome. Guidelines for outcome assessment were developed on the basis of the available evidence and consensus of expert opinion.


Coordination-Driven Syntheses of Compact Supramolecular Metallacycles toward Extended Metallo-organic Stacked Supramolecular Assemblies.
One important concept associated with supramolecular chemistry is supramolecular self-assembly, which deals with the way discrete individual components interact via intermolecular interactions in order to build, upon their spontaneous association, high order functional assemblies. The accumulation of these very simple and localized noncovalent interactions (such as H-bonding, dipole-dipole, hydrophobic/hydrophilic, van der Waals, π-π, π-CH, etc.) is ubiquitous in the complexity of natural systems (such as DNA, proteins, membranes, micelles, etc.). It can also be transposed to the directed synthesis of intricate artificial scaffolds, which have anticipated geometries and properties. Among the synthetic strategies based on this concept, coordination-driven supramolecular chemistry uses the robust, reversible, and directional metal-to-ligand coordinative bond to build discrete metallo-supramolecular architectures. Within the last two decades, coordination-driven supramolecular chemistry has proved to be one of the most powerful contemporary synthetic approaches and has provided a significant number of increasingly complex supramolecular assemblies, which have predetermined sizes and geometries. While much focus has been devoted to architectures bearing internal cavities for host-guest chemistry or to generate specific reactivity, particular attention can also be paid to compact supramolecular assemblies given that their specific structures are characterized by peculiar synthetic guiding rules as well as by alternative long-range self-assembling properties. This Account describes how a preassembled CuI bimetallic clip bearing short intermetallic distances can be used as a U-shaped molecular clip to give general and versatile access to a large variety of original compact supramolecular metallacycles. When this CuI precursor is reacted with various cyano-capped ditopic linkers that have increasing lengths and complexities, specific effects guiding the selective and straightforward syntheses of such compact supramolecular objects are highlighted. Whereas a subtle compromise between the length of the ditopic linkers and the steric bulk of the molecular clip appears to be a purely stereogeometric preliminary parameter to master, lateral interlinker interactions (π-π stacking interactions or aurophilic interactions depending on the nature of the internal cores of the linkers) can circumvent these constraints regardless of the length of the linkers and allow the selective formation of new compact supramolecular structures. Generally, such derivatives presented a strong tendency to self-assemble in the solid state due to inter-supramolecule interactions. This approach thus opens a new door toward molecular materials having an attractive solid state structure for potential applications related to charge carrier mobility and luminescence properties. These compact supramolecular assemblies can therefore be considered as original secondary binding units directing the predictive preparation of such extended networks. The on-purpose design of original building blocks bearing specific cores allowed the formation of new compact supramolecular metallacycles such as "U-shaped" π-stacked assemblies or "pseudodouble paracyclophanes". Similarly, the control of the secondary structure of one-dimensional coordination polymers alternating π-stacked compact supramolecular metallacycles was also conducted. The results that are discussed in this Account illustrate how the rational design of both preassembled polymetallic precursors bearing short intermetallic distances and ditopic linkers able to induce cumulative lateral weak interactions can implement the general synthetic guiding rules of coordination driven supramolecular chemistry. This opens perspectives to use such compact supramolecular assemblies as secondary building blocks for the design of long-range organized functional molecular materials that have predictable architectures and targeted properties.


Persistent rotavirus infection in mice with severe combined immunodeficiency.
Rotaviruses are important pathogens of human infants and the infants of many animal species. The disease produced by these viruses can be described as an acute, self-limiting diarrheal disease, with virus replication localized to the differentiated epithelial enterocytes of the small intestine. Immunologically normal infants shed virus for approximately 5 to 12 days after the onset of infection. Recently, it has been shown that rotavirus can produce a chronic infection in severely immunocompromised children, with virus shedding and intermittent diarrhea lasting from 6 weeks to 2 years (G. A. Losonsky, J. P. Johnson, J. A. Winkelstein, and R. H. Yolken, J. Clin. Invest. 76:2362-2367, 1985; F. T. Saulsbury, J. A. Winkelstein, and R. H. Yolken, J. Pediatr. 97:61-65, 1980). These findings point to an important role for the immune system in recovery from the disease. The study described here examined the outcome of murine rotavirus infection in mice with severe combined B- and T-cell immunodeficiency (SCID) and in immunologically normal seronegative BALB/c mice. Persistent rotavirus infection was established in all mice with SCID which had been inoculated orally as pups. Low levels of virus replication and constant fecal virus shedding characterized the chronic infection. This is the first report of a persistent rotavirus infection in an animal model.


Identification of Duchenne/Becker muscular dystrophy mosaic carriers through a combined DNA/RNA analysis.
The Duchenne/Becker muscular dystrophy (DMD) carrier screening includes the evaluation of mutations in DMD gene, and the most widely used analysis is the multiplex ligation-dependent probe amplification (MLPA) for the DMD deletions/duplications detection. The high frequency of de novo mutations permits to estimate a risk up to 20% of mosaicisms for mothers of sporadic DMD children. The purpose of this study is to evaluate alternative analytical strategy for the detection of mosaics carrier women, in order to improve the recurrence risk estimation. Different DNA and RNA analyses were conducted on samples from a woman that conceived a DMD fetus without previous family history of dystrophynopathy. Standard MLPA analysis failed to identify mosaicism, even if MLPA doses suggested it. Electrophoresis and direct sequencing conducted on RNA permitted to detect two different amplicons of cDNAs, demonstrating the presence of somatic mosaicism. Subsequent detection of a second affected fetus confirmed the mosaic status on the mother. The implementation of RNA analysis in diagnostic algorithm can increase the sensitivity of carrier test for mothers of sporadic affected patients, permitting detection of mosaic status. A revision of analytical guidelines is needed in order to improve the recurrence risk estimation and support prenatal genetic counseling.


Utility of the 3-Vessel and 3-Vessel and Trachea Views in Prenatal Diagnosis of a Pulmonary Artery Sling.
A pulmonary artery (PA) sling is a very rare congenital cardiovascular anomaly, and only a few studies have reported PA slings in fetuses. The relationship of the PA, aorta, ductus arteriosus, and trachea can be evaluated in the 3-vessel and 3-vessel and trachea views during fetal echocardiography. A PA sling can be detected by abnormal positioning of the left PA in relation to the trachea when sweeping from the 3-vessel view cranially to the 3-vessel and trachea view. Here we report 3 cases of fetal PA slings and their follow-ups. Two cases were confirmed by postnatal echocardiography, and the other case was confirmed by a cardiovascular cast after pregnancy termination. We emphasize that the 3-vessel and 3-vessel and trachea views are of crucial importance in the prenatal diagnosis of a PA sling.


Conserved cysteine residues are critical for the enzymatic function of the lymphocyte-specific tyrosine protein kinase p56lck.
We have evaluated the possibility that conserved cysteine residues are critical for the enzymatic function of p56lck. Through oligonucleotide-directed mutagenesis, 5 Lck residues (cysteines 217, 224, 378, 464, and 475) were individually mutated to alanines, and the effects of these substitutions were tested in various in vitro and in vivo assays. We found that mutation of either of 2 cysteines located in the carboxyl portion of the kinase domain (cysteines 464 and 475) abolished the catalytic function of Lck. In addition, it was noted that alteration of cysteine 475 resulted in a dramatic reduction of the half-life of p56lck. These cysteine residues are highly conserved throughout the tyrosine protein kinase family, suggesting that they may play important functions in catalysis and/or substrate recognition.


A randomised crossover simulation study comparing the impact of chemical, biological, radiological or nuclear substance personal protection equipment on the performance of advanced life support interventions.
Recent incidents involving chemical, biological, radiological and nuclear substances have stressed the importance of sufficient personal protection equipment for medical first-responders. Modern lightweight, battery-independent, suit ensembles may prove superior to the current protective suit used in the UK. This study compared the powered respiratory protective suit (PRPS ensemble) with a lightweight suit consisting of a SARATOGA® Multipurpose CBRN Protective Coverall Polyprotect 12 in conjunction with the Avon C50 Respirator/Avon CBRNF12CE filter canister and butyl rubber protective gloves (Polyprotect 12 ensemble). Thirty anaesthetists carried out a standardised resuscitation scenario either unprotected (control) or wearing the PRPS or Polyprotect 12 ensembles in a randomised, crossover simulation study. Treatment times for five simulated advanced life support interventions (application of monitoring; bag/mask ventilation; tracheal intubation; drug and fluid administration; and external pacing) were measured. Wearer comfort was also assessed for the two protective suits by questionnaire. All participants accomplished the treatment objectives of all study arms without adverse events. Total mean (SD) completion time for the five interventions was significantly longer for the PRPS compared with the Polyprotect 12 ensemble (204 (53) s vs. 149 (36) s, respectively; p < 0.0001). Participants rated mobility, noise, heat, vision, dexterity and speech intelligibility significantly better in the Polyprotect 12 ensemble compared with the PRPS ensemble. The combination of a lightweight Polyprotect 12 suit and an Avon C50 air-purifying respirator is preferable to the powered respiratory protective suit during simulated emergency life support, due to a combination of shorter task completion times and improved mobility, communication and dexterity.


Ocular emergencies.
General practitioners, especially those located in country areas, are commonly the first contact for many patients with an ocular emergency. This article aims to provide the GP with a guide to several relatively common ocular emergencies relevant to the general practice setting. It details assessment, initial management and referral planning. Successful patient outcomes in the setting of ocular emergencies depends on correct recognition and assessment as well as appropriate initial management and referral. Assessment involves a concise history, observation, pupil examination; ocular movements should be tested in patients with a suspected orbital wall fracture. Documentation of visual acuity or subjective vision is vital. Important conditions to recognise include penetrating injuries, nonpenetrating injuries, chemical burns, acute angle closure glaucoma, orbital cellulitis and retinal detachment.


Isolation and Characterization of Avian Chlamydia psittaci from Symptomatic Pet Birds in Southern Hunan, China.
Chlamydia psittaci is a zoonotic pathogen with multiple hosts, especially avian, and can be transmitted to humans, causing psittacosis or ornithosis. No effective vaccines have been developed. We therefore isolate and genotype avian C. psittaci strains and investigate the pathogenicity of isolates in the southern Hunan area of China. Among 200 suspicious avian specimens, eight were positive for the C. psittaci outer membrane protein A (ompA) gene (4%), and seven were successfully cultured in human epithelial type 2 and Vero cells (87.5%). Genotyping of the ompA gene of the eight PCR-positive samples revealed that all of the cultured strains, except for the E9 strain, belonged to genotype A. Pathologic changes in the mice infected with C. psittaci via intranasal inoculation showed severe pneumonia and intense infiltration of inflammatory cells in the lung in a dose-dependent manner, and immunohistochemical staining displayed different levels of infiltration of C. psittaci inclusions in the heart, liver, spleen, kidney, and, especially, lung. Our findings demonstrate that genotype A dominates all C. psittaci genotypes in the southern Hunan area and that the C. psittaci avian isolates in this region possess dose-dependent pathogenicity.


Retroperitoneal imaging with third and fourth generation computed axial tomography in clinical stage I nonseminomatous germ cell tumors.
To examine the accuracy rate of abdominal staging using third and fourth generation computed tomography (CT) scanning in clinical Stage I testicular nonseminoma patients. Between January 1985 and August 1993, 57 patients presented to our center with clinical Stage I testicular nonseminoma. Retroperitoneal computed tomographic staging studies were interpreted to be normal preoperatively in the entire group. In addition, tumor marker values were normal or returned to normal postorchiectomy within the appropriate half-life intervals. All patients were subjected to radical or modified retroperitoneal lymph node dissection (19% and 72%, respectively). Original abdominal CT scans (preretroperitoneal lymph node dissection) were available for blinded retrospective re-review in 16 cases (7 pathologic Stage II, 9 pathologic Stage I). Nineteen of 57 (33%) patients were upstaged at surgery including 6 patients (11%) who demonstrated II B volume disease. Third and fourth generation CT scanning of the retroperitoneum yielded a 66% accuracy rate in this population. Six out of 7 pathologic Stage II pre-lymph node dissection abdominal CT scans that were available for blinded re-review revealed nonpathologic nodes by size criteria in the primary landing zone for the corresponding original tumor. Our data suggests that for clinical Stage I nonseminoma in the 1985 to 1993 era, undue reliance was placed on a less than ideal staging test. The 33% false-negative rate reported showed no improvement over earlier reports and reaffirms concern for relying solely on third or fourth generation CT imaging of the retroperitoneum in the staging of clinical Stage I nonseminomatous germ cell tumor (NSGCT) patients. The presence of any number of lymph nodes in the expected primary landing zone, regardless of size, should raise serious suspicion for occult nodal disease.


5-Heptadecylresorcinol Ameliorates Obesity-Associated Skeletal Muscle Mitochondrial Dysfunction through SIRT3-Mediated Mitophagy.
Skeletal muscle dysfunction caused by obesity is characterized by the decline in mitochondrial content and function. 5-Heptadecylresorcinol (AR-C17) is a specific bioactive component derived from whole wheat and rye, which has been evidenced to improve obesity-associated skeletal muscle dysregulation. However, the mechanism underlying its protective activity requires further exploration. Herein, we found that AR-C17 (5, 10, and 20 μM) intervention reversed PA-induced (0.5 mM) reduction in mitochondrial content, mitochondrial membrane potential, and mitochondrial energy metabolism in C2C12 cells. Meanwhile, AR-C17 evidently alleviated PA-mediated myotube mitochondrial dysfunction via elevating mitochondria autophagy flux and upregulating the expression level of autophagy-related protein, while this effect was abolished by an autophagy inhibitor (3-MA). Further analysis showed that SIRT3-FOXO3A-PINK-Parkin-mediated mitophagy was involved in the modulation of myocyte mitochondrial dysfunction by AR-C17. In addition, AR-C17 administration (30 and 150 mg/kg/day) significantly improved high-fat-diet-induced mitochondrial dysregulation in mice skeletal muscle tissue via SIRT3-dependent mitophagy. Our findings indicate that skeletal muscle cells are responsive to AR-C17, which improves myogenesis and mitophagy in vitro and in vivo.


Role of diffusion in excitation energy transfer and migration.
Effect of diffusion on excitation energy transfer and migration in a dye pair sodium fluorescein (donor) and Rhodamine-6G (acceptor) has been studied for different viscosities by both steady state and time domain fluorescence spectroscopic measurements. The donor-donor interaction appears to be weaker as compared to donor-acceptor interaction and thus favors direct Forster-type energy transfer. Interestingly, at low viscosity (water in this case) transfer appears to be controlled by material diffusion/energy migration. Further, acceptor dynamics reveals the fact that direct Forster transfer dominates in viscous media.


Intractable epilepsy of infancy due to homozygous mutation in the EFHC1 gene.
The molecular etiology of primary intractable epilepsy in infancy is largely unknown. We studied a nonconsanguineous Moroccan-Jewish family, where three of their seven children presented with intractable seizures and died at 18-36 months. Homozygous regions were searched using 250 K DNA single nucleotide polymorphism (SNP) array. The sequence of 50 Mb exome of a single patient was determined using SOLiD 5500XL deep sequencing analyzer. A single homozygous 11.3 Mb genomic region on chromosome 6 was linked to the disease in this family. This region contained 110 genes encoding a total of 1,000 exons. Whole exome sequencing revealed a single pathogenic homozygous variant within the critical region. The mutation, Phe229Leu in the EFHC1 gene was previously shown, in a carrier state, to be associated with juvenile myoclonic epilepsy. Although heterozygosity for the Phe229Leu mutation is known to be associated with a relatively benign form of epilepsy in adolescence; homozygosity for the same mutation is associated with lethal epilepsy of infancy. Given the considerable carrier rate of this mutation worldwide, the sequence of the EFHC1 gene should be determined in all patients with primary intractable epilepsy in infancy.


[Neurological disorders in preterm children with neuropathy].
To establish the correlation between the frequency and severity of hypoxic CNS lesions in preterm children with neuropathy and improve the early diagnosis of lesions of the brain structures based on clinical ophthalmologic results. The authors examined 712 premature infants with body mass <1500 g born before 30 weeks of gestation during 2006-2016. Ophthalmological monitoring of retinopathy (RP), an analysis of medical history, neurological examination and neurosonography were performed. RP was found in 367 (51.5%) children. In 255 children, the disease regressed naturally. One hundred and twelve (15.7%) children, underwent laser coagulation of the avascular retina due to the severity of RP. Signs of intraventricular hemorrhages (IVH) were noted in 434 (61%) children in the neonatal period. IVH were found in 285 (77.6%) children with RP. RP with the regression after laser coagulation was combined with IVH in 98% of cases, with the higher frequency (55.3%) of IVH, 3rd degree. Periventricular leucomalation (PVL) was found in 10% of children without RP, in 22.3% of children with RP with naturally regression and in 51,7% of children with RP with laser coagulation of the retina. In 70 children, neurosonographic signs of ischemia of the head of caudate nucleus were identified on the 14-15th days of life. In this group, RP developed in 54 (77%) children, 27 (38.5%) children needed laser coagulation of the retina. The correlation found between the severity of RP and hypoxic CNS lesions in highly preterm infants might allow the prognosis of visual and neurosomatic disturbances in the early age and timely effective rehabilitation.


Direct Monofluoromethylthiolation with S-(Fluoromethyl) Benzenesulfonothioate.
An electrophilic shelf-stable monofluoromethylthiolating reagent S-(fluoromethyl) benezenesulfonothioate (1) was developed. In the presence of a copper catalyst, reagent 1 coupled with a variety of aryl boronic acids to give the corresponding monofluoromethylthiolated arenes in high yields. In addition, addition of reagent 1 to alkyl alkenes in the presence of a silver catalyst gave alkyl monofluoromethylthioethers in high yields.


[Percutaneous ethanol and acetic acid injection for liver metastasis from colon cancer--two case reports].
Two cases of liver metastasis from colon cancer were treated by percutaneous ethanol (PEI) and acetic acid (PAI) injection for the recurrent lesion after surgery. Case 1 was a 60-year-old female who received sigmoidectomy with partial hepatectomy, and intraarterial 5-FU infusion was done after surgery. One year later, recurrence of liver tumor was detected, and PEI and PAI were performed for the metastatic lesions of the liver. Tumor regression and histopathological examination revealed coagulative necrosis. The patient died of lung metastasis 2 years and 10 months after treatment. Case 2 was a 58-year-old-male with ascending colon cancer and liver metastasis, who received surgery, and chemotherapy with intraarterial 5-FU infusion was continued. Four months later, recurrence of liver metastasis with elevation of serum CEA was noted. The patient received PEI three times and CEA decreased. Re-operation of hepatectomy revealed complete necrosis at the site of PEI. The patient has been alive for 1 year and 6 months with a new recurrence in the liver and is receiving repeated PEI therapy. PEI and PAI seem to be useful for the treatment of unresectable liver metastasis.


The analysis of hand movement distinction based on relative frequency band energy method.
For the purpose of successfully developing a prosthetic control system, many attempts have been made to improve the classification accuracy of surface electromyographic (SEMG) signals. Nevertheless, the effective feature extraction is still a paramount challenge for the classification of SEMG signals. The relative frequency band energy (RFBE) method based on wavelet packet decomposition was proposed for the prosthetic pattern recognition of multichannel SEMG signals. Firstly, the wavelet packet energy of SEMG signals in each subspace was calculated by using wavelet packet decomposition and the RFBE of each frequency band was obtained by the wavelet packet energy. Then, the principal component analysis (PCA) and the Davies-Bouldin (DB) index were used to perform the feature selection. Lastly, the support vector machine (SVM) was applied for the classification of SEMG signals. Our results demonstrated that the RFBE approach was suitable for identifying different types of forearm movements. By comparing with other classification methods, the proposed method achieved higher classification accuracy in terms of the classification of SEMG signals.


Physical activity, mental and physical health during the Covid-19 outbreak: longitudinal predictors of suicide ideation in Germany.
Suicide ideation has increased since the outbreak of Covid-19 in many countries. The present longitudinal study investigated potential predictors of suicide ideation. Data of 406 participants from Germany (age M = 27.69, SD = 6.88) were assessed via online surveys in spring 2020 (baseline, BL) and in spring 2021 (follow-up, FU). The current results reveal a significant increase in symptoms of depression, anxiety, and stress between 2020 and 2021. Positive mental health (PMH), sense of control, and physical health significantly decreased. Depression symptoms (BL), PMH (BL), and consciously enhanced physical activity since the pandemic outbreak (FU) significantly predicted 12-month suicide ideation (FU). In a moderated mediation analysis, the positive relationship between depression and suicide ideation was significantly mediated by PMH. Consciously enhanced physical activity significantly moderated the negative association between PMH and suicide ideation. The context of Covid-19 could negatively impact mental health and physical health. This might increase the risk for suicide ideation. However, PMH and physical activity might serve as protective factors. The protective effect of physical activity could be especially important in people with high depression symptoms and low PMH, such as clinical patients. Potential ways of how PMH and physical activity could be enhanced in the Covid-19 context to prevent suicide ideation are discussed.


A subtelomeric non-LTR retrotransposon Hebe in the bdelloid rotifer Adineta vaga is subject to inactivation by deletions but not 5' truncations.
Rotifers of the class Bdelloidea are microscopic freshwater invertebrates best known for: their capacity for anhydrobiosis; the lack of males and meiosis; and for the ability to capture genes from other non-metazoan species. Although genetic exchange between these animals might take place by non-canonical means, the overall lack of meiosis and syngamy should greatly impair the ability of transposable elements (TEs) to spread in bdelloid populations. Previous studies demonstrated that bdelloid chromosome ends, in contrast to gene-rich regions, harbour various kinds of TEs, including specialized telomere-associated retroelements, as well as DNA TEs and retrovirus-like retrotransposons which are prone to horizontal transmission. Vertically-transmitted retrotransposons have not previously been reported in bdelloids and their identification and studies of the patterns of their distribution and evolution could help in the understanding of the high degree of TE compartmentalization within bdelloid genomes. We identified and characterized a non-long terminal repeat (LTR) retrotransposon residing primarily in subtelomeric regions of the genome in the bdelloid rotifer Adineta vaga. Contrary to the currently prevailing views on the mode of proliferation of non-LTR retrotransposons, which results in frequent formation of 5'-truncated ('dead-on-arrival') copies due to the premature disengagement of the element-encoded reverse transcriptase from its template, this non-LTR element, Hebe, is represented only by non-5'-truncated copies. Most of these copies, however, were subject to internal deletions associated with microhomologies, a hallmark of non-homologous end-joining events. The non-LTR retrotransposon Hebe from the bdelloid rotifer A. vaga was found to undergo frequent microhomology-associated deletions, rather than 5'-terminal truncations characteristic of this class of retrotransposons, and to exhibit preference for telomeric localization. These findings represent the first example of a vertically transmitted putatively deleterious TE in bdelloids, and may indicate the involvement of microhomology-mediated non-homologous end-joining in desiccation-induced double-strand break repair at the genome periphery.


Patient Turnover: A Concept Analysis.
Patient turnover influences the quality and safety of patient care. However, variations in the conceptual underpinnings of patient turnover limit the understanding of the phenomenon. A concept analysis was completed to clarify the role of patient turnover in relation to outcomes in the acute care hospital setting. The defining attributes, antecedents, consequences, and empirical referents of patient turnover were proposed. Nursing leaders should account for patient turnover in workload and staffing calculations. Further research is needed to clarify the influence of patient turnover on the quality and safety of nursing care using a unified understanding of the phenomenon.


Iron-Binding Capacity of Defatted Rice Bran Hydrolysate and Bioavailability of Iron in Caco-2 Cells.
The present study was aimed at utilizing defatted rice bran (DRB) protein as an iron-binding peptide to enhance iron uptake in humans. DRB samples were treated with Alcalase and Flavourzyme, and the total extractable peptides were determined. Furthermore, the iron-binding capacities of the DRB protein hydrolysates were determined, whereas iron bioavailability studies were conducted using an in vitro digestion and absorption model (Caco-2 cells). The results showed that the DRB protein hydrolysates produced by combined Alcalase and Flavourzyme hydrolysis had the best iron-binding capacity (83%) after 90 min of hydrolysis. The optimal hydrolysis time to produce the best iron-uptake in Caco-2 cells was found to be 180 min. The results suggested that DRB protein hydrolysates have potent iron-binding capacities and may enhance the bioavailability of iron, hence their suitability for use as iron-fortified supplements.


[Clinical observation on electroacupuncture combined with rehabilitation techniques for treatment of shoulder subluxation after stroke].
To compare the therapeutic effect difference between electroacupuncture combined with rehabilitation techniques and simple rehabilitation techniques for treatment of the shoulder subluxation after stroke. Sixty cases of shoulder subluxation patients were divided into an acupuncture combined with rehabilitation group (30 cases) and a rehabilitation group (30 cases). The acupuncture combined with rehabilitation group was treated with electroacupuncture at Jianyu (LI 15), Jianliao (TE 14) and Jianzhen (SI 9) etc. combined with rehabilitation treatment, the rehabilitation group was treated with rehabilitation only. The therapeutic effects were evaluated and the pain score of the shoulder joint, passive range of motion of shoulder joint, the muscle strength of middle group of the deltoid and the function grade of the upper limbs motion of the two groups were evaluated before and after treatment. The acupuncture combined with rehabilitation group was superior to that of the rehabilitation group in pain score of the shoulder joint, passive range of motion of shoulder joint, the muscle strength of middle group of the deltoid and the function grade of the upper limbs motion with significant differences between the two groups (all P<0.05); the total effective rate of 93. 3% in the acupuncture combined with rehabilitation group was superior to that of 73.3% in the rehabilitation group (P<0.05). The therapy of electroacupuncture combined with rehabilitation techniques can act a good regulating role on muscle strength and the muscular tension of shoulder joint and the muscles around the scapula through many steps, and can effectively improve the shoulder subluxation.


[Severe encephalopathy in Schönlein-Henoch purpura (case report)].
We are reporting on the case of a 7-year old, previously healthy boy with Schönlein-Henoch Purpura complicated by a severe encephalopathy: Two days after the initial abdominal symptoms he showed a prolonged cerebral convulsion followed by a 6-day semicomatous state, right-sided paralysis and aphasia. The typical skin lesions and joint swellings only appeared two days after the seizure. He was treated with acyclovir, followed by beta-interferon, both without apparent improvement. After initiating corticosteroids improvement of the neurological deficits was noticed. After 3 1/2 months he was still left with a right-sided spastic paralysis and mild aphasia. The cranial CT demonstrated initially a left-sided cerebral edema, later developing into a cerebral atrophy involving the periventricular structures as well as cortical regions predominantly of the left side.


Predicting critical temperatures of ionic and non-ionic fluids from thermophysical data obtained near the melting point.
In the correlation and prediction of thermophysical data of fluids based on a corresponding-states approach, the critical temperature Tc plays a central role. For some fluids, in particular ionic ones, however, the critical region is difficult or even impossible to access experimentally. For molten salts, Tc is on the order of 3000 K, which makes accurate measurements a challenging task. Room temperature ionic liquids (RTILs) decompose thermally between 400 K and 600 K due to their organic constituents; this range of temperatures is hundreds of degrees below recent estimates of their Tc. In both cases, reliable methods to deduce Tc based on extrapolations of experimental data recorded at much lower temperatures near the triple or melting points are needed and useful because the critical point influences the fluid's behavior in the entire liquid region. Here, we propose to employ the scaling approach leading to universal fluid behavior [Román et al., J. Chem. Phys. 123, 124512 (2005)] to derive a very simple expression that allows one to estimate Tc from the density of the liquid, the surface tension, or the enthalpy of vaporization measured in a very narrow range of low temperatures. We demonstrate the validity of the approach for simple and polar neutral fluids, for which Tc is known, and then use the methodology to obtain estimates of Tc for ionic fluids. When comparing these estimates to those reported in the literature, good agreement is found for RTILs, whereas the ones for the molten salts NaCl and KCl are lower than previous estimates by 10%. The coexistence curve for ionic fluids is found to be more adequately described by an effective exponent of βeff = 0.5 than by βeff = 0.33.


New species of the spider genera Aysenia and Aysenoides from Chile and Argentina: description and phylogenetic relationships (Araneae: Anyphaenidae, Amaurobioidinae).
New spider species of the genera Aysenia Tullgren and Aysenoides Ramírez are described and their phylogenetic relationships discussed. The new species Aysenia paposo, from the coastal desert in northern Chile is sister to Aysenia araucana Ramírez. The diagnosis of Aysenia araucana is updated and new somatic variability is reported for the species. We present new records for other species of Aysenia and Aysenoides. The new species Aysenoides simoi, from temperate forests in Chile and adjacent Argentina is sister to Aysenoides nahuel. The phylogenetic analysis confirmed the monophyly of both genera. The support values of the genera are relatively high, but some internal branches show low support values. The genus Aysenia is supported by three synapomorphies, two of these from leg spination and one from the male genitalia. Aysenoides is supported by three synapomorphies from male and female genitalia.


Interference study of the chi c0(13P0) in the reaction -pp-->pi0pi0.
Fermilab experiment E835 has observed (-)pp annihilation production of the charmonium state chi(c0) and its subsequent decay into pi(0)pi(0). Although the resonant amplitude is an order of magnitude smaller than that of the nonresonant continuum production of pi(0)pi(0), an enhanced interference signal is evident. A partial wave expansion is used to extract physics parameters. The amplitudes J=0 and 2, of comparable strength, dominate the expansion. Both are accessed by L=1 in the entrance (-)pp channel. The product of the input and output branching fractions is determined to be B((-)pp-->chi(c0))xB(chi(c0)-->pi(0)pi(0))=(5.09+/-0.81+/-0.25)x10(-7).


Crossmodal Connections of Primary Sensory Cortices Largely Vanish During Normal Aging.
During aging, human response times (RTs) to unisensory and crossmodal stimuli decrease. However, the elderly benefit more from crossmodal stimulus representations than younger people. The underlying short-latency multisensory integration process is mediated by direct crossmodal connections at the level of primary sensory cortices. We investigate the age-related changes of these connections using a rodent model (Mongolian gerbil), retrograde tracer injections into the primary auditory (A1), somatosensory (S1), and visual cortex (V1), and immunohistochemistry for markers of apoptosis (Caspase-3), axonal plasticity (Growth associated protein 43, GAP 43), and a calcium-binding protein (Parvalbumin, PV). In adult animals, primary sensory cortices receive a substantial number of direct thalamic inputs from nuclei of their matched, but also from nuclei of non-matched sensory modalities. There are also direct intracortical connections among primary sensory cortices and connections with secondary sensory cortices of other modalities. In very old animals, the crossmodal connections strongly decrease in number or vanish entirely. This is likely due to a retraction of the projection neuron axonal branches rather than ongoing programmed cell death. The loss of crossmodal connections is also accompanied by changes in anatomical correlates of inhibition and excitation in the sensory thalamus and cortex. Together, the loss and restructuring of crossmodal connections during aging suggest a shift of multisensory processing from primary cortices towards other sensory brain areas in elderly individuals.


Lipoma of the transverse colon covered by tubulovillous adenoma: a rare indication for surgical treatment.
Lipomas of the digestive tract are rare benign tumours which, in most cases, are totally asymptomatic. Because of their localization within the intestinal wall, endoscopy may be completely negative so contrast-enhanced computed tomography (CT) is very important for detecting and typing these lesions. The case of a 49-year-old man with abdominal pain is presented. Colonoscopy and biopsy of a polypoid lesion on the right colonic flexure concluded for tubulovillous adenoma. The subsequent CT showed a polylobate lesion of 5 cm in diameter with predominant fat density causing luminal sub-stenosis. Histological examination of the surgical specimen confirmed the presence of a voluminous submucosal lipoma. CT allows to diagnose lipomas of the large bowel thanks to the density measurement (between -40 and -120 Hunsfield Units) with an accurate detection of the site and nature of lumen stenosis.


Composite Lymphoma's Ability to Adapt and Change Through Plasticity: A Case Report and Literature Review.
Composite/synchronous lymphoma is an uncommon condition that presents a challenge in histopathological diagnosis due to the simultaneous appearance of two or more lymphomas, including instances where they are intermixed within the same location. Performing a core needle tissue biopsy adds a challenge for pathologists when trying to diagnose a condition that requires excisional tissue for a thorough evaluation. This report highlights a distinctive instance of composite lymphoma in which classical Hodgkin lymphoma (cHL) is combined with diffuse large B-cell lymphoma (DLBCL). All pertinent information including clinical, histopathological, and immunohistochemical data for each of these composite lymphomas is provided. In addition, we conducted a literature review of the published data. The findings from these data further support the theory of a shared clonal origin and transdifferentiation occurrence in the process of lymphoma development.


Effects of plasma from patients with fulminant hepatic failure on function of primary rat hepatocytes in three-dimensional culture.
As biotechnology continues to advance, a bioartificial liver is expected to be developed for the treatment of patients with fulminant hepatic failure (FHF) whose liver dysfunction is potentially reversible or for providing liver support as a bridge to liver transplantation. While monolayer-cultured hepatocytes rapidly lose their capacity to express many liver-specific functions over time when cultured, spherical-shaped hepatocytes in three-dimensional culture with the use of extracellular matrix components sustain long-term survival by maintaining differentiated hepatocyte functions. The aim of this study was to investigate whether sufficient functions of viable spherical-shaped hepatocytes could be maintained in plasma of patients with FHF in order to use these cells in an extracorporeal system. Hepatocyte functions were evaluated under monolayer or three-dimensional culture in FHF plasma. Primary rat hepatocytes on poly-N-p-vinylbenzyl-D-lactonamide (PVLA) formed spheroids even in FHF plasma and maintained their spherical shapes in FHF plasma as long as in medium. Spherical-shaped hepatocytes on PVLA cultured in FHF plasma showed higher activity in albumin secretion, urea formation, and gluconeogenesis than those in normal human plasma or medium. As being cultured in medium, hepatocytes on PVLA cultured in plasma were also superior to cells on collagen in regard to albumin secretion, amino acid metabolism, and gluconeogenesis. These findings demonstrated that FHF plasma is not toxic to rat hepatocyte spheroids and that hepatocyte spheroids have potential use in the development of a bioartificial liver.


Vaccines against Infections Caused by Salmonella, Shigella, and Pathogenic Escherichia coli.
Infectious diseases represent one of the most common causes of death worldwide, with the enteropathogenic bacteria Salmonella and Shigella and pathogenic Escherichia coli being among the most detrimental. Currently, vaccination represents the preferred method of preventing such infections. For stimulating the adaptive immune response, immunizations are frequently based on formulations which include inactivated whole-cell vaccines, live attenuated vaccines, or subunit vaccines. These can be administered via a parenteral or mucosal route, the latter having the advantage that it most closely mimics the actual course of infection. In addition to the type of vaccine and method of application, important consideration needs to be paid to safety, efficacy, and cost, which are often major bottlenecks in the successful implementation of vaccines. In this chapter we take a limited look at the history surrounding vaccinations involving Salmonella, Shigella, and pathogenic E. coli. Salmonella infections, which can lead to typhoid fever, are becoming increasing difficult to treat with antibiotics due to multi-drug-resistant strains. At present, the parenteral Vi-based subunit vaccines and the live attenuated oral vaccine Ty21a have proven to be the vaccines of choice, with high levels of protective efficacy and limited side effects. Shigella infections are responsible for the diarrheal disease shigellosis. Various live and nonliving mucosal and parenteral vaccines have been tested, with the most promising candidates evolving around those that stimulate the production of O-antigen-specific antibodies. Pathogenic Escherichia coli infections can lead to severe diseases due to the bacterium's production of several specific toxins. Vaccines against this bacterium target its toxins, as well as surface-exposed antigens, all of which have been found to be effective as immunogens.


Biochemical Characterisation of Phage Pseudomurein Endoisopeptidases PeiW and PeiP Using Synthetic Peptides.
Pseudomurein endoisopeptidases cause lysis of the cell walls of methanogens by cleaving the isopeptide bond Ala-ε-Lys in the peptide chain of pseudomurein. PeiW and PeiP are two thermostable pseudomurein endoisopeptidases encoded by phage ΨM100 of Methanothermobacter wolfei and phages ΨM1 and ΨM2 of Methanothermobacter marburgensis, respectively. A continuous assay using synthetic peptide substrates was developed and used in the biochemical characterisation of recombinant PeiW and PeiP. The advantages of these synthetic peptide substrates over natural substrates are sensitivity, high purity, and characterisation and the fact that they are more easily obtained than natural substrates. In the presence of a reducing agent, purified PeiW and PeiP each showed similar activity under aerobic and anaerobic conditions. Both enzymes required a divalent metal for activity and showed greater thermostability in the presence of Ca(2+). PeiW and PeiP involve a cysteine residue in catalysis and have a monomeric native conformation. The kinetic parameters, K(M) and k(cat), were determined, and the ε-isopeptide bond between alanine and lysine was confirmed as the bond lysed by these enzymes in pseudomurein. The new assay may have wider applications for the general study of peptidases and the identification of specific methanogens susceptible to lysis by specific pseudomurein endoisopeptidases.


Evidence for the evolutionary nascence of a novel sex determination pathway in honeybees.
Sex determination in honeybees (Apis mellifera) is governed by heterozygosity at a single locus harbouring the complementary sex determiner (csd) gene, in contrast to the well-studied sex chromosome system of Drosophila melanogaster. Bees heterozygous at csd are females, whereas homozygotes and hemizygotes (haploid individuals) are males. Although at least 15 different csd alleles are known among natural bee populations, the mechanisms linking allelic interactions to switching of the sexual development programme are still obscure. Here we report a new component of the sex-determining pathway in honeybees, encoded 12 kilobases upstream of csd. The gene feminizer (fem) is the ancestrally conserved progenitor gene from which csd arose and encodes an SR-type protein, harbouring an Arg/Ser-rich domain. Fem shares the same arrangement of Arg/Ser- and proline-rich-domain with the Drosophila principal sex-determining gene transformer (tra), but lacks conserved motifs except for a 30-amino-acid motif that Fem shares only with Tra of another fly, Ceratitis capitata. Like tra, the fem transcript is alternatively spliced. The male-specific splice variant contains a premature stop codon and yields no functional product, whereas the female-specific splice variant encodes the functional protein. We show that RNA interference (RNAi)-induced knockdowns of the female-specific fem splice variant result in male bees, indicating that the fem product is required for entire female development. Furthermore, RNAi-induced knockdowns of female allelic csd transcripts result in the male-specific fem splice variant, suggesting that the fem gene implements the switch of developmental pathways controlled by heterozygosity at csd. Comparative analysis of fem and csd coding sequences from five bee species indicates a recent origin of csd in the honeybee lineage from the fem progenitor and provides evidence for positive selection at csd accompanied by purifying selection at fem. The fem locus in bees uncovers gene duplication and positive selection as evolutionary mechanisms underlying the origin of a novel sex determination pathway.


Parasocial Interactions and Relationships in Early Adolescence.
Parasocial interactions and relationships, one-sided connections imagined with celebrities and media figures, are common in adolescence and might play a role in adolescent identity formation and autonomy development. We asked 151 early adolescents (Mage = 14.8 years) to identify a famous individual of whom they are fond; we examined the type of celebrities chosen and why they admired them, and the relationships imagined with these figures across the entire sample and by gender. Adolescents emphasized highly salient media figures, such as actors, for parasocial attention. While different categories of celebrities were appreciated equally for their talent and personality, actors/singers were endorsed for their attractiveness more so than other celebrity types. Most adolescents (61.1%) thought of their favorite media figures as relationship partners, and those who did reported more parasocial involvement and emotional intensity than those who did not. Gender differences emerged in that boys chose more athletes than girls and were more likely to imagine celebrities as authority figures or mentors than friends. Celebrities afforded friendship for girls, who overwhelmingly focused on actresses. Hierarchical parasocial relationships may be linked to processes of identity formation as adolescents, particularly boys, imagine media figures as role models. In contrast, egalitarian parasocial relationships might be associated with autonomy development via an imagined affiliation with an attractive and admirable media figure.