Clarifying the morphology of the ostium primum defect.
The 'ostium primum' defect is still frequently considered to be the consequence of deficient atrial septation, although the key feature is a common atrioventricular junction. The bridging leaflets of the common atrioventricular valve, which are joined to each other, are depressed distal to the atrioventricular junction, and fused to the crest of the muscular ventricular septum, which is bowed in the concave direction towards the ventricular apex. As a result, shunting across the defect occurs between the atrial chambers. These observations suggest that the basic deficiency in the 'ostium primum' defect is best understood as a product of defective atrioventricular septation, rather than an atrial septal defect. We have now encountered four examples of 'ostium primum' defects in mouse embryos that support this view. These were identified from a large number of mouse embryo hearts collected from a normal, outbred mouse colony and analysed by episcopic microscopy as part of an ongoing study of normal mouse cardiac development. The abnormal hearts were identified from embryos collected at embryonic days 15.5, 16.5 and 18.5 (two cases). We have analysed the features of the abnormal hearts, and compared the findings with those obtained in the large number of normally developed embryos. Our data show that the key feature of normal atrioventricular septation is the ventral growth through the right pulmonary ridge of a protrusion from the dorsal pharyngeal mesenchyme, confirming previous findings. This protrusion, known as the vestibular spine, or the dorsal mesenchymal protrusion, reinforces the closure of the primary atrial foramen, and muscularises along with the mesenchymal cap of the primary atrial septum to form the ventro-caudal buttress of the oval foramen, identified by some as the 'canal septum'. Detailed analysis of the four abnormal hearts suggests that in each case there has been failure of growth of the vestibular spine, with the result that the common atrioventricular junction found earlier during normal development now persists during cardiac development. Failure of separation of the common junction also accounts for the trifoliate arrangement of the left atrioventricular valve in the abnormal hearts. Analysis of the episcopic datasets also permits recognition of the location of the atrioventricular conduction axis. Comparison of the location of this tract in the normal and abnormal hearts shows that there is no separate formation of a ventricular component of the 'canal septum' as part of normal development. We conclude that it is abnormal formation of the primary atrial septum that is the cause of so-called 'secundum' atrial septal defects, whereas it is the failure to produce a second contribution to atrial septation (via growth of the vestibular spine) that results in the 'ostium primum' defect.


Seasonal inactivated influenza virus vaccines.
Inactivated influenza virus vaccines are the primary modality used for prevention of influenza. A system of annual identification of new strains causing illnesses, selections for vaccines, chick embryo growth, inactivation, processing, packaging, distribution and usage has been in place for decades. Current vaccines contain 15 microg of the HA of an A/H1N1, A/H3N2 and B strain and are given parenterally to induce serum anti-HA antibody for prevention of subsequent infection and illness from natural influenza. Reactogenicity is low and protection among healthy older children and adults is good; protection levels are generally lower in young children and the elderly. Needs include ensuring antigenic matches of vaccine and epidemic viruses each season, enhancing immunization rates, and providing new and improved vaccines and immunization approaches for the varied populations and circumstances globally.


Determining Membrane Protein-Lipid Binding Thermodynamics Using Native Mass Spectrometry.
Membrane proteins are embedded in the biological membrane where the chemically diverse lipid environment can modulate their structure and function. However, the thermodynamics governing the molecular recognition and interaction of lipids with membrane proteins is poorly understood. Here, we report a method using native mass spectrometry (MS), to determine thermodynamics of individual ligand binding events to proteins. Unlike conventional methods, native MS can resolve individual ligand binding events and, coupled with an apparatus to control the temperature, determine binding thermodynamic parameters, such as for protein-lipid interactions. We validated our approach using three soluble protein-ligand systems (maltose binding protein, lysozyme, and nitrogen regulatory protein) and obtained similar results to those using isothermal titration calorimetry and surface plasmon resonance. We also determined for the first time the thermodynamics of individual lipid binding to the ammonia channel (AmtB), an integral membrane protein from Escherichia coli. Remarkably, we observed distinct thermodynamic signatures for the binding of different lipids and entropy-enthalpy compensation for binding lipids of variable chain length. Additionally, using a mutant form of AmtB that abolishes a specific phosphatidylglycerol (PG) binding site, we observed distinct changes in the thermodynamic signatures for binding PG, implying these signatures can identify key residues involved in specific lipid binding and potentially differentiate between specific lipid binding sites.


Effects of capsicine on rat cytochrome P450 isoforms CYP1A2, CYP2C19, and CYP3A4.
Due to the frequent consumption of capsaicin (CAP) and its current therapeutic application, the correct assessment of this compound is important from a public health standpoint. The purpose of this study was to find out whether CAP affects rat cytochrome P450 (CYP) enzymes (CYP1A2, CYP2C19, and CYP3A4) by using cocktail probe drugs in vivo. A cocktail solution at a dose of 5 mL/kg, which contained phenacetin (15 mg/kg), omeprazole (15 mg/kg), and midazolam (10 mg/kg), was given orally to rats treated for 7 d with oral administration of CAP. Blood samples were collected at a series of time-points and the concentrations of probe drugs in plasma were determined by HPLC-MS. The results showed that treatment with multiple doses of CAP had no significant effect on rat CYP1A2. However, CAP had a significant inhibitory effect on CYP2C19 and an inductive effect on CYP3A4. Therefore, caution is needed when CAP is co-administered with some CYP substrates clinically because of potential drug-CAP interactions.


An experimental methodology to relate local strain to microstructural texture.
This paper introduces an experimental methodology for obtaining high resolution full-field strain measurements in polycrystalline metals. The (sub)grain level resolution of these measurements was indispensable for relating measured strain fields to observed microstructure in the material. Microstructural information was obtained through electron backscatter diffraction and the optical technique of digital image correlation (DIC) was used to acquire full-field deformation measurements. By spatially overlaying both sets of results, the effects of different microstructural features such as orientation, grain boundary character, misorientation between grains, and twin boundaries on material response can be quantitatively studied. To obtain the necessary resolution for such measurements, the images used in DIC had to be captured at high magnifications. This necessity reduces the field of view and constrains the area of interest that can be monitored. To address this issue, results from adjacent measurement areas are combined together to create a data set with high spatial strain resolution over a larger region than can otherwise be observed. The procedure for performing this technique is outlined here, along with benefits, drawbacks, possible modifications, and example applications of the technique to cyclic plasticity and fatigue crack growth.


CCN3/nephroblastoma overexpressed matricellular protein regulates integrin expression, adhesion, and dissemination in melanoma.
CCN3/nephroblastoma overexpressed belongs to the CCN family of genes that encode secreted proteins associated with the extracellular matrix (ECM) and exert regulatory effects at the cellular level. Overexpression of CCN3 was shown in metastatic melanoma cells compared with cells of the primary tumor from the same patient. Analysis of short-term cultures from 50 primary and metastatic melanomas revealed a heterogeneous expression pattern of both the 46-kDa full-length cytoplasmic/secreted protein and the 32-kDa nuclear-truncated form. The different protein expression patterns were not associated with gene alterations or polymorphisms. Like the metastatic cells expressing high levels of the 46-kDa CCN3, cells transfected to overexpress CCN3 showed increased adhesion to ECM proteins, whereas inhibition of CCN3 expression by small interfering RNA decreased adhesion to laminin and vitronectin. CCN3 overexpression induced increased expression of laminin and vitronectin integrin receptors alpha 7 beta 1 and alpha v beta 5 by increasing their mRNA production. Moreover, CCN3 secreted by melanoma cells acted as an adhesion matrix protein for melanoma cells themselves. Analysis of CCN3 protein expression with respect to melanoma progression detected the protein in all visceral metastases tested and in most nodal metastases from relapsing patients but in only a few nodal metastases from nonrelapsing patients and cutaneous metastases. Consistently, xenotransplantation in immunodeficient mice showed a higher metastatic potential of melanoma cells overexpressing CCN3. Together, these data indicate a role for CCN3 in melanoma cell interaction with the ECM by regulating integrin expression, resulting in altered cell adhesion and leading melanoma progression to aggressive disease.


Nutritional chemistry of foods eaten by gorillas in Bwindi Impenetrable National Park, Uganda.
Foods eaten by gorillas (Gorilla beringei) in Bwindi Impenetrable National Park (BINP), Uganda, were analyzed for their nutrient content. The goal of the study was to assess the amounts of fiber, protein, and sugars in the foods eaten by the Bwindi gorillas, and to determine whether condensed tannins and cyanide are present in these foods. A total of 127 food plant parts representing 84 plant species eaten by two groups of Bwindi gorillas were collected, processed, and analyzed for their chemical contents. The Bwindi gorilla ate foods that contain 2-28% crude protein (CP), 21-88% neutral detergent fiber (NDF), 14-60% acid detergent fiber (ADF), 2-42% acid detergent lignin (ADL), and </=1-50% water-soluble carbohydrates (WSC) on a dry matter (DM) basis. Moisture in these foods ranged from 7% to 96%. Approximately 35% of the foods analyzed contained condensed tannins, and two foods contained cyanogenic glycosides. This is the first detailed report regarding the nutritional chemistry of gorilla foods in Bwindi, many of which are also eaten by other primates. This unique data set adds to our knowledge about the nutritional composition of foods eaten by gorillas across habitats, is useful for understanding aspects of feeding behavior, and provides valuable comparative data for optimizing the diets of gorillas ex situ.


[Ultrasound-guided caudal block with ropivacaine in infraumbilical surgeries in pediatric patients].
The use of local anesthetics as part of multimodal analgesia is an attractive concept in pediatric patients, since the evaluation and management of pain is a challenge in children under 7 years of age. Despite having guidelines and multiple studies on volume calculation, no safe and effective recommendation has been issued. To demonstrate that ultrasound-guided caudal block with a dose of 0.75 mL/kg of 0.2% ropivacaine has the same analgesic effect as a dose of 1.2 mL/kg. Randomized, prospective, longitudinal, comparative equivalence clinical trial. One hundred patients 0-7 years of age scheduled for elective or emergency infraumbilical surgery were enrolled between April 2021 and January 2022. Children were randomized 1:1 to be assigned to ultrasound-guided caudal block. 100 patients divided into two groups with 0.2% ropivacaine volumes (0.75 mL vs. 1.2 mL). Both groups demonstrated the trans-anesthetic and post-anesthetic sedoanalgesia variables without significant differences for both groups on the FLACC pain scale after surgery and in recovery (p > 0.5), in the pain reassessment on the FLACC scale in the office called chronic pain (p > 0.5) in both groups. No complications were reported in the follow-up consultations and no arrhythmias were reported in both groups during surgery. the results of both groups did not show differences between a volume of 0.75 mL and 1.2 mL, the administration of 0.2% ropivacaine is favored with the use of ultrasound, which allows effective administration of lower doses of local anesthetic with reduced risk of complications. It is necessary to carry out studies in other types of surgery to compare the use of less volume of local anesthetic compared to the Melman formula used in this study.


A novel microbial habitat in the mid-ocean ridge subseafloor.
The subseafloor at the mid-ocean ridge is predicted to be an excellent microbial habitat, because there is abundant space, fluid flow, and geochemical energy in the porous, hydrothermally influenced oceanic crust. These characteristics also make it a good analog for potential subsurface extraterrestrial habitats. Subseafloor environments created by the mixing of hot hydrothermal fluids and seawater are predicted to be particularly energy-rich, and hyperthermophilic microorganisms that broadly reflect such predictions are ejected from these systems in low-temperature ( approximately 15 degrees C), basalt-hosted diffuse effluents. Seven hyperthermophilic heterotrophs isolated from low-temperature diffuse fluids exiting the basaltic crust in and near two hydrothermal vent fields on the Endeavour Segment, Juan de Fuca Ridge, were compared phylogenetically and physiologically to six similarly enriched hyperthermophiles from samples associated with seafloor metal sulfide structures. The 13 organisms fell into four distinct groups: one group of two organisms corresponding to the genus Pyrococcus and three groups corresponding to the genus Thermococcus. Of these three groups, one was composed solely of sulfide-derived organisms, and the other two related groups were composed of subseafloor organisms. There was no evidence of restricted exchange of organisms between sulfide and subseafloor habitats, and therefore this phylogenetic distinction indicates a selective force operating between the two habitats. Hypotheses regarding the habitat differences were generated through comparison of the physiology of the two groups of hyperthermophiles; some potential differences between these habitats include fluid flow stability, metal ion concentrations, and sources of complex organic matter.


Active relearning for robust supervised training of emphysema patterns.
Radiologists are adept at recognizing the character and extent of lung parenchymal abnormalities in computed tomography (CT) scans. However, the inconsistent differential diagnosis due to subjective aggregation necessitates the exploration of automated classification based on supervised or unsupervised learning. The robustness of supervised learning depends on the training samples. Towards optimizing emphysema classification, we introduce a physician-in-the-loop feedback approach to minimize ambiguity in the selected training samples. An experienced thoracic radiologist selected 412 regions of interest (ROIs) across 15 datasets to represent 124, 129, 139 and 20 training samples of mild, moderate, severe emphysema and normal appearance, respectively. Using multi-view (multiple metrics to capture complementary features) inductive learning, an ensemble of seven un-optimized support vector models (SVM) each based on a specific metric was constructed in less than 6 s. The training samples were classified using seven SVM models and consensus labels were created using majority voting. In the active relearning phase, the ensemble-expert label conflicts were resolved by the expert. The efficacy and generality of active relearning feedback was assessed in the optimized parameter space of six general purpose classifiers across the seven dissimilarity metrics. The proposed just-in-time active relearning feedback with un-optimized SVMs yielded 15 % increase in classification accuracy and 25 % reduction in the number of support vectors. The average improvement in accuracy of six classifiers in their optimized parameter space was 21 %. The proposed cooperative feedback method enhances the quality of training samples used to construct automated classification of emphysematous CT scans. Such an approach could lead to substantial improvement in quantification of emphysema.


Optimization of microwave-assisted extraction followed by RP-HPLC for the simultaneous determination of oleanolic acid and ursolic acid in the fruits of Chaenomeles sinensis.
An optimized microwave-assisted extraction (MAE) method and an efficient HPLC analysis method were developed for fast extraction and simultaneous determination of oleanolic acid and ursolic acid in the fruit of Chaenomeles sinensis. The open vessel MAE process was optimized by using a central composite experimental design. The optimal conditions identified were microwave power 600 W, temperature 52 degrees C, solvent to material ratio 32 mL/g and extraction time 7 min. The results showed that MAE is a more rapid extraction method with higher yield and lower solvent consumption. The HPLC-photodiode array detection analysis method was validated to have good linearity, precision, reproduction and accuracy. Compared with conventional extraction and analysis methods, MAE-HPLC-photodiode array detection is a faster, convenient and appropriate method for determination of oleanolic acid and ursolic acid in the fruits of C. sinensis.


A New Approach for Multimodal Usage of Gene Expression and Its Image Representation for the Detection of Alzheimer's Disease.
Alzheimer's disease (AD) is a complex neurodegenerative disorder and the multifaceted nature of it requires innovative approaches that integrate various data modalities to enhance its detection. However, due to the cost of collecting multimodal data, multimodal datasets suffer from an insufficient number of samples. To mitigate the impact of a limited sample size on classification, we introduce a novel deep learning method (One2MFusion) which combines gene expression data with their corresponding 2D representation as a new modality. The gene vectors were first mapped to a discriminative 2D image for training a convolutional neural network (CNN). In parallel, the gene sequences were used to train a feed forward neural network (FNN) and the outputs of the FNN and CNN were merged, and a joint deep network was trained for the binary classification of AD, normal control (NC), and mild cognitive impairment (MCI) samples. The fusion of the gene expression data and gene-originated 2D image increased the accuracy (area under the curve) from 0.86 (obtained using a 2D image) to 0.91 for AD vs. NC and from 0.76 (obtained using a 2D image) to 0.88 for MCI vs. NC. The results show that representing gene expression data in another discriminative form increases the classification accuracy when fused with base data.


Effect of physical exercise on secretion of growth hormone, glucagon, and cortisol in obese and diabetic children.
The effect of muscular exertion of moderate intensity on blood sugar (BS), plasma levels of growth hormone (GH), glucagon, and cortisol (F) has been studied in endocrinologically normal children with short stature and compared with children with clinical diabetes mellitus and obese children with normal and diminished carbohydrate tolerance. In diabetic children, physical exertion induces a rise in plasma GH levels comparable to that in controls; in obese children with normal or with diminished glucose tolerance, the rise is considerably smaller. Physical exertion caused no change in F levels in the groups tested, although basal level in the obese children was significantly higher than in the controls. Basal glucagon levels were similar in all groups and showed no change on physical exertion. The behavior of GH and glucagon in diabetic children was comparable to that in the controls even where blood sugar level was high.


Capsule polysaccharide mediates bacterial resistance to antimicrobial peptides.
The innate immune system plays a critical role in the defense of areas exposed to microorganisms. There is an increasing body of evidence indicating that antimicrobial peptides and proteins (APs) are one of the most important weapons of this system and that they make up the protective front for the respiratory tract. On the other hand, it is known that pathogenic organisms have developed countermeasures to resist these agents such as reducing the net negative charge of the bacterial membranes. Here we report the characterization of a novel mechanism of resistance to APs that is dependent on the bacterial capsule polysaccharide (CPS). Klebsiella pneumoniae CPS mutant was more sensitive than the wild type to human neutrophil defensin 1, beta-defensin 1, lactoferrin, protamine sulfate, and polymyxin B. K. pneumoniae lipopolysaccharide O antigen did not play an important role in AP resistance, and CPS was the only factor conferring protection against polymyxin B in strains lacking O antigen. In addition, we found a significant correlation between the amount of CPS expressed by a given strain and the resistance to polymyxin B. We also showed that K. pneumoniae CPS mutant bound more polymyxin B than the wild-type strain with a concomitant increased in the self-promoted pathway. Taken together, our results suggest that CPS protects bacteria by limiting the interaction of APs with the surface. Finally, we report that K. pneumoniae increased the amount of CPS and upregulated cps transcription when grown in the presence of polymyxin B and lactoferrin.


A reexamination of the role of oxygen in retrolental fibroplasia.
A critical review of the literature of retrolental fibroplasia indicates that the cause of this disease is not yet known. Oxygen is certainly a critical factor but it is still not possible to make precise recommendations as to the amount or the duration of therapy that is safe. We have overemphasized the role of oxygen in the past, and as a result of this the false impression has been created that RLF is a disease that can be prevented. This gross oversimplification of a complex disease with multiple causes has resulted in many unjustified malpractice claims. A study of the present epidemic indicates that excessive oxygen administration probably plays a minor role, in contrast to the first epidemic in which prolonged oxygen administration was clearly a major factor. A reasonable working hypothesis is that the developing retina is highly sensitive to any disturbance in its oxygen supply, either hyperoxemic or hypoxemic. The retinal circulation is subject to the same wide fluctuations as the cerebral circulation in newborn infants. The very low-birth-weight, sick premature infant suffers from a number of conditions, many of which can seriously disturb the retinal circulation, resulting in hypoperfusion and ischemia. These factors (immaturity, hyperoxia, hypoxia, blood transfusions, intraventricular hemorrhage, apnea, infection, hypercarbia, hypocarbia, patent ductus arteriosus, prostaglandin synthetase inhibitors, vitamin E deficiency, lactic acidosis, prenatal complications, genetic factors) may all be present in an infant. They may interact to produce various degrees of retinal damage. Nearly all of these factors cannot be prevented or controlled by our present methods of care. Unfortunately, this means that RLF is an extremely difficult disease to prevent, treat, or investigate. A disease of this complexity with multiple causes will require very large numbers of infants in any controlled study of a therapy. Retrolental fibroplasia should not be considered an avoidable iatrogenic disease in very low-birth-weight infants. Its cause in these infants is not known.


Semi-quantitative contrast-enhanced MR analysis of indeterminate ovarian tumours: when to say malignancy?
To evaluate the ability of dynamic post-contrast sequence to specify indeterminate ovarian masses with inconclusive MR features of malignancy. Since management is dramatically different, special focus on the ability to differentiate borderline from invasive malignancy was considered. 150 ovarian masses were detected by pelvic ultrasound in 124 patients. Masses had been considered for dynamic post-contrast MRI. We expressed the kinetic parameters (i.e. enhancement amplitude, time peak of maximal uptake and maximal slope) in the form of maximum relative enhancement percentage (MRE%), time of maximal peak of contrast uptake (Tmax) and slope enhancement ratio (SER) curves. Histological findings were the gold standard of reference. Malignant ovarian masses showed higher MRE% than benign and borderline masses (p < 0.001). Tmax was shorter for malignant than benign (p < 0.01) and borderline (p < 0.001) ovarian masses. SER curves were the most suggestive of malignancy with a specificity and accuracy of 85.7% and 84.7%, respectively. Dynamic contrast-enhanced MRI could be a specific sequence to differentiate ovarian masses with indeterminate MR morphology with a special discrimination for low potential from invasive ovarian malignancy. The study evaluated the diagnostic performance of the individual parameters of dynamic post-contrast MR sequence in evaluating ovarian masses. Management divert between benign, borderline and invasive malignant masses; our work presented a cut-off value for the peak of contrast uptake of 120%, which helped in the differentiation between benign and malignant tumours; the SER curves with Type III (early washout) pattern that was indicative of invasive malignancy was more specific than borderline malignancy.


[Surgical gloves--how well do the protect against infections?].
Health care workers (HCW) in surgery are at high risk for bloodborne infections (BBI) e.g. by hepatitis-B(HB)-, hepatitis-C(HC)- and HI-virus. On the other hand, infectious medical staff can cause nosocomial BBI in patients, too. Intact gloves provide an efficient barrier against BBI but glove perforations are common during several surgical procedures. A review of studies on glove perforations during different surgical operations was carried out with special regard to user, number and location of perforations, duration and kind of operation. We compared the results with the frequency of glove perforation during operations in 1938 single used gloves. They were collected after different surgical procedures in a department for general surgery and tested for perforation by 1-liter-water-filling method according to DIN 455/1. The product used during the period of this investigation was Sempermed sterile latex surgical glove. Most perforations were found on the index finger and thumb of the non-dominant hand. Duration of operation, role of the user (primary surgeon) and kind of operation are predictors for the incidence of glove perforations. Double gloving, endoscopic and no-touch techniques decrease the possibility of blood contact during operation. Indicator systems are useful for detection of the loss of glove integrity. Due to the high perforation rate found in this study glove change as a routine during surgical procedures should be discussed (e.g. every 30 minutes).


F1 alpha: a novel mucin antigen associated with gastric carcinogenesis.
In a previous study, we obtained a novel monoclonal antibody (F1 alpha-75) directed against a synthetic mucin antigen termed F1 alpha, and demonstrated that this antigen was expressed in a high percentage (80.2%; 89/111) of gastric carcinomas. In the present study, we compared the expression of F1 alpha with that of sialyl-Tn antigen, a mucin antigen similar to F1 alpha, in 54 human early gastric carcinomas, intestinal metaplasia and adenomatous polyps of the stomach to determine how differences in the expression of these antigens correlated with gastric carcinogenesis. The rate of expression of F1 alpha in early gastric carcinoma tissues, 81.5%, was higher than that of sialyl-Tn antigen, 57.4%. No correlation was found between the rate of expression and histological type, depth of cancerous invasion or lymph node metastasis. Sialyl-Tn antigen was extensively expressed in 77.5% of specimens of complete intestinal metaplasia and in 78.6% of those of incomplete intestinal metaplasia; however, the expression of F1 alpha in those specimens was rare and sporadic, with rates of only 25.0 and 27.7%, respectively. In adenomatous polyps, the rate of expression of F1 alpha is 6.25% and that of sialyl-Tn antigen is 43.8%. Our findings indicate that F1 alpha is a more specific antigen for gastric cancer than sialyl-Tn antigen, and that F1 alpha is an antigen associated with carcinogenesis of the stomach.


Dimerization and oxidation of tryptophan in UV-A photolysis sensitized by kynurenic acid.
Photoinduced generation of radicals in the eye lens may play an important role in the modification of proteins leading to their coloration, aggregation, and insolubilization. The radicals can be formed via the reactions of photoexcited endogenous chromophores of the human lens with lens proteins, in particular with tryptophan residues. In the present work we studied the reactions induced by UV-A (315-400nm) light between kynurenic acid (KNA), an effective photosensitizer present in the human lens, and N-acetyl-L-tryptophan (NTrpH) under aerobic and anaerobic conditions. Our results show that the reaction mechanism strongly depends on the presence of oxygen in solution. Under aerobic conditions, the generation of singlet oxygen is the major channel of the effective NTrpH oxidation. In argon-bubbled solutions, the quenching of triplet KNA by NTrpH results in the formation of KNA•- and NTrp• radicals. Under laser pulse irradiation, when the radical concentration is high, the main pathway of the radical decay is the back electron transfer with the restoration of initial reagents. Other reactions include (i) the radical combination yielding NTrp dimers and (ii) the oxygen atom transfer from KNA•- to NTrp• with the formation of oxidized NTrp species and deoxygenated KNA products. In continuous-wave photolysis, even trace amounts of molecular oxygen are sufficient to oxidize the majority of KNA•- radicals with the rate constant of (2.0 ± 0.2) × 109M-1s-1, leading to the restoration of KNA and the formation of superoxide radical O2•-. The latter reacts with NTrp• via either the radical combination to form oxidized NTrp (minor pathway), or the electron transfer to restore NTrpH in the ground state (major pathway). As the result, the quantum yields of the starting compound decomposition under continuous-wave anaerobic photolysis are rather low: 1.6% for NTrpH and 0.02% for KNA. The photolysis of KNA with alpha-crystallin yields the same deoxygenated KNA products as the photolysis of KNA with NTrpH, indicating the similarity of the photolysis mechanisms. Thus, inside the eye lens KNA can sensitize both protein photooxidation and protein covalent cross-linking with the minor self-degradation. This may play an important role in the lens protein modifications during the normal aging and cataract development.


Genital Gender-Affirming Surgery in Transgender Men in The Netherlands from 1989 to 2018: The Evolution of Surgical Care.
The number of individuals with gender identity incongruence seeking treatment, including genital gender-affirming surgery, has increased over the past decade. Surgical techniques for genital gender-affirming surgery in transgender men have evolved considerably. The aim of this study was to present an overview of the evolution of genital gender-affirming surgery for transgender men in a single, high-volume center. The authors performed a retrospective analysis of medical records of all transgender men who underwent genital gender-affirming surgery from January of 1989 to January of 2018 at their institution. Subject demographics, type of surgery, and surgical techniques were recorded. Over time, four hundred transgender men underwent genital gender-affirming surgery. Metoidioplasty was performed in 222 patients (56 percent) and phalloplasty in 178 patients (44 percent). Before 2010, the majority of patients underwent metoidioplasty, whereas since 2010, phalloplasty has been performed in most patients. Of 400 patients, 332 (83 percent) underwent urethral lengthening. The option to undergo genital surgery without urethral lengthening was offered from 2004. Since then, 68 (34 percent) patients chose to undergo genital gender-affirming surgery without urethral lengthening. The choice of surgical options for genital gender-affirming surgery in transgender men has grown from five in 1989 to 12 techniques currently. The variety and combination of flaps used for phalloplasty has increased over time. Conversely, the use of some flaps has been abandoned, whereas other flaps have reemerged. Genital gender-affirming surgery has developed over time in line with reconstructive surgical innovations. Despite technical advancements, there is no one ideal technique, and every technique has specific advantages and disadvantages. With the increase in surgical options, shared decision-making and a shift from surgeon-centered toward more patient-centered care are required.


Discovery of the Migraine Prevention Therapeutic Aimovig (Erenumab), the First FDA-Approved Antibody against a G-Protein-Coupled Receptor.
In 2018, the United States Food and Drug Administration (FDA) approved Aimovig (erenumab) for the prevention of migraine. Erenumab is the first FDA approved antibody therapeutic against a G-protein-coupled receptor, the canonical receptor of calcitonin gene related peptide (CGRP-R). A novel, epitope-focused antigen was created to reconstruct the extracellular domains of the CGRP-R in a stable conformation. Successful inoculation of XenoMouse animals and careful screening yielded multiple candidate molecules for high potency and exquisite selectivity toward the CGRP-R over related receptors. These efforts led to the discovery of erenumab which has demonstrated the desired efficacy and safety profiles in multiple clinical studies for the prevention of migraine. The innovation developed in the discovery of erenumab furthers the ability to target G-coupled protein receptors using antibody approaches.


Electronic brachytherapy as adjuvant therapy for early stage breast cancer: a retrospective analysis.
This multicenter, retrospective study evaluated treatment and clinical outcomes of patients with early stage breast cancer who received adjuvant high-dose rate (HDR) electronic brachytherapy (EBT) treatment post-lumpectomy using the Axxent(®) EBT system. Dosimetric data from the EBT treatment plans were compared with those based on iridium-192 HDR brachytherapy. Medical records of 63 patients with early stage breast cancer (Tis, T1a, T1b, T1c, and T2) who were treated post-lumpectomy with EBT alone or in combination with external beam radiation therapy were reviewed. The prescribed EBT dose was 34 Gy (10 fractions over 5 days, 3.4 Gy each) to 1 cm from the balloon surface. Dosimetry data from 12 patients were compared with these of treatment plans using an iridium-192 source prepared for the same 12 patients. The majority of patients (90.5%) were older than 50 years and had one or more risk factors for breast cancer (80.6%). Tumor sizes were 0.1 cm to 3.5 cm (mean 1.3 cm). Median follow-up was 7 months (1 to 18 months) post-EBT. Balloon applicators were implanted 0 to 85 days (mean 13.4 days) post-lumpectomy/re-excision. The most common adverse events were erythema, rash dermatitis, and pain or breast tenderness. No recurrences were reported. Dosimetric analyses demonstrated comparable target coverage, increased high-dose regions, and a significantly reduced dose to the ipsilateral breast and lungs as well as the heart with EBT as compared with the iridium-192 treatment plans. This retrospective, multicenter study showed that postsurgical adjuvant radiation therapy for early stage breast cancer can be administered using the EBT system with similar toxicity outcomes to those reported with iridium-192 brachytherapy. EBT offers a convenient, portable, nonisotope alternative to HDR brachytherapy using iridium-192.


Highly (H5N1) and low (H7N2) pathogenic avian influenza virus infection in falcons via nasochoanal route and ingestion of experimentally infected prey.
An experimental infection with highly pathogenic avian influenza (HPAI) and low pathogenic avian influenza (LPAI) viruses was carried out on falcons in order to examine the effects of these viruses in terms of pathogenesis, viral distribution in tissues and viral shedding. The distribution pattern of influenza virus receptors was also assessed. Captive-reared gyr-saker (Falco rusticolus x Falco cherrug) hybrid falcons were challenged with a HPAI H5N1 virus (A/Great crested grebe/Basque Country/06.03249/2006) or a LPAI H7N2 virus (A/Anas plathyrhynchos/Spain/1877/2009), both via the nasochoanal route and by ingestion of previously infected specific pathogen free chicks. Infected falcons exhibited similar infection dynamics despite the different routes of exposure, demonstrating the effectiveness of in vivo feeding route. H5N1 infected falcons died, or were euthanized, between 5-7 days post-infection (dpi) after showing acute severe neurological signs. Presence of viral antigen in several tissues was confirmed by immunohistochemistry and real time RT-PCR (RRT-PCR), which were generally associated with significant microscopical lesions, mostly in the brain. Neither clinical signs, nor histopathological findings were observed in any of the H7N2 LPAI infected falcons, although all of them had seroconverted by 11 dpi. Avian receptors were strongly present in the upper respiratory tract of the falcons, in accordance with the consistent oral viral shedding detected by RRT-PCR in both H5N1 HPAI and H7N2 LPAI infected falcons. The present study demonstrates that gyr-saker hybrid falcons are highly susceptible to H5N1 HPAI virus infection, as previously observed, and that they may play a major role in the spreading of both HPAI and LPAI viruses. For the first time in raptors, natural infection by feeding on infected prey was successfully reproduced. The use of avian prey species in falconry husbandry and wildlife rehabilitation facilities could put valuable birds of prey and humans at risk and, therefore, this practice should be closely monitored.


Protective effect of Garcinia kola (kolaviron) extract on predisposition of rats to cardiovascular diseases following separate administration of amodiaquine and artesunate.
This study was carried out to investigate the cardiovascular effect of administration of antimalarial drugs amodiaquine and artesunate and the efficacy of Garcinia kola extract (kolaviron) in protecting against such possible effect. Thirty (30) adult male albino rats divided into six (6) groups were used in this study. Groups D, E and F were treated with 100 mg/Kg b. w. of the extract twice daily for the first one week and 200 mg/Kg b. w./day for the subsequent three (3) weeks. Amodiaquine (10 mg/Kg b. w./day) was administered orally for four (4) days into rats in groups A and E while rats in groups B and F were treated with artesunate (5 mg/Kg b. w./day) for four (4) days. Group C rats (normal control) were treated with normal saline. All the rats were sacrificed after four (4) weeks treatment period. Blood was withdrawn by cardiac puncture while the liver, kidney, stomach and heart were removed, cleansed and weighed. Total cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides were measured in the serum, while total fibrinogen, platelet count, red blood cell and white blood cell count were measured in the whole blood. The artherogenic and coronary risk index were also determined. Results indicate that both amodiaquine and artesunate predispose to cardiovascular disease, however the effect was more pronounced with artesunate than amodiaquine. The result also suggests that both drugs could increase the risk of coronary and artherogenic diseases and that Garcinia kola do not prevent the cardiotoxicity and coronary risk effect.


The substrate of Greatwall kinase, Arpp19, controls mitosis by inhibiting protein phosphatase 2A.
Initiation and maintenance of mitosis require the activation of protein kinase cyclin B-Cdc2 and the inhibition of protein phosphatase 2A (PP2A), which, respectively, phosphorylate and dephosphorylate mitotic substrates. The protein kinase Greatwall (Gwl) is required to maintain mitosis through PP2A inhibition. We describe how Gwl activation results in PP2A inhibition. We identified cyclic adenosine monophosphate-regulated phosphoprotein 19 (Arpp19) and α-Endosulfine as two substrates of Gwl that, when phosphorylated by this kinase, associate with and inhibit PP2A, thus promoting mitotic entry. Conversely, in the absence of Gwl activity, Arpp19 and α-Endosulfine are dephosphorylated and lose their capacity to bind and inhibit PP2A. Although both proteins can inhibit PP2A, endogenous Arpp19, but not α-Endosulfine, is responsible for PP2A inhibition at mitotic entry in Xenopus egg extracts.


Role of endogenous TRPV1 agonists in a postburn pain model of partial-thickness injury.
Oxidized linoleic acid metabolites (OLAMs) are a class of endogenous transient receptor potential vanilloid 1 (TRPV1) channel agonists released on exposure of tissue to transient noxious temperatures. These lipid compounds also contribute to inflammatory and heat allodynia. Because persistent pain after a burn injury represents a significant clinical challenge for treatment, we developed an in vivo rat model of partial-thickness cutaneous thermal injury and examined whether TRPV1 and specific OLAM metabolites play a role in mediating postburn pain injury. This peripheral model of burn injury had marked thermal allodynia peaking at 24h after thermal injury, with allodynia being maintained for up to 7d. Immunohistochemical characterization of tissue taken from injury sites revealed an increase in leukocyte/macrophage infiltration that was colocalized with TRPV1-positive fibers. Using this peripheral thermal injury model, we found that pharmacological blockade of peripheral TRPV1 receptors reduced thermal allodynia by about 98%. Moreover, there was a significant increase in OLAM levels compared to naive controls in hind paw skin biopsies. Additional studies of the metabolism of [C(14)]-linoleic acid in skin biopsies revealed the role of the cytochrome P450 (CYP) system in mediating the metabolism of linoleic acid after thermal injury. Finally, we demonstrated that direct inhibition of OLAMs using OLAM antibodies and indirect inhibition using the CYP inhibitor ketoconazole significantly reduced postburn thermal allodynia. Collectively, these findings point to a novel role of the OLAMs and CYP-related enzymes in generating postburn allodynia via activation of peripheral TRPV1.


When tuberculosis comes back: who develops recurrent tuberculosis in california?
Recurrent tuberculosis suggests potentially modifiable gaps in tuberculosis treatment and control activities. The frequency of late recurrences following treatment completion has not been well-studied. We determined the frequency of, and risk factors associated with, tuberculosis that recurs at least one year after completion of anti-tuberculosis therapy in California. The study population included culture-positive, pulmonary tuberculosis patients reported to the California tuberculosis case registry from 1993 to 2007 who completed anti-tuberculosis therapy. A person with late recurrent tuberculosis was defined as an individual that appeared in the registry more than once, determined by match on name and date-of-birth, with at least one year between treatment completion of the first episode and treatment initiation of the second episode. Among 23,517 tuberculosis patients, 148 (0.63%) had a late recurrence. Independent risk factors for recurrence included: infection with a pyrazinamide mono-resistant isolate (adjusted hazard ratio, 2.93; p = 0.019); initiation of an isoniazid- and rifampin-only treatment regimen (adjusted hazard ratio, 2.55; p = 0.0412); sputum smear-positive disease (adjusted hazard ratio, 1.96; p = 0.0003); human immunodeficiency virus infection (adjusted hazard ratio, 1.81; p = 0.0149); and birth in the United States (adjusted hazard ratio, 1.88; p = 0.0002). Infection with an isoniazid mono-resistant isolate was protective (adjusted hazard ratio, 0.25; p = 0.0171). The low frequency of late recurrent tuberculosis in California suggests that local TB control programs are largely successful at preventing this adverse outcome. Nonetheless, we identified subpopulations at increased risk of late tuberculosis recurrence that may benefit from additional medical or public health interventions.


Morphological analysis of pancreatic adenocarcinoma on multidetector row computed tomography: implications for treatment response evaluation.
Response Evaluation Criteria in Solid Tumors (RECIST) guidelines assume spherical shape of tumors. Morphology of pancreatic adenocarcinoma (PAC) on multidetector row computed tomography was investigated to evaluate the applicability of RECIST guidelines. Study population comprised 16 patients with histologically confirmed localized PAC enrolled in a phase II clinical trial of chemoradiation. Pancreatic adenocarcinomas were segmented on baseline and follow-up multidetector row computed tomography with commercially available software. Tumor volumes (mL), RECIST diameter (mm), volume equivalent sphere diameter (VESD, mm), maximum 3-dimensional diameter (M3DD, mm), and elongation value were obtained. RECIST diameter, VESD and M3DD of the tumors at baseline and follow-up were compared to determine differences. Elongation values were analyzed. The significance level was set at P less than 0.05. Mean volume, RECIST diameter, VESD, M3DD, and elongation for baseline versus follow-up studies were 23.12 mL versus 19.43 mL (P > 0.05), 41.86 mm versus 39.35 mm (P > 0.05), 33.14 mm versus 32.1 mm (P > 0.05), 51.76 mm versus 51.73 mm (P > 0.05), and 0.67 versus 0.76 (P > 0.05), respectively. There was a significant difference at baseline and follow-up between RECIST diameter, VESD, and M3DD (P < 0.05, in all instances). Our results suggest that PACs are not spherical in shape. Evaluation of PAC treatment response based on RECIST guidelines may not be accurate.


Deficient lung surfactant apoproteins in amniotic fluid with mature phospholipid profile from diabetic pregnancies.
An enzyme-linked immunoassay to quantitate lung surfactant apoproteins (15 to 250 ng/ml) in human amniotic fluid is described. The immunoassay was used to quantify lung surfactant in 72 samples of amniotic fluid, for which lecithin/sphingomyelin (L/S) ratios, lecithin and phosphatidylglycerol concentrations, and foam stability indices were also available. The results obtained with the immunoassay were in general agreement with those of the other methods. Measurement of the apoproteins, however, may be a better predictor of fetal lung immaturity and of respiratory distress syndrome than the L/S ratio and the concentration of lecithin. This conclusion is based on the data obtained in the analyses of samples of amniotic fluid from four diabetic and five nondiabetic pregnancies, the infants of which developed respiratory distress. In all cases, the apoprotein concentration was less than 2.1 micrograms/ml, which indicated fetal lung immaturity. In six of these cases (one diabetic and five nondiabetic pregnancies), lung immaturity was also predicted on the basis of other tests. However, in three other cases of diabetic pregnancy, the L/S ratio and lecithin concentration falsely indicated lung maturity. In addition to its being an effective predictor of fetal lung maturity in diabetic, as well as nondiabetic, pregnancies, the immunoassay is better suited for clinical use because of its high specificity, sensitivity, and ease of performance.


Pain as the Perception of Someone: An Analysis of the Interface Between Pain Medicine and Philosophy.
Based largely on the so-called problem of "asymmetry in concept application", philosopher Murat Aydede has argued for a non-perceptual view of pain. Aydede is of course not denying basic neurobiological facts about neurons, action potentials, and the like, but he nonetheless makes a strong philosophical case for pain not being the perception of something extramental. In the present paper, after having stated some of the presuppositions I hold as a physician and pain researcher, and after having shortly described Aydede's critique of perceptual theories of pain, I make a constructive proposal centred around the concept of pain as the perception of some-one, not some-thing. In doing so, I propose that there often is a problematic duality at work when we think about pain, namely the mental/extramental duality. This pre-reflective mindset creates difficulties when reflecting over pain. Instead, I propose the body/world duality as being more helpful. Two neologisms, cosmoception and egoception, are presented as an alternative to the twin concepts of exteroception and interoception. It is argued that the new concepts have the advantage of not pushing our thought into a mental/extra-mental dichotomy. Hence, when in pain (which is an instance of egoception), I get epistemic access to the body that is I, to how I fare in this world. From that perspective, pain is not the perception of something, but of someone-namely, the self. In the final part of the paper, this proposal is discussed in dialogue with a paper from phenomenological thinker Jennifer Bullington.


Superiority of transcriptional profiling over procalcitonin for distinguishing bacterial from viral lower respiratory tract infections in hospitalized adults.
Distinguishing between bacterial and viral lower respiratory tract infection (LRTI) remains challenging. Transcriptional profiling is a promising tool for improving diagnosis in LRTI. We performed whole blood transcriptional analysis in 118 patients (median age [interquartile range], 61 [50-76] years) hospitalized with LRTI and 40 age-matched healthy controls (median age, 60 [46-70] years). We applied class comparisons, modular analysis, and class prediction algorithms to identify and validate diagnostic biosignatures for bacterial and viral LRTI. Patients were classified as having bacterial (n = 22), viral (n = 71), or bacterial-viral LRTI (n = 25) based on comprehensive microbiologic testing. Compared with healthy controls, statistical group comparisons (P < .01; multiple-test corrections) identified 3376 differentially expressed genes in patients with bacterial LRTI, 2391 in viral LRTI, and 2628 in bacterial-viral LRTI. Patients with bacterial LRTI showed significant overexpression of inflammation and neutrophil genes (bacterial > bacterial-viral > viral), and those with viral LRTI displayed significantly greater overexpression of interferon genes (viral > bacterial-viral > bacterial). The K-nearest neighbors algorithm identified 10 classifier genes that discriminated between bacterial and viral LRTI with a 95% sensitivity (95% confidence interval, 77%-100%) and 92% specificity (77%-98%), compared with a sensitivity of 38% (18%-62%) and a specificity of 91% (76%-98%) for procalcitonin. Transcriptional profiling is a helpful tool for diagnosis of LRTI.


Gender Typicality, Pressure to Conform to Gender Norms, and Anti-Fat and Appearance Stereotypes in Girls.
This mixed-methods study investigated the relation among gender identity (i.e., self-perceived gender similarity to girls and boys; self-perceived parental and peer pressure to conform to gender norms) and stereotyping about weight (i.e., anti-fat stereotypes), weight change, and appearance in 83 girls ages 6-9 (Mage = 7.60 years, SD = .85; 65% White, 16% Mixed/Other, 11% Black, 8% Latina) in the U.S. Stereotypes about weight change were assessed with open-ended responses (i.e., qualitatively), and the rest of the constructs were assessed with closed-ended responses (i.e., quantitatively). There was a positive association between pressure from parents to conform to gender norms and appearance stereotypes, and between pressure from peers and negative stereotypes about the fat and thin figures. Girls who were more gender typical, or more similar to girls, were more likely to endorse appearance stereotypes. There was no significant relation among stereotypes about weight change and gender identity. Many girls deemed the fat figure as unattractive, physically restrained, unhealthy, and likely to be bullied and believed that the fat figure should change back to look like a thinner figure. Most girls were aware that exercising, eating healthy, and dietary restriction can lead to weight loss and that food consumption and lack of exercise can lead to weight gain. Findings suggest that certain aspects of weight (anti-fat) and appearance stereotypes are closely linked to gender identity in girls, whereas other aspects of weight stereotypes (e.g., stereotypes about weight change) are so pervasive that they are common in most girls, regardless of their gender identity.


Medicaid beneficiaries' experiences in HMO and fee-for-service health care.
How do HMO-enrolled Medicaid beneficiaries' ratings of access to, and satisfaction with, their health care compare with the ratings of those beneficiaries receiving care in fee-for-service settings? Do poor single mothers report differences in access to, and satisfaction with, their HMO health care compared with those living in other family structures? These questions were examined with survey data from 961 California Medicaid recipients in 1991. Medicaid recipients enrolled in HMOs reported more difficulty gaining access to, and less satisfaction with, various aspects of the health care system. HMO-enrolled single mothers reported particularly negative experiences with their health care. The findings suggest a potential lack of fit between the health needs of the poor and the aims of managed health care.


The role of angiotensin receptor blockers in the management of chronic heart failure.
Clinical and basic science research has repeatedly confirmed the importance of the renin-angiotensin-aldosterone system in the pathophysiology of chronic heart failure. Accordingly, blockade of this system by angiotensin-converting enzyme (ACE) inhibitors has assumed a central role in the treatment of heart failure. Recently, angiotensin II receptor blockers (ARBs) have gained prominence as a possible substitute for ACE inhibitors in therapy for heart failure. However, clinical data compiled on this use of ARBs have shown them to be useful only as alternative therapy in ACE inhibitor-intolerant patients. Continuing large-scale clinical investigations may lead to an expansion of their role in therapy for various cardiovascular diseases.


Electron paramagnetic resonance detectable states of cytochrome P-450cam.
Cytochrome P-450cam is a low-spin Fe3+hemoprotein (g = 2.45, 2.26, and 1.91) which is made 60% high spin (g = 7.85, 3.97, and 1.78) at 12 K by the addition of 1 mol of substrate per mol of enzyme. Low-temperature EPR spectra show that the low-spin fraction of substrate-bound P-450cam contains two magnetic species. The majority species has an unusual EPR spectrum (g = 2.42, 2.24, and 1.97) which connot be simulated by using the range of crystal field parameters known for other heme proteins. The minority species has the same g values as substrate-free enzyme. Both low-spin species show Curie law temperature dependence below 50 K and have similar saturation behavior. Above 50 K the g = 2.42, 2.24, and 1.97 species rapidly loses signal intensity. The distribution of low-spin species is pH dependent (apparent pKa = 6.2) with the g = 2.42, 2.24, and 1.97 magnetic species favored at high pH. The substrate binding stoichiometry and the equilibria observed in the low-spin fraction suggest that there are not multiple protein forms of cytochrome P-450cam. Putidaredoxin and other effector molecules which specifically catalyze hydroxylation convert either the high-spin or the g = 2.42, 2.24, and 1.97 low-spin species to another new magnetic species (g = 2.47, 2.26, and 1.91). This species is only seen in the presence of substrate, and its stability reflects the catalytic potency of the effector molecule. The EPR and UV-visible spectra of cytochrome P-420 depend upon the manner in which the P-420 is generated. Incubation with acetone or reaction with N-ethylmaleimide or diethyl pyrocarbonate generates P-420 with different spectral characteristics. Through identification of active-site amino acids by chemical modification and comparison with porphyrin model complexes, the range of ligands likely to participate in each of the EPR detectable species is assigned. Mechanisms of interconversion of these species and their bearing on catalysis are discussed.


Self-assembly-induced luminescence of Eu3+-complexes and application in bioimaging.
Design and engineering of highly efficient emitting materials with assembly-induced luminescence, such as room-temperature phosphorescence (RTP) and aggregation-induced emission (AIE), have stimulated extensive efforts. Here, we propose a new strategy to obtain size-controlled Eu3+-complex nanoparticles (Eu-NPs) with self-assembly-induced luminescence (SAIL) characteristics without encapsulation or hybridization. Compared with previous RTP or AIE materials, the SAIL phenomena of increased luminescence intensity and lifetime in aqueous solution for the proposed Eu-NPs are due to the combined effect of self-assembly in confining the molecular motion and shielding the water quenching. As proof of concept, we also show that this system can be further applied in bioimaging, temperature measurement and HClO sensing. The SAIL activity of the rare-earth (RE) system proposed here offers a further step forward on the roadmap for the development of RE light conversion systems and their integration in bioimaging and therapy applications.


Industrial Trans Fatty Acid and Serum Cholesterol: The Allowable Dietary Level.
Trans fatty acid (TFA) from partially hydrogenated oil is regarded as the worst dietary fatty acid per gram due to its role in coronary heart disease. TFA consumption is decreasing worldwide, but some but not all observational studies indicate that TFA intake has little relevance to serum cholesterol levels in populations with low TFA intake (<1% E [percentage of total energy intake], <approximately 2 g/day). Few intervention trials examined the effect of TFAs on blood cholesterol at relatively low levels (<2% E); no definite evidence is available on the tolerable upper level of the intake. A series of our intervention studies in Japanese suggested that an industrial TFA intake at <1% E does not influence the serum cholesterol level. To establish allowable level, we must consider not only the dietary level of TFAs, but also the composition of dietary fats simultaneously consumed, that is, saturated and unsaturated fatty acids. These fatty acids strengthen or counteract the adverse effect of TFAs on serum cholesterol levels. In this review we describe the complex situation of the cardiovascular effects of industrial TFAs. The relationship between dietary industrial TFAs and concentration of plasma cholesterol should be evaluated from the viewpoint of dietary patterns rather than TFAs alone.


A cross-scale trophic cascade from large predatory fish to algae in coastal ecosystems.
Trophic cascades occur in many ecosystems, but the factors regulating them are still elusive. We suggest that an overlooked factor is that trophic interactions (TIs) are often scale-dependent and possibly interact across spatial scales. To explore the role of spatial scale for trophic cascades, and particularly the occurrence of cross-scale interactions (CSIs), we collected and analysed food-web data from 139 stations across 32 bays in the Baltic Sea. We found evidence of a four-level trophic cascade linking TIs across two spatial scales: at bay scale, piscivores (perch and pike) controlled mesopredators (three-spined stickleback), which in turn negatively affected epifaunal grazers. At station scale (within bays), grazers on average suppressed epiphytic algae, and indirectly benefitted habitat-forming vegetation. Moreover, the direction and strength of the grazer-algae relationship at station scale depended on the piscivore biomass at bay scale, indicating a cross-scale interaction effect, potentially caused by a shift in grazer assemblage composition. In summary, the trophic cascade from piscivores to algae appears to involve TIs that occur at, but also interact across, different spatial scales. Considering scale-dependence in general, and CSIs in particular, could therefore enhance our understanding of trophic cascades.


Ehlers-Danlos syndrome: surgical management of mitral regurgitation and atrial fibrillation.
Ehlers-Danlos syndrome is associated with fatal cardiovascular complications and intraoperative death. The lack of literature on cardiac surgery in the syndrome makes it difficult for surgeons to assess the risk of an operation and to choose optimal treatment. We describe the case of a 63-year-old man with Ehlers-Danlos syndrome type I or II, mitral regurgitation and atrial fibrillation who was managed successfully with mitral valve repair and surgical cardioversion.


Approach to the patient with unexplained chest pain.
Patients with unexplained or noncardiac chest pain continue to present a difficult challenge to the gastroenterologist. Cardiac disease must be ruled out first as the history will not distinguish between coronary artery disease and other causes of substernal chest pain. A systematic approach to evaluation should include reassurance that the heart is normal and attempts to confirm an esophageal etiology. Gastroesophageal reflux disease is the most common esophageal abnormality associated with unexplained chest pain and may be identified by an aggressive trial of anti-reflux therapy or an abnormal prolonged ambulatory pH monitoring study. Endoscopy is almost always normal and of less use in this population than in those with heartburn as the presenting symptom. Judicious use of manometry with provocative testing to evaluate for esophageal motility abnormalities or esophageal sensitivity allows for optimal evaluation of those who do not have gastroesophageal reflux disease. This article reviews the clinical presentation, differential diagnosis, and approach to evaluation and therapy of this complex group of patients.


Identification and Characterization of TALE Homeobox Genes in the Endangered Fern Vandenboschia speciosa.
We report and discuss the results of a quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis of the expression patterns of seven three amino acid loop extension (TALE) homeobox genes (four KNOTTED-like homeobox (KNOX) and three BEL1-like homeobox (BELL) genes) identified after next generation sequencing (NGS) and assembly of the sporophyte and gametophyte transcriptomes of the endangered fern species Vandenboschia speciosa. Among the four KNOX genes, two belonged to the KNOX1 class and the other two belonged to the KNOX2 class. Analysis of the deduced amino acid sequences supported the typical domain structure of both types of TALE proteins, and the homology to TALE proteins of mosses, lycophytes, and seed plant species. The expression analyses demonstrate that these homeodomain proteins appear to have a key role in the establishment and development of the gametophyte and sporophyte phases of V. speciosa lifecycle, as well as in the control of the transition between both phases. Vandenboschia speciosa VsKNAT3 (a KNOX2 class protein) as well as VsBELL4 and VsBELL10 proteins have higher expression levels during the sporophyte program. On the contrary, one V. speciosa KNOX1 protein (VsKNAT6) and one KNOX2 protein (VsKNAT4) seem important during the development of the gametophyte phase. TALE homeobox genes might be among the key regulators in the gametophyte-to-sporophyte developmental transition in regular populations that show alternation of generations, since some of the genes analyzed here (VsKNAT3, VsKNAT6, VsBELL4, and VsBELL6) are upregulated in a non-alternating population in which only independent gametophytes are found (they grow by vegetative reproduction outside of the range of sporophyte distribution). Thus, these four genes might trigger the vegetative propagation of the gametophyte and the repression of the sexual development in populations composed of independent gametophytes. This study represents a comprehensive identification and characterization of TALE homeobox genes in V. speciosa, and gives novel insights about the role of these genes in fern development.


Comprehensive investigation into the effects of yeast dietary fiber and temperature on konjac glucomannan/kappa-carrageenan for the development of fat analogs.
In this study, yeast dietary fiber (YDF) was incorporated into konjac glucomannan/kappa-carrageenan (KGM/κ-KC) for the development of fat analogs, and the impact of YDF on the gelation properties and behavior of KGM/κ-KC composite gels was assessed. YDF improved the composite gel whiteness value, and affected the mechanical properties of the composite gel, especially enhancing its hardness, and decreasing its chewiness, elasticity, and gel strength, making it more similar to porcine back fat. When the yeast dietary fiber content was 0.033 g/mL and the heating temperature was 80 °C (T80-2), the textural properties of the composite gel were closest to porcine back fat. The frequency sweep results suggested that YDF incorporation led to enhancement of the intermolecular interaction and intermixing and interaction among more easily at higher processing temperatures (80 °C and 90 °C). By scanning electron microscopy, the fatty surface of porcine back fat was flat and covered with a large amount of oil, while KGM/κ-KC/YDF composite gels developed a dense, stacked network structure. YDF caused more fragmented, folded, and uneven structures to emerge. Overall, YDF could influence the gel behavior of KGM/κ-KC composite gels, and change their colors and mechanical properties. This work could serve as a guide for preparing fat analogs with KGM/κ-KC composite gels.


Detection and Analysis of the Hedgehog Signaling Pathway-Related Long Non-Coding RNA (lncRNA) Expression Profiles in Keloid.
BACKGROUND Hedgehog (Hh) signaling pathway-related genes have important roles in several physiological and disease processes that involve cell proliferation. Long non-coding region RNAs (lncRNAs) have a regulatory role on gene expression. Keloid is characterized by excessive proliferation of scar tissue following trauma. The aims of this study were to evaluate the Hh signaling pathway in keloid skin tissues and its downstream gene expression and lncRNAs, compared with normal skin. MATERIAL AND METHODS Four pairs of keloids and adjacent normal skin epidermis underwent total RNA extraction. Gene chip high-throughput real-time quantitative polymerase chain reaction (qPCR) was used to examine the differential expression profiles of the Hh signaling pathway-related lncRNAs and mRNAs in the human keloid and normal skin. The differentially expressed mRNAs were analyzed by Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) to identify their biological roles. RESULTS In keloid tissue, differential expression of 33 mRNAs and 30 lncRNAs relating to the Hh pathway, were verified by gene chip qPCR. The results of GO and KEGG analysis showed that the upregulated mRNAs were involved in cell proliferation, cell growth, and tissue repair, and down-regulated mRNAs were involved in apoptosis. The lncRNA, AC073257.2, affected cell keloid growth and proliferation by its upstream target the GLI2 gene at the transcriptional level. The lncRNA, HNF1A-AS1, affected cell keloid growth and proliferation by its neighboring target gene, HNF1A. CONCLUSIONS Differential expression occurred in Hh signaling pathway-related lncRNAs and mRNAs, which may provide further insight into the development of keloid.


Morphological and functional effects of extracellular matrix on pancreatic islet cell cultures.
Extracellular matrix (ECM) has been reported to enhance epithelial cell attachment and proliferation as well as to induce differentiation in vitro. In an attempt to determine the benefits of culturing pancreatic islet cells on ECM, we studied the morphological and functional patterns of rat islet cells and an insulin-secreting tumor cell line. ECM enhanced islet cell attachment and proliferation when compared to plastic, as suggested by a higher specific activity of DNA synthesis and a higher mitotic index. Cells on ECM were heterogeneous in size and insulin content. They showed extended areas of confluence. Cultures on plastic demonstrated an organisation in clusters and low mitotic activity. However, ECM did not allow for reconstitution of an islet-like structure. When compared to plastic, an initial decrease in basal and stimulated insulin secretion per million cells was observed on ECM, but B-cell activity was restored after 6 days of culture. Glucagon and somatostatin secretion were similar on both substrates. These data suggest that ECM enhances markedly islet cells attachment and proliferation, as well as long-term culture maintenance.


Design and Implementation of a Portable Knee Actuator for the Improvement of Crouch Gait in Children with Cerebral Palsy.
Common manifestation of spastic Cerebral Palsy (CP) are abnormal gait pathologies. These conditions require greater energy expenditure to successfully ambulate and are linked with significant deterioration in joint health and childhood musculoskeletal development. Crouch gait presents with knee hyperflexion throughout stance due to extensor muscle weakness and spasticity in flexor muscles stemming from neurological damage. The goal of this study was to develop a wearable cable-driven robotic system that applies controlled perturbation to the knee joint during overground walking in children with CP. Two children with spastic CP were recruited in this pilot study. They were tested in two conditions, i.e., applying knee resistance vs. knee assistance during overground walking. Kinematic and EMG data were recorded during overground walking. Data indicated that it was feasible to apply controlled knee perturbation torque during overground walking in children with crouch and preliminary results showed an improvement in crouch gait pattern in children with CP after one session of walking with the robotic system.Clinical Relevance- This study might have a potential clinical significance modifying neuromuscular control of CP patients with Crouch Gait.


Xenoreactive natural antibodies in the world of natural antibodies: typical or unique?
Xenoreactive natural antibodies are thought to be responsible for initiating the hyperacute rejection of porcine organs transplanted into primates. Progress has been made in recent years in the characterization of the specificity and functions of these antibodies. Xenoreactive natural antibodies recognize Gal alpha 1-3Gal, a carbohydrate related to the blood group A and blood group B antigens. The presence of Gal alpha 1-3Gal may not be sufficient to allow the binding of xenoreactive natural antibodies under physiological conditions; rather, the clustering of Gal alpha 1-3Gal determinants may dictate the extent to which xenoreactive natural antibodies attach to surfaces on which Gal alpha 1-3Gal is expressed. The predominant role of xenoreactive natural antibodies in the pathogenesis of hyperacute rejection in porcine-to-primate xenotransplantation involves the activation of complement. Complement activation is mediated by IgM, not IgG xenoreactive natural antibodies. Based on functional avidity, thermal binding optima, thermal liability and homogeneity of binding interactions, xenoreactive natural IgM appear to be members of a family of natural antibodies which includes isohaemagglutinins. The classification of xenoreactive antibodies and isohaemagglutinins into a family of antibodies may provide further insight into the nature and physiological functions of xenoreactive natural antibodies.


Psammaplin A inhibits hepatitis C virus NS3 helicase.
Hepatitis C virus (HCV) is the causative agent of hepatitis C, a chronic infectious disease that can lead to development of hepatocellular carcinoma. The NS3 nucleoside triphosphatase (NTPase)/helicase has an essential role in HCV replication, and is therefore an attractive target for direct-acting antiviral strategies. In this study, we employed high-throughput screening using a photo-induced electron transfer (PET) system to identify an inhibitor of NS3 helicase from marine organism extracts. We successfully identified psammaplin A as a novel NS3 inhibitor. The dose-response relationship clearly demonstrates the inhibition of NS3 RNA helicase and ATPase activities by psammaplin A, with IC₅₀ values of 17 and 32 μM, respectively. Psammaplin A has no influence on the apparent Km value (0.4 mM) of NS3 ATPase activity, and acts as a non-competitive inhibitor. Additionally, it inhibits the binding of NS3 to single-stranded RNA in a dose-dependent manner. Furthermore, psammaplin A shows an inhibitory effect on viral replication, with EC₅₀ values of 6.1 and 6.3 μM in subgenomic replicon cells derived from genotypes 1b and 2a, respectively. We postulate that psammaplin A is a potential anti-viral agent through the inhibition of ATPase, RNA binding and helicase activities of NS3.


Symmetry and coplanarity of organic molecules affect their packing and photovoltaic properties in solution-processed solar cells.
In this study we synthesized three acceptor-donor-acceptor (A-D-A) organic molecules, TB3t-BT, TB3t-BTT, and TB3t-BDT, comprising 2,2'-bithiophene (BT), benzo[1,2-b:3,4-b':5,6-d″]trithiophene (BTT), and benzo[1,2-b;4,5-b']dithiophene (BDT) units, respectively, as central cores (donors), terthiophene (3t) as π-conjugated spacers, and thiobarbituric acid (TB) units as acceptors. These molecules display different degrees of coplanarity as evidenced by the differences in dihedral angles calculated from density functional theory. By using differential scanning calorimetry and X-ray diffractions for probing their crystallization characteristics and molecular packing in active layers, we found that the symmetry and coplanarity of molecules would significantly affect the melting/crystallization behavior and the formation of crystalline domains in the blend film with fullerene, PC61BM. TB3t-BT and TB3t-BDT, which each possess an inversion center and display high crystallinity in their pristine state, but they have different driving forces in crystallization, presumably because of different degrees of coplanarity. On the other hand, the asymmetrical TB3t-BTT behaved as an amorphous material even though it possesses a coplanar structure. Among our tested systems, the device comprising as-spun TB3t-BDT/PC61BM (6:4, w/w) active layer featured crystalline domains and displayed the highest power conversion efficiency (PCE) of 4.1%. In contrast, the as-spun TB3t-BT/PC61BM (6:4, w/w) active layer showed well-mixed morphology and with a device PCE of 0.2%; it increased to 3.9% after annealing the active layer at 150 °C for 15 min. As for TB3t-BTT, it required a higher content of fullerene in the TB3t-BTT/PC61BM (4:6, w/w) active layer to optimize its device PCE to 1.6%.


[Appraisal of Lyme borreliosis].
Lyme borreliosis is due to infection with a tick-borne spirochete. The variety of signs and symptoms and also the laboratory tests of this multisystem illness often cause great problems in the appraisal of this disease. Frequently indispositions are associated with positive antibody tests for Borrelia burgdorferi. Terms as fibromyalgia or chronic fatigue syndrome are often connected with the diagnosis of Lyme disease. Outdoor workers such as farmers, foresters, hunters, woodcutters and gamekeepers in areas of endemic disease take a great occupational risk of infection with borreliosis. In the German health and social insurance the appraisal of this disease is of great importance. Affected working people can receive financial compensation. Not only serological investigations with the presence of specific antibodies, but also clinical findings must be considered.


Paeoniflorin on Rat Myocardial Ischemia Reperfusion Injury of Protection and Mechanism Research.
To study the myocardial benefit effect and mechanism of paeoniflorin on myocardial ischemia reperfusion injury (MIRI) in rats. Hundred SD rats were randomly divided into 5 groups: sham group, model group, Paeoniflorin (15 mg/kg) group, Paeoniflorin (30 mg/kg) group, and Paeoniflorin (60 mg/kg) group. Myocardial ischemia reperfusion model was established in each group except the sham group. The myocardial infarction and morphological changes were measured by the TTC staining and HE staining respectively. Myocardial caspase-3 was detected by immunohistochemistry. In addition, the protein levels of Bcl-2 and Bax and the expression ratio of p-erk, p-jnk, and p-p38 were detected by Western blot. Myocardial superoxide dismutase (SOD) activity and malondialdehyde (MDA) level were measured by the assay kit. Paeoniflorin (30 mg/kg) and Paeoniflorin (60 mg/kg) can obviously alleviate myocardial infarction caused by MIRI (p < 0.05). HE staining showed that the myocardial morphology in the treatment group was obviously better than that in the model group. WB and immunohistochemistry showed that Paeoniflorin (30 mg/kg) and Paeoniflorin (60 mg/kg) can significantly increase the reduced protein level of bcl-2 (p < 0.05) and reduce the increased protein level of caspase3, bax -p-erk, p-jnk, and p-p38 caused by MIRI (p < 0.05). The activity of SOD was increased and the level of MDA was decreased after Paeoniflorin treatment. Paeoniflorin preconditioning has a protective effect on MIRI in rats. Its mechanism is related to reducing oxidative stress and apoptosis by inhibiting the expression of apoptosis-related signaling pathway.


Surgical considerations in aortitis. Part II: Mycotic aneurysms.
Mycotic aneurysms were first described as the result of nonsyphilitic bacterial infection of the arterial wall. They were often caused by septic emboli from bacterial endocarditis and were much more common prior to the era of antibiotics. Although the aorta is most often affected, such aneurysms may arise in any artery. Classified according to their etiology as primary, secondary, and cryptic mycotic aneurysms, it is imperative that immediate treatment be instituted once the diagnosis is confirmed by angiography. The patient should be placed on high doses of antibiotics appropriate for the offending pathogens. The goal of surgical treatment is the complete removal of the infected aneurysm and preservation of the distal circulation. Alternate methods of treating these lesions include the use of bypass grafts and interposition grafts. The virulence of the offending pathogen and the sensitivity to antibiotic therapy play a significant role in the success of these procedures.


Photothermal Circular Dichroism Measurements of Single Chiral Gold Nanoparticles Correlated with Electron Tomography.
Chemically synthesized metal nanoparticles with morphological chiral features are known to exhibit strong circular dichroism. However, we still lack understanding of the correlation between morphological and chiroptical features of plasmonic nanoparticles. To shed light on that question, single nanoparticle experiments are required. We performed photothermal circular dichroism measurements of single chiral and achiral gold nanoparticles and correlated the chiroptical response to the 3D morphology of the same nanoparticles retrieved by electron tomography. In contrast to an ensemble measurement, we show that individual particles within the ensemble display a broad distribution of strength and handedness of circular dichroism signals. Whereas obvious structural chiral features, such as helical wrinkles, translate into chiroptical ones, nanoparticles with less obvious chiral morphological features can also display strong circular dichroism signals. Interestingly, we find that even seemingly achiral nanoparticles can display large g-factors. The origin of this circular dichroism signal is discussed in terms of plasmonics and other potentially relevant factors.


[A new interferon inducer active via peroral administration].
Biological activities of a new oral interferon inducer, Tash-4, were studied. In maximal soluble doses (2000 mg/kg body weight) the inducer was shown to have no toxicity. An important advantage of Tash-4 over other known interferon inducers consists in its capacity to induce interferon in high concentrations in the intestinal tract. Upon repeated administration of Tash-4 hyporeactivity is detectable within 3 days and persists for up to 6 days. A change of inducers may be used to overcome the hyporeactivity. Sequential administration of Tash-4 and Tash-3 induced in animals interferon production in response first to one inducer, then to the other. It is assumed that the mechanisms of induction and suppression of interferon induced by these preparations differ.


Body composition and pulmonary function in cystic fibrosis.
Lower body mass index (BMI) is associated with worse pulmonary function in cystic fibrosis (CF). lean body mass (LBM) is more strongly associated with pulmonary function than BMI is. Anthropometrics, body composition by dual x-ray absorptiometry, and pulmonary function were determined in pancreatic insufficient CF (PI-CF) youth. Sex and age-adjusted Z-scores (BMI-Z, LBMI-Z, FMI-Z) were generated for CF and controls. (1) Associations of BMI-Z with LBMI-Z and FMI-Z and (2) age-adjusted associations of BMI-Z, LBMI-Z, and FMI-Z with FEV1%-predicted were tested. Two hundred eight PI-CF subjects had lower BMI-Z, LBMI-Z, and FMI-Z compared to 390 controls. BMI-Z was associated with lower LBMI-Z (p < 0.0001) in PI-CF. In females, LBMI-Z and BMI-Z were positively associated with FEV1%-predicted; this relationship did not persist for FMI-Z after adjustment for LBMI-Z. In males, only LBMI-Z and BMI-Z were associated with FEV1%-predicted. In PI-CF youth, deficits in LBM were apparent. At lower BMI percentiles, BMI may not accurately depict LBM in PI-CF. In under-nourished PI-CF youth, this preservation of FM in preference to LBM is relevant since LBMI-Z, but not FMI-Z, is positively associated with FEV1%-predicted. Lean body mass index is more strongly associated with lung function compared to BMI, especially in the under-nourished child and adolescent with PI-CF.


Krüppel-like transcription factors KLF1 and KLF2 have unique and coordinate roles in regulating embryonic erythroid precursor maturation.
The Krüppel-like transcription factors KLF1 and KLF2 are essential for embryonic erythropoiesis. They can partially compensate for each other during mouse development, and coordinately regulate numerous erythroid genes, including the β-like globins. Simultaneous ablation of KLF1 and KLF2 results in earlier embryonic lethality and severe anemia. In this study, we determine that this anemia is caused by a paucity of blood cells, and exacerbated by diminished β-like globin gene expression. The anemia phenotype is dose-dependent, and, interestingly, can be ameliorated by a single copy of the KLF2, but not the KLF1 gene. The roles of KLF1 and KLF2 in maintaining normal peripheral blood cell numbers and globin mRNA amounts are erythroid cell-specific. Mechanistic studies led to the discovery that KLF2 has an essential function in erythroid precursor maintenance. KLF1 can partially compensate for KLF2 in this role, but is uniquely crucial for erythroid precursor proliferation through its regulation of G1- to S-phase cell cycle transition. A more drastic impairment of primitive erythroid colony formation from embryonic progenitor cells occurs with simultaneous loss of KLF1 and KLF2 than with loss of a single factor. KLF1 and KLF2 coordinately regulate several proliferation-associated genes, including Foxm1. Differential expression of FoxM1, in particular, correlates with the observed KLF1 and KLF2 gene dosage effects on anemia. Furthermore, KLF1 binds to the FoxM1 gene promoter in blood cells. Thus KLF1 and KLF2 coordinately regulate embryonic erythroid precursor maturation through the regulation of multiple homeostasis-associated genes, and KLF2 has a novel and essential role in this process.


Endogenous CD4+ T Cells Recognize Neoantigens in Lung Cancer Patients, Including Recurrent Oncogenic KRAS and ERBB2 (Her2) Driver Mutations.
T cells specific for neoantigens encoded by mutated genes in cancers are increasingly recognized as mediators of tumor destruction after immune-checkpoint inhibitor therapy or adoptive cell transfer. Much of the focus has been on identifying epitopes presented to CD8+ T cells by class I MHC. However, CD4+ class II MHC-restricted T cells have been shown to have an important role in antitumor immunity. Unfortunately, the vast majority of neoantigens recognized by CD8+ or CD4+ T cells in cancer patients result from random mutations and are patient-specific. Here, we screened the blood of 5 non-small cell lung cancer (NSCLC) patients for T-cell responses to candidate mutation-encoded neoepitopes. T-cell responses were detected to 8.8% of screened antigens, with 1 to 7 antigens identified per patient. A majority of responses were to random, patient-specific mutations. However, CD4+ T cells that recognized the recurrent KRAS G12V and the ERBB2 (Her2) internal tandem duplication (ITD) oncogenic driver mutations, but not the corresponding wild-type sequences, were identified in two patients. Two different T-cell receptors (TCR) specific for KRAS G12V and one T-cell receptor specific for Her2-ITD were isolated and conferred antigen specificity when transfected into T cells. Deep sequencing identified the Her2-ITD-specific TCR in the tumor but not nonadjacent lung. Our results showed that CD4+ T-cell responses to neoantigens, including recurrent driver mutations, can be derived from the blood of NSCLC patients. These data support the use of adoptive transfer or vaccination to augment CD4+ neoantigen-specific T cells and elucidate their role in human antitumor immunity.


Cutting costs: the impact of price lists on the cost development at the emergency department.
It was shown that physicians working at the Swedish emergency department (ED) are unaware of the costs for investigations performed. This study evaluated the possible impact of price lists on the overall laboratory and radiology costs at the ED of a Swedish university hospital. Price lists including the most common laboratory analyses and radiological investigations at the ED were created. The lists were distributed to all internal medicine physicians by e-mail and exposed above their working stations continually. No lists were provided for the orthopaedic control group. The average costs for laboratory and radiological investigations during the months of June and July 2007 and 2008 were calculated. Neither clinical nor admission procedures were changed. The physicians were blinded towards the study. Statistical analysis was performed using the Student's t-test. A total of 1442 orthopaedic and 1585 medical patients were attended to in 2007. In 2008, 1467 orthopaedic and 1637 medical patients required emergency service. The average costs per patient were 980.27 SKR (98€)/999.41 SKR (100€, +1.95%) for orthopaedic and 1081.36 SKR (108€)/877.3 SKR (88€, -18.8%) for medical patients. Laboratory costs decreased by 9% in orthopaedic and 21.4% in medical patients. Radiology costs changed +5.4% in orthopaedic and -20.59% in medical patients. The distribution and promotion of price lists as a tool at the ED to heighten cost awareness resulted in a major decrease in the investigation costs. A significant decrease in radiological costs could be observed. It can be concluded that price lists are an effective tool to cut costs in public healthcare.


Association between fertility and HIV status: what implications for HIV estimates?
Most estimates of HIV prevalence have been based on sentinel surveillance of pregnant women which may either under-estimate or over-estimate the actual prevalence in adult female population. One situation which can lead to either an underestimate or an overestimate of the actual HIV prevalence is where there is a significant difference in fertility rates between HIV-positive and HIV-negative women. Our aim was to compare the fertility rates of HIV-infected and HIV-uninfected women in Cameroon in order to make recommendations on the appropriate adjustments when using antenatal sentinel data to estimate HIV prevalence. Cross-sectional, population-based study using data from 4493 sexually active women aged 15 to 49 years who participated in the 2004 Cameroon Demographic and Health Survey. In the rural area, the age-specific fertility rates in both HIV positive and HIV negative women increased from 15-19 years age bracket to a maximum at 20-24 years and then decreased monotonically till 35-49 years. Similar trends were observed in the urban area. The overall fertility rate for HIV positive women was 118.7 births per 1000 woman-years (95% Confidence Interval [CI] 98.4 to 142.0) compared to 171.3 births per 1000 woman-years (95% CI 164.5 to 178.2) for HIV negative women. The ratio of the fertility rate in HIV positive women to the fertility rate of HIV negative women (called the relative inclusion ratio) was 0.69 (95% CI 0.62 to 0.75). Fertility rates are lower in HIV-positive than HIV-negative women in Cameroon. The findings of this study support the use of summary RIR for the adjustment of HIV prevalence (among adult female population) obtained from sentinel surveillance in antenatal clinics.


Guanosine 5'-triphosphate and guanosine 5'-[beta gamma-imido]triphosphate effect a collision coupling mechanism between the glucagon receptor and catalytic unit of adenylate cyclase.
1. GTP, but not p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate), abolishes the sensitivity of glucagon-stimulated adenylate cyclase to the lipid-phase separations occurring in the outer half of the bilayer in liver plasma membranes from rat. 2. When either GTP or p[NH]ppG alone stimulate adenylate cyclase, the enzyme senses only those lipid-phase separations occurring in the inner half of the bilayer. 3. Trypsin treatment of intact hepatocytes has no effect on the basal, fluoride-, GTP- or p[NH]ppG-stimulated adenylate cyclase activity. However, (125)I-labelled-glucagon specific binding decays with a half-life matching that of the decay of glucagon-stimulated adenylate cyclase activity. 4. When GTP or p[NH]ppG are added to assays of glucagon-stimulated activity, the half-life of the trypsin-mediated decay of activity is substantially increased and the decay plots are no longer first-order. 5. Trypsin treatment of purified rat liver plasma membranes abolishes basal and all ligand-stimulated adenylate cyclase activity, and (125)I-labelled-glucagon specific binding. 6. Benzyl alcohol activates the GTP- and p[NH]ppG-stimulated activities in an identical fashion, whereas these activities are affected differently when glucagon is present in the assays. 7. We suggest that guanine nucleotides alter the mode of coupling between the receptor and catalytic unit. In the presence of glucagon and GTP, a complex of receptor, catalytic unit and nucleotide regulatory protein occurs as a transient intermediate, releasing a free unstable active catalytic unit. In the presence of p[NH]ppG and glucagon, the transient complex yields a relatively stable complex of the catalytic unit associated with a p[NH]ppG-bound nucleotide-regulatory protein.


Acquisition of concomitant oral and genital infection with herpes simplex virus type 2.
Primary oral and genital infections caused by herpes simplex virus type 2 were diagnosed in an 18-year-old female. A history of sexual practices was critical in determining the anatomic sites of infection. Restriction endonuclease analysis of viral DNAs helped to identify the male sexual partner from whom the virus had been acquired. He had been infected recently by a previous sexual partner but had not yet developed lesions. Clinicians should obtain a history of sexual practices from patients with newly acquired genital herpes and should advise patients with genital herpes that transmission of virus to sexual partners can occur in the absence of overt lesions.


Pleomorphic adenoma of the breast. A case report and distinction from mucinous carcinoma.
Pleomorphic adenoma of the breast is a rare, benign tumor accounting for 68 cases in the literature. It is most commonly seen in postmenopausal women and is characterized by an admixture of epithelial and myoepithelial cells embedded in abundant myxomatous stroma. Its clinical and histologic appearance can be challenging and may lead to a misdiagnosis of invasive carcinoma. We report a case of mammary pleomorphic adenoma in an asymptomatic 59-year-old woman and briefly discuss its distinction from mucinous carcinoma through the use of special stains.


Doppler echocardiographic study of right ventricular systolic performance in inferior myocardial infarction.
Twenty-four male patients with myocardial infarction (MI) without clinical and electrocardiographic signs of right ventricular (RV) involvement were selected to enter the study. All the patients were divided into two groups: Group I consisted of 12 patients with anterior MI and infarct-related left anterior descending artery and Group II included 12 patients with inferior MI and infarct-related right coronary artery. Patients of Group II had higher right atrial pressure and right atrial pressure/pulmonary capillary wedge pressure ratio (p<0.01, p<0.01) and lower values of pulmonary flow velocity, mean acceleration, and pulmonary flow velocity2/acceleration time ratio than patients of Group I (p<0.01, p<0.01, p<0.01, respectively). Pulmonary flow indices correlated inversely and significantly with hemodynamic dysfunction in patients with inferior myocardial infarction and right coronary proximal lesions (p<0.01).


Formate-induced inhibition of photoreceptor function in methanol intoxication.
Formic acid is the toxic metabolite responsible for the retinal and optic nerve toxicity produced in methanol intoxication. Previous studies in our laboratory have documented formate-induced retinal dysfunction and histopathology in a rodent model of methanol intoxication. The present studies define the time and concentration dependence of formate-induced retinal toxicity in methanol-intoxicated rats. Retinal function was assessed 24, 48, and 72 h after the initial dose of methanol by flicker electroretinographic measurements. Retinal histopathology was assessed at the same time intervals. Rod- and cone-mediated electroretinogram (ERG) responses were attenuated in a formate concentration- and time-dependent manner, and both retinal sensitivity and maximal responsiveness to light were diminished. Attenuation of UV-cone-mediated responses was temporally delayed in comparison to the functional deficits observed in the 15 Hz/510 nm responses, which have a rod-mediated component and occurred at significantly higher formate concentrations. Both 15 Hz/510 nm and UV-cone-mediated ERG responses were undetectable by 72 h; however, if light intensity was increased, a retinal ERG response could be recorded, indicating that photoreceptor function was profoundly attenuated, but not abolished, under these intoxication conditions. Functional changes preceded structural alterations. Histopathological changes were most pronounced in the outer retina with evidence of inner segment swelling, photoreceptor mitochondrial disruption, and the appearance of fragmented photoreceptor nuclei in the outer nuclear layer. The nature of both the functional and structural alterations observed are consistent with formate-induced inhibition of mitochondrial energy production, resulting in photoreceptor dysfunction and pathology.


Sequence and structure classification of kinases.
Kinases are a ubiquitous group of enzymes that catalyze the phosphoryl transfer reaction from a phosphate donor (usually ATP) to a receptor substrate. Although all kinases catalyze essentially the same phosphoryl transfer reaction, they display remarkable diversity in their substrate specificity, structure, and the pathways in which they participate. In order to learn the relationship between structural fold and functional specificities in kinases, we have done a comprehensive survey of all available kinase sequences (>17,000) and classified them into 30 distinct families based on sequence similarities. Of these families, 19, covering nearly 98% of all sequences, fall into seven general structural folds for which three-dimensional structures are known. These fold groups include some of the most widespread protein folds, such as Rossmann fold, ferredoxin fold, ribonuclease H fold, and TIM beta/alpha-barrel. On the basis of this classification system, we examined the shared substrate binding and catalytic mechanisms as well as variations of these mechanisms in the same fold groups. Cases of convergent evolution of identical kinase activities occurring in different folds are discussed.