[Economic assessment of the use of biological medications in military medicine].
By the example of rheumatoid arthritis was considered the methods of economic assessment of advisability of modern biological medication usage in military medicine. The usage of these medications leads to quick clinical response, decrease of the additional medication intake, decrease of the average duration of treatment, "move aside" time of permanent disablement. Modern active therapy can increase the effectiveness of treatment; improve the quality of life, decrease the average duration of hospital stay, following treatment expense and welfare payment for this category of ill.


High-voltage leak detection of a parenteral proteinaceous solution product packaged in form-fill-seal plastic laminate bags. Part 2. Method performance as a function of heat seal defects, product-package refrigeration, and package plastic laminate lot.
Part 1 of this three-part research series detailed the development and validation of a high-voltage leak detection test (HVLD, also known as an electrical conductivity and capacitance test) for verifying the container-closure integrity of a small-volume laminate plastic bag containing an aqueous solution formulation of the rapid-acting insulin analogue, insulin aspart (NovoRapid®/NovoLog®) by Novo Nordisk A/S, Bagsværd, Denmark. Leak detection capability was verified using positive controls each with a single laser-drilled hole in the bag film face. In this Part 2, HVLD leak detection capability was further explored in four separate studies. Study 1 investigated the ability of HVLD to detect weaknesses and/or gaps in the bag heat seal. Study 2 checked the HVLD detection of bag holes in packages stored 4 days at ambient conditions followed by 17 days at refrigeration. Study 3 examined HVLD test results for packages tested when cold. Study 4 compared HVLD test results as a function of bag plastic film lots. The final Part 3 of this series will report the impact of HVLD exposure on product visual appearance and chemical stability. In Part 1 of this three-part series, a leak test method based on electrical conductivity and capacitance, also called high-voltage leak detection (HVLD), was used to find leaks in small plastic bags filled with a solution for injection of the rapid-acting insulin analogue, insulin aspart (NovoRapid®/NovoLog®) by Novo Nordisk A/S, Bagsværd, Denmark. In this Part 2, HVLD leak detection capability was further explored in four separate studies. Study 1 investigated the ability of HVLD to detect bag heat seal leaks. Study 2 checked HVLD's ability to detect bag holes after a total of 21 days at ambient plus refrigerated temperatures. Study 3 looked to see if HVLD results changed for packages tested when still cold. Study 4 compared HVLD results for multiple bag plastic film lots. The final Part 3 of this series will report any evidence of drug component degradation caused by HVLD exposure.


Isoflavone composition, phenol content, and antioxidant activity of soybean seeds from India and Bulgaria.
Isoflavone levels and isoflavone chemical composition in 11 cultivars of soybean, including 4 Indian and 7 genotypes of soybean grown in Bulgaria, were analyzed as determined by C 18 reversed phase high-performance liquid chromatography coupled with a photodiode array detector. Antioxidant activity of soybean extracts was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and total phenolic compounds (TPC) were determined by using Folin-Ciocalteu reagent. The range of total isoflavones (TI) was 558.2-1048.6 microg g (-1) of soy in Indian cultivars, and it was 627.9-1716.9 microg g (-1) of soy in the case of Bulgarian cultivars. The highest and lowest total isoflavone contents were observed for Maus-2 (1048.6 microg g (-1) of soy) and Hardee (558.2 microg g (-1) of soy), respectively, for the Indian cultivars, and they were observed for Boryara (1716.9 microg g (-1) of soy) and Line 5 (627.9 microg g (-1) of soy) for the Bulgarian genotypes. DPPH radical scavenging activity did not differ significantly among the cultivars and did not correlate with TI, whereas TPC correlated well with TI and weakly with DPPH. Malonylglucoside of all the aglycones, total genistein (TGin), and total daidzein (TDin) showed strong correlation with total isoflavones, whereas acetylglucoside and aglycone levels did not significantly correlate with total isoflavone. Profiling of soybean isoflavone is helpful in understanding the regulation of isoflavone biosynthesis for greater improved resistance of crops to disease and greater health benefits for humans. This comparative study of soybean cultivars grown in India and Bulgaria throws light on their composition and nutraceutical value.


Comparing the health status of low-income children in and out of foster care.
Children in foster care face poverty, family dysfunction, neglect, and abuse, with high rates of chronic health, emotional, and developmental problems. This study compared the overall health status of a group of children entering foster care with a group of Medicaid-eligible children living with their parents, matched for age and gender. It identified significantly more health and developmental problems in children in foster care than in the comparison group. Possible contributors to the higher percentage of problems among foster care children may be that the foster children have more problems related to the underlying risk factors resulting in placement, or that the foster care physicians conducted a more comprehensive assessment or had lower clinical thresholds. Further research is necessary to identify and treat the problems of this high-risk group.


Poison ivy/oak dermatitis. Use of polyamine salts of a linoleic acid dimer for topical prophylaxis.
Closed patch tests were used to evaluate the ability of 156 different preparations (based on 22 different chemicals) to prevent poison ivy dermatitis. Several polyamine salts of a linoleic acid dimer were identified that were totally able to prevent the usual dermatitis in approximately 70% of subjects. The effectiveness of the preparations improved when the antigen and the protectant were washed off within eight to 12 hours, instead of remaining on the skin for 48 hours. When washed off, and depending on the protectant, concentration, and vehicle used, several of the preparations were totally able to prevent a dermatitis in a range of 56% to 100% of subjects tested. Further work with these compounds may greatly benefit the many people currently plagued by their allergy to poison ivy and poison oak.


Dual-pulse frequency compounded superharmonic imaging.
Tissue second-harmonic imaging is currently the default mode in commercial diagnostic ultrasound systems. A new modality, superharmonic imaging (SHI), combines the third through fifth harmonics originating from nonlinear wave propagation through tissue. SHI could further improve the resolution and quality of echographic images. The superharmonics have gaps between the harmonics because the transducer has a limited bandwidth of about 70% to 80%. This causes ghost reflection artifacts in the superharmonic echo image. In this work, a new dual-pulse frequency compounding (DPFC) method to eliminate these artifacts is introduced. In the DPFC SHI method, each trace is constructed by summing two firings with slightly different center frequencies. The feasibility of the method was established using a single-element transducer. Its acoustic field was modeled in KZK simulations and compared with the corresponding measurements obtained with a hydrophone apparatus. Subsequently, the method was implemented on and optimized for a setup consisting of an interleaved phased-array transducer (44 elements at 1 MHz and 44 elements at 3.7 MHz, optimized for echocardiography) and a programmable ultrasound system. DPFC SHI effectively suppresses the ghost reflection artifacts associated with imaging using multiple harmonics. Moreover, compared with the single-pulse third harmonic, DPFC SHI improved the axial resolution by 3.1 and 1.6 times at the -6-dB and -20-dB levels, respectively. Hence, DPFC offers the possibility of generating harmonic images of a higher quality at a cost of a moderate frame rate reduction.


Production of Dextran from Locally Lactobacillus Spp. Isolates.
Dextran is a commercially available bacterial exopolysaccharide (EPS) with several industrial applications in the food industry and in the biomedical industry as an adjuvant, emulsifier, carrier, and stabilizer. The production of dextran at the industrial level occurs through the fermentation of a sucrose-rich medium. Research to optimize dextran production has found that the yield of dextran varies depending the specific conditions for production. The aim of this study was to produce dextran and establish the optimal conditions for dextran biosynthesis from different Lactobacillus species isolated from healthy vaginal and infant stool samples. Lactobacillus spp. were isolated and identified from vaginal and infant stool samples via the VITEK 2 system. The presence of dextran biosynthesis from the different Lactobacillus spp. isolates was determined by a screening test for mucoid colonies and confirmed via the ethanol precipitation method. To optimize for the maximum yield of dextran, the effects of various parameters such as temperature, incubation time, pH, inoculum size, aeration, and sucrose concentration were examined. All Lactobacillus spp. isolates were able to produce dextran. The optimal conditions for dextran production was at 24 hours of incubation at 30 °C with 15% sucrose, 4% inoculation size at pH 7.0 in aerobic conditions. This yielded a dextran dry weight of 580 mg/100 mL. Dextran production from Lactobacillus species isolates vagina and infant stool had the ability to produce dextran.


[The relationship of the larval behavioral traits of the malarial mosquito Anopheles messeae (Diptera: Culicidae) to its sensitivity to the entomopathogenic bacterium Bacillus thuringiensis subspecies israelensis].
Behavioural actions of the malarial mosquito Anopheles messeae larvae and their correlations with the susceptibility to the entomopathogenic bacterium Bacillus thuringiensis israelensis (Bti) were studied. The larvae were differentiated according to their feeding mode ("filter-feeders" and "scraper-feeders") and to their avoidance reaction on a concussion of water surface by water drops ("divers" and "undivers"). The "scraper-feeders" and "divers" showed a tendency of decreased susceptibility to Bti, while the "filter-feeders" and "undivers" did not. The presence of the white pigment (striped genotypes) on a dorsum of the larvae correlated with the reaction of avoidance and phyllotaxis but not with the susceptibility to Bti.


[Sensory-motor neuropathy: a slow and misleading case of leprosy].
The diagnostic process of sensory-motor neuropathies is difficult. Atypical variants and rare etiologies also contribute to delay the diagnosis. We report the case of a 70-year-old woman with slowly progressive asymmetric axonal sensory-motor neuropathy. Leprosy was identified after an eight-year delay. Nerve biopsy was required to establish the diagnosis: electron microscopy revealed debris of Hansen's bacillus in the nerve. Treatment was fully curative after several months. Leprosy is a rare cause of neuropathy in Europeans. Systematic inquiry about travel to endemic areas would be helpful in establishing the diagnosis. In such cases, nerve biopsy is crucial.


A process evaluation of medical ethics education in the first year of a new medical curriculum.
Despite the recent increase in activity in the field of medical ethics education, few evaluative studies have been carried out. Most studies have taken place in North America, in curricula where teaching is discipline-based, and have concentrated on outcome rather than on the curricular processes adopted. To evaluate the process of medical ethics education in the first year of a new learner-centred, problem-based, integrated medical curriculum. A qualitative, multi-method approach was adopted using open questionnaires, focus groups and tutor evaluation rating scales. The study involved all 238 students in the first year of the new medical curriculum, and the 30 clinical tutors who facilitated ethics learning. A stratified sampling technique was used to choose focus group participants. Small group teaching proved highly acceptable to both students and tutors. Tutors' teaching skills were central to its effectiveness. Tutors played an important role in promoting students' appreciation of the relevance of medical ethics to clinical practice, and in establishing a climate where constructive criticism of colleagues' actions is acceptable. Course integration, including the provision for students of clinical experiences on which to reflect, was an important aid to learning. Students and tutors were noted to be driving the ethics curriculum towards having a contextual rather than theoretical base. This evaluation identified those aspects of the medical ethics course which contributed to its effectiveness and those which detracted from it. This information will be used to inform future development.


The Solanum lycopersicum auxin response factor 7 (SlARF7) regulates auxin signaling during tomato fruit set and development.
Auxin response factors (ARFs) are encoded by a gene family of transcription factors that specifically control auxin-dependent developmental processes. A tomato ARF gene, homologous to Arabidopsis NPH4/ARF7 and therefore designated as Solanum lycopersicum ARF7 (SlARF7), was found to be expressed at a high level in unpollinated mature ovaries. More detailed analysis of tomato ovaries showed that the level of SlARF7 transcript increases during flower development, remains at a constant high level in mature flowers, and is down-regulated within 48 h after pollination. Transgenic plants with decreased SlARF7 mRNA levels formed seedless (parthenocarpic) fruits. These fruits were heart-shaped and had a rather thick pericarp due to increased cell expansion, compared with the pericarp of wild-type fruits. The expression analysis, together with the parthenocarpic fruit phenotype of the transgenic lines, suggests that, in tomato, SlARF7 acts as a negative regulator of fruit set until pollination and fertilization have taken place, and moderates the auxin response during fruit growth.


Effect of horn fly (Haematobia irritans) control on growth and reproduction of beef heifers.
The influence of horn fly control with commercially available ear tags was studied on beef replacement heifers (n = 670) for growth and reproductive performance. The study was conducted at five sites in Louisiana over 3 yr. Heifers used were yearling replacement females that were exposed to fertile bulls during a limited spring breeding season that coincided with the horn fly season. In mid to late May of each year, heifers were randomly assigned to one of two horn fly treatments: untreated and treated for horn fly control. The trial continued each year until September or October at the end of fly season. Pregnancy status was determined by rectal palpation. Horn fly populations were controlled on the treated heifers at moderate levels (84%). Total weight gain of treated heifers was 14% greater than for untreated heifers. Horn fly treatment had no effect on pregnancy rate (78% and 75% for untreated and treated heifers, respectively). Treatment differences for weight gain were of greater magnitude for heifers failing to conceive than for heifers that became pregnant. Weight gains of nonpregnant treated heifers were 33% greater than for nonpregnant untreated heifers, whereas weight gains of pregnant treated heifers were 8% greater than for pregnant untreated heifers. In conclusion, horn fly control on yearling beef replacement heifers improved weight gain but had no effect on first exposure reproduction.


Variant antigen gene expression in malaria.
Pathogens of the genus Plasmodium are unicellular parasites that infect a variety of animals, including reptiles, birds and mammals. All Plasmodium species target host erythrocytes and replicate asexually within this niche. In humans, proliferation within erythrocytes causes disease symptoms ranging from asymtomatic infection to severe disease, including mild to severe febrile and respiratory symptoms, profound anaemia and obstruction of blood flow. The most serious form of human malaria is caused by Plasmodium falciparum, a pathogen that is responsible for several million deaths annually throughout the developing world. Malaria parasites succeed in evading the host immune response to establish long-term, persistent infections, thus increasing the efficiency by which they are transmitted to the mosquito vector. The ability to evade the host immune system, in particular the avoidance of antibody-mediated immunity against parasite-encoded surface proteins, is the result of amplification of extensive repertoires of multicopy, hypervariable gene families that encode infected erythrocyte or merozoite surface proteins. Via switching between antigenically diverse genes within these large families, populations of parasites have the capacity for rapid variation in antigenicity and virulence over the course of an infection. Here we review the amplification and generation of antigenic diversity within the Plasmodium variant gene families, as well as discuss the mechanisms underlying their tightly controlled gene expression and antigenic switching.


Do traditional Gail model risk factors account for increased breast cancer in women with lupus?
To determine to what extent the observed experience of breast cancer in a combined cohort of patients with systemic lupus erythematosus (SLE) could be explained by the profile of breast cancer risk factors. Data were pooled from 2 centers, the Montreal General Hospital and the Feinberg School of Medicine at Northwestern University in Chicago. For each female cohort member, the probability of developing breast cancer during followup was estimated based on factors (including the individual's age, parity, age at first live birth, age of menarche, personal history of benign breast disease, and family history) using the Gail model, an established model for predicting breast cancer risk. The actual occurrence of cancer cases was determined by linkage with regional cancer registries. Of the 583 women in the combined cohort, 5 had been diagnosed with breast cancer prior to cohort entry, and 14 declined participation. In those remaining, 12 cases of breast cancer occurred compared to 5.6 predicted by the Gail model (standardized incidence ratio 2.1, 95% confidence interval: 1.1, 3.7). Thus, after controlling for risk factors, the incidence of breast cancer was elevated. Our data suggest that the risk of breast cancer in our SLE cohort is not completely explained by traditional factors found in the Gail model. Other factors, such as carcinogenic exposures (i.e., alkylating agents and immunosuppressive drugs) or the immunologic dysregulation of SLE itself, may be contributory.


Meta-analysis of GWAS on two Chinese populations followed by replication identifies novel genetic variants on the X chromosome associated with systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease that affects mainly females. What role the X chromosome plays in the disease has always been an intriguing question. In this study, we examined the genetic variants on the X chromosome through meta-analysis of two genome-wide association studies (GWAS) on SLE on Chinese Han populations. Prominent association signals from the meta-analysis were replicated in 4 additional Asian cohorts, with a total of 5373 cases and 9166 matched controls. We identified a novel variant in PRPS2 on Xp22.3 as associated with SLE with genome-wide significance (rs7062536, OR = 0.84, P = 1.00E-08). Association of the L1CAM-MECP2 region with SLE was reported previously. In this study, we identified independent contributors in this region in NAA10 (rs2071128, OR = 0.81, P = 2.19E-13) and TMEM187 (rs17422, OR = 0.75, P = 1.47E-15), in addition to replicating the association from IRAK1-MECP2 region (rs1059702, OR = 0.71, P = 2.40E-18) in Asian cohorts. The X-linked susceptibility variants showed higher effect size in males than that in females, similar to results from a genome-wide survey of associated SNPs on the autosomes. These results suggest that susceptibility genes identified on the X chromosome, while contributing to disease predisposition, might not contribute significantly to the female predominance of this prototype autoimmune disease.


Stability and Safety of an AAV Vector for Treating RPGR-ORF15 X-Linked Retinitis Pigmentosa.
Our collaborative successful gene replacement therapy using AAV vectors expressing a variant of human RPGR-ORF15 in two canine models provided therapeutic proof of concept for translation into human treatment. The ORF15 sequence contained within this AAV vector, however, has ORF15 DNA sequence variations compared to the published sequence that are likely due to its unusual composition of repetitive purine nucleotides. This mutability is a concern for AAV vector production and safety when contemplating a human trial. In this study, we establish the safety profile of AAV-hIRBP-hRPGR and AAV-hGRK1-hRPGR vectors used in the initial canine proof-of-principle experiments by demonstrating hRPGR-ORF15 sequence stability during all phases of manipulation, from plasmid propagation to vector production to its stability in vivo after subretinal administration to animals. We also evaluate potential toxicity in vivo by investigating protein expression, retinal structure and function, and vector biodistribution. Expression of hRPGR is detected in the inner segments and synaptic terminals of photoreceptors and is restricted to the connecting cilium when the vector is further diluted. Treated eyes exhibit no toxicity as assessed by retinal histopathology, immunocytochemistry, optical coherence tomography, fundoscopy, electroretinogram, and vector biodistribution. Therefore, the hRPGR-ORF15 variant in our AAV vectors appears to be a more stable form than the endogenous hRPGR cDNA when propagated in vitro. Its safety profile presented here in combination with its proven efficacy supports future gene therapy clinical trials.


Outcome of lower-limb preservation with an expandable endoprosthesis after bone tumor resection in children.
The optimal treatment of malignant pediatric lower-extremity bone tumors is controversial. Expandable endoprostheses allow limb preservation, but the revision rate and limited function are considered barriers to their use. This study investigated the functional, emotional, and oncologic outcomes of thirty-eight patients treated with an expandable endoprosthesis. A retrospective chart review was performed, and surviving patients were asked to complete the Musculoskeletal Tumor Society (MSTS) outcomes instrument and the Pediatric Outcomes Data Collection Instrument (PODCI). Additional data including the range of hip and knee motion, limb-length discrepancy, and total lengthening were also obtained. Thirty-eight patients were treated with an expandable endoprosthesis, and twenty-six of these patients were alive at the time of the study. The mean global MSTS score was 26.1, and the mean global PODCI score was 85.8. The mean emotional acceptance and happiness subscores were high. The mean sagittal-plane hip motion in patients who had undergone replacement of the proximal aspect of the femur was 103°. The mean knee motion in patients who had undergone replacement of the proximal aspect of the femur, the distal aspect of the femur, or the proximal aspect of the tibia was 127°, 97°, and 107°, respectively. The mean lengthening at the time of skeletal maturity was 4.5 cm, and the mean limb-length discrepancy was 0.7 cm. Forty-two percent of the patients experienced complications, with ten patients requiring prosthesis revision and two of these patients requiring amputation. Current technology does not offer a single best reconstruction option for children. Previous studies and the present series have indicated that physical and emotional functioning in patients treated with an expandable endoprosthesis are good but that complication rates remain high. Amputation and rotationplasty are alternative treatments if patients and their families are amenable to these procedures. The literature supports no single superior treatment among these three options with regard to physical or emotional health.


The management of imaging dose during image-guided radiotherapy: report of the AAPM Task Group 75.
Radiographic image guidance has emerged as the new paradigm for patient positioning, target localization, and external beam alignment in radiotherapy. Although widely varied in modality and method, all radiographic guidance techniques have one thing in common--they can give a significant radiation dose to the patient. As with all medical uses of ionizing radiation, the general view is that this exposure should be carefully managed. The philosophy for dose management adopted by the diagnostic imaging community is summarized by the acronym ALARA, i.e., as low as reasonably achievable. But unlike the general situation with diagnostic imaging and image-guided surgery, image-guided radiotherapy (IGRT) adds the imaging dose to an already high level of therapeutic radiation. There is furthermore an interplay between increased imaging and improved therapeutic dose conformity that suggests the possibility of optimizing rather than simply minimizing the imaging dose. For this reason, the management of imaging dose during radiotherapy is a different problem than its management during routine diagnostic or image-guided surgical procedures. The imaging dose received as part of a radiotherapy treatment has long been regarded as negligible and thus has been quantified in a fairly loose manner. On the other hand, radiation oncologists examine the therapy dose distribution in minute detail. The introduction of more intensive imaging procedures for IGRT now obligates the clinician to evaluate therapeutic and imaging doses in a more balanced manner. This task group is charged with addressing the issue of radiation dose delivered via image guidance techniques during radiotherapy. The group has developed this charge into three objectives: (1) Compile an overview of image-guidance techniques and their associated radiation dose levels, to provide the clinician using a particular set of image guidance techniques with enough data to estimate the total diagnostic dose for a specific treatment scenario, (2) identify ways to reduce the total imaging dose without sacrificing essential imaging information, and (3) recommend optimization strategies to trade off imaging dose with improvements in therapeutic dose delivery. The end goal is to enable the design of image guidance regimens that are as effective and efficient as possible.


Phospholipase D1 activity is crucial for cytosolic phospholipase A2 -dependent prostaglandin E2 formation in murine osteoblastic MC3T3-E1 cells.
In bone, prostaglandin E2 (PGE2) is highly osteogenic and formed by osteoblasts, a key modulatory event in the regulation of bone cell activity. MC3T3-E1 cells are widely used as an in vitro model of osteoblast function. It is still not clear which pathways contribute to the release of AA in these cells. In this study we have focussed on the contribution of phospholipase D (PLD) enzymes to osteoblastic PGE2 formation after stimulation with endothelin-1 (ET-1). Using specific inhibitors of PLD1 and PLD2 we could show that PGE2 formation was strictly dependent on PLD1 but not PLD2 activity and cytosolic phospholipase A2 (cPLA2) was activated by triggering through PLD1. We have identified diacyl glycerol (DAG) as a possible effector molecule which may serve as a triggering signal for PKC activation and subsequent cPLA2 phosphorylation.


Diagnosis-wide analysis of COVID-19 complications: an exposure-crossover study.
Many studies reporting coronavirus disease 2019 (COVID-19) complications have involved case series or small cohorts that could not establish a causal association with COVID-19 or provide risk estimates in different care settings. We sought to study all possible complications of COVID-19 to confirm previously reported complications and to identify potential complications not yet known. Using United States health claims data, we compared the frequency of all International Classification of Diseases, 10th Revision, Clinical Modification (ICD-10-CM) diagnosis codes occurring before and after the onset of the COVID-19 pandemic in an exposure-crossover design. We included patients who received a diagnosis of COVID-19 between Mar. 1, 2020, and Apr. 30, 2020, and computed risk estimates and odds ratios (ORs) of association with COVID-19 for every ICD-10-CM diagnosis code. Among 70 288 patients with COVID-19, 69 of 1724 analyzed ICD-10-CM diagnosis codes were significantly associated with COVID-19. Disorders showing both strong association with COVID-19 and high absolute risk included viral pneumonia (OR 177.63, 95% confidence interval [CI] 147.19-214.37, absolute risk 27.6%), respiratory failure (OR 11.36, 95% CI 10.74-12.02, absolute risk 22.6%), acute kidney failure (OR 3.50, 95% CI 3.34-3.68, absolute risk 11.8%) and sepsis (OR 4.23, 95% CI 4.01-4.46, absolute risk 10.4%). Disorders showing strong associations with COVID-19 but low absolute risk included myocarditis (OR 8.17, 95% CI 3.58-18.62, absolute risk 0.1%), disseminated intravascular coagulation (OR 11.83, 95% CI 5.26-26.62, absolute risk 0.1%) and pneumothorax (OR 3.38, 95% CI 2.68-4.26, absolute risk 0.4%). We confirmed and provided risk estimates for numerous complications of COVID-19. These results may guide prognosis, treatment decisions and patient counselling.


The effects of knee joint kinematics on anterior cruciate ligament injury and articular cartilage damage.
This study determined which knee joint motions lead to anterior cruciate ligament (ACL) rupture with the knee at 25° of flexion. The knee was subjected to internal and external rotations, as well as varus and valgus motions. A failure locus representing the relationship between these motions and ACL rupture was established using finite element simulations. This study also considered possible concomitant injuries to the tibial articular cartilage prior to ACL injury. The posterolateral bundle of the ACL demonstrated higher rupture susceptibility than the anteromedial bundle. The average varus angular displacement required for ACL failure was 46.6% lower compared to the average valgus angular displacement. Femoral external rotation decreased the frontal plane angle required for ACL failure by 27.5% compared to internal rotation. Tibial articular cartilage damage initiated prior to ACL failure in all valgus simulations. The results from this investigation agreed well with other experimental and analytical investigations. This study provides a greater understanding of the various knee joint motion combinations leading to ACL injury and articular cartilage damage.


Polymorphism of genes encoding PmrAB in colistin-resistant strains of Escherichia coli and Salmonella enterica isolated from poultry and swine.
To detect the occurrence of low susceptibility to colistin (polymyxin E), a last-resort antimicrobial, among enterobacteria isolated from samples of animal origin (poultry and swine) and to find out the molecular basis of colistin resistance. Salmonella enterica and Escherichia coli were isolated from eggs and swine samples. Bacterial strains were screened for colistin resistance by using MIC determinations interpreted according to EUCAST recommendations. pmrAB genes were amplified by PCR from bacterial isolates and their sequences were characterized. Nine colistin-resistant strains were detected in a collection of 739 enterobacteria (S. enterica and E. coli) isolated from animal samples taken in different environments. Sequences encoding the PmrAB two-component sensor-regulator from two colistin-resistant E. coli strains isolated from swine faeces presented three non-synonymous polymorphisms, producing the variants 39S → I and 81R → S of PmrA and 161V → G of PmrB, among which the involvement of mutations in PmrA-81 and PmrB-161 in resistance to the antimicrobial had been previously shown. No variation at the protein level was detected after analysis of PmrAB sequences from seven colistin-resistant S. enterica strains. E. coli strains carrying mutations in PmrAB that confer resistance to polymyxins, which might have evolved in vivo and have been rarely detected, are described for the first time in enterobacteria isolated from animals.


Frequency-domain optical image reconstruction in turbid media: an experimental study of single-target detectability.
An experimental study of the detectability of an object embedded in optically tissue-equivalent media by frequency-domain image reconstruction is presented. The experiments were performed in an 86-mm-diameter cylindrical phantom containing an optically homogeneous cylindrical target whose absorption and scattering properties presented a 2:1 contrast with the background medium. The parameter space explored during experimentation involved object size (15-, 8-, and 4-mm targets) and location (centered, 20-mm off-centered, and 35-mm off-centered) variations. Image reconstruction was achieved with a previously reported regularized least-squares approach that incorporates finite-element solutions of the diffusion equation and Newton's method solutions of the nonlinear minimization problem. Also included during image formation were image enhancement schemes-(1) total variation minimization, (2) dual meshing, and (3) spatial low-pass filtering-which have recently been added. Quantitative measures of image quality including the size, location, and shape of the heterogeneity along with errors in its recovered optical property values are used to quantify the image reconstructions. The results show that a near 22:1 ratio of tissue thickness relative to detectable object size has been achieved with this approach in the laboratory conditions and parameter space that have been investigated.


Neural mechanisms of spatial tactile discrimination: neural patterns evoked by braille-like dot patterns in the monkey.
1. The experiments reported here were designed to investigate the responses of cutaneous mechanoreceptive afferents to spatially configured dot patterns scanned across the skin. Braille-like patterns were selected because the discrimination of Braille characters must depend on spatial patterning rather than some other facet of the afferent discharge. 2. A multifactorial experimental design was used in which each afferent fibre was studied using every combination of six dot patterns, two dot sizes, two dot spacings, two contact forces and two scanning velocities. Two other factors, scanning direction relative to the skin ridges and intermittent versus continuous scanning, were studied. 3. Beside the general question concerning the response properties of the mechanoreceptive afferents, three major questions were addressed here. (i) What is the critical spatial dimension at which neural spatial patterning breaks down and below which tactual discrimination must depend on facets of the afferent discharge other than spatial neural patterning? (ii) Which mechanoreceptive population sets this critical dimension? (iii) Why is tactual discrimination enhanced by lateral scanning? 4. The results presented here suggest that the critical dimension, below which spatial neural patterning breaks down, is of the order of 1.0 mm and that the slowly adapting (SA) afferent fibres are responsible for this limit. 5. At dimensions above approximately 1.0 mm the spatial contrast between peaks and troughs in the SA discharge is markedly enhanced during scanning. When the skin is stationary the discharge rates in the SA population drop rapidly to low levels. A second possible reason for enhanced tactual discrimination during scanning is related to the increased spatiotemporal information in a coherent pattern of neural activity moving across a discrete population of afferent fibres. 6. The effects of variations in conduction velocity are analysed and it is shown that they place serious constraints on the transmission of spatiotemporal information.


Humoral and cardiac effects of TIPS in cirrhotic patients with different "effective" blood volume.
The aim of this study was to evaluate the cardiac effects of transjugular intrahepatic portosystemic shunts (TIPS) in cirrhotic patients with different effective blood volume. Two-dimensional echocardiography was performed before and 7 and 28 days after TIPS insertion in 7 cirrhotic patients with PRA <4 ng/mL/h (group A, normal effective blood volume) and 15 with PRA >4 ng/mL/h (group B, reduced effective blood volume). Before TIPS, most cirrhotic patients showed diastolic dysfunction as indicated by reduced early maximal ventricular filling velocity (E)/late filling velocity (A) ratio. Patients of group B differed from patients of group A because of smaller left ventricular volumes and stroke volume, indicating central underfilling. After TIPS insertion, portal decompression was associated with a significant increase of cardiac output (CO) and a decrease of peripheral resistances. The most important changes were recorded in patients of group B, who showed a significant increase of both the end-diastolic left ventricular volumes and the E/A ratio and a significant decrease of PRA. In conclusion, these results show that the hemodynamic effects of TIPS differ according to the pre-TIPS effective blood volume. Furthermore, TIPS improves the diastolic cardiac function of cirrhotic patients with effective hypovolemia. This result is likely due to a TIPS-related improvement of the fullness of central blood volume.


Modification of membrane permeability in poliovirus-infected HeLa cells: effect of guanidine.
A drastic modification in permeability to monovalent ions occurred in HeLa cells infected with poliovirus, starting in the period from the third to the fourth h post-infection. The bulk of poliovirus protein synthesis took place from the third to the sixth h in cells in which the concentration of monovalent ions in the cytoplasm had changed considerably compared to uninfected cells. Under our conditions of infection (HeLa cells grown in monolayer), poliovirus translation continued beyond the eighth h. Modification of permeability to 86Rb+ ions induced by poliovirus infection was prevented by the addition of 2 mM-guanidine at the time of infection. However, when poliovirus replication was allowed to take place for 1 h before addition of guanidine, the membrane did become modified to some extent. The degree of leakiness to 86Rb+ ions increased when guanidine was added later. The blockage of membrane leakiness by early addition of guanidine was overcome by the addition of choline. The inhibition of cellular protein synthesis, which occurred early in infection or in the presence of guanidine, did not coincide with the modification of permeability to 86Rb+ ions. Viral protein synthesis was necessary in order to modify the membrane late in infection, since addition of 10(-5) M-cycloheximide during the first 2 h of infection prevented the leakiness to 86Rb+ ions observed from the fourth h after infection. Membrane potential, as measured by the lipophilic cation TPP+ (tetraphenylphosphonium), dropped from the fourth h post-infection. This change in the membrane was also prevented when viral gene expression was inhibited by the presence of guanidine.


T cell chemo-vaccination effects after repeated mucosal SHIV exposures and oral pre-exposure prophylaxis.
Pre-exposure prophylaxis (PrEP) with anti-viral drugs is currently in clinical trials for the prevention of HIV infection. Induction of adaptive immune responses to virus exposures during anti-viral drug administration, i.e., a "chemo-vaccination" effect, could contribute to PrEP efficacy. To study possible chemo-vaccination, we monitored humoral and cellular immune responses in nine rhesus macaques undergoing up to 14 weekly, low-dose SHIV(SF162P3) rectal exposures. Six macaques concurrently received PrEP with intermittent, oral Truvada; three were no-PrEP controls. PrEP protected 4 macaques from infection. Two of the four showed evidence of chemo-vaccination, because they developed anti-SHIV CD4(+) and CD8(+) T cells; SHIV-specific antibodies were not detected. Control macaques showed no anti-SHIV immune responses before infection. Chemo-vaccination-induced T cell responses were robust (up to 3,940 SFU/10(6) PBMCs), predominantly central memory cells, short-lived (≤22 weeks), and appeared intermittently and with changing specificities. The two chemo-vaccinated macaques were virus-challenged again after 28 weeks of rest, after T cell responses had waned. One macaque was not protected from infection. The other macaque concurrently received additional PrEP. It remained uninfected and T cell responses were boosted during the additional virus exposures. In summary, we document and characterize PrEP-induced T cell chemo-vaccination. Although not protective after subsiding in one macaque, chemo-vaccination-induced T cells warrant more comprehensive analysis during peak responses for their ability to prevent or to control infections after additional exposures. Our findings highlight the importance of monitoring these responses in clinical PrEP trials and suggest that a combination of vaccines and PrEP potentially might enhance efficacy.


[Simple extension of an ultrasound B image device to a cw duplex scanner for carotid artery studies].
In contrast to cw-Doppler sonography, ultrasound B-mode imaging is rarely applied in routine examinations for carotid artery stenosis. This may be due to the fact that B-mode imaging without accompanying Doppler sonography lacks reliability, and that Duplex scanners with pulsed Doppler modules are expensive. In this paper a method is described for constructing a cw-Duplex scanner by slight modification and combination of conventional B-mode and cw-Doppler devices. Our experience with this equipment shows that it facilitates clear distinction of the carotid branches on the B-mode image, and that it is capable of visualising carotid occlusions as well as overlapping and atypical vascular courses, all of which frequently limit the diagnostic potential of Doppler sonography. For routine examinations of the carotid bifurcation, ease of application and considerably lower costs make the cw-Duplex scanner preferable to pulsed Duplex devices.


Microalgal biofactories: a promising approach towards sustainable omega-3 fatty acid production.
Omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) provide significant health benefits and this has led to an increased consumption as dietary supplements. Omega-3 fatty acids EPA and DHA are found in animals, transgenic plants, fungi and many microorganisms but are typically extracted from fatty fish, putting additional pressures on global fish stocks. As primary producers, many marine microalgae are rich in EPA (C20:5) and DHA (C22:6) and present a promising source of omega-3 fatty acids. Several heterotrophic microalgae have been used as biofactories for omega-3 fatty acids commercially, but a strong interest in autotrophic microalgae has emerged in recent years as microalgae are being developed as biofuel crops. This paper provides an overview of microalgal biotechnology and production platforms for the development of omega-3 fatty acids EPA and DHA. It refers to implications in current biotechnological uses of microalgae as aquaculture feed and future biofuel crops and explores potential applications of metabolic engineering and selective breeding to accumulate large amounts of omega-3 fatty acids in autotrophic microalgae.


No effect of blood sampling or phytohaemagglutinin injection on postfledging survival in a wild songbird.
The injection of phytohaemagglutinin (PHA) and sampling of blood are widely used in studies of wild vertebrates to assess components of immune and endocrine function and health state and to obtain genetic material. Despite the pervasive use of these techniques in the life sciences, their potential effects on survival are rarely considered. For example, whether injection of the immunogen PHA into body parts critical for locomotion (e.g., the prepatagium, or wing web, in birds) affects survival has not been tested. Here, we test whether injection of PHA into the wing web and blood sampling from nestling house wrens affects their subsequent recruitment and survival as breeding adults. Capture-mark-recapture analysis on a large sample of young (N = 20,152 fledglings from 3959 broods) treated over 10 years revealed that neither PHA injection nor blood sampling affected individual survival and detection probability. Recruitment as a breeder varied among years, but this variation was not attributable to sampling effort, or the percent of all adults identified at the nest during a given year. Variation in the percent of adults identified was primarily attributable to the effect of nest depredation on our ability to capture nesting pairs. Our results indicating lack of an effect of blood sampling and immune stimulation on survival are encouraging, but we recommend further work to assess the potential negative effects of all commonly used techniques on the survival of study subjects in the wild, including the potential costs associated with mounting various immunological responses.


Interferon-tau upregulates prolactin receptor mRNA in the ovine endometrium during the peri-implantation period.
Our objective was to determine the effect of ovine interferon-tau (IFN-tau) on prolactin receptor (PRL-R) gene expression in the ovine endometrium. IFN-tau is an embryonic cytokine which, via its paracrine anti-luteolytic activity, plays a critical role in maternal recognition of pregnancy in ruminants. Using ribonuclease protection assay procedures, we compared endometrial PRL-R mRNA levels in ewes that were intrauterine injected with either 2 mg bovine serum albumin or 2 mg recombinant ovine IFN-tau on day 10 of the oestrous cycle (day 0 = day of oestrus). IFN treatment significantly increased the abundance of both the long and short forms of PRL-R mRNA in the ovine uterus, but had no effect on the long:short form ratio. In situ hybridization experiments revealed that the increase in abundance of PRL-R mRNA in the uterus was localized to the glandular compartment of the endometrium. In pregnant ewes, a similar increase in PRL-R mRNA abundance was found to occur in ovine endometrium on days 14-15 post conception. Collectively, these data provided strong evidence that IFN-tau modulates the level of lactogenic hormone receptor mRNA in the ovine uterus. Whether the effect of IFN-tau on PRL-R expression is mediated directly or influenced, at least in part, by progesterone remains to be elucidated.


Influence of wet heating and autoclaving on chemical composition and standardized ileal crude protein and amino acid digestibility in full-fat soybeans for pigs.
One batch each of eight full-fat soybeans (FFSB) was used to determine the effect of different heat treatments including wet heating (WH) and autoclaving (AC) on chemical composition and standardized ileal digestibility (SID) of CP and AA in growing pigs. The raw FFSB (K0) were either treated by WH at 80°C for 1 min (K1), at 100°C for 6 min (K2), or at 100°C for 16 min (K3). Thereafter, these batches were expanded at 125°C for 15 s. A further heat treatment included AC at 110°C for 15 (Z1), 30 (Z2), 45 (Z3), or 60 (Z4) min of FFSB that were subjected to the same WH treatment as K3. Diets were formulated to contain the respective FFSB as the sole source of CP and AA. A N-free diet was used to measure basal endogenous losses of CP and AA in an additional period at the end of the experiment. Eight ileally cannulated pigs (28 ± 1 kg) were allocated to a row-column design with 8 diets and 6 periods of 7 d each. An increase in the duration of WH had no effect on contents of AA (% of CP) and NDF, but NDIN contents linearly increased ( < 0.05) with increasing time for WH. Autoclaving resulted in a linear decrease ( < 0.05) of trypsin inhibitor activity (TIA), contents of Arg, Leu, Ala, Asp, Cys, and Gly as well as Lys to CP ratio (Lys:CP), reactive Lys to CP ratio (rLys:CP), and in an increase ( < 0.05) in contents of NDF and NDIN. There was a quadratic response ( < 0.05) of SID of CP and AA as time for WH at 100°C increased from 0 (K0) to 6 (K2) up to 16 (K3) min. Moreover, a quadratic response ( < 0.05) to increasing time of AC was observed for SID of Arg, Phe, and Pro. The SID of CP and all indispensable AA showed a quadratic response ( < 0.05) to decreasing TIA, urease activity, protein solubility in 0.2% potassium hydroxide, protein dispersibility index, Lys:CP, (lightness), and to increasing NDIN and (redness). In addition, there was a linear increase ( < 0.05) in SID values with decreasing rLys:CP and increasing NDF contents. In conclusion, WH proved to be suitable for increasing SID values. Further improvement of SID of most AA could be achieved on additional AC treatment from Z1 to Z3, however, it needs to be considered, if the observed increase due to AC is cost effective in view of the additional production costs. Several chemical and physical parameters can be used in the feed industry for quality control purposes to predict the extent of heat damage on SID of CP and indispensable AA in FFSB.


Comparison of school day eating behaviours of 8-11 year old children from Adelaide, South Australia, and London, England: Child eating behaviours in South Australia and England.
School food intake makes a considerable contribution to children's overall diet, especially fruit and vegetable intake. Comparing differing school food provision systems can provide novel insights for intervention and improved nutrition policy. This study compared school day food in children from Australia and England. Children completed food frequency questionnaires reporting school day food intake, breakfast intake and family evening meals. Differences in food consumed over the school day between Australian and English children were evaluated. Multinomial logistic regressions compared fruit and vegetable intake, family dinner frequency and breakfast in Australian and English children adjusting for confounders: age, sex, ethnicity and parent education. 27 Primary schools in Adelaide, Australia and 32 in London, England. N = 772 children aged 8-11 years from the Australian REACH study (n = 347) and UK RHS School Gardening Trial in England (n = 425). Considerably more English children reported consuming vegetables at school than Australian children (recess/lunchtime Australian children 3.4%/6.1%; English children recess/lunctime 3.6/51.1%). However, Australian children were more likely to consume vegetables daily (OR = 4.1; 1.3, 12.5), and have family evening meals everyday [OR = 4.01; 1.88, 8.55], and were less likely to consume breakfast (OR = 0.26; 0.08, 0.79) than English children. Findings indicate that provision of a school lunch meal, compared to a packed lunch from home, may be more supportive of children's vegetable intake. However, without a supportive home environment that encourages vegetable intake, children will not be able to consume sufficient amounts of vegetables.


Analysis of a retrieved delta III total shoulder prosthesis.
A reversed Delta III total shoulder prosthesis was retrieved post-mortem, eight months after implantation. A significant notch was evident at the inferior pole of the scapular neck which extended beyond the inferior fixation screw. This bone loss was associated with a corresponding, erosive defect of the polyethylene cup. Histological examination revealed a chronic foreign-body reaction in the joint capsule. There were, however, no histological signs of loosening of the glenoid base plate and the stability of the prosthetic articulation was only slightly reduced by the eroded rim of the cup.


The mitochondrial proteome and human disease.
For nearly three decades, the sequence of the human mitochondrial genome (mtDNA) has provided a molecular framework for understanding maternally inherited diseases. However, the vast majority of human mitochondrial disorders are caused by nuclear genome defects, which is not surprising since the mtDNA encodes only 13 proteins. Advances in genomics, mass spectrometry, and computation have only recently made it possible to systematically identify the complement of over 1,000 proteins that comprise the mammalian mitochondrial proteome. Here, we review recent progress in characterizing the mitochondrial proteome and highlight insights into its complexity, tissue heterogeneity, evolutionary origins, and biochemical versatility. We then discuss how this proteome is being used to discover the genetic basis of respiratory chain disorders as well as to expand our definition of mitochondrial disease. Finally, we explore future prospects and challenges for using the mitochondrial proteome as a foundation for systems analysis of the organelle.


Treatment of severely ill COVID-19 patients with anti-interleukin drugs (COV-AID): A structured summary of a study protocol for a randomised controlled trial.
The purpose of this study is to test the safety and effectiveness of individually or simultaneously blocking IL-6, IL-6 receptor and IL-1 versus standard of care on blood oxygenation and systemic cytokine release syndrome in patients with COVID-19 coronavirus infection and acute hypoxic respiratory failure and systemic cytokine release syndrome. A phase 3 prospective, multi-center, interventional, open label, 6-arm 2x2 factorial design study. Subjects will be recruited at the specialized COVID-19 wards and/or ICUs at 16 Belgian participating hospitals. Only adult (≥18y old) patients will be recruited with recent (≤16 days) COVID-19 infection and acute hypoxia (defined as PaO2/FiO2 below 350mmHg or PaO2/FiO2 below 280 on supplemental oxygen and immediately requiring high flow oxygen device or mechanical ventilation) and signs of systemic cytokine release syndrome characterized by high serum ferritin, or high D-dimers, or high LDH or deep lymphopenia or a combination of those, who have not been on mechanical ventilation for more than 24 hours before randomisation. Patients should have had a chest X-ray and/or CT scan showing bilateral infiltrates within the last 2 days before randomisation. Patients with active bacterial or fungal infection will be excluded. Patients will be randomized to 1 of 5 experimental arms versus usual care. The experimental arms consist of Anakinra alone (anti-IL-1 binding the IL-1 receptor), Siltuximab alone (anti-IL-6 chimeric antibody), a combination of Siltuximab and Anakinra, Tocilizumab alone (humanised anti-IL-6 receptor antibody) or a combination of Anakinra with Tocilizumab in addition to standard care. Patients treated with Anakinra will receive a daily subcutaneous injection of 100mg for a maximum of 28 days or until hospital discharge, whichever comes first. Siltuximab (11mg/kg) or Tocilizumab (8mg/kg, with a maximum dose of 800mg) are administered as a single intravenous injection immediately after randomization. The primary end point is the time to clinical improvement defined as the time from randomization to either an improvement of two points on a six-category ordinal scale measured daily till day 28 or discharge from the hospital or death. This ordinal scale is composed of (1) Death; (2) Hospitalized, on invasive mechanical ventilation or ECMO; (3) Hospitalized, on non-invasive ventilation or high flow oxygen devices; (4) Hospitalized, requiring supplemental oxygen; (5) Hospitalized, not requiring supplemental oxygen; (6) Not hospitalized. Patients will be randomized using an Interactive Web Response System (REDCap). A 2x2 factorial design was selected with a 2:1 randomization regarding the IL-1 blockade (Anakinra) and a 1:2 randomization regarding the IL-6 blockade (Siltuximab and Tocilizumab). In this open-label trial neither participants, caregivers, nor those assessing the outcomes are blinded to group assignment. A total of 342 participants will be enrolled: 76 patients will receive usual care, 76 patients will receive Siltuximab alone, 76 patients will receive Tocilizumab alone, 38 will receive Anakinra alone, 38 patients will receive Anakinra and Siltuximab and 38 patients will receive Anakinra and Tocilizumab. COV-AID protocol version 3.0 (15 Apr 2020). Participant recruitment is ongoing and started on April 4th 2020. Given the current decline of the COVID-19 pandemic in Belgium, it is difficult to anticipate the rate of participant recruitment. The trial was registered on Clinical Trials.gov on April 1st, 2020 (ClinicalTrials.gov Identifier: NCT04330638) and on EudraCT on April 3rd 2020 (Identifier: 2020-001500-41). The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Granulocyte progenitor cell (CFUC) harvest by continuous apheresis in dogs. Effects of blood volume and lithium on yields.
The ability to harvest large amounts of hematopoietic stem cells from blood would eliminate the more difficult approach of bone marrow harvest. Unfortunately, concentration of stem cells in the blood compartment is less than 1% of their concentration in bone marrow. Attempts to increase harvest of blood stem cells, as assayed by granulocyte progenitor cells (CFUC), have been only partially successful. Our study confirms previous reports that CFUC can be mobilized into the blood compartment in dogs, but this mobilization is rate-limited. Unlike platelets and granulocytes that are effectively harvested during the first blood volume processed by continuous apheresis, effective CFUC harvest begins during the second blood volume (606 +/- 97.9 CFUC/ml), peaks by the third (740 +/- 30 CFUC/ml), and remains constant through five blood volumes processed (700 +/- 272 CFUC/ml). Since blood CFUC concentration falls at the end of five blood volumes processed (40% of initial values), further continuous apheresis would not be effective. Treatment of animals with lithium did not improve CFUC harvest. These results show that apheresis procedures can be developed to a limited extent to increase the harvest of hematopoietic progenitor and stem cells.


Base-editing mutagenesis maps alleles to tune human T cell functions.
CRISPR-enabled screening is a powerful tool for the discovery of genes that control T cell function and has nominated candidate targets for immunotherapies1-6. However, new approaches are required to probe specific nucleotide sequences within key genes. Systematic mutagenesis in primary human T cells could reveal alleles that tune specific phenotypes. DNA base editors are powerful tools for introducing targeted mutations with high efficiency7,8. Here we develop a large-scale base-editing mutagenesis platform with the goal of pinpointing nucleotides that encode amino acid residues that tune primary human T cell activation responses. We generated a library of around 117,000 single guide RNA molecules targeting base editors to protein-coding sites across 385 genes implicated in T cell function and systematically identified protein domains and specific amino acid residues that regulate T cell activation and cytokine production. We found a broad spectrum of alleles with variants encoding critical residues in proteins including PIK3CD, VAV1, LCP2, PLCG1 and DGKZ, including both gain-of-function and loss-of-function mutations. We validated the functional effects of many alleles and further demonstrated that base-editing hits could positively and negatively tune T cell cytotoxic function. Finally, higher-resolution screening using a base editor with relaxed protospacer-adjacent motif requirements9 (NG versus NGG) revealed specific structural domains and protein-protein interaction sites that can be targeted to tune T cell functions. Base-editing screens in primary immune cells thus provide biochemical insights with the potential to accelerate immunotherapy design.


Contaminant trends in reservoir sediment cores as records of influent stream quality.
When reconstructing water-quality histories from lake and reservoir cores, it is sometimes assumed that the chemical signatures in the cores reflect historical water quality in the influent streams. To investigate this assumption, concentrations of metals, PAHs, and organochlorine compounds in sediment cores were compared to those associated with an influent-stream suspended sediment for three reservoirs in Fort Worth, TX, and two reservoirs in Boston, MA, U.S.A., and interpreted in light of land-use and regulation histories. In evaluating relations between suspended sediments and cores, three levels of preservation were indicated: (1) influent concentrations and historical trends are preserved in cores (metals at all sites; some organic contaminants at some sites); (2) some loss occurs during transport and initial deposition but relative historical trends are preserved in cores (some organic contaminants at some sites); and (3) neither stream concentrations nor relative historical trends are preserved (dieldrin and p,p'-DDT). The degree of preservation of influent concentration histories varied between lakes, particularly for PAHs. The results support the use of sediment cores to infer streamwater-quality histories for many contaminants but indicate that reservoir-bottom sediment samples might underestimate concentrations of organic contaminants in some streams.


Redesigning trypsin: alteration of substrate specificity.
A general method for modifying eukaryotic genes by site-specific mutagenesis and subsequent expression in mammalian cells was developed to study the relation between structure and function of the proteolytic enzyme trypsin. Glycine residues at positions 216 and 226 in the binding cavity of trypsin were replaced by alanine residues, resulting in three trypsin mutants. Computer graphic analysis suggested that these substitutions would differentially affect arginine and lysine substrate binding of the enzyme. Although the mutant enzymes were reduced in catalytic rate, they showed enhanced substrate specificity relative to the native enzyme. This increased specificity was achieved by the unexpected differential effects on the catalytic activity toward arginine and lysine substrates. Mutants containing alanine at position 226 exhibited an altered conformation that may be converted to a trypsin-like structure upon binding of a substrate analog.


Teaching reading to children with neurofibromatosis type 1: a clinical trial with random assignment to different approaches.
Neurofibromatosis type 1 (NF1) is a genetic disorder with a cognitive profile that includes visual-spatial perception deficits and a high incidence of reading disability. There is a paucity of information about how this cognitively complex population responds to mainstream reading interventions. The clinical trial goals were to determine whether children and adolescents with NF1 and reading deficits (NF+RD) benefit from mainstream remedial reading programs and whether responsiveness varies with differences in program-related visual-spatial demands. Forty-nine participants (28 males, 21 females; aged 8-14y) with either NF+RD (n=17, 11 males, six females) or idiopathic reading deficit (IRD) (n=32, 17 males, 15 females) were randomly assigned to intensive remedial teaching using one of two multisensory reading programs: one with greater kinesthetic demands and the other with greater visual-spatial demands. Two control groups - wait-list IRD (n=14, 11 males, three females) and typically developing readers (n=26, 13 males, 13 females) - received no treatment. Repeated measures and multivariate ANOVA analyses compared each group's growth in reading achievement from pre- to post-testing. Treated groups showed significant growth whereas untreated groups did not. Comparing treated groups, the IRD group responded equally well to both interventions, whereas the NF+RD group showed a better response to the more kinesthetic approach. Results suggest that multisensory remedial reading teaching that emphasizes kinesthetic demands more than visual-spatial demands is suitable for students with NF+RD.


Role of growth hormone, insulin-like growth factors (IGFs) and IGF-binding proteins in the renal complications of diabetes.
This review has updated recent facets of evidence for the significance of the GH/IGF system in the development of diabetic kidney disease. It seems evident, however, that there is still an extensive number of questions that need to be answered before diabetic kidney disease is fully understood. The knowledge we have today indicates that GH/IGF axis, through a complex system comprising GHR, GHBP, IGFs, IGF receptors and IGFBPs may be responsible for both early and late renal changes in experimental diabetes (Fig. 3). In view of the complexity of the GH/IGF system, it will be a challenge to fully characterize the renal effects of GH/IGFs in diabetic kidney disease. There is no doubt that information on this topic will occur with increasing pace in the near future and that a understanding of the above-mentioned mechanisms will allow the further development of existing antagonists and design of new drugs, which may prove to be useful for therapeutic manipulation in the treatment of diabetic nephropathy. The development of long-acting somatostatin analogues and GH antagonists, both with a specific action on the GH/IGF axis, seems to be one important step ahead. The combined administration of one of these antagonists with other drugs with a well described renoprotective action (such as ACE inhibitors) opens an interesting new dimension.


Study of the cytotoxic effects of the new synthetic Isothiocyanate CM9 and its fullerene derivative on human T-leukemia cells.
One important strategy to develop effective anticancer agents is based on natural products. Many active phytochemicals are in human clinical trials and have been used for a long time, alone and in association with conventional anticancer drugs, for the treatment of various types of cancers. A great number of in vitro, in vivo and clinical reports document the multi-target anticancer activities of isothiocyanates and of compounds characterized by a naphthalenetetracarboxylic diimide scaffold. In order to search for new anticancer agents with a better pharmaco-toxicological profile, we investigated hybrid compounds obtained by inserting isothiocyanate group(s) on a naphthalenetetracarboxylic diimide scaffold. Moreover, since water-soluble fullerene derivatives can cross cell membranes thus favoring the delivery of anticancer therapeutics, we explored the cytostatic and cytotoxic activity of hybrid compounds conjugated with fullerene. We studied their cytostatic and cytotoxic effects on a human T-lymphoblastoid cell line by using different flow cytometric assays. In order to better understand their pharmaco-toxicological potential, we also analyzed their genotoxicity. Our global results show that the synthesized compounds reduced significantly the viability of leukemia cells. However, the conjugation with a non-toxic vector did not increase their anticancer potential. This opens an interesting research pattern for certain fullerene properties.


The role of the occupational therapist-work evaluator.
This paper presents the role of the occupational therapist as that of a work evaluator who provides services to injured workers. Therapists, by virtue of their professional knowledge, have a frame of reference for work evaluation and understand the factors that interrupt the work process. In work evaluation, the therapist assists the injured worker to develop work readiness and the physical capacities necessary for working productively. This development takes place in a therapeutic milieu where the "worker" role is regained. The role of the occupational therapist-work evaluator includes work-oriented treatment in the acute care setting, job analysis, work tolerance screening, work capacity evaluation, work hardening, and job market reentry management in the work evaluation and community settings.


Cancer of the proximal colon after a "normal" colonoscopy.
In common with other diagnostic tests, colonoscopy has a false negative rate which is infrequently assessed. The available literature suggests that lesion miss rate is higher for proximal colonic tumors. A total of 367 patients were diagnosed with cancer of the colon and rectum over a period of 2 years. Ninety-two of these patients had tumors proximal to the splenic flexure. Their 5-year pre-diagnosis colonoscopic exposure was analyzed. The primary end-point of this study was to confirm the false negative colonoscopy rate in patients subsequently diagnosed with cancer of the proximal colon. The secondary endpoint was to assess the effects of diagnostic delay on tumor stage and presentation. In the group of patients with proximal colon cancer (n = 92) we identified 10 patients (11%) who, as a result of incomplete (2 cases) or falsely negative (8 cases) colonoscopies, suffered a median diagnostic delay of 17 months (range 3-60). At diagnosis, 4 of these patients had Dukes' D caecal cancer, 4 had Dukes' C caecal cancer and 2 had Dukes' B transverse colon cancer; 3 presented with perforated tumours and 1 with intestinal obstruction. In this small subgroup of patients therefore 40% presented with emergency complications compared to 8% in the rest of the group with proximal cancers (p < 0.01). Missed cancers are more likely to present with complications. This study highlights the importance of recognition of an incomplete examination and the adverse impact of missed diagnosis on subsequent presentation.


Effects of Echinacea extract on the performance, antibody titres, and intestinal histology of layer chicks.
1. This research was conducted to determine the effect of diet supplementation with Echinacea extract (cichoric acid) on the growth performance, antibody titres and intestinal tissue histology of layer chicks. 2. White, 1-d-old, Hy-Line hybrid chicks (n = 540) were divided into three treatments, each consisting of 6 groups of 30 chicks (n = 180): (1) control; (2) 2·5 mg/kg cichoric-acid-fed; and (3) 5 mg/kg cichoric-acid-fed. The trial lasted 60 d. 3. While the growth performance of the chicks was depressed between d 1 and 45, it was found to improve between d 45 and 60. 4. Feed consumption was lower in both of the cichoric-acid-fed groups than in the control group between d 1-15 and 15-30, but was higher between d 30 and 45. Overall, mean feed consumption did not differ between the control and cichoric-acid-fed groups during the 60 d study period. 5. During the 60 d evaluation period, live weight gain, feed utilisation rate and final live weight were higher in the control group than in both of the cichoric-acid-fed groups. 6. Antibody titres against infectious bronchitis and infectious bursal disease did not differ between the three groups, but those for Newcastle disease were higher in the 2·5 mg/kg cichoric-acid-fed group than in the control group after 45 d. 7. Height and width of the jejunal villus and the thickness of the muscle layer were lower in the 5 mg/kg cichoric-acid-fed group than in both the control and the 2·5 mg/kg cichoric-acid-fed groups. The height of the ileal villus was also lower in the 5 mg/kg cichoric-acid-fed group than in the other two groups. 8. Echinacea extract supplementation for layer chicks appears not to benefit growth performance and intestinal histology during the growing period.


Analytical methods for the detection and characterization of unapproved phosphodiesterase type 5 inhibitors (PDE-5i) used in adulteration of dietary supplements- a review.
This article is an up-to-date review of 112 unapproved phosphodiesterase type 5 inhibitors (PDE-5i) found as adulterants in sexual enhancement dietary supplements and other products from 2003 to July 2023. Seventy-five of these unapproved PDE-5i are analogues of sildenafil (67%), followed by 26 analogues of tadalafil (23%), 9 analogues of vardenafil (8%) and 2 other type of compounds (2%). The products have been formulated in various packaging, primarily in capsule, tablet, and powder forms. Common screening techniques allowing detection of such analogues include high performance or ultra-high performance liquid chromatography in tandem with ultra-violet detector (HPLC-UV or UPLC-UV) (50%) and thin-layer chromatography in tandem with ultra-violet detection (TLC-UV) (7%). Screening by mass spectrometry (MS) is relatively less common with the use of single-, triple-quadrupole or time-of-flight (TOF) mass spectrometers (9%). Meanwhile, the combined detection by UV-MS has been recorded at 10% usage. Screening by proton nuclear magnetic resonance spectroscopy (NMR) (11%) has also been applied. For compound characterization, i.e. structural elucidation, NMR spectroscopy has been preferred (100 out of 112 compounds), followed by high-resolution mass spectrometry (HRMS) (74 out of 112 compounds) and Fourier-transform infrared spectroscopy (FTIR) (44 out of 112 compounds). Over the past two decades, analytical technology has been evolving with enhanced sensitivity and resolution. Despite this, structural elucidation of the new emerging analogues in adulterated dietary supplements remains a challenge, especially when the analogues involve complex structural modification. Therefore, the above-mentioned techniques may not be adequate to characterize the analogues. Additional work involving chiroptical methods, two-dimensional (2D) NMR experiments and X-ray crystallography are likely to be required in the future.


PGC-1β promotes enterocyte lifespan and tumorigenesis in the intestine.
The mucosa of the small intestine is renewed completely every 3-5 d throughout the entire lifetime by small populations of adult stem cells that are believed to reside in the bottom of the crypts and to migrate and differentiate into all the different populations of intestinal cells. When the cells reach the apex of the villi and are fully differentiated, they undergo cell death and are shed into the lumen. Reactive oxygen species (ROS) production is proportional to the electron transfer activity of the mitochondrial respiration chain. ROS homeostasis is maintained to control cell death and is finely tuned by an inducible antioxidant program. Here we show that peroxisome proliferator-activated receptor-γ coactivator-1β (PGC-1β) is highly expressed in the intestinal epithelium and possesses dual activity, stimulating mitochondrial biogenesis and oxygen consumption while inducing antioxidant enzymes. To study the role of PGC-1β gain and loss of function in the gut, we generated both intestinal-specific PGC-1β transgenic and PGC-1β knockout mice. Mice overexpressing PGC-1β present a peculiar intestinal morphology with very long villi resulting from increased enterocyte lifespan and also demonstrate greater tumor susceptibility, with increased tumor number and size when exposed to carcinogens. PGC-1β knockout mice are protected from carcinogenesis. We show that PGC-1β triggers mitochondrial respiration while protecting enterocytes from ROS-driven macromolecule damage and consequent apoptosis in both normal and dysplastic mucosa. Therefore, PGC-1β in the gut acts as an adaptive self-point regulator, capable of providing a balance between enhanced mitochondrial activity and protection from increased ROS production.


Associations of Elements of Parental Social Integration with Migrant Children's Vaccination: An Epidemiological Analysis of National Survey Data in China.
Our study explored the effects of parental social integration on migrant children's vaccination status in China. Using data obtained from the 2014 China Migrants Dynamic Survey, a total of 4915 participants were included in this study. Social integration was measured by economic, social, cultural, and internal identity. Univariate chi-square testing was used to calculate associations between all variables and migrant children's vaccination status. Binary logistic regression was employed to calculate the impacts of social integration on migrant children's vaccination status. In total, 94.7% of migrant children had complete vaccinations for their age. Migrants who had medical insurance, spoke the native language when communicating with locals, lived mainly with locals, and did not perceive discrimination were more likely to have their children completely vaccinated. Social integration was positively associated with migrant children's vaccination status. Our study indicated that to improve vaccination coverage of migrant children, more policy support for migrant employment and housing, promotion of health services for migrants, and language support in health institutions is needed.


Piriformis pyomyositis, a cause of piriformis syndrome-a systematic search and review.
Piriformis pyomyositis is a rare form of purulent skeletal myositis. As previous studies concerning piriformis pyomyositis had lower level of evidence and no systematic review has been published yet, we performed a systematic search to review and describe causes, symptoms, red flags, and available treatment options for piriformis pyomyositis. Using PubMed and PubMed Central databases, we found 21 articles describing 23 cases of piriformis pyomyositis. Based on the retrieved information, alongside acute sciatica like buttock and/or hip pain, high-grade fever, aggressive deep seated gluteal pain, neurological deficit of sciatic nerve distribution, positive straight leg raising test, and raised inflammatory biomarkers (erythrocyte sedimentation rate, ESR, C-reactive protein, CRP) provide clues for diagnosis of piriformis pyomyositis. Some cases were very ill but no death was documented. Staphylococcus aureus was the most common pathogen, but Group A as well as Group β Streptococcus, Salmonella typhi, Proteus mirabilis, Brucella melitensis, and Escherichia coli were also involved in the disorder. To treat the piriformis pyomyositis, broad-spectrum antibiotics were found to be useful; however, sometimes, antibiotic switching was warranted based on blood and tissue aspirate reports. Drainage and/or surgical exploration of the affected piriformis muscle were required in cases where antibiotics appeared ineffective. Piriformis pyomyositis is a rara avis and performing of prospective studies will hardly be feasible.


Benefits of sodium glucose cotransporter 2 inhibitors across the spectrum of cardiovascular diseases.
Sodium glucose cotransporter 2 inhibitors (SGLT2i) have emerged as a class of medications with positive cardiovascular (CV) effects across a spectrum of patients with and without type 2 diabetes (T2D). In heart failure with reduced ejection fraction, there is clear evidence that SGLT2i reduce hospitalisations and mortality regardless of the presence of diabetes, and they are now recognised as the fourth pillar of pharmacological management. Recent trial data also indicate promising effects in heart failure with preserved ejection fraction. In patients with T2D and atherosclerotic CV diseases, multiple CV outcomes trials have shown reductions in major adverse CV events. Meta-analysis of these trials also shows lower rates of incident and recurrent atrial fibrillation with SGLT2i. Concerns regarding utilisation in patients with chronic kidney disease have been allayed in trials showing SGLT2i in fact have renoprotective effects. Questions still remain regarding the safety of SGLT2i in the acute heart failure setting and immediately post myocardial infarction, as well as in patients with more advanced stages of chronic kidney disease. Furthermore, studies are underway evaluating SGLT2i in patients with heart valve disease, where positive effects on left ventricular remodelling may, for example, improve functional mitral regurgitation. In this review, we summarise the available evidence of recent CV outcomes trials of SGLT2i, focusing particularly on the application of these agents across various CV diseases. We detail evidence to support increased utilisation of these drugs, which in many cases will reduce mortality and improve quality of life in patients routinely encountered by the CV specialist physician.


A Competitive binding mechanism between Skp1 and exportin 1 (CRM1) controls the localization of a subset of F-box proteins.
SCF-type E3 ubiquitin ligases are crucial regulators of cell cycle progression. As the F-box protein is the substrate-specifying subunit of this family of ligases, their availability dictates the timing and the location of the ubiquitination of substrates. We report here our investigation into the regulation of the localization of F-box proteins, in particular Fbxo7, whose mislocalization is associated with human disease. We identified a motif in Fbxo7 that we have characterized as a functional leucine-rich nuclear export sequence (NES), and which allowed binding to the nuclear export protein, exportin 1 (CRM1). Unusually, the NES was embedded within the F-box domain, which is bound by Skp1 and enables the F-box protein to form part of an E3 ubiquitin ligase. The NES of Fbxo7 controlled its localization and was conserved in Fbxo7 homologues in other species. Skp1 binding prevented Fbxo7 from contacting CRM1. We propose that this competitive binding allowed Fbxo7 to accumulate within the nucleus starting at the G1/S transition. More than ten other F-box proteins also contain an NES at the same location in their F-box domains, indicating that this competitive binding mechanism may contribute to the regulation of a sixth of the known F-box proteins.


In silico identification of Corynebacterium pseudotuberculosis antigenic targets and application in immunodiagnosis.
Caseous lymphadenitis (CLA) is a disease caused by Corynebacterium pseudotuberculosis. It affects mainly small ruminants and causes significant economic losses worldwide. Because symptoms are not immediately noticeable, CLA clinical diagnosis is not effective. Numerous serological tests are being developed to detect the disease in asymptomatic animals, but currently available immunoassays have problems with sensitivity. Current ELISA formats use native bacterial antigens, and recombinant proteins could be useful for improving the immunoassay parameters. The C. pseudotuberculosis proteins CP0126a, CP0369 and CP1957 were identified from 2097 candidate proteins by mature epitope density (MED) analysis, expressed in Escherichia coli and evaluated in an indirect immunoenzymic system. The CP0126a, CP0369 and CP1957 ELISAs showed 77.5 %, 92.5 % and 92.5 % specificity and 95 %, 90 % and 85 % sensitivity, respectively. Receiver operating characteristic (ROC) curve analysis showed an area under the curve of 0.874, 0.951 and 0.881, respectively. The proteins identified in silico were recognized by antibodies in the sera from infected animals without being recognized in negative samples. The ELISA assay using the rCP0369 protein as antigen had the greatest specificity and sensitivity values, followed by rCP1957. This is an interesting strategy for seroepidemiological investigations in sheep flocks due to its significant specificity and sensitivity.


Covered Self-expandable Metallic Stent Insertion as a Rescue Procedure for Postoperative Leakage after Primary Repair of Perforated Duodenal Ulcer.
Surgery has been the standard treatment for perforated duodenal ulcers, with mostly good results. However, the resolution of postoperative leakage after primary repair of perforated duodenal ulcer remains challenging. There are several choices for re-operation required in persistent leakage from perforated duodenal ulcers. However, many of these choices are complicated surgical procedures requiring prolonged general anesthesia that may increase the chances of morbidity and mortality. Several recent reports have demonstrated postoperative leakage after primary repair of a perforated duodenal ulcer treated with endoscopic insertion using a covered self-expandable metallic stent, with good clinical results. We report a case with postoperative leakage after primary repair of a perforated duodenal ulcer treated using a covered self-expandable metallic stent.


Movements of the eyes and hands are coordinated by a common predictive strategy.
Although attempts to intercept a ball in flight are often preceded by predictive gaze behavior, the relationship between the predictive control of gaze and the effector is largely unexplored. The present study was designed to investigate the influence of the spatiotemporal demands of the task on a switch to the predictive control. Ten subjects immersed in a virtual environment attempted to intercept a ball that disappeared for 500 ms of its parabolic approach. The timing of the blank was varied through manipulation of the post-blank duration prior to the ball's arrival, and the shape of the trajectory was manipulated through variation of the pre-blank duration. Results reveal that the gaze movement trajectory during the blank was curvilinear, appropriately scaled to the curvature of the invisible moving ball, and the gaze vector was within 4° of the ball upon reappearance, despite 10° to 13° of ball movement. The timing of the blank did not influence the accuracy of predictive positioning of the paddle at the time of ball reappearance, indicated by the distance of the paddle relative to the ball's eventual passing location. However, analysis of trial-by-trial covariations revealed that, when the gaze vector more accurately predicted the ball's trajectory at reappearance, the paddle was also held closer to the ball's eventual passing location. This suggests that predictive strategies for paddle placement are more strongly mediated by the accuracy of gaze behavior than by the observed range of trajectories, or the timing of the blank.


Neuroradiology and art: a review and personal contribution.
Radiology has attracted the world of art with the esthetic value of its images, and as a new medium for the artistic expression. In order to investigate the links between neuroradiology and art, we examined 12,763 artworks presented in corresponding publications and in Google images on the Internet. The selected artworks were created by 1,964 authors. To give our own contribution to this field, we produced several artful radiological images using the X-ray of 4 cerebral hemispheres, one dissected brain, serial sections of one head and brain, the vascular casts of 2 brains, the magnetic resonance imaging (MRI) scans of one volunteer, and various options in Photoshop. Among the examined artworks, neuroradiological images were used in 129 artworks (1.01%) that were created by 31 artists (1.58%). The artists applied different radiological techniques: X-ray, angiography, computed tomography (CT), multislice CT, MRI, functional MRI, positron emission tomography (PET) or single-photon emission computed tomography (SPECT), either alone or in various combinations. They used the original images, i.e. radiographs or scans, or their electronic modifications in Photoshop or three-dimensional (3D) software. Some artworks presented the skull, yet others the brain, and still others both, either with or without a head image. The neuroradiological artworks were created as paintings, photographs, digital works and sculptures. Their authors were professional artists, designers, amateurs and radiologists. In conclusion, thanks to the esthetics of some radiological images and the valuable creations of certain artists, neuroradiology has become an important field of contemporary art.


Pathological correlates of late-onset dementia in a multicentre, community-based population in England and Wales. Neuropathology Group of the Medical Research Council Cognitive Function and Ageing Study (MRC CFAS).
We have undertaken a large unselected, community-based neuropathology study in an elderly (70-103 years) UK population in relation to prospectively evaluated dementia status. The study tests the assumption that dementing disorders as defined by current diagnostic protocols underlie this syndrome in the community at large. Respondents in the Medical Research Council Cognitive Function and Ageing Study were approached for consent to examine the brain at necropsy. Dementia status was assigned by use of the automated geriatric examination for computer-assisted taxonomy algorithm. Neuropathological features were standardised by use of the protocol of the Consortium to Establish a Registry of Alzheimer's Disease, which assesses the severity and distribution of Alzheimer-type pathology, vascular lesions, and other potential causes of dementia. A statistical model of dementia risk related predominantly to Alzheimer-type and vascular pathology was developed by multivariate logistic regression. We report on the first 209 individuals who have come to necropsy. The median age at death was 85 years for men, and 86 years for women. Cerebrovascular (78%) and Alzheimer-type (70%) pathology were common. Dementia was present in 100 (48%), of whom 64% had features indicating probable or definite Alzheimer's disease. However, 33% of the 109 non-demented people had equivalent densities of neocortical neuritic plaques. Some degree of neocortical neurofibrillary pathology was found in 61% of demented and 34% of non-demented individuals. Vascular lesions were equally common in both groups, although the proportion with multiple vascular pathology was higher in the demented group (46% vs 33%). Alzheimer-type and vascular pathology were the major pathological correlates of cognitive decline in this elderly sample, as expected, but most patients had mixed disease. There were no clear thresholds of these features that predicted dementia status. The findings therefore challenge conventional dementia diagnostic criteria in this setting. Additional factors must determine whether moderate burdens of cerebral Alzheimer-type pathology and vascular lesions are associated with cognitive failure.


[Bettolepsy and chronic pleurisy].
The author submits a clinical observation of a patient who had chronic tuberculous pleurisy complicated by constriction of the root of the lung having resulted in bettolepsy. The paper contains theorization concerning its pathogenesis. An outline of differential diagnosis is given together with surgery indications.


Enhanced trimethylamine metabolism and gut dysbiosis in type 2 diabetes mellitus with microalbumin.
Abnormal gut microbiota and blood trimethylamine-N-oxide (TMAO) metabolome have been reported in patients with type 2 diabetes mellitus (T2DM) and advanced diabetic nephropathy. This study aimed to investigate the gut microbiota profiles and a group of targeted urine metabolic characteristics in T2DM patients with or without microalbuminuria, to determine the correlation between the gut microbiota composition, trimethylamine (TMA) metabolism, and the clinical features during progression of diabetic kidney disease (DKD). This study included 26 T2DM patients with microalbuminuria (Micro), 26 T2DM patients with normoalbuminuria (Normo), and 15 healthy controls (HC). Urine and Fecal samples were detected using ultra performance liquid chromatography tandem mass spectrometry and 16S ribosomal DNA gene sequencing, respectively. The TMAO/TMA ratio decreased gradually during the HC-Normo-Micro transition. The levels of TMA, choline and betaine were significantly different between the HC group and the T2DM patients belonging to both Normo and Micro groups. At the operational taxonomic unit (OTU) level, the gut microflora diversity was significantly reduced in the Micro groups compared to the HC groups and the Normo groups. Taxonomic analyses revealed significant consumption in the relative abundances of eight bacterial genera and significant enrichment of two bacterial genera during the HC-Normo-Micro transition. Furthermore, the relative abundances of six bacterial genera, namely, Ruminococcus_1, [Eubacterium]_ruminantium_group, Roseburia, Faecalibacterium, Fusicatenibacter and Coprococcus_3 exhibited significant differences, and were associated with elevated urinary albumin creatinine ratio (UACR), TMAO/TMA, TMA and its precursors in the Micro group compared with the other groups. The imbalance of gut microbiota has occurred in patients with early-stage DKD, and the consumption of short-chain fatty acid-producing bacteria were associated with the accumulation of TMA and UACR.


Genomic characterization and expression analysis of the first nonmammalian renin genes from zebrafish and pufferfish.
Renin is a key enzyme in the renin-angiotensin system (RAS), a pathway which plays an important physiological role in blood pressure and electrolyte homeostasis. The origin of the RAS is believed to have accompanied early evolution of vertebrates. However, renin genes have so far only been unequivocally identified in mammals. Whether or not a bona fide renin gene exists in nonmammalian vertebrates has been an intriguing question of physiological and evolutionary interest. Using a genomic analytical approach, we identified renin genes in two nonmammalian vertebrates, zebrafish (Danio rerio) and pufferfish (Takifugu rubripes). Phylogenetic analysis demonstrates that the predicted fish renins cluster together with mammalian renins to form a distinct subclass of vertebrate aspartyl proteases. RT-PCR results confirm generation of the predicted zebrafish mRNA and its expression in association with the opisthonephric kidney of adult zebrafish. Comparative in situ hybridization analysis of wild-type and developmental mutants indicates that renin expression is first detected bilaterally in cells of the interrenal primordia at 24 h postfertilization, which subsequently migrate to lie adjacent to, but distinct from, the glomerulus of the developing pronephric kidney. Our report provides the first molecular evidence for the existence of renin genes in lower vertebrates. The observation that the earliest renin-expressing cells, arising during ontogeny of this teleost vertebrate, are of adrenocortical lineage raises an interesting hypothesis as regards the origin of renin-expressing cells in the metanephric kidney of higher vertebrates.


A genome-wide association study identifies genetic loci associated with specific lobar brain volumes.
Brain lobar volumes are heritable but genetic studies are limited. We performed genome-wide association studies of frontal, occipital, parietal and temporal lobe volumes in 16,016 individuals, and replicated our findings in 8,789 individuals. We identified six genetic loci associated with specific lobar volumes independent of intracranial volume. Two loci, associated with occipital (6q22.32) and temporal lobe volume (12q14.3), were previously reported to associate with intracranial and hippocampal volume, respectively. We identified four loci previously unknown to affect brain volumes: 3q24 for parietal lobe volume, and 1q22, 4p16.3 and 14q23.1 for occipital lobe volume. The associated variants were located in regions enriched for histone modifications (DAAM1 and THBS3), or close to genes causing Mendelian brain-related diseases (ZIC4 and FGFRL1). No genetic overlap between lobar volumes and neurological or psychiatric diseases was observed. Our findings reveal part of the complex genetics underlying brain development and suggest a role for regulatory regions in determining brain volumes.


Examining the characteristics of social and behavior change communication intervention costs in low- and middle-income countries: A hedonic method approach.
Understanding the costs of health interventions is critical for generating budgets, planning and managing programs, and conducting economic evaluations to use when allocating scarce resources. Here, we utilize techniques from the hedonic pricing literature to estimate the characteristics of the costs of social and behavior change communication (SBCC) interventions, which aim to improve health-seeking behaviors and important intermediate determinants to behavior change. SBCC encompasses a wide range of interventions including mass media (e.g., radio, television), mid media (e.g., community announcements, live dramas), digital media (e.g., short message service/phone reminders, social media), interpersonal communication (e.g., individual or group counseling), and provider-based SBCC interventions focused on improving provider attitudes and provider-client communication. While studies have reported on the costs of specific SBCC interventions in low- and middle-income countries, little has been done to examine SBCC costs across multiple studies and interventions. We use compiled data across multiple SBCC intervention types, health areas, and low- and middle-income countries to explore the characteristics of the costs of SBCC interventions. Despite the wide variation seen in the unit cost data, we can explain between 63 and 97 percent of total variance and identify a statistically significant set of characteristics (e.g., health area) for media and interpersonal communication interventions. Intervention intensity is an important determinant for both media and interpersonal communication, with costs increasing as intervention intensity increases; other important characteristics for media interventions include intervention subtype, target population group, and country income as measured by per capita Gross National Income. Important characteristics for interpersonal communication interventions include health area, intervention subtype, target population group and geographic scope.