Screening of plant toxins in food, feed and botanicals using full-scan high-resolution (Orbitrap) mass spectrometry.
A generic method based on LC with full-scan high-resolution (Orbitrap) mass spectrometry (MS) was systematically investigated for the simultaneous detection of a wide range of plant toxins in a variety of food and feed matrices. For a selection of 150 substances, representing various chemical classes, the limit of detection was established using fixed LC-MS conditions. Ion suppression effects and selectivity were evaluated using generic extracts from representative and relevant matrices (food supplement, honey, silage, compound feed). The majority of the substances could be measured as positive ions after electrospray ionisation (ESI(+)). Using a mass resolving power of 100,000 a reliable high mass accuracy was obtained despite the high abundance of co-extractants in the sample extracts. This enabled the use of ±5 ppm mass extraction windows, which in turn resulted in a high degree of selectivity. On the other hand, except for honey, strong ion suppression effects were frequently observed which adversely affected the detection limits. Nevertheless, for the majority of the substances the detection limits were in the range 0.01-0.05 mg kg(-1). Since non-selective sample preparation and non-targeted data acquisition were performed, the presence of plant toxins initially not targeted for during data review can be subsequently investigated, which is a very useful option because for many known toxins no analytical reference standards are yet available. The applicability of the method was demonstrated by analysis of a variety of real-life samples purchased on the market or from cases of intoxication. These included honey, herbal tea, food supplements, poppy seeds, traditional Chinese medicines, compound feed, silage and herb-based feed additives. Plant toxins that were detected included various pyrrolizidine alkaloids, grayanotoxins, opium alkaloids, strychnine, ricinine (a marker for ricin), aconitine, aristolochic acid and cardiac glycosides (e.g. digitoxin, digoxin).


Taming the elephant: Salmonella biology, pathogenesis, and prevention.
Salmonella infections continue to cause substantial morbidity and mortality throughout the world. However, recent discoveries and new paradigms promise to lead to novel strategies to diagnose, treat, and prevent Salmonella infections. This review provides an update of the Salmonella field based on oral presentations given at the recent 3rd ASM Conference on Salmonella: Biology, Pathogenesis and Prevention.


Subcellular compartmentalization of lead in the earthworm, Eisenia fetida: Relationship to survival and reproduction.
Metals are detoxified and sequestered into subcellular compartments when accumulated by earthworms. Differential centrifugation was used to quantify subcellular Pb in three separate studies to measure 14-day acute toxicity (lethality), 28/56-day reproductive effects, and 90-day bioaccumulation in spiked-soil exposed earthworms, Eisenia fetida. Observed toxicity and total body Pb was consistent with published work of others. Pb showed concentration-dependent toxicity relationships (lethality and reproduction) for total and subcellular Pb. Toxic fraction and total Pb showed similar concentration-response patterns in the 14-day and 28/56-day studies and tended to increase towards a plateau at higher concentrations. Linear correlations of subcellular to total Pb was observed in all studies except the 90-day bioaccumulation study in which toxic fraction Pb appeared to approach a maximum over the period between Day 56 and Day 90. In a follow-on study using two different contaminated soil types, toxic fraction and total Pb concentrations as related to reproductive effects were consistent with data from our spiked soil studies, and this suggests it may be possible to use such values to "factor out" matrix-specific influences that otherwise skew toxicity values when expressed relative to soil concentrations. Our findings, however, suggest the subcellular fractionation approach may not offer advantages over total Pb determination in short-term exposure studies but may become important when longer exposure periods (greater than 90 days) are considered. In this respect, the technique we describe has the potential to provide valuable information for assessing and interpreting Pb toxicity as a function of earthworm body burden.


Identification and Localization of Cell Types in the Mouse Olfactory Bulb Using Slide-SeqV2.
Spatial transcriptomics maps RNA molecules to the location in a tissue where they are expressed. Here we document the use of Slide-SeqV2 to visualize gene expression in the mouse olfactory bulb (OB). This approach relies on spatially identified beads to locate and quantify individual transcripts. The expression profiles associated with the beads are used to identify and localize individual cell types in an unbiased manner. We demonstrate the various cell types and subtypes with distinct spatial locations in the olfactory bulb that are identified using Slide-SeqV2.


Exposure of adolescent girls to cigar images in women's magazines, 1992-1998.
This study assessed the exposure of adolescent girls to cigar images in women's magazines from 1992 to 1998. Data were obtained from the 5 women's magazines with the highest readership of adolescent girls. We found a significant upward trend in cigar images portrayed in women's magazines and a significant increase in the portrayal of women cigar smokers over the period observed. Cigar images were less likely than cigarette images to promote tobacco or nontobacco products. Among advertisements, nearly all those that featured cigars promoted nontobacco products; advertisements featuring cigarettes overwhelmingly promoted tobacco products. Between 1992 and 1998, adolescent readers of women's magazines were increasingly exposed to images of cigars.


The initial Trinidad experience with Cine MRI in clinical cardiology.
We describe the initial Trinidad experience with Magnetic Resonance Imaging (MRI) and Cine MRI as a diagnostic tool in clinical cardiology. Six patients from the following categories were referred for Cine MRI evaluation: congenital heart disease, valvular heart disease, aortic diseases, cardiomyopathy and intracardiac mass. All patients underwent echocardiography. MRI and Cine MRI were performed on all patients using a Siemens Magnetom 1.0 Tesla MR system at MRI Trinidad and Tobago Ltd. Selected patients underwent Angiography and/or computed tomography (CT) scanning. Clinical data and images of the six patients evaluated are described. MRI and Cine MRI provided excellent anatomical and functional details of the heart and aorta in five patients with dissection of the aorta, aneurysm of the ascending aorta, suspected left ventricular apical thrombus, infiltrative cardiomyopathy and arrhythmogenic right ventricular dysplasia. Technical difficulty was experienced with one patient who had a congenital defect (common atrium). In this study, Cine MRI provided excellent images in all but one patient. This new noninvasive technique enhanced diagnostic capabilities and facilitated management in patients with certain cardiovascular diseases.


Interactive Effects of Perinatal BPA or DES and Adult Testosterone and Estradiol Exposure on Adult Urethral Obstruction and Bladder, Kidney, and Prostate Pathology in Male Mice.
Obstructive voiding disorder (OVD) occurs during aging in men and is often, but not always, associated with increased prostate size, due to benign prostatic hyperplasia (BPH), prostatitis, or prostate cancer. Estrogens are known to impact the development of both OVD and prostate diseases, either during early urogenital tract development in fetal-neonatal life or later in adulthood. To examine the potential interaction between developmental and adult estrogen exposure on the adult urogenital tract, male CD-1 mice were perinatally exposed to bisphenol A (BPA), diethylstilbestrol (DES) as a positive control, or vehicle negative control, and in adulthood were treated for 4 months with Silastic capsules containing testosterone and estradiol (T+E2) or empty capsules. Animals exposed to BPA or DES during perinatal development were more likely than negative controls to have urine flow/kidney problems and enlarged bladders, as well as enlarged prostates. OVD in adult T+E2-treated perinatal BPA and DES animals was associated with dorsal prostate hyperplasia and prostatitis. The results demonstrate a relationship between elevated exogenous estrogen levels during urogenital system development and elevated estradiol in adulthood and OVD in male mice. These findings support the two-hit hypothesis for the development of OVD and prostate diseases.


Confusing thoughts and speech: source monitoring and psychosis.
To explore the idea that deficits in source monitoring may underlie positive symptoms of schizophrenia, the current study compared schizophrenic patients' performance (n=15) on an internal source-monitoring task with that of normal controls (n=15). On the basis of a source-monitoring task in which participants had to recall whether they had verbalized answers or merely thought about these answers, overall source monitoring performance, discrimination index, and response bias were calculated. In addition, participants completed cognitive tests and symptomatology questionnaires. Relative to controls, patients had significantly more difficulties with monitoring their own actions and showed a tendency towards misclassifying imagined thoughts as verbalized thoughts. Source-monitoring performance was related to selective attention, but not to other cognitive domains. No relationship was found between source-monitoring and symptomatology. Failures in internal source monitoring are a prominent feature of schizophrenia, and our results suggest that they form a more enduring characteristic of this disorder than has previously been assumed.


Comparative Analysis of Cytokines of Tumor Cell Lines, Malignant and Benign Effusions Around Breast Implants.
More than 700 women have developed an anaplastic large T cell lymphoma (ALCL) surrounding textured surface breast implants, termed breast implant-associated ALCL (BIA-ALCL). Most patients with BIA-ALCL present with an accumulation of fluid (delayed seroma) around the implant. However, benign seromas without malignant cells complicating scar contracture, implant rupture, trauma, infection, and other causes are more common. For proper patient management and to avoid unnecessary surgery, a simple diagnostic test to identify malignant seromas is desirable. The aim of this study was to develop an ancillary test for the diagnosis of malignant seromas and to gain insight into the nature of the malignant cells and their microenvironment. We employed an immunologic assay on only 50 µL of aspirated seroma fluid. The assay measures 13 cytokines simultaneously by flow cytometry. To establish a baseline for clinical studies we measured cytokines secreted by BIA-ALCL and cutaneous ALCL lines. Our study of cell line culture supernatants, and 8 malignant compared with 9 benign seromas indicates that interleukin 9 (IL-9), IL-10, IL-13, IL-22, and/or interferon γ concentrations >1000 pg/mL distinguish malignant seromas from benign seromas. IL-6, known to be a driver of malignant cells, is also elevated in benign seromas and does not distinguish them from malignant seromas. The cytokine assay introduced in this study can be used together with levels of soluble CD30 to identify malignant seromas. Validation of these findings in a larger prospective patient cohort is warranted. The unique pattern of cytokine expression in malignant effusions surrounding breast implants gives further insight into the pathogenesis and cells of origin of BIA-ALCL. Level of Evidence: 5.


Gold-silica quantum rattles for multimodal imaging and therapy.
Gold quantum dots exhibit distinctive optical and magnetic behaviors compared with larger gold nanoparticles. However, their unfavorable interaction with living systems and lack of stability in aqueous solvents has so far prevented their adoption in biology and medicine. Here, a simple synthetic pathway integrates gold quantum dots within a mesoporous silica shell, alongside larger gold nanoparticles within the shell's central cavity. This "quantum rattle" structure is stable in aqueous solutions, does not elicit cell toxicity, preserves the attractive near-infrared photonics and paramagnetism of gold quantum dots, and enhances the drug-carrier performance of the silica shell. In vivo, the quantum rattles reduced tumor burden in a single course of photothermal therapy while coupling three complementary imaging modalities: near-infrared fluorescence, photoacoustic, and magnetic resonance imaging. The incorporation of gold within the quantum rattles significantly enhanced the drug-carrier performance of the silica shell. This innovative material design based on the mutually beneficial interaction of gold and silica introduces the use of gold quantum dots for imaging and therapeutic applications.


Mitochondrial retrograde signaling mediated by UCP2 inhibits cancer cell proliferation and tumorigenesis.
Cancer cells tilt their energy production away from oxidative phosphorylation (OXPHOS) toward glycolysis during malignant progression, even when aerobic metabolism is available. Reversing this phenomenon, known as the Warburg effect, may offer a generalized anticancer strategy. In this study, we show that overexpression of the mitochondrial membrane transport protein UCP2 in cancer cells is sufficient to restore a balance toward oxidative phosphorylation and to repress malignant phenotypes. Altered expression of glycolytic and oxidative enzymes mediated the effects of this metabolic shift. Notably, UCP2 overexpression increased signaling from the master energy-regulating kinase, adenosine monophosphate-activated protein kinase, while downregulating expression of hypoxia-induced factor. In support of recent new evidence about UCP2 function, we found that UCP2 did not function in this setting as a membrane potential uncoupling protein, but instead acted to control routing of mitochondria substrates. Taken together, our results define a strategy to reorient mitochondrial function in cancer cells toward OXPHOS that restricts their malignant phenotype.


Body mass index: comparing mean values and prevalence rates from telephone and examination surveys.
Cost effective means of assessing the levels of risk factors in the population have to be defined in order to monitor these factors over time and across populations. This study is aimed at analyzing the difference in population estimates of the mean levels of body mass index (BMI) and the prevalences of overweight, between health examination survey and telephone survey. The study compares the results of two health surveys, one by telephone (N=820) and the other by physical examination (N=1318). The two surveys, based on independent random samples of the population, were carried out over the same period (1992-1993) in the same population (canton of Vaud, Switzerland). Overall participation rates were 67% and 53% for the health interview survey (HIS) and the health examination survey (HES) respectively. In the HIS, the reporting rate was over 98% for weight and height values. Self-reported weight was on average lower than measured weight, by 2.2 kg in men and 3.5 kg in women, while self-reported height was on average greater than measured height, by 1.2 cm in men and 1.9 cm in women. As a result, in comparison to HES, HIS led to substantially lower mean levels of BMI, and to a reduction of the prevalence rates of obesity (BMI>30 kg/m(2)) by more than a half. These differences are larger for women than for men. The two surveys were based on different sampling procedures. However, this difference in design is unlikely to explain the systematic bias observed between self-reported and measured values for height and weight. This bias entails the overall validity of BMI assessment from telephone surveys.


Gene expression plasticity in response to salinity acclimation in threespine stickleback ecotypes from different salinity habitats.
Phenotypic plasticity is thought to facilitate the colonization of novel environments and shape the direction of evolution in colonizing populations. However, the relative prevalence of various predicted patterns of changes in phenotypic plasticity following colonization remains unclear. Here, we use a whole-transcriptome approach to characterize patterns of gene expression plasticity in the gills of a freshwater-adapted and a saltwater-adapted ecotype of threespine stickleback (Gasterosteus aculeatus) exposed to a range of salinities. The response of the gill transcriptome to environmental salinity had a large shared component common to both ecotypes (2159 genes) with significant enrichment of genes involved in transmembrane ion transport and the restructuring of the gill epithelium. This transcriptional response to freshwater acclimation is induced at salinities below two parts per thousand. There was also differentiation in gene expression patterns between ecotypes (2515 genes), particularly in processes important for changes in the gill structure and permeability. Only 508 genes that differed between ecotypes also responded to salinity and no specific processes were enriched among this gene set, and an even smaller number (87 genes) showed evidence of changes in the extent of the response to salinity acclimation between ecotypes. No pattern of relative expression dominated among these genes, suggesting that neither gains nor losses of plasticity dominated the changes in expression patterns between the ecotypes. These data demonstrate that multiple patterns of changes in gene expression plasticity can occur following colonization of novel habitats.


A case of heterozygous familial hypercholesterolemia associated with hyperthyroidism: effects of triiodothyronine on low-density lipoprotein receptor and cholesterol synthesis.
A 58-year-old patient with heterozygous familial hypercholesterolemia (FH) showed normal levels of serum cholesterol (193 mg/dL) in coexistence with hyperthyroidism. After hyperthyroidism therapy with radioiodine and methimazole, the patient's lipid profile showed high concentrations of cholesterol (whole serum 318 mg/dL, VLDL 35 mg/dL, LDL 217 mg/dL, HDL 44 mg/dL). There was a significant inverse correlation between serum cholesterol levels and serum thyroxine levels (r = -0.815, p less than 0.01). Effects of triiodothyronine on LDL degradation and cholesterol synthesis from 14C-labeled acetate were studied in cultured skin fibroblasts. Triiodothyronine (T3) stimulated both LDL degradation and cholesterol synthesis in the cells from normal subjects and patients with heterozygous FH. The T3 increased cellular cholesterol synthesis markedly in the cells from patients with homozygous FH but did not increase LDL receptor activity. These results suggest that normal serum cholesterol levels in our case result in part from an enhancement of LDL receptors by thyroid hormone.


Primary atypical teratoid rhabdoid tumor in the adult spine.
Atypical teratoid/rhabdoid tumor (ATRT) is a highly aggressive tumor of the central nervous system (WHO grade IV), which is most frequently found intracranially in young children and infants. Only three prior cases of primary ATRT involving the adult spine were found following a literature review, and the average survival for these patients was only 20 postoperative months. A 43 year-old female presented with an acute exacerbation of chronic neck pain. While awaiting magnetic resonance (MR) studies of the cervical spine, she was found pulseless in her room. Although cardiopulmonary resuscitation was successful, she was found to be quadriplegic. The subsequent cervical MR imaging revealed a C1-3 intradural, extramedullary ventrolateral mass, markedly compressing the upper cervical spinal cord. Following successful surgical resection of the lesion, which proved pathologically to be an ATRT, she was treated with a full course of fractionated radiation therapy. Over the successive 6-month period, her neurological examination continued to improve to 4-/5 functional strength in her upper extremities, however, remained with 2/5 nonfunctional strength in her legs. ATRT involving the adult spine are rare and may often be misdiagnosed. This study points out that aggressive surgery followed by radiation therapy may improve outcome.


Contributions of thalamic nuclei to declarative memory functioning.
In spite of the acknowledged role that the thalamus plays in declarative memory, details about the precise memory processes it is involved in and which are the structures of the thalamus that contribute to these processes remain unknown. An overview is presented of human clinical and animal experimental findings showing the involvement of the thalamus, at the level of white matter tracts and separate nuclei, in aspects of memory functioning. The region in the thalamus that contributes to declarative memory is the anterior and medial division, containing the anterior nuclei, the medial dorsal nucleus and the intralaminar and midline nuclei. A lesion to the anterior nuclei or their afferent white matter tract, the mammillothalamic tract, results in deficits of encoding of new stimuli. Lesions to the medial dorsal nucleus affect executive processes pertaining to declarative memory, such as the use of memory strategies for retrieval; damage to the intralaminar and midline nuclei results in decreased arousal and thus affects the declarative memory process. Based on anatomical and functional data, a theory is proposed of how the thalamus might play a role at different levels of declarative memory functioning. Firstly, the anterior and mediodorsal nucleus are involved in processing the contents of the stimuli for storage and recall. The anterior nuclei influence the selection of material to be stored and remembered, whereas the mediodorsal nucleus is involved in the coordination and selection of the strategies used to retrieve material. Secondly, the intralaminar and midline nuclei and specifically the lateral and ventral components, maintain a necessary state of the cortical regions involved in the ongoing memory processes. The two types of function subserved by these groups of thalamic nuclei, focussing on contents vs. state, need to work in parallel to mediate and allow memory functioning, respectively.


Regulation and function of inhibins in the normal menstrual cycle.
The development of assays specific for dimeric inhibin A and inhibin B defined the distinct physiology of these two hormones in the normal menstrual cycle. Inhibin A and inhibin B expression and secretion along with their differential regulation by gonadotropins explain their unique serum patterns and their potential endocrine and ovarian autocrine-paracrine functions. There is evidence that inhibin A and inhibin B play an endocrine role in the negative regulation of follicle-stimulating hormone (FSH) in nonhuman primates and humans. However, some studies suggest that estradiol is a more important, if not the only, negative feedback regulator of FSH in women. There is also evidence from animal models that inhibins and activins play a critical role in follicle development. Future work will be necessary to define further the relative role of the inhibins, estradiol, and other autocrine-paracrine factors in these important reproductive functions.


Aspergillus fumigatus Cell Wall α-(1,3)-Glucan Stimulates Regulatory T-Cell Polarization by Inducing PD-L1 Expression on Human Dendritic Cells.
Human dendritic cell (DC) response to α-(1,3)-glucan polysaccharide of Aspergillus fumigatus and ensuing CD4+ T-cell polarization are poorly characterized. α-(1,3)-Glucan was isolated from A. fumigatus conidia and mycelia cell wall. For the analysis of polarization, DCs and autologous naive CD4+ T cells were cocultured. Phenotype of immune cells was analyzed by flow cytometry, and cytokines by enzyme-linked immunosorbent assay (ELISA). Blocking antibodies were used to dissect the role of Toll-like receptor 2 (TLR2) and programmed death-ligand 1 (PD-L1) in regulating α-(1,3)-glucan-mediated DC activation and T-cell responses. DCs from TLR2-deficient mice were additionally used to consolidate the findings. α-(1,3)-Glucan induced the maturation of DCs and was dependent in part on TLR2. "α-(1,3)-Glucan-educated" DCs stimulated the activation of naive T cells and polarized a subset of these cells into CD4+CD25+FoxP3+ regulatory T cells (Tregs). Mechanistically, Treg stimulation by α-(1,3)-glucan was dependent on the PD-L1 pathway that negatively regulated interferon-gamma (IFN-γ) secretion. Short α-(1,3)-oligosaccharides lacked the capacity to induce maturation of DCs but significantly blocked α-(1,3)-glucan-induced Treg polarization. PD-L1 dictates the balance between Treg and IFN-γ responses induced by α-(1,3)-glucan. Our data provide a rationale for the exploitation of immunotherapeutic approaches that target PD-1-PD-L1 to enhance protective immune responses to A. fumigatus infections.


Assessing commercial feasibility: a practical and ethical prerequisite for human clinical testing.
This article proposes that an assessment of commercial feasibility should be integrated as a prerequisite for human clinical testing to improve the quality and relevance of materials being investigated, as an ethical aspect for human subject protection, and as a means of improving accountability where clinical development is funded on promises of successful translational research. A commercial feasibility analysis is not currently required to justify human clinical testing, but is assumed to have been conducted by industry participants, and use of public funds for clinical trials should be defensible in the same manner. Plant-made vaccines (PMVs) are offered in this discussion as a model for evaluating the relevance of commercial feasibility before human clinical testing. PMVs have been proposed as a potential solution for global health, based on a vision of immunizing the world against many infectious diseases. Such a vision depends on translating current knowledge in plant science and immunology into a potent vaccine that can be readily manufactured and distributed to those in need. But new biologics such as PMVs may fail to be manufactured due to financial or logistical reasons--particularly for orphan diseases without sufficient revenue incentive for industry investment--regardless of the effectiveness which might be demonstrated in human clinical testing. Moreover, all potential instruments of global health depend on translational agents well beyond the lab in order to reach those in need. A model compromising five criteria for commercial feasibility is suggested for inclusion by regulators and ethics review boards as part of the review process prior to approval of human clinical testing. Use of this model may help to facilitate safe and appropriate translational research and bring more immediate benefits to those in need.


The RISE FOR HEALTH study: Methods for in-person assessment and biospecimen collection.
To describe the methods for the in-person assessment of the RISE FOR HEALTH (RISE) study, a population-based multicenter prospective cohort study designed to identify factors that promote bladder health and/or prevent lower urinary tract symptoms in adult women, conducted by the Prevention of Lower Urinary Tract Symptoms Research Consortium (PLUS). A subset of RISE participants who express interest in the in-person assessment will be screened to ensure eligibility (planned n = 525). Eligible consenting participants are asked to complete 15 physical assessments in addition to height and weight, to assess pelvic floor muscle function, musculoskeletal (MSK) status, and pain, and to provide urogenital microbiome samples. Pelvic floor muscle assessments include presence of prolapse, strength, levator attachment integrity (tear) and myofascial pain. MSK tests evaluate core stability, lumbar spine, pelvic girdle and hip pain and function. Participants are asked to complete the Short Physical Performance Battery to measure balance, lower extremity strength, and functional capacity. All participants are asked to provide a voided urine sample and a vaginal swab for microbiome analyses; a subset of 100 are asked to contribute additional samples for feasibility and validation of a home collection of urinary, vaginal, and fecal biospecimens. Online and in-person training sessions were used to certify research staff at each clinical center before the start of RISE in-person assessments. Standardized protocols and data collection methods are employed uniformly across sites. The RISE in-person assessment is an integral portion of the overall population-based RISE study and represents an innovative approach to assessing factors hypothesized to promote bladder health and/or prevent lower urinary tract symptoms. Data collected from this assessment will be used to prioritize future research questions and prevention strategies and interventions. This description of the assessment methods is intended to provide methodologic transparency and inform other researchers who join efforts to understand and improve bladder health.


Ornithinibacillus contaminans sp. nov., an endospore-forming species.
A Gram-stain-positive, endospore-forming rod, designated CCUG 53201(T), was isolated from a human blood sample of a 75-year-old woman. 16S rRNA gene sequence-based phylogenetic analysis showed that strain CCUG 53201(T) clustered with the type strains of species of the genus Ornithinibacillus. Strain CCUG 53201(T) was most closely related to Ornithinibacillus bavariensis WSBC 24001(T) and Ornithinibacillus californiensis DSM 16628(T) (97.9 and 98.7 % 16S rRNA gene sequence similarity, respectively). Strain CCUG 53201(T) contained a peptidoglycan of type A4β l-Orn-d-Asp. The quinone system was composed of the menaquinone MK-7 and small amounts of MK-6. The polar lipid profile of strain CCUG 53201(T) consisted of major amounts of diphosphatidylglycerol and an unidentified phospholipid, moderate amounts of phosphatidylglycerol and another two unidentified phospholipids and minor amounts of several other components. The fatty acid profile comprised mainly anteiso- and iso-branched fatty acids and was in accordance with those of members of the genus Ornithinibacillus. The polyamine pattern exhibited the major compounds spermidine and spermine. The results of physiological and biochemical tests and DNA-DNA hybridization allowed the phenotypic and genotypic differentiation of strain CCUG 53201(T) from its closest phylogenetic neighbours. We propose a novel species with the name Ornithinibacillus contaminans sp. nov., with type strain CCUG 53201(T) (=DSM 22953(T)).


Bioengineering of Fetal Skin: Differentiation of Amniotic Fluid Stem Cells into Keratinocytes.
Open fetal spina bifida repair has become a novel clinical standard of care. In very large spina bifida lesions, the skin defect cannot be covered primarily, asking for alternative solutions. We hypothesize that amniotic fluid stem cells (AFSC) could be differentiated into keratinocytes that could then be used to bioengineer autologous skin usable for in utero back coverage. To obtain human AFSC, amniotic fluid samples obtained from fetal surgeries were subjected to immunoselection for c-kit. C-kit-positive samples and controls were cultured with the additives morphogenetic protein 4 and vitamin C to induce differentiation towards keratinocytes. This process was monitored by measuring the expression of K8 and K14 via immunohistochemical staining, flow cytometry, and polymerase chain reaction. After immunoselection and expansion, most cells were positive for K8, but none for K14. After completion of the differentiation protocol, cell colonies with keratinocyte-like appearance could be observed, but cells remained positive for K8 and negative for K14, indicating failed differentiation into keratinocytes. Culturing of keratinocyte-like cells from AFSC, harvested intraoperatively, was not feasible in this setting. The reasons for failure must be investigated and eliminated, as bioengineering of fetal skin for clinical use during fetal surgery for spina bifida remains an attractive goal.


Phospho-mTOR is not upregulated in metastatic SDHB paragangliomas.
Pheochromocytomas (PCCs)/paragangliomas (PGLs) are neuroendocrine tumours that may cause arrhythmia and death if untreated. Treatment for patients with metastatic tumours is lacking. As new PCC/PGL susceptibility genes are discovered that are associated with the mTOR pathway, treatment targets focusing on this pathway are being intensively explored. Twenty-one human PCC/PGLs were analysed from two tertiary care centres. Immunohistochemistry (IHC) analysis was performed for phospho-mTOR (pmTOR), phospho-S6K (pS6K), phosphoinositide 3-kinase (PI3K), phospho-4EBP1 (p4EBP1), HIF1α and MIB-1 in 6 metastatic SDHB PCC/PGLs, 15 nonmetastatic PCC/PGLs, (including 1 TMEM127 PCC and 1 nonmetastatic SDHB PGL) and 6 normal adrenal medullas. The product of the intensity of stain and percentage of cells stained was calculated as an H score. Using a two-sample t-test and paired t-test, pmTOR and pS6K had significantly higher H scores in nonmetastatic PCC/PGLs than in metastatic SDHB PCC/PGLs. HIF1α had significantly higher H scores in metastatic SDHB PCC/PGLs compared with nonmetastatic PCC/PGLs and normal adrenal medulla. No difference in H scores was seen with p4EBP1, PI3K and MIB-1 when comparing metastatic SDHB PCC/PGLs and nonmetastatic PCC/PGLs. Significantly higher difference in pS6K was seen in normal adrenal medullas compared to nonmetastatic PCC/PGLs and metastatic SDHB PCC/PGLs. The present results suggest that the use of mTOR inhibitors alone for metastatic SDHB PCC/PGLs may not achieve good therapeutic efficacy in patients.


Prognostic factors for locoregional recurrence and survival in stage IIIC breast carcinoma: impact of adjuvant radiotherapy.
The aims of the present study were to define the prognostic factors for locoregional recurrence (LRR) and survival in stage IIIC breast carcinoma as well as to examine the impact of adjuvant radiotherapy on the outcome of the disease. The records of 586 consecutive patients with stage IIIC breast carcinoma who underwent modified radical mastectomy were evaluated, and the prognostic factors for LRR and survival were analysed. Survival curves were generated using the Kaplan-Meier method, and multivariate analysis was performed using the Cox proportional hazard model. Five-year LRR and survival of stage IIIC breast carcinoma were 15 percent and 41.3 percent, respectively. Five-year LRR was significantly lower and five-year survival was significantly higher for all patients as well as for T1-2 patients with one to three apical node involvements who were treated with adjuvant radiotherapy. In multivariate analysis, apical node involvement, age below 35 years, T4 tumour, grade 3, extracapsular extension and lymphovascular invasion decreased survival, whereas adjuvant tamoxifen and adjuvant radiotherapy (risk ratio [RR] 0.51, 95 percent confidence interval [CI] 0.39-0.67) increased survival. Adjuvant radiotherapy was the sole independent factor that was found to be significantly associated with decreased LRR (RR 0.25, 95 percent CI 0.16-0.38). Radiotherapy decreased LRR and increased survival significantly in all stage IIIC patients and in the subgroup of T1-2 patients with one to three apical node involvements. Thus, it should be considered in the treatment of stage IIIC breast carcinoma.


Pediatric Femoral Shaft Fractures: A Multicenter Review of the AAOS Clinical Practice Guidelines Before and After 2009.
To determine if the AAOS clinical practice guidelines (CPG) for the treatment of pediatric femoral shaft fractures (2009) changed treatment, we analyzed pediatric femoral shaft fractures at 4 high-volume, geographically separated, level-1 pediatric trauma centers over a 10-year period (2004 to 2013). Consecutive series of pediatric femoral shaft fractures (ages, birth to 18 y) treated at the 4 centers were reviewed. Treatment methods were analyzed by age and treatment method for each center and in aggregate. Of 2646 fractures, 1476 (55.8%) were treated nonoperatively and 1170 fractures operatively. Of the operative group, flexible intramedullary nails (IMN) were used for 568 patients (21.5%), locked intramedullary nails (LIMNs) for 309 (11.7%), and plating for 188 (7.1%). In total, 105 fractures were treated with external fixation or skeletal traction. Analysis before and after the CPG publication revealed a significant increase in the use of interlocked IMNs in patients younger than 11 years (0.5% before, 3.8% after; P<0.001). Over the same time period there was an increase in surgical management, regardless of technique, for patients younger than 5 years (6.4% before, 8.4% after; P=0.206). There were considerable differences in treatment among centers: 74% of fractures treated with plating were from a single center (center A), which also contributed 68% of patients younger than 5 years treated with plating; center B had the highest rate (41%) of flexible IMN in children younger than 5 years; center C had the highest rate (63%) of LIMN in children younger than 11 years; and center D treated the fewest patients outside the CPG guidelines. Following publication of the AAOS CPG, there was a significant increase in the use of LIMNs in patients younger than 11 years old and a trend toward surgical treatment in patients younger than 5 years. The considerable variability among centers in treatment methods and adherence to the CPG highlights the need for further outcome studies to better define optimal treatment methods and perhaps update the AAOS CPG guidelines. Level III-therapeutic.


Research trends on immunotherapy for pancreatic cancer: A bibliometric analysis.
This study aims to summarize and visually analyze the current research status in pancreatic cancer immunotherapy during the past two decades by bibliometrics and explore the current research hotspots and future development directions. The literature related to pancreatic cancer immunotherapy from 2002 to 2021 was downloaded from the core database of the Web of Science. VOSviewer and CiteSpace software were used to visualize the included literature. A total of 2528 articles were included. In the past two decades, publications in the pancreatic cancer immunotherapy field have increased almost annually. As the country with the largest publications, the United States has various research institutions dedicated to pancreatic cancer immunotherapy. Jaffee EM and Zheng L from Johns Hopkins University and Vonderheide RH from the University of Pennsylvania have published the most articles in this field. The current research hotspots of pancreatic cancer immunotherapy include the tumor microenvironment, immune cells, immune checkpoint blockade, and combination therapy. The study of novel immunotherapies and combination therapy may become the primary focus of future research on pancreatic cancer immunotherapy. More prospective clinical studies with high evidence levels should be conducted.


An account of 335 cases of megaloblastic anaemia of pregnancy and the puerperium.
The incidence of megaloblastic anaemia in pregnancy and the puerperium in north Staffordshire has steadily declined as a result of prophylaxis with folic acid. In the presence of advanced folic acid deficiency and with a florid megaloblastic marrow, the anaemia is usually severe, but in many patients the disease is relatively mild and the degree of anaemia is determined more by blood loss or associated iron deficiency than by the megaloblastosis. Microscopic examination of marrow films is still the most reliable method of diagnosis, although estimation of the labile serum folate has produced a 95% correlation with the marrow findings. There are three main factors which operate in the pathogenesis of megaloblastic anaemia in pregnancy and the puerperium. First, the maternal stores of folic acid are used up by the growing foetus, and this process is accelerated in multiple pregnancies, after haemorrhage, or in women with haemolytic anaemia. Secondly, an insufficient intake of folic acid, due to poor diet in pregnancy, plays a part in many cases. The third, and possibly the most important, factor is an absorption defect. Folic acid absorption is usually impaired in established cases, and this can still be demonstrated years later in a majority of patients, when they are neither pregnant nor anaemic. More than 20% of all cases also show abnormal fat absorption. An inherited defect in folic acid absorption may also explain why certain women appear to be constitutionally predisposed to megaloblastic anaemia of pregnancy and the puerperium, as shown by the abnormal blood group distribution in these patients and by the tendency of megaloblastic anaemia to recur not only in subsequent pregnancies, but, as in six of our cases, following other kinds of stress. The significance of commonly associated conditions like pre-eclampsia and infection is still incompletely understood. Although the treatment of megaloblastic anaemia is simple and effective, the main emphasis should be placed on prophylaxis by administering folic acid to all pregnant women.


[The effect produced by vacuum compression created by micro-keratome in keratomileusis procedure on the condition of the retina].
Described are the results of a clinical experiment involving 15 patients (30 eyes) with myopia ranging from 3.25 to 14.0 diopters. Vacuum compression (application of the vacuum ring from the "Moria" microkeratotome) was preliminarily made on one or both eyes for 30 sec. Computer perimetry and fluorescence angiography of the retina were performed before and after the procedure with an interval of 24 hours. Laser keratomylosis was subsequently made in all patients. According to fluorescence angiography, there were no differences before and after vacuum fixation. According to computer perimetry, the number of relative scotomas was higher, in 20% of cases, in high myopia after the procedure than before it; however this number was within the mean parameters.


Lipid consumption in coral larvae differs among sites: a consideration of environmental history in a global ocean change scenario.
The success of early life-history stages is an environmentally sensitive bottleneck for many marine invertebrates. Responses of larvae to environmental stress may vary due to differences in maternal investment of energy stores and acclimatization/adaptation of a population to local environmental conditions. In this study, we compared two populations from sites with different environmental regimes (Moorea and Taiwan). We assessed the responses of Pocillopora damicornis larvae to two future co-occurring environmental stressors: elevated temperature and ocean acidification. Larvae from Taiwan were more sensitive to temperature, producing fewer energy-storage lipids under high temperature. In general, planulae in Moorea and Taiwan responded similarly to pCO2 Additionally, corals in the study sites with different environments produced larvae with different initial traits, which may have shaped the different physiological responses observed. Notably, under ambient conditions, planulae in Taiwan increased their stores of wax ester and triacylglycerol in general over the first 24 h of their dispersal, whereas planulae from Moorea consumed energy-storage lipids in all cases. Comparisons of physiological responses of P. damicornis larvae to conditions of ocean acidification and warming between sites across the species' biogeographic range illuminates the variety of physiological responses maintained within P. damicornis, which may enhance the overall persistence of this species in the light of global climate change.


Catalytic Antibodies May Contribute to Demyelination in Myalgic Encephalomyelitis/Chronic Fatigue Syndrome.
Here we report preliminary data demonstrating that some patients with myalgic encephalomyelitis/chronic fatiguesyndrome (ME/CFS) may have catalytic autoantibodies that cause the breakdown of myelin basic protein (MBP). We propose that these MBP-degradative antibodies are important to the pathophysiology of ME/CFS, particularly in the occurrence of white matter disease/demyelination. This is supported by magnetic resonance imagining studies that show these findings in patients with ME/CFS and could explain symptoms of nerve pain and muscle weakness. In this work, we performed a series of experiments on patient plasma samples where we isolated and characterized substrate-specific antibodies that digest MBP. We also tested glatiramer acetate (copaxone), an FDA approved immunomodulator to treat multiple sclerosis, and found that it inhibits ME/CFS antibody digestion of MBP. Furthermore, we found that aprotinin, which is a specific serine protease inhibitor, specifically prevents breakdown of MBP while the other classes of protease inhibitors had no effect. This coincides with the published literature describing catalytic antibodies as having serine protease-like activity. Postpandemic research has also provided several reports of demyelination in COVID-19. Because COVID-19 has been described as a trigger for ME/CFS, demyelination could play a bigger role in patient symptoms for those recently diagnosed with ME/CFS. Therefore, by studying proteolytic antibodies in ME/CFS, their target substrates, and inhibitors, a new mechanism of action could lead to better treatment and a possible cure for the disease.


Dementia and depression with ischemic heart disease: a population-based longitudinal study comparing interventional approaches to medical management.
We compared the proportion of ischemic heart disease (IHD) patients newly diagnosed with dementia and depression across three treatment groups: percutaneous coronary intervention (PCI), coronary artery bypass grafting (CABG) and medical management alone (IHD-medical). De-identified, individual-level administrative records of health service use for the population of Manitoba, Canada (approximately 1.1 million) were examined. From April 1, 1993 to March 31, 1998, patients were identified with a diagnosis of IHD (ICD-9-CM codes). Index events of CABG or PCI were identified from April 1, 1998 to March 31, 2003. Outcomes were depression or dementia after the index event. Patients were followed forward to March 31, 2006 or until censored. Proportional hazards regression analysis was undertaken. Independent variables examined were age, sex, diabetes, hypertension and income quintile, medical management alone for IHD, or intervention by PCI or CABG. Age, sex, diabetes, and presence of hypertension were all strongly associated with the diagnosis of depression and dementia. There was no association with income quintile. Dementia was less frequent with PCI compared to medical management; (HR = 0.65; p = 0.017). CABG did not provide the same protective effect compared to medical management (HR = 0.90; p = 0.372). New diagnosis depression was more frequent with interventional approaches: PCI (n = 626; hazard ratio = 1.25; p = 0.028) and CABG (n = 1124, HR = 1.32; p = 0.0001) than non-interventional patients (n = 34,508). Subsequent CABG was nearly 16-fold higher (p<0.0001) and subsequent PCI was 22-fold higher (p<0.0001) for PCI-managed than CABG-managed patients. Patients managed with PCI had the lowest likelihood of dementia-only 65% of the risk for medical management alone. Both interventional approaches were associated with a higher risk of new diagnosed depression compared to medical management. Long-term myocardial revascularization was superior with CABG. These findings suggest that PCI may confer a long-term protective effect from dementia. The mechanism(s) of dementia protection requires elucidation.


Description of illness: limitations and approaches.
The collection of health information by diaries has raised questions about the limitations of existing diagnostic terms and taxonomies in describing ill health in an holistic fashion. Despite the advantage of the problem oriented medical record system in recognizing the social and psychological dimensions of illness, problem lists do not communicate the unique mix of problems for any individual. An argument is made for more anecdotal description of illness in medical records and more research to develop a systems approach to describe ill health.


Postmating-prezygotic isolation is not an important source of selection for reinforcement within and between species in Drosophila pseudoobscura and D. persimilis.
Most work on adaptive speciation to date has focused on the role of low hybrid fitness as the force driving reinforcement (the evolution of premating isolation after secondary contact that reduces the likelihood of matings between populations). However, recent theoretical work has shown that postmating, prezygotic incompatibilities may also be important in driving premating isolation. We quantified premating, postmating-prezygotic, and early postzygotic fitness effects in crosses among three populations: Drosophila persimilis, D. pseudoobscura USA (sympatric to D. persimilis), and D. pseudoobscura Bogotá (allopatric to D. persimilis). Interspecific matings were more likely to fail when they involved the sympatric populations than when they involved the allopatric populations, consistent with reinforcement. We also found that failure rate in sympatric mating trials depended on whether D. persimilis females were paired with D. pseudoobscura males or the reverse. This asymmetry most likely indicates differences in discrimination against heterospecific males by females. By measuring egg laying rate, fertilization success and hatching success, we also compared components of postmating-prezygotic and early postzygotic isolation. Postmating-prezygotic fitness costs were small and not distinguishable between hetero- and conspecific crosses. Early postzygotic fitness effects due to hatching success differences were also small in between-population crosses. There was, however, a postzygotic fitness effect that may have resulted from an X-linked allele found in one of the two strains of D. pseudoobscura USA. We conclude that the postmating-prezygotic fitness costs we measured probably did not drive premating isolation in these species. Premating isolation is most likely driven in sympatric populations by previously known hybrid male sterility.


Use of endoscopic transorbital and endonasal approaches for 360° circumferential access to orbital tumors.
Orbital tumors are often surgically challenging because they require an extensive fronto-temporo-orbital zygomatic approach (FTOZ) and a multidisciplinary team approach to provide the best outcomes. Recently, minimally invasive endoscopic techniques via a transorbital superior eyelid approach (ETOA) or endoscopic endonasal approach (EEA) have been proposed as viable alternatives to transcranial approaches for orbital tumors. In this study, the authors investigated the feasibility of 360° circumferential access to orbital tumors via both ETOA and EEA. Between April 2014 and June 2019, 16 patients with orbital tumors underwent either ETOA or EEA at the authors' institution. Based on the neuro-topographic "four-zone model" of the orbit with its tumor epicenter around the optic nerve in the coronal plane, ETOA (n = 10, 62.5%) was performed for tumors located predominantly superolateral to the nerve and EEA (n = 6, 37.5%) for those located predominantly inferomedial to the nerve. Eight patients (50%) presented with intraconal tumors and 8 (50%) with extraconal ones. The orbital tumors included orbital schwannoma (n = 6), cavernous hemangioma (n = 2), olfactory groove meningioma (n = 1), sphenoorbital meningioma (n = 1), chondrosarcoma (n = 1), trigeminal schwannoma (n = 1), metastatic osteosarcoma (n = 1), mature cystic teratoma (n = 1), sebaceous carcinoma (n = 1), and ethmoid sinus osteoma (n = 1). The clinical outcomes and details of surgical techniques were reviewed. Gross-total resection was achieved in 12 patients (75%), near-total resection in 3 (18.8%), and subtotal resection in 1 (6.2%). Eight (88.9%) of the 9 patients with preoperative proptosis showed improvement after surgery, and 4 (66.7%) of the 6 patients with visual symptoms demonstrated improvement. Four (40%) of the 10 patients treated with ETOA experienced partial third nerve palsy immediately after surgery (3 transient and 1 persistent). There have been no postoperative CSF leaks or infections in this series. Without transcranial approaches requiring temporalis muscle dissection and orbitozygomatic osteotomy, the selection of ETOA or EEA based on a concept of a four-zone model with its epicenter around the optic nerve successfully provides a minimally invasive 360° circumferential access to the entire orbit with acceptable morbidity.


Prednisolone and Prednisone Pharmacokinetics in Pediatric Renal Transplant Recipients-A Prospective Study.
Prednisolone is a standard component of immunosuppressive protocols in renal transplantation (Tx) and despite standardized treatment regimens, adverse side effects are still frequent. The aim of this study was to characterize the pharmacokinetics of prednisolone and prednisone in pediatric renal transplant recipients in the first 52 weeks post Tx, to describe the relationship between prednisolone and prednisone, and to investigate a possible relationship between the development of new-onset diabetes after Tx (NODAT) and glucocorticoid exposure. Renal transplant recipients receiving prednisolone (n = 11, age 1-15 years) were included in this prospective open-label, descriptive, nonrandomized, and noninterventional study. Blood samples were drawn pre-Tx and during selected dose intervals (0, 1, 2, 4, 6, and 12 hours postdose; less frequent in children <10 kg) at 1, 2, 3, 4, 12, and 52 weeks post-Tx. Concentrations of prednisolone and cortisol, their inactive keto forms, plus methylprednisolone, were measured using a validated LC-MS/MS method. Genetic variants in the CYP3A4, CYP3A5, ABCB1, and HSD11B2 genes were analyzed using real-time polymerase chain reaction and Sanger sequencing. Correlation with NODAT was investigated. The patients displayed considerable intra- and inter-individual variability in prednisolone exposure, with up to 5-fold differences in the area under the concentration-time curve (AUC). There were up to 7-fold differences in prednisolone/prednisone AUC ratio between patients, and patients experiencing NODAT tended to have a higher ratio (>12) compared with patients without NODAT (<12). Genetic variants in CYP3A5 and ABCB1 were found, but due to the limited study population causality cannot be definitive. The study suggests that a high prednisolone/prednisone AUC ratio may be a possible risk factor for NODAT. Further studies of individualization of glucocorticoid treatment in pediatric organ Tx are warranted.


Caring for patients at the end of life in an HMO.
Increasing numbers of Americans are receiving care within managed care organizations (MCOs), and the ability of MCOs to provide high-quality end-of-life care is under greater scrutiny. Although many anecdotal reports have been published, almost no research has been published on patient/family experiences with end-of-life care in MCOs. We undertook a qualitative study of end-of-life care during 1996 and 1997 in the closed-panel staff-model, not-for-profit, health center division of Harvard Pilgrim Health Care. We collected and analyzed data from five focus groups composed of family members of patients who had died during the preceding year (n = 22) and four focus groups composed of clinicians and administrators who routinely care for dying patients and/or have responsibility for managing end-of-life care resources and systems (n = 31). Problems noted in this setting that are also reported in other health care settings included divergent expectations and unclear primary care/specialist roles and hospice/primary physician roles, difficulties in transitions across care settings, ineffective communication between physicians and patients/families about end-of-life care preferences, and lack of systematic attention to bereavement care. More unique to managed care settings were problems related to the absence of a consistently applied hospice benefit and concerns about the impact of cost containment on the quality of care, reflecting the conflict between the dual roles of MCOs as both insurers and providers of care. Our findings highlight specific issues involved in providing quality end-of-life care in MCOs and suggest areas for further exploration.


Deprived yet healthy: neighbourhood-level resilience in New Zealand.
Geographical inequalities in health are omnipresent with health and related behaviours typically worse in socioeconomically deprived places. However, this is not always true. Deprived places with unexpectedly good health outcomes, or what might be considered 'resilient' places, have been noted. Few studies have quantitatively examined resilience in neighbourhoods or investigated potential explanations for this resilience. This paper examines the paradox of low mortality despite high social deprivation in New Zealand neighbourhoods and considers possible neighbourhood characteristics that contribute to unanticipated positive health outcomes. Using area-level mortality (2005-2007) and socioeconomic data, we developed the Resilience Index New Zealand to quantify neighbourhood levels of resilience across the country. We then examined relationships between this measure and a suite of built, physical and social characteristics. We found that resilient places tended to be densely populated, urban areas. We observed gradients and increases/decreases in the most resilient groups in access to or levels of physical environment factors (environmental deprivation, safe drinking water, air quality) and unhealthy living infrastructure (alcohol and gambling outlets). Since these factors are amenable to change, these findings are the strongest evidence that such improvements may lower mortality in similarly deprived places. The social environment of resilient areas was characterised by high levels of incoming residents. We also found some surprising associations and observed U-shaped relationships for a number of the neighbourhood factors. Such findings suggest the need to develop a better proxy of community cohesion and a better understanding of the interactions between people and their neighbourhoods, rather than simply the presence of certain factors. We argue that this study has identified amenable neighbourhood characteristics and highlighted the importance of 'place-specific' resilience factors that may be effective in reducing mortality in some neighbourhoods, but be less effective in others.


Integrating GHS into the Ghrelin System.
Oligopeptide derivatives of metenkephalin were found to stimulate growth-hormone (GH) release directly by pituitary somatotrope cells in vitro in 1977. Members of this class of peptides and nonpeptidyl mimetics are referred to as GH secretagogues (GHSs). A specific guanosine triphosphatate-binding protein-associated heptahelical transmembrane receptor for GHS was cloned in 1996. An endogenous ligand for the GHS receptor, acylghrelin, was identified in 1999. Expression of ghrelin and homonymous receptor occurs in the brain, pituitary gland, stomach, endothelium/vascular smooth muscle, pancreas, placenta, intestine, heart, bone, and other tissues. Principal actions of this peptidergic system include stimulation of GH release via combined hypothalamopituitary mechanisms, orexigenesis (appetitive enhancement), insulinostasis (inhibition of insulin secretion), cardiovascular effects (decreased mean arterial pressure and vasodilation), stimulation of gastric motility and acid secretion, adipogenesis with repression of fat oxidation, and antiapoptosis (antagonism of endothelial, neuronal, and cardiomyocyte death). The array of known and proposed interactions of ghrelin with key metabolic signals makes ghrelin and its receptor prime targets for drug development.


Identification of potential biophysical and molecular signalling mechanisms underlying hyaluronic acid enhancement of cartilage formation.
This study determined the effects of exogenous hyaluronic acid (HA) on the biomechanical and biochemical properties of self-assembled bovine chondrocytes, and investigated biophysical and genetic mechanisms underlying these effects. The effects of HA commencement time, concentration, application duration and molecular weight were examined using histology, biomechanics and biochemistry. Additionally, the effects of HA application on sulphated glycosaminoglycan (GAG) retention were assessed. To investigate the influence of HA on gene expression, microarray analysis was conducted. HA treatment of developing neocartilage increased compressive stiffness onefold and increased sulphated GAG content by 35 per cent. These effects were dependent on HA molecular weight, concentration and application commencement time. Additionally, applying HA increased sulphated GAG retention within self-assembled neotissue. HA administration also upregulated 503 genes, including multiple genes associated with TGF-β1 signalling. Increased sulphated GAG retention indicated that HA could enhance compressive stiffness by increasing the osmotic pressure that negatively charged GAGs create. The gene expression data demonstrate that HA treatment differentially regulates genes related to TGF-β1 signalling, revealing a potential mechanism for altering matrix composition. These results illustrate the potential use of HA to improve cartilage regeneration efforts and better understand cartilage development.


The distribution of sexually-transmitted Human Papillomaviruses in HIV positive and negative patients in Zambia, Africa.
Human Papillomaviruses (HPV) are double-stranded DNA viruses, considered to be the primary etiological agents in cervical intraepithelial neoplasias and cancers. Approximately 15-20 of the 40 mucosal HPVs confer a high-risk of progression of lesions to invasive cancer. In this study, we investigated the prevalence of sexually transmitted HPVs in Human Immunodeficiency Virus (HIV) positive and negative patients in Zambia, Africa. The rate of high-risk HPV genotypes worldwide varies within each country. Thus, we sought to investigate the rates of HPV infection in sub-Saharan Africa and the potential role of HIV in affecting the HPV genotype distribution. This retrospective cross-sectional study reports findings on the association and effects of HIV on HPV infections in an existing cohort of patients at University Teaching Hospital (UTH) Lusaka, Zambia. The objective of this study was to assess HPV prevalence, genotype distribution and to identify co-factors that influence HPV infection. Polymerase chain reaction (PCR) with two standard consensus primer sets (CpI/II and GP5+/6+) was used to test for the presence of HPV DNA. Primers specific for beta-actin were used to monitor DNA quality. Vaginal lavage samples, collected between 1998-1999 from a total of 70 women, were part of a larger cohort that was also analyzed for HIV and human herpesvirus infection. Seventy of the samples yielded usable DNA. HIV status was determined by two rapid assays, Capillus and Determine. The incidence of HIV and HPV infections and HPV genotype distributions were calculated and statistical significance was determined by Chi-Squared test. We determined that most common HPV genotypes detected among these Zambian patients were types 16 and 18 (21.6% each), which is approximately three-fold greater than the rates for HPV16, and ten-fold greater than the rates for HPV18 in the United States. The worldwide prevalence of HPV16 is approximately 14% and HPV18 is 5%. The overall ratio of high-risk (HR) to low-risk (LR) HPVs in the patient cohort was 69% and 31% respectively; essentially identical to that for the HR and LR distributions worldwide. However, we discovered that HIV positive patients were two-times as likely to have an HR HPV as HIV negative individuals, while the distribution of LR HPVs was unaffected by HIV status. Interestingly, we observed a nine-fold increase in HPV18 infection frequency in HIV positive versus HIV negative individuals. The rate of oncogenic HPVs (type 16 and 18) in Zambia was much higher than in the U.S., potentially providing an explanation for the high-rates of cervical cancer in Zambia. Surprisingly, we discovered a strong association between positive HIV status and the prevalence of HR HPVs, and specifically HPV18.


Factors governing the Na(+) vs K(+) selectivity in sodium ion channels.
Monovalent Na(+) and K(+) ion channels, specialized pore-forming proteins that play crucial biological roles such as controlling cardiac, skeletal, and smooth muscle contraction, are characterized by a remarkable metal selectivity, conducting the native cation while rejecting its monovalent contender and other ions present in the cellular/extracellular milieu. Compared to K(+) channels, the principles governing Na(+) vs K(+) selectivity in both epithelial and voltage-gated Na(+) channels are much less well understood due mainly to the lack of high-resolution 3D structures. Thus, many questions remain. It is not clear if the serines lining the pore of epithelial Na(+) channel bind to the metal cation via their backbone or side chain O atoms and why substituting the Lys lining the pore of voltage-gated Na(+) channels to another residue such as Arg drastically reduces or even reverses the Na(+)/K(+) selectivity. This work systematically evaluates the effects of various factors such as (i) the number, chemical type, and charge of the pore's coordinating groups, (ii) the hydration number and coordination number of the metal cation, and (iii) the solvent exposure and the size/rigidity of the pore on the Na(+) vs K(+) selectivity in model Na(+) channel selectivity filters (the narrowest part of the pore) using a combined density functional theory/continuum dielectric approach. The results reveal that the Na(+) channel's selectivity for Na(+) over K(+) increases if (1) the pore provides three rather than four protein ligands to coordinate to the metal ion, (2) the protein ligands have strong charge-donating ability such as Asp/Glu carboxylate or backbone carbonyl groups, (3) the passing Na(+) is bare or less well hydrated inside the filter than the competing K(+), and (4) the pore is relatively rigid, constricted, and solvent exposed. They also reveal that factors favoring Na(+)/K(+) selectivity in Na(+) channels generally disfavor K(+)/Na(+) selectivity in K(+) channels and vice versa. The different selectivity principles for the K(+) and Na(+) channels are consistent with the different architecture, composition, and properties of their selectivity filters. They provide clues to the metal-binding site structure in the selectivity filters of epithelial and voltage-gated Na(+) channels.


A multicenter study to evaluate oxidative stress by oxidation-reduction potential, a reliable and reproducible method.
Seminal oxidative stress (OS) is well-known to affect male fertility status. The discrepancy in OS measurement has hindered its clinical use as a quality indicator for semen. Some tests measured single markers of oxidants or reductants, leading to lack of standardization of results. Oxidation-reduction potential (ORP) is a better representative for OS as it provides an overall measure of the activity of both oxidants and reductants. ORP assessment by MiOXSYS has been introduced as a measure of OS with high specificity in differentiating fertile from infertile semen samples. This is a retrospective study comparing data from semen analysis and ORP measurements between two andrology laboratories in the USA and Qatar over a period of 12 months. The same protocol was followed by both laboratories. The USA dataset contained 194 patients and 51 fertile donors, while the Qatar dataset contained 400 patients and 50 fertile donors. In both datasets and in the combined dataset, the infertile group had significantly lower sperm concentration, total and progressive motility, and normal morphology as well as higher ORP levels compared to fertile men (p < 0.05). When comparing data from both centers, the infertile group showed significant difference between both datasets regarding progressive motility and morphology (p < 0.001). Also, the percentage of patients with abnormal semen volume, sperm count, total and progressive motility were significantly different between both datasets (p < 0.05). ORP levels showed no significant difference between both datasets (p < 0.08). ROC analysis indicated that ORP cutoff value of 1.42 mV/106 /mL in the USA group, Qatar group, and combined dataset can accurately differentiate fertile from infertile semen groups. Although other semen parameters showed significant differences between the two centers, ORP remained consistent in both datasets individually or in combined data. This proves its reproducibility and reliability.


A multicenter evaluation of Copan's Colibrí™, an automated instrument for MALDI TOF MS target application for bacterial identification.
The Colibrí™ is a new instrument that automates picking and placement of colonies on target plates for MALDI identification. This study compared the performance of the Colibrí™ to standard manual spotting using the VITEK® MS for bacterial identification. Colonies were selected from cultures of urine, wound, respiratory, and positive blood cultures. The Colibrí™ sampled the colonies, transferred them to a MALDI target slide, and overlayed each spot with matrix. Manual spotting was then performed using the same or similar colonies. A total of 444 bacteria were compared. Identification was achieved with both methods for 432 organisms with only 2 discrepant results, overall agreement of 99.54%. Twelve organisms (2.70%) gave no identification using Colibrí™. The Colibrí™ provides automation to a manual process with a high accuracy. Use of the Colibrí™ instrumentation provides an opportunity to reallocate technologist time to more complicated tasks and provides complete traceability from plating to organism identification.


Chirality in antirheumatic drugs.
Use of chiral molecules in clinical practice may cause problems because different chiral forms of a drug (enantiomers) may have different biological activities--yet clinicians have little awareness of these risks. After discussion of the chemical conventions used to describe chirality, examples of the influence of chirality on the efficacy and toxicity of antirheumatic drugs are given. It is recommended that single enantiomers should be used in biological experiments and clinical trials.


Plasma and Erytrocyte Oxidative Stress Markers in Children with Frequent Breath-Holding Spells.
Breath-holding spells (BHS) are common non-paroxysmal events with unknown pathophysiology. BHS have been associated not only with iron deficiency anemia (IDA) but also with oxidant/antioxidant imbalance and erythrocyte injury induced by hypoxia. The present study was designed to investigate the contribution of IDA in BHS and the oxidant/antioxidant balance in children with or without IDA in BHS and compare them with healthy controls.Additionally, the study also aimed to examine the effect of the frequency of BHS attacks (mild or severe) on the oxidant/antioxidant balance and to determine the best predictive oxidant and antioxidant markers. The study included 66 children with BHS aged 6-48 months who had been followed up for a minimum period of one year between 2014 and 2018. A control group of 30 age- and gender-matched healthy children was included in the study. The patient group was divided into 2 groups (IDA and non-IDA) and these groups were compared between each other and also with the control group. The IDA group was divided into subgroups based on the frequency of BHS attacks. Blood samples were obtained within a maximum period of 24 h following the spell. Levels of protein carbonyl, nitrite, nitrate, TOS, TAS, OSI, MDA, enzyme activities of GPx, CAT,enzyme activities of erythrocyte SOD, CAT, and GPx, and the level of MDA were measured. In patients with IDA, the oxidant levels increased while the antioxidant enzyme activities decreased. In all patients, the levels of MDA, carbonyl, TOS, OSI increased and the levels of TAS, activities SOD, and CAT decreased, whereas the enzyme activities of erythrocyte SOD, CAT, GPx decreased significantly compared to those of control group. Increased of erythrocyte MDA levels had 10.32, decreased enzyme activities of erythrocyte SOD levels had a 10.25, and decreased enzyme activities of erythrocyte CAT had a 5.33 times greater risk for spell. The results indicated that the oxidant/antioxidant balance in children with BHS was impaired in favor of oxidants at both levels, regardless of the presence of IDA and the increased frequency of BHS attacks per day. Moreover, the presence of IDA was found to be associated with increased oxidative stress in children with BHS, particularly at the erythrocyte level. Erythrocyte level; among the erythrocyte MDA oxidant parameters, erythrocyte SOD and antioxidant parameters, they are the biomarkers that show the best probability of having a BHS attack and an increase in the frequency of apnea attacks.


Readiness for living technology: a comparative study of the uptake of robot technology in the Danish health-care sector.
This article is a comparative analysis between workers in health care with high and low degree of readiness for living technology such as robotics. To explore the differences among workers' readiness, statistical analysis was conducted in a data set obtained from 200 respondents. The results showed important differences between high- and low-readiness types on issues such as staff security, documentation, autonomy, and future challenges.


Short stature, unusual face, delta phalanx, and abnormal vertebrae and ribs in a girl born to half-siblings.
Delta phalanx is a rare abnormality typically associated with additional features. We describe a patient with a phenotype resembling Catel-Manzke syndrome, but with delta phalanx and abnormal vertebrae and ribs. The patient was the only child of half siblings born with a marked prenatal growth deficiency. At 10 years of age, she had a short stature, long face, long and tubular nose with small alae nasi, high palate, short and broad thorax, and short index fingers with radial deviation. There were hyperpigmentations following Blaschko's lines. Radiology showed a proximal delta phalanx in the index finger of hands, abnormal vertebrae, and fused and small ribs. GTG-Banding karyotype and microarray analysis yielded normal results. Exome sequencing identified 25 genes that harbored homozygous variants, but none of these is assumed to be a good candidate to explain (part of) the phenotype. The here described patient may have a new condition, possibly following an autosomal recessive pattern of inheritance, although due to the high degree of consanguinity a compound etiology of the phenotype by variants in various genes may be present as well.


Scalp localization using a simple algorithm.
We have developed a simple algorithm for scalp localization for craniotomy. Using a series of CT/MRI data, the point on the scalp closest to the center of the lesion (T) is determined by our program. A plane (plane R) is defined by three points: bilateral auditory meati and T. Point S is the intersection of three planes: the plane R, the scalp surface and the midsagittal plane. The distance from either S or the ipsilateral external auditory meatus to T is measured along the scalp surface on the plane R. The distance from the nasion to S along the scalp surface on the midsagittal plane is also measured. To determine the craniotomy site, these distances are measured directly on a patient's scalp in the operating room. This simple and accurate method for scalp localization could be used by installing our program in conventional CT/MR scanners.


Bacteroides fragilis ubiquitin homologue drives intraspecies bacterial competition in the gut microbiome.
Interbacterial antagonism and associated defensive strategies are both essential during bacterial competition. The human gut symbiont Bacteroides fragilis secretes a ubiquitin homologue (BfUbb) that is toxic to a subset of B. fragilis strains in vitro. In the present study, we demonstrate that BfUbb lyses certain B. fragilis strains by non-covalently binding and inactivating an essential peptidyl-prolyl isomerase (PPIase). BfUbb-sensitivity profiling of B. fragilis strains revealed a key tyrosine residue (Tyr119) in the PPIase and strains that encode a glutamic acid residue at Tyr119 are resistant to BfUbb. Crystal structural analysis and functional studies of BfUbb and the BfUbb-PPIase complex uncover a unique disulfide bond at the carboxy terminus of BfUbb to mediate the interaction with Tyr119 of the PPIase. In vitro coculture assays and mouse studies show that BfUbb confers a competitive advantage for encoding strains and this is further supported by human gut metagenome analyses. Our findings reveal a previously undescribed mechanism of bacterial intraspecies competition.


Unveiling Cellular Diversity in the Buffalo Corneal Stroma: Insights into Telocytes and Keratocytes Using Light Microscope, Transmission Electron Microscope, and Immunofluorescence Analysis.
Telocytes and keratocytes are important cells that maintain the structure and function of the cornea. The buffalo cornea, known for its resilience in harsh conditions, has not been extensively studied regarding the presence and role of telocytes and keratocytes. We used light microscopy, transmission electron microscopy (TEM), and immunofluorescence assays with platelet-derived growth factor receptor alpha (PDGFRα), CD34, and Vimentin markers to investigate their expression and localization in the cornea. TEM analysis confirmed the presence of spindle-shaped keratocytes with intercellular connections, while telocytes exhibited small spindle-shaped bodies with long, thin branches connecting to corneal keratocytes. Immunofluorescence findings showed that CD34 was more abundant near the endothelium, Vimentin was prominently expressed near the epithelium, and PDGFRα was uniformly distributed throughout the corneal stroma. Co-expression of CD34 and Vimentin, PDGFRα and Vimentin, as well as CD34 and PDGFRα, was observed in keratocytes and telocytes within the stroma, indicating the potential presence of mesenchymal cells. These results suggest the involvement of telocytes and keratocytes in corneal wound healing, transparency maintenance, and homeostasis. The co-expression of these markers highlights the critical role of telocytes and keratocytes in regulating corneal physiological functions, further enhancing our understanding of corneal biology in the buffalo model.


Topological nature of the genetic code.
A model for topological coding of proteins is proposed. The model is based on the capacity of hydrogen bonds (property of connectivity) to fix conformations of protein molecules. The protein chain is modeled by an n -arc graph with the following elements: vertices (alpha -carbon atoms), structural edges (peptide bonds) and connectivity edges (virtual edges connecting non-adjacent atoms). It was shown that 64 conformations of the 4-arc graph can be described in the binary system by matrices of six variables which form a supermatrix containing four blocks. On the basis of correspondences between the pairs of variables in matrices and four letters of the genetic code matrices and supermatrix are converted, respectively, into the triplets and the table of the genetic code. An algorithm admitting computer programming is proposed for coding the n -arc graph and protein chain. Connectivity operators (polar amino acids) are assigned to blocks of triplets coding for cyclic conformations (G, A-in the second position), while anti-connectivity operators (non-polar amino acids) correspond to blocks of triplets coding for open conformations (C, U-in the second position). Amino acids coded by triplets differing by the first base have different structures. The third base for C, U and G, A is degenerated. Properties of the real genetic code are in full agreement with the model. The model provides an insight into the topological nature of the genetic code and can be used for development of algorithms for the prediction of the protein structure.


Tumor sprouting in papillary thyroid carcinoma is correlated with lymph node metastasis and recurrence.
Identification of poor prognostic factors in papillary thyroid carcinoma (PTC) patients is important for the patients' care and follow-up. We can sometimes see small tumor clusters without desmoplasia and no evidence of lymphatic emboli around the main tumor mass of PTC. We termed this form of tumor clustering, 'tumor sprouting,' and determined whether these tumors correlate with lymphovascular invasion, lymph node metastasis, and recurrence. We analyzed a total of 204 cases of papillary thyroid macrocarcinoma. Number, size and distance from the main tumor of the tumor sprouting were observed and analyzed with clinicopathologic characteristics. Tumor sprouting was observed in 101 patients. Presence of tumor sprouting was significantly associated with positive resection margin (p=.002), lymphovascular invasion (p=.001), lymph node metastasis (p<.001), and recurrence (p=.004). Univariate analysis of recurrence-free survival revealed that tumor multiplicity (p=.037), positive resection margin (p=.007), lymphovascular invasion (p=.004), lymph node metastasis (p<.001), and tumor sprouting (p=.004) were poor prognostic factors. In multivariate analysis, positive resection margin was an independent poor prognostic factor of recurrence. In conclusion, tumor sprouting is significantly correlated with lymph node metastasis and recurrence. Evaluation of tumor sprouting in PTC patients could be helpful in predicting tumor recurrence or lymph node metastasis.


Metabolism and chemotherapeutic activity of 9-beta-D-arabinofuranosyl-2-fluoroadenine against murine leukemia L1210 and evidence for its phosphorylation by deoxycytidine kinase.
The 2-fluoro derivative of 9-beta-D-arabinofuranosyladenine (2-F-ara-A) and its soluble 5'-formate and 5'-phosphate derivatives were therapeutically effective against the parent leukemia L1210 (L1210/0). 2-F-ara-A and 9-beta-D-arabinofuranosyladenine 5'-formate were inactive aginst a 1-beta-D-arabinofuranosylcytosine-resistant subline (L1210/ara-C) that was deficient in deoxycytidine kinase. Deoxycytidine prevented 2-F-ara-A-induced inhibition of proliferation of L1210/0 cells in culture and alleviated 2-F-ara-a inhibition of DNA synthesis. After treatment of mice with 9-beta-D-arabinofuranosyladenine 5'-formate, intracellular levels of the 5'-triphosphate of 9-beta-D-arabinofuranosylfluoroadenine in leukemia cells were more than 10 times higher in L1210/0 cells than in L1210/ara-C cells. Similar results were obtained in this line of leukemia cells from mice treated with the 5'-monophosphate of 9-beta-D-arabinofuranosyl-2-fluoroadenine. Thus, L1210/ara-C cells deficient in deoxycytidine kinase activity were also deficient in capacity to phosphorylate 2-F-ara-A. Kinase activity from L1210/0 cells for deoxycytidine and for 2-F-ara-A coeluted from calcium phosphate cellulose and from diethylaminoethyl cellulose columns and had similar mobility on gel electrophoresis. Deoxyadenosine kinase was clearly separated from deoxycytidine kinase. Deoxycytidine competed with 2-F-ara-A for phosphorylation by the partially purified enzyme from L1210 cells. These results indicate that 2-F-ara-A is phosphorylated to the 5'-monophosphate by deoxycytidine kinase of leukemia L1210 cells.


Taxonomic assessment of Alligator Snapping Turtles (Chelydridae: Macrochelys), with the description of two new species from the southeastern United States.
The Alligator Snapping Turtle, Macrochelys temminckii, is a large, aquatic turtle limited to river systems that drain into the Gulf of Mexico. Previous molecular analyses using both mitochondrial and nuclear DNA suggested that Macrochelys exhibits significant genetic variation across its range that includes three distinct genetic assemblages (western, central, and eastern = Suwannee). However, no taxonomic revision or morphological analyses have been conducted previously. In this study, we test previous hypotheses of distinct geographic assemblages by examining morphology, reanalyzing phylogeographic genetic structure, and estimating divergence dating among lineages in a coalescent framework using Bayesian inference. We reviewed the fossil record and discuss phylogeographic and taxonomic implications of the existence of three distinct evolutionary lineages. We measured cranial (n=145) and post-cranial (n=104) material on field-captured individuals and museum specimens. We analyzed 420 base pairs (bp) of mitochondrial DNA sequence data for 158 Macrochelys. We examined fossil Macrochelys from ca. 15-16 million years ago (Ma) to the present to better assess historical distributions and evaluate named fossil taxa. The morphological and molecular data both indicate significant geographical variation and suggest three species-level breaks among genetic lineages that correspond to previously hypothesized genetic assemblages. The holotype of Macrochelys temminckii is from the western lineage. Therefore, we describe two new species as Macrochelys apalachicolae sp. nov. from the central lineage and Macrochelys suwanniensis sp. nov. from the eastern lineage (Suwannee River drainage). Our estimates of divergence times suggest that the most recent common ancestor (MRCA) of M. temminckii (western) and M. apalachicolae (central) existed 3.2-8.9 Ma during the late Miocene to late Pliocene, whereas M. temminckii-M. apalachicolae and M. suwanniensis last shared a MRCA 5.5-13.4 Ma during the mid-Miocene to early Pliocene. Examination of fossil material revealed that the fossil taxon M. floridana is actually a large Chelydra. Our taxonomic revision of Macrochelys has conservation and management implications in Florida, Georgia, and Alabama.


Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) composite biomaterials for bone tissue regeneration: in vitro performance assessed by osteoblast proliferation, osteoclast adhesion and resorption, and macrophage proinflammatory response.
The efficacy of composite materials for bone tissue engineering is dependent on the materials' ability to support bone regeneration whilst inducing a minimal inflammatory response. In this study we examined the in vitro osteogenic and inflammatory properties of poly(3-hydroxybutyrate-co-3-valerate) (PHBV) with various calcium phosphate-reinforcing phases: nano-sized hydroxyapatite (HA); submicron-sized calcined hydroxyapatite (cHA); and submicron-sized beta-tricalcium phosphate (beta-TCP), using bioassays of cultured osteoblasts, osteoclasts, and macrophages. Our study showed that the addition of a nano-sized reinforcing phase to PHBV, whilst improving osteogenic properties, also reduces the proinflammatory response. Proinflammatory responses of RAW264.7/ELAM-eGFP macrophages to PHBV were shown to be markedly reduced by the introduction of a reinforcing phase, with HA/PHBV composites having the lowest inflammatory response. Osteoclasts, whilst able to attach to all the materials, failed to form functional actin rings or resorption pits on any of the materials under investigation. Cultures of osteoblasts (MC3T3-E1) readily attached and mineralised on all the materials, with HA/PHBV inducing the highest levels of mineralization. The improved biological performance of HA/PHBV composites when compared with cHA/PHBV and beta-TCP/PHBV composites is most likely a result of the nano-sized reinforcing phase of HA/PHBV and the greater surface presentation of mineral in these composites. Our results provide a new strategy for improving the suitability of PHBV-based materials for bone tissue regeneration.


Results from ACROCOVID: an international survey on the care of acromegaly during the COVID-19 era.
With most global healthcare resources focused on COVID-19, altered resource allocation is disrupting the continuum of care for chronic endocrine conditions, including acromegaly. In order to assess the effects of COVID-19 on care of patients with acromegaly, we conducted an international online survey of endocrinologists. The questionnaire was drafted by a Steering Committee of acromegaly experts and covered both respondent demographics, contact and communication with patients, and patient care. Respondent awareness was generated through social media campaigns and the survey completed online using Google forms. The majority of endocrinologists who responded (N = 84) were based in Europe (67.9%) and were female (58.3%). Slightly more than half of respondents worked in a specialized pituitary center (52.4%) and a large minority cared for more than 50 acromegaly patients (47.6%). Most respondents (85.7%) indicated surgery was their first-line treatment, with only 14.3% indicating medical therapy as a first-choice treatment option. One-third (33.3%) of respondents reported having delayed a planned surgery due to a lack of COVID-19 testing provision and 54.8% due to a lack of surgical provision; 19.1% indicated that a lack of personal protective equipment had reduced their ability to treat patients with acromegaly. Just 21.4% of respondents reported no negative effects from the pandemic on diagnostic practice patterns, and just 19.1% reported no negative effect on patient follow-up practices. Many respondents (55.9%) indicated that remote methods had improved their ability to communicate with their patients and 69.0% indicated that they would continue to use methods of consultation necessitated by the COVID-19 pandemic. Our data suggest the COVID-19 pandemic is substantially affecting the care of acromegaly. However, these results also suggest that endocrinologists are embracing aspects of the 'new normal' to create a novel continuum of care better suited to the presumed post-COVID-19 environment. The goal of these changes must be both to improve care while shielding patients from more severe involvement in concomitant acute illnesses such COVID-19.


Experimental realization of a four-photon seven-qubit graph state for one-way quantum computation.
We propose and demonstrate the scaling up of photonic graph states through path qubit fusion. Two path qubits from separate two-photon four-qubit states are fused to generate a two-dimensional seven-qubit graph state composed of polarization and path qubits. Genuine seven-qubit entanglement is verified by evaluating the witness operator. Six qubits from the graph state are used to demonstrate the Deutsch-Jozsa algorithm for general two-bit functions with a success probability greater than 90%.


Effects of corn straw on meat characteristics of lambs in the Brazilian semi-arid region.
The present study aimed to evaluate the influence of increasing levels of corn straw replacement of sorghum silage on average daily gain, loin eye area, subcutaneous fat thickness, weight of commercial cuts, leg tissue composition, and physico-chemical characteristics of the meat from F1 Santa Inês × Dorper lambs maintained in a feedlot system. Treatments consisted of 0, 33, 66 and 100% corn straw replacement of sorghum silage. There were thirty-six 150-day-old male lambs. Corn straw replacement of sorghum silage in lamb feed reduced the average daily gain and weight of the ribs, besides promoting a quadratic influence on leg fat content. However, there was no influence on commercial carcass cuts, leg muscle index, leg tissue composition, and physico-chemical characteristics of the meat. Thus, corn straw can be used as alternative feedstuff for sheep rations to improve the production of high-quality sheep meat in semi-arid regions.


[Uteroplacental insufficiency in chorangiomatosis].
Chorangioma is a benign angiomatous tumor of the placenta occurring with a frequency of approximately 1 % of all examined placentas. Pregnancies especially with large chorangiomas are frequently complicated by abruptio placentae, premature labour, fetal anaemia, hydrops, cardiomegaly and polyhydramnios. We here report a case of an extremely rare chorangiomatosis. Multiple chorangiomas measuring up to 1.5 cm comprised approximately one third of the placenta. A caesarean section had to be performed at 36 weeks of gestation because of uteroplacental insufficiency with oligohydramnios. Displacement of normal placental parenchyma by chorangiomatosis as a cause of uteroplacental insufficiency is very likely. Colour doppler imaging allows early diagnosis and management of chorangiomas. Thus, in our case early diagnosis of uteroplacental insufficiency was rendered. Principally, early diagnosis of fetal hazard due to myocardial insufficiency of the fetus is feasible. In-utero-endoscopy permits prenatal laser devascularisation of large chorangiomas. In the reported case the pregnancy was closely monitored with colour flow imaging. The diagnosis of uteroplacental insufficiency was established because of abnormal blood flow of the uterine arteries and abnormal fetal heart rate (FHR). At the same time fetal cardiomegaly and pathologic blood flow of the ductus venosus in terms of beginning fetal myocardial insufficiency was recognized. Consequently, caesarean section could be performed opportune.


Factors influencing initiation of breast-feeding.
We used the critical incidence method to study factors motivating 33 primigravidas and 39 multigravidas to initiate breast-feeding of their infants. Women chose breast-feeding because they believed that it would provide protection to the infant against infection, establish maternal-infant bonding, was convenient, provided better nutrition than cow's milk formula, was emotionally satisfying, and was the natural way to feed infants. The decision to breast-feed was made well in advance of pregnancy by primigravidas and shortly before pregnancy by multigravidas. Friends who had successfully nursed infants were as influential as immediate family members in influencing our study subjects in their decision to breast-feed. Prenatal counseling, though important, may not be the optimal period for motivating women to breast-feed.


Haematopoietic cancer and medical history: a multicentre case control study.
Viruses (such as Epstein-Barr virus) and pathological conditions (mainly involving immunosuppression) have been shown to increase the risk of haematolymphopoietic malignancies. Other associations (diabetes, tonsillectomy, autoimmune diseases) have been inconsistently reported. The association between different haematolymphopoietic malignancies (lymphomas, myelomas and leukaemias) and the previous medical history has been studied in a population-based case-control investigation conducted in Italy, based on face to face interviews to 2669 cases and 1718 population controls (refusal rates 10% and 19%, respectively). Controls were a random sample of the general population. Previous findings were confirmed concerning the association between non-Hodgkin's lymphoma (NHL) and lupus erythematosus (odds ratio, OR=8.4; 95% CI 1. 6, 45), tuberculosis (OR=1.6; 1.05, 2.5) and hepatitis (1.8; 1.4, 2. 3). An association was found also between NHL and maternal (OR=2.8; 1.1, 6.9) or paternal tuberculosis (OR=1.7; 0.7, 3.9). Odds ratios of 4.0 (1.4, 11.8) and 4.4 (1.1, 6.6) were detected for the association between NHL and Hodgkin's disease, respectively, and previous infectious mononucleosis, but recall bias cannot be ruled out. No association was found with diabetes, tonsillectomy and adenoidectomy. An association with malaria at young age and "low grade" lymphatic malignancies is suggested. One interesting finding was the observation of four cases of poliomyelitis among NHL patients, one among Hodgkin's disease and one among myeloid leukaemia patients, compared with none among the controls (Fisher's exact test for NHL and Hodgkin's disease, p= 0.03, one tail). Some of these findings are confirmatory of previous evidence. Other observations, such as the putative role of the polio virus and of malaria are new. A unifying theory on the mechanisms by which previous medical history may increase the risk of haematolymphopoietic malignancies is still lacking.


Intestinal maturation: calcium transport by basolateral membranes.
The overall characteristics of calcium transport across the intestine have been defined using in vivo perfusion techniques and in vitro everted gut sacs. However, calcium transport represents three separate processes: entry at the brush border membranes, movement across the cytoplasm, and exit at the basolateral membranes (BLM). Studies in adult animals indicated that the active step in calcium transport is located at the BLM. The present studies describe for the first time the maturational aspects of calcium transport across the BLM of the enterocyte. We utilized a percoll density gradient to prepare enriched BLMs from suckling and adolescent rats. Calcium uptake into BLMs represented mainly transport into the intravesicular space. Calcium transport in both age groups was driven by ATP; the calcium transport in the presence and absence of ATP was significantly greater in suckling rats compared to adolescent rats. Kinetics of calcium uptake calculated from uptake values in the presence of ATP minus no ATP conditions showed a Km of 0.06 +/- 0.01 and 0.03 +/- 0.01 microM for adolescent and suckling rats respectively (p less than 0.05). Vmax was 1.5 +/- 0.1 and 0.8 +/- 0.08 nmol/mg protein/min for adolescent and suckling rats respectively (p less than 0.01). Calcium/sodium exchange mechanisms was also present in both age groups. However, the magnitude of sodium-dependent calcium exchange was smaller in suckling rats compared to adolescent rats. These data suggest that calcium exit at the BLMs of enterocytes of suckling and adolescent rats occurs by an ATP-dependent and a calcium/sodium exchange mechanism. Both mechanisms exhibit maturational changes.