[Morphine in the treatment of acute pulmonary oedema].
Morphine is still used in Norway and the rest of Europe as part of the treatment for pulmonary oedema, but the scientific basis for this is tenuous. In this article we assess the literature that supports and challenges the use of morphine in cases of pulmonary oedema. The article is based on a literature search in Medline and EMBASE and on the articles which form the basis of Norwegian and international guidelines. Morphine has been used for several decades in cases of pulmonary oedema due to the anxiolytic and vasodilatory properties of the drug. Vasodilation caused by morphine has been described in other patient groups, but there is little evidence in the literature to suggest that morphine causes vasodilation in patients with pulmonary oedema. Non-specific depression of the central nervous system is probably the most significant factor for the changes in haemodynamics in pulmonary oedema. Retrospective studies have shown both negative and neutral effects in acute decompensated heart failure. There are no reliable clinical studies that document better prognosis from the use of morphine. Based on the available studies, the possibility cannot be excluded that the use of morphine results in increased mortality among patients with acute pulmonary oedema. In addition, there is little evidence that the vasodilatory properties of morphine are of any significance for this condition. The benefits and risks of using morphine in cases of acute pulmonary oedema are still unclear, but so far there is little evidence to support the beneficial use of the drug.


Intrarenal solitary fibrous tumor of the kidney report of a case with emphasis on the differential diagnosis in the wide spectrum of monomorphous spindle cell tumors of the kidney.
Solitary fibrous tumor (SFT) is a neoplasm that can occur in the urogenital tract, and is also reported occurring in the spermatic cord, seminal vesicles, urinary bladder, prostate, and kidney. Furthermore, it is most important to consider its existence in the kidney, because it is usually diagnosed as renal cell carcinoma pre-operatively. To our knowledge, only 10 cases of SFT have been reported in the kidney to date. We report the clinico-pathological features of an intrarenal SFT occurring in a 31-year-old woman. The tumor, measuring 8.6 cm in its greatest diameter, completely replaced the cortex and the medulla of the middle region of the right kidney, compressing the pelvis. Radiological imaging was consistent with a renal cell carcinoma. Histologically, the tumor was composed of a proliferation of bland-looking vimentin+, CD34+, bcl2+ and CD99+ spindle cells exhibiting a haphazard to storiform growth pattern, pushing borders, and a low mitotic rate (2 mitoses x 10 HPF). We placed emphasis on the differential diagnostic problems, i.e., its differentiation from other primary monomorphous benign and malignant spindle cell tumors of the kidney, such as fibroma, benign fibrous histiocytoma, hemangiopericytoma, inflammatory myofibroblastic (pseudo-)tumor, leiomyoma, angiomyolipoma with predominant spindle cell smooth muscle component, benign peripheral nerve sheath tumors, renal mixed epithelial/stromal tumors, adult type mesoblastic nephroma, fibrous type monophasic synovial sarcoma, malignant peripheral nerve sheath tumors, fibrosarcoma, and low-grade fibromyxoid sarcoma.


Psychological, cognitive and quality of life features in the elderly with chronic headache.
Chronic headache is common in the elderly, but there is little specific research on the impact on quality of life of headache and beliefs about pain in this age group. This study investigated the influence of headache type as well as headache frequency (> or =15 headache days/month vs. <15 headache days/month) on quality of life and determined the relationships between elder's well-being, pain beliefs and related headache features including commonly reported neck pain. 118 headache subjects and 44 non-headache controls, aged 60-75 years, were recruited from the community. Subjects completed a headache questionnaire for classification purposes, the SF-36, the Geriatric Depression Scale-short form (GDS-S), the Survey of Pain Attitudes (SOPA-35) and the Neck Disability Index (NDI). The results revealed that elders with headache compared to the control group scored lower on most SF-36 domains, higher on the GDS-S (p < 0.05) and comparably on the SOPA-35 (p > 0.05), noting that the GDS score was below the threshold value for depression. These measures were not different between the headache types but were influenced by headache frequency. Subjects with headaches > or =15 days/month scored lowest on SF-36 domains and highest on GDS-S questionnaire compared to those with headache <15 days/month and controls (all p < 0.05). The mean NDI score in the headache subjects indicated the presence of mild to moderate neck pain and disability. There were no strong relationships between well-being and headache frequency, intensity and length of headache history. The NDI score had the greatest influence on physical well-being and GDS-S score on mental well-being (p < 0.001). The results suggest that frequency of headache has an impact on health-related quality of life in elders. Notably, the level of neck pain and disability is an important factor influencing well-being and may warrant attention in the management of elders with chronic headache.


Development of reverse genetics systems and investigation of host response antagonism and reassortment potential for Cache Valley and Kairi viruses, two emerging orthobunyaviruses of the Americas.
Orthobunyaviruses such as Cache Valley virus (CVV) and Kairi virus (KRIV) are important animal pathogens. Periodic outbreaks of CVV have resulted in the significant loss of lambs on North American farms, whilst KRIV has mainly been detected in South and Central America with little overlap in geographical range. Vaccines or treatments for these viruses are unavailable. One approach to develop novel vaccine candidates is based on the use of reverse genetics to produce attenuated viruses that elicit immune responses but cannot revert to full virulence. The full genomes of both viruses were sequenced to obtain up to date genome sequence information. Following sequencing, minigenome systems and reverse genetics systems for both CVV and KRIV were developed. Both CVV and KRIV showed a wide in vitro cell host range, with BHK-21 cells a suitable host cell line for virus propagation and titration. To develop attenuated viruses, the open reading frames of the NSs proteins were disrupted. The recombinant viruses with no NSs protein expression induced the production of type I interferon (IFN), indicating that for both viruses NSs functions as an IFN antagonist and that such attenuated viruses could form the basis for attenuated viral vaccines. To assess the potential for reassortment between CVV and KRIV, which could be relevant during vaccination campaigns in areas of overlap, we attempted to produce M segment reassortants by reverse genetics. We were unable to obtain such viruses, suggesting that it is an unlikely event.


Long-term survival after sporadic and delayed metastases of conventional osteosarcoma: A case report.
Histologically conventional osteosarcoma, once metastasized to the lung, generally causes a rapid and fatal outcome. Osteosarcoma metastasis to the gastrointestinal tract is extremely rare.We report herein a case of osteoblastic osteosarcoma with exceptionally unique features: sporadic lung metastases and delayed metastases to the stomach and the jejunum with long-term survival. She received multiple operations and chemotherapies, but consequently died of peritoneal dissemination. A review of the literature on osteosarcoma metastasis to the gastrointestinal tract is presented.This patient was very unusual in terms of a long-term survival and metastatic sites, suggesting the importance of vigilance and thorough follow-up for patients with conventional osteosarcoma.


Glucose turnover during insulin-induced hypoglycemia in liver-denervated rats.
The role of hepatic autonomic nerves in glucose production during hypoglycemia was studied. Selective, surgical denervation of the liver was performed in rats, which reduced hepatic norepinephrine concentrations by 96%. Hypoglycemia was induced by 250 mU of insulin intra-arterially in anesthetized as well as in chronically catheterized, awake rats. Half of the anesthetized denervated or sham-operated rats had previously been adrenodemedullated. Glucose turnover was measured by primed, constant intravenous infusion of [3-3H]glucose. Before as well as during hypoglycemia the arterial glucose concentration and rates of production and utilization of glucose were similar in denervated rats and control rats. Also hepatic glycogen depletion was similar in the groups. The lack of effect of denervation could not be ascribed to compensating changes in hormone or substrate levels. In adrenodemedullated rats lack of glucose recovery from hypoglycemia was accompanied by delayed normalization of glucose clearance. In fed rats, activity in hepatic autonomic nerves is not a primary mechanism increasing glucose production during acute hypoglycemia. Epinephrine enhances glucose recovery by decreasing glucose clearance rather than by increasing glucose production, at least when glucagon is present.


Decreased membrane fluidity and beta-adrenergic responsiveness in atherosclerotic quail.
The effects of increased cholesterol on erythrocyte membrane fluidity and beta-adrenergic function were studied in a quail model of atherosclerosis. Birds fed a cholesterol-supplemented diet developed severe atherosclerosis and hypercholesterolemia after 6 wk. This cholesterol-enriched diet led to a markedly elevated serum cholesterol and a 26% increase in the cholesterol-to-phospholipid ratio in erythrocyte membranes. Electron paramagnetic resonance spectra measured with 5- and 12-doxyl-stearic acid spin-label probes were used to estimate the order of quail erythrocyte membranes. Membrane preparations from cholesterol-fed birds were more highly ordered near the membrane leaflet surface, as well as deeper in the membrane interior, compared with controls. beta-Adrenergic receptor stimulation of adenosine 3',5'-cyclic monophosphate accumulation was blunted in erythrocytes from the hypercholesterolemic quail. There was no change in beta-receptor density or affinity in the cholesterol-enriched membranes. These studies demonstrate that cholesterol incorporation into erythrocyte membranes in vivo is associated with decreased membrane fluidity and decreased beta-adrenergic responsiveness. The atherosclerotic quail may serve as a useful model to further probe the sequelae of hypercholesterolemia on the function of integral membrane proteins.


Pentobarbital-like discriminative stimulus properties of halothane, 1,1,1-trichloroethane, isoamyl nitrite, flurothyl and oxazepam in mice.
Volatile inhalants represent a diverse group of chemicals which pose a public health problem because of their abuse potential and neurobehavioral toxicity. Although relatively little is known about their pharmacology, they share certain pharmacological properties with classic central nervous system depressants. Drug discrimination procedures were used to compare the effects produced by pentobarbital (PB), oxazepam and several inhalants. Mice were trained to discriminate PB from saline injections in a two-lever operant task. Stimulus generalization was examined after 20-min inhalation exposures to halothane (500-8000 ppm), 1,1,1-trichloroethane (500-12000 ppm), isoamyl nitrite (150-1050 ppm), flurothyl (562-1300 ppm) and injections of oxazepam (0.1-20 mg/kg). Halothane, 1,1,1-trichloroethane and oxazepam produced discriminative stimulus effects similar to those produced by PB. Isoamyl nitrite and flurothyl, while decreasing rates of responding, were not consistently generalized from PB. Differences are apparent in the PB-like discriminative effects of inhalants. Toluene, as shown in a previous study, as well as halothane and 1,1,1-trichloroethane have PB-like discriminative effects which are also shared by oxazepam and may be another common effect of central nervous system depressants. Other chemicals, like isoamyl nitrite and flurothyl, are representative of qualitatively different classes of behaviorally active inhalants.


The occurrence of cis-octadec-15-enoic acid as a major biohydrogenation product from methyl linolenate in bovine rumen liquor.
The biohydrogenation products from methyl linolenate varied according to its concentration in the rumen. At low concentrations, over 60% of the biohydrogenated products was stearic acid, but when the concentration was raised above 100 mg/100 ml of rumen liquor, monoenoic acids, including cis-octadec-15-enoic acid, were the major products.


Clinicopathologic implications of tissue inhibitor of metalloproteinase-1-positive diffuse large B-cell lymphoma.
The tissue inhibitor of metalloproteinase-1 (TIMP-1) is a stromal factor that promotes plasmablastic differentiation, and the survival of germinal center B-cells. The expression of TIMP-1 is known to be correlated with a subset of non-Hodgkin lymphoma at the mRNA level, and Epstein-Barr virus infection in vitro. To characterize TIMP-1(+) diffuse large B-cell lymphoma, TIMP-1 expression was investigated in tissue microarrays from 182 cases of de novo diffuse large B-cell lymphoma and compared with prognostic factors, immunophenotypes, and Epstein-Barr virus infection status. TIMP-1 was expressed not only in tumor cells themselves, in 14 of 182 cases (8%), designated as TIMP-1(+) diffuse large B-cell lymphoma, but also in stromal cells like fibroblasts and endothelial cells. In univariate analysis and hierarchical clustering, our findings suggest that TIMP-1 expression may represent a distinct subgroup. In multivariate analysis, TIMP-1(+) diffuse large B-cell lymphoma (n=14) was associated with unfavorable outcomes compared to TIMP-1(-) diffuse large B-cell lymphoma (n=168) (odds ratio=2.5, P=0.049). Together with TIMP-1 expression, age (greater than 60 years), the presence of B-symptoms, abnormal lactate dehydrogenase level, or more advanced stage (III/IV) was correlated with a poor overall survival. However, TIMP-1 expression in diffuse large B-cell lymphoma was not correlated with other prognostic factors including: clinical stage, international prognostic index score, and nongerminal center B-cell phenotype, as well as Epstein-Barr virus infection. Our results suggest that TIMP-1 expression may be an independent negative prognostic factor in patients with diffuse large B-cell lymphoma.


Secondary prevention of stroke--results from the Southern Africa Stroke Prevention Initiative (SASPI) study.
To describe the prevalence of risk factors and experience of preventive interventions in stroke survivors, and identilfy barriers to secondary prevention in rural South Africa. A clinician visited individuals in the Agincourt field site (in South Africa's rural north east) who were identified in a census as possible stroke victims to confirm the diagnosis of stroke. We explored the impact of stroke on the individual's family, and health-seeking behaviour following stroke by conducting in-depth interviews in the households of 35 stroke survivors. We held two workshops to understand the knowledge, experience, and views of primary care nurses, who provide the bulk of professional health care. We identified 103 stroke survivors (37 men), 73 (71%) of whom had hypertension, but only 8 (8%) were taking anti-hypertensive treatment. Smoking was uncommon; 8 men and 1 woman smoked a maximum of ten cigarettes daily. 94 (91%) stroke survivors had sought help, which involved allopathic health care for most of them (81; 79%). 42 had also sought help from traditional healers and churches, while another 13 people had sought help only from those sources. Of the 35 survivors who were interviewed, 29 reported having been prescribed anti-hypertensive pills after their stroke. Barriers to secondary prevention included cost of treatment, reluctance to use pills, difficulties with access to drugs, and lack of equipment to measure blood pressure. A negative attitude to allopathic care was not an important factor. In this rural area hypertension is the most important modifiable risk factor in stroke survivors. Effective secondary prevention may reduce the incidence of recurrent strokes, but there is no system to deliver such care. New strategies for care are needed involving both allopathic and non-allopathic-health care providers.


Clinical outcomes of TAVI with the Myval balloon-expandable valve for non-calcified aortic regurgitation.
Transcatheter aortic valve replacement (TAVR) in non-calcified aortic regurgitation (NCAR) is an off-label procedure. The balloon-expandable Myval includes extra-large sizes (30.5 mm and 32 mm) of interest in this setting. We aimed to evaluate the safety and feasibility of Myval in NCAR. This was an international, multicentre, observational study that enrolled all consecutive patients with symptomatic severe NCAR undergoing TAVR with the Myval device. The images were centrally analysed. A total of 113 patients were recruited, 64.6% were men, the mean age was 78.4±7.5 years, and the Society of Thoracic Surgeons score was 2.7±1.7%. Aortic root dilatation was present in 59.3% of patients, 7.1% were bicuspid, and the mean annular area was 638.6±106.0 mm2. The annular area was beyond the recommended range for extra-large sizes in 2.6% of cases, and additional volume was added in 92% (median 4 cc, up to 9 cc). The extra-large sizes were used in 95 patients (84.1%), and the mean oversizing was 17.9±11.0%. The technical success rate was 94.7%; the rate of residual ≥moderate aortic regurgitation was 8.9%, and the pacemaker rate was 22.2%. There were no cases of annular rupture, cardiac tamponade, or aortic dissection, but in 4 patients (3.5%) valve embolisation occurred (1 antegrade and 3 ventricular), all in cases with a tapered left ventricle outflow tract (p=0.007). Thirty-day and 1-year mortality were 5.3% and 9.7%, respectively. Technical success was associated with better survival (97.1% vs 72.7%; p=0.012), and valve embolisation was the main determinant of mortality (p=0.047). Myval is a feasible and safe option for selected non-operable patients with NCAR and demonstrated good midterm outcomes and lack of impact of oversizing on device durability.


Off-label and unlicensed drug use in a Spanish Neonatal Intensive Care Unit.
Objetive: To describe off-label and unlicensed drugs in clinical practice in a Neonatal Intensive Care Unit (NICU). The design of the study was a descriptive and retrospective three months research in a NICU of a Spanish University Hospital. All prescriptions were classified as approved, off-label and unlicensed drug used, according to the information available on the Summary of Product Characteristics. Off-label drugs prescriptions were divided into four groups (dose, frecuency, age and indication of use) depending on the reason of disconformity. Forty-one neonates were included (46.3% premature patients) and a total of 273 drug prescriptions were evaluated. Of them, 53.1% (145) were classified as an approved drug prescriptions, 41.4% (113) were off-label use and 5.5% (15) were unlicensed drugs. 90.2% of the patients (37/41) received at least one off label prescription, with a median of 3 (range 1-7). Age was the foremost reason for off-label use (42.5%), followed by dose (31.0%), frecuency (16.8%) and dose-frecuency off-label drug use (8.8%). The last place was reserved for off-label drug use by indication (0.9%). Group J (corresponding with antiinfectives for systemic use) was the most frecuently prescribed, with ampicillin (18.6%) and gentamicin (16.8%) as the drugs most commonly use as off-label prescriptions. Finally, caffeine citrate was the unlicensed drug most frecuently prescribed. Off-label and unlicensed drug use prescriptions in our NICU clinical practice are highly frecuent as other authors have previously published in other countries where they were studied. Although there are few studies in newborn patients, both off-label and unlicensed drug prescriptions are commonly recomended in Paediatrics Guidelines.


Adsorption and Diffusion of Lithium and Sodium on Defective Rhenium Disulfide: A First Principles Study.
Single-layer rhenium disulfide (ReS2) is a unique material with distinctive, anisotropic electronic, mechanical, and optical properties and has the potential to be used as an anode in alkali-metal-ion batteries. In this work, first principles calculations were performed to systematically evaluate the potential of monolayer pristine and defective ReS2 as anodes in lithium (Li)- and sodium (Na)-ion batteries. Our calculations suggest that there are several potential adsorption sites for Li and Na on pristine ReS2, owing to its low-symmetry structure. Additionally, the adsorption of Li and Na over pristine ReS2 is very strong with adsorption energies of -2.28 and -1.71 eV, respectively. Interestingly, the presence of point defects causes significantly stronger binding of the alkali-metal atoms with adsorption energies in the range -2.98 to -3.17 eV for Li and -2.66 to -2.92 eV for Na. Re single vacancy was found to be the strongest binding defect for Li adsorption, whereas S single vacancy was found to be the strongest for Na. The diffusion of these two alkali atoms over pristine ReS2 is anisotropic, with an energy barrier of 0.33 eV for Li and 0.16 eV for Na. The energy barriers associated with escaping a double vacancy and single vacancy for Li atoms are significantly large at 0.60 eV for the double-vacancy case and 0.51 eV for the single-vacancy case. Similarly, for Na, they are 0.59 and 0.47 eV, respectively, which indicates slower migration and sluggish charging/discharging. However, the diffusion energy barrier over a Re single vacancy is found to be merely 0.42 eV for a Li atom and 0.28 eV for Na. Overall, S single and double vacancies can reduce the diffusion rate by 103-105 times for Li and Na ions, respectively. These results suggest that monolayer ReS2 with a Re single vacancy adsorbs Li and Na stronger than pristine ReS2, with negligible negotiation with the charging/discharging rate of the battery, and therefore they can be used as an anode in Li- and Na-ion batteries.


The year's new drugs and biologics 2020.
2020 will go down in history as a year marked in every respect by the emergence and astonishingly rapid spread of the first major global viral pandemic in a century. It seems like nearly every event or story of the year was influenced in some way by COVID-19, and in that respect, the year ended on a high note with the authorization for emergency use of the first vaccines to prevent SARS-CoV-2 infection and drugs to treat COVID-19. Despite the pandemic's dominance of the 2020 headlines, productivity was at a record high level across all therapeutic areas, as seen by the number of products in this year's review: approximately 50% more than the previous year. Notable achievements include the launch of the first treatment for hepatitis D; regulatory decisions on a suite of biologics for the prevention and treatment of Ebola virus disease, fruit of the 2016-2018 outbreak in the Democratic Republic of Congo; the approval of the first-ever drug to treat Hutchinson-Gilford progeria syndrome, a rare genetic disorder that leads to premature aging; the first treatment developed specifically for thyroid eye disease, also known as Graves' ophthalmopathy; the first nonhormonal, on-demand, vaginal pH-regulating contraceptive; and the first oral allergen immunotherapy for peanut allergy.


The role of BTA stat in clinical practice.
BTA stat is a rapid, urine-based test for bladder cancer that detects human complement factor H related protein (HCFHrp) by monoclonal assay. This aim of this study was to assess the efficacy of BTA stat as a diagnostic tool for bladder cancer in symptomatic patients suspected of bladder cancer and in the surveillance of patients with a history of treated bladder cancer. One hundred and six patients presenting with haematuria (gross or microscopic) or irritative bladder symptoms presenting to the urology outpatient clinic of Changi General Hospital and 13 patients under bladder cancer surveillance were recruited for this prospective study. All underwent voided urine cytology (VUC), urine culture, urine BTA stat, intravenous urogram and cystoscopy. Sensitivity, specificity, positive and negative predictive values were calculated for both tests. The stage and grade of bladder tumours detected were also correlated with both test results. Causes of false positives and specificity in different patient groups were analysed. BTA stat is more sensitive than VUC in detecting primary and recurrent bladder tumours (85% versus 55%) but is less specific (62.6% versus 100%). Urinary tract infections and urinary calculi accounted for 62% of false positives with BTA stat. When patients with positive urine cultures and benign IVU abnormalities were excluded, specificity of BTA stat improved (93.9% cf. 62.6%). BTA stat was highly specific (100%) and more sensitive than VUC (75% versus 25%) in detecting recurrent tumours in asymptomatic patients on cancer surveillance. A high false positive rate and low predictive value limits the use BTA stat in screening symptomatic patients. However, it has a role in cancer surveillance and in the screening of high-risk asymptomatic individuals. Further prospective trials should be performed to better assess its role in this respect.


Hepatocyte Growth Factor Modulates MET Receptor Tyrosine Kinase and β-Catenin Functional Interactions to Enhance Synapse Formation.
MET, a pleiotropic receptor tyrosine kinase implicated in autism risk, influences multiple neurodevelopmental processes. There is a knowledge gap, however, in the molecular mechanism through which MET mediates developmental events related to disorder risk. In the neocortex, MET is expressed transiently during periods of peak dendritic outgrowth and synaptogenesis, with expression enriched at developing synapses, consistent with demonstrated roles in dendritic morphogenesis, modulation of spine volume, and excitatory synapse development. In a recent coimmunoprecipitation/mass spectrometry screen, β-catenin was identified as part of the MET interactome in developing neocortical synaptosomes. Here, we investigated the influence of the MET/β-catenin complex in mouse neocortical synaptogenesis. Western blot analysis confirms that MET and β-catenin coimmunoprecipitate, but N-cadherin is not associated with the MET complex. Following stimulation with hepatocyte growth factor (HGF), β-catenin is phosphorylated at tyrosine(142) (Y142) and dissociates from MET, accompanied by an increase in β-catenin/N-cadherin and MET/synapsin 1 protein complexes. In neocortical neurons in vitro, proximity ligation assays confirmed the close proximity of these proteins. Moreover, in neurons transfected with synaptophysin-GFP, HGF stimulation increases the density of synaptophysin/bassoon (a presynaptic marker) and synaptophysin/PSD-95 (a postsynaptic marker) clusters. Mutation of β-catenin at Y142 disrupts the dissociation of the MET/β-catenin complex and prevents the increase in clusters in response to HGF. The data demonstrate a new mechanism for the modulation of synapse formation, whereby MET activation induces an alignment of presynaptic and postsynaptic elements that are necessary for assembly and formation of functional synapses by subsets of neocortical neurons that express MET/β-catenin complex.


T cell and interferon-gamma involvement in the adjuvant action of a detoxified endotoxin.
The adjuvant activities of a detoxified derivative of endotoxic lipopolysaccharides, isolated from the outer membrane of gram-negative bacteria, were evaluated in aging mice. This monophosphoryl lipid A (MPL) (Ribi) was capable of enhancing antibody production in vitro in splenic cultures from 2-3-month-old male Balb/c mice as well as cultures from 22-24-month-old Balb/c mice. Separation of spleen cells from MPL and phosphate-buffered saline-injected mice into adherent and nonadherent populations and subsequent mixing of populations and culture with antigen implicated an adherent cell as being involved in the enhancement of antibody formation induced by MPL. However, separation of normal spleen cells into purified populations of adherent cells, T-lymphocytes, and B-lymphocytes, followed by in vitro stimulation of the individual populations with MPL and subsequent transfer into cultures of normal spleen cells, revealed only the T cell as capable of transferring the enhancement of antibody formation. In addition, culture filtrates from MPL-stimulated T cells were able to enhance antibody production by spleen cell cultures from aging mice twofold above that of filtrates from unstimulated T cells. The enhancement of antibody formation induced by such filtrates and also by MPL in spleen cell cultures from young and aging mice was inhibited by a monoclonal antibody (MAb) to recombinant interferon-gamma (rIFN-gamma) as well as antiserum against IFN-alpha, -beta, and -gamma, but not by an antiserum to IFN-alpha/beta. Enhancement of antibody formation correlated well with an increase in interleukin-1 (IL-1) but not with an increase in IL-2 production. Addition of anti-asialo-GM1 MAb plus complement to the effective spleen populations did not diminish the adjuvant action.


Comparison of disk excision and combined disk excision and spinal fusion for lumbar disk ruptures.
In a series of private patients 77 per cent over-all satisfactory results were obtained from surgical disk excision performed without spinal fusion. Patients in whom spinal fusion combined with discectomy has been recommended by other authors are not much more satisfied than patients with discectomy alone without fusion (81% versus 74% satisfactory). Patients in whom the third party influence prevails were significantly more dissatisfied (only 59% satisfactory). Findings of a definite disk rupture at the time of surgery result in a higher incidence of satisfactory rating than those with degenerative changes or with normal findings.


Investigation into suitable carrier molecules for use in an anti-gonadotrophin releasing hormone vaccine.
Gonadal function can be controlled through immunoneutralisation of gonadotrophin releasing hormone (GnRH), with an analogue, GnRH-glycys, linked to a carrier molecule and an appropriate adjuvant. In this study, four different types of carrier molecule were investigated: (a) single and branched amino acid polymers--[poly-(D-glu, D-lys) and poly-(phe, glu)-poly(DL-ala)-poly(lys)]; (b) bacterial toxoids--diphtheria (DT) and tetanus (TT); (c) synthetic T-helper epitopes--derived from malarial circumsporozite protein (CS) and measles virus fusion protein (MVF); and (d) thyroglobulin (Thy)--a large protein. The effect of non-ionic surfactant vesicles (NISV) and an aluminum hydroxide based adjuvant (alum), was also examined. Although good antibody responses were achieved with GnRH-glycys-DT, GnRH-glycys-TT and GnRH-glycys-Thy, adsorbed onto alum and the dimerised synthetic T-helper epitope constructs, incorporated into NISV, a critical antibody titre was necessary to result in morphological changes in the gonads and complete suppression of spermatogenesis. This was only achieved with tetanus toxoid and the dimerised T-helper epitopes.


Estrogen Receptor Alpha and its Ubiquitination in Breast Cancer Cells.
More than 70% of all breast cancer cases are estrogen receptor alpha-positive (ERα). ERα is a member of the nuclear receptor family, and its activity is implicated in the gene transcription linked to the proliferation of breast cancer cells, as well as in extranuclear signaling pathways related to the development of resistance to endocrine therapy. Protein-protein interactions and posttranslational modifications of ERα underlie critical mechanisms that modulate its activity. In this review, the relationship between ERα and ubiquitin protein (Ub), was investigated in the context of breast cancer cells. Interestingly, Ub can bind covalently or non-covalently to ERα resulting in either a proteolytic or non-proteolytic fate for this receptor. Thereby, Ub-dependent molecular pathways that modulate ERα signaling may play a central role in breast cancer progression, and consequently, present critical targets for treatment of this disease.


Giant osteoclast formation and long-term oral bisphosphonate therapy.
Bisphosphonates decrease bone resorption and are commonly used to treat or prevent osteoporosis. However, the effect of bisphosphonates on their target cells remains enigmatic, since in patients benefiting from therapy, little change, if any, has been observed in the number of osteoclasts, which are the cells responsible for bone resorption. We examined 51 bone-biopsy specimens obtained after a 3-year, double-blind, randomized, placebo-controlled, dose-ranging trial of oral alendronate to prevent bone resorption among healthy postmenopausal women 40 through 59 years of age. The patients were assigned to one of five groups: those receiving placebo for 3 years; alendronate at a dose of 1, 5, or 10 mg per day for 3 years; or alendronate at a dose of 20 mg per day for 2 years, followed by placebo for 1 year. Formalin-fixed, undecalcified planar sections were assessed by bone histomorphometric methods. The number of osteoclasts was increased by a factor of 2.6 in patients receiving 10 mg of alendronate per day for 3 years as compared with the placebo group (P<0.01). Moreover, the number of osteoclasts increased as the cumulative dose of the drug increased (r=0.50, P<0.001). Twenty-seven percent of these osteoclasts were giant cells with pyknotic nuclei that were adjacent to superficial resorption cavities. Furthermore, giant, hypernucleated, detached osteoclasts with 20 to 40 nuclei were found after alendronate treatment had been discontinued for 1 year. Of these large cells, 20 to 37% were apoptotic, according to both their morphologic features and positive findings from in situ end labeling. Long-term alendronate treatment is associated with an increase in the number of osteoclasts, which include distinctive giant, hypernucleated, detached osteoclasts that are undergoing protracted apoptosis.


Metanephric transforming growth factor-alpha is required for renal organogenesis in vitro.
The role of transforming growth factor-alpha (TGF-alpha) in metanephric development was examined. Metanephroi were removed from 13-day-old rat embryos and grown in organ culture for up to 6 days using serum-free chemically defined media. During this time the metanephroi increased in size and morphological complexity. Messenger RNA for TGF-alpha was present in the renal anlage. Immunoreactive TGF-alpha was produced by metanephroi in vitro and released into culture media. TGF-alpha of metanephric origin coeluted with recombinant human 125I-TGF-alpha after high-performance liquid chromatography of media. In contrast, epidermal growth factor was not detectable. Levels of TGF-alpha were relatively constant during 4 days in culture and averaged approximately 10(-10) M. Growth of the metanephric anlage, arborization of the ureteric bud, and tubulogenesis within the metanephric blastema were inhibited by the addition of anti-TGF-alpha antibodies to organ cultures. These data demonstrate production of TGF-alpha by developing rat metanephroi in organ culture. The peptide is necessary for growth and development in vitro. Our findings suggest a necessary role for TGF-alpha of metanephric origin as a promoter of renal organogenesis in vivo.


Is 18F-fluorocholine-positron emission tomography/computerized tomography a new imaging tool for detecting hyperfunctioning parathyroid glands in primary or secondary hyperparathyroidism?
Preoperative ultrasonography and scintigraphy using (99m)Tc-sestamibi are commonly used to localize abnormal parathyroid glands. In cases of discrepant results between scintigraphy and ultrasonography, it is important to rely on another diagnostic imaging modality. (18)F-fluorodeoxyglucose (FDG) and (11)C-methionine positron emission tomography (PET) have been studied, but are imperfect to detect abnormal parathyroid glands. Recently, first cases of abnormal parathyroid glands taking-up radiolabelled choline were discovered incidentally in men referred to (11)C-choline or (18)F-fluorocholine (FCH)-PET/CT for prostate cancer. We checked if FCH uptake was a general feature of adenomatous or hyperplastic parathyroid glands. FCH-PET/CT was performed in 12 patients with primary (n = 8) or secondary hyperparathyroidism (1 dialyzed, 3 grafted) and with discordant or equivocal results on preoperative ultrasonography (US) and/or (123)I/(99m)Tc-sestamibi dual-phase scintigraphy. The results of the FCH-PET/CT were evaluated, with surgical exploration and histopathologic examination as the standard of truth. On a per-patient level, the detection rate of FCH-PET/CT (at least one FCH focus corresponding to an abnormal parathyroid gland in a given patient) was 11/12 = 92%. FCH-PET/CT detected 18 foci interpreted as parathyroid glands and correctly localized 17 abnormal parathyroid glands (7 adenomas and 10 hyperplasias). On a per-lesion level, FCH-PET/CT results were 17 TP, 2 false negative ie, a lesion-based sensitivity of 89%, and 1 false positive. As the main result of this pilot study, we show that in patients with hyperparathyroidism and with discordant or equivocal results on scintigraphy or on ultrasonography, adenomatous or hyperplastic parathyroid glands can be localized by FCH-PET/CT with good accuracy. Furthermore, FCH-PET/CT can solve discrepant results between preoperative ultrasonography and scintigraphy and has thus a potential as a functional imaging modality in the detection of abnormal parathyroid glands. Our preliminary results are encouraging and prompt us to further evaluate FCH-PET/CT as a functional imaging agent in patients with biochemical hyperparathyroidism.


Inhalation poisoning with palytoxin from aquarium coral: case description and safety advice.
Palythoa spp. corals and some other marine organisms contain one of the most poisonous substances ever known - palytoxin (PTX). Due to their modest life requirements and ease of breeding, these corals are popular in home aquariums. Here we refer to a case of PTX poisoning of a middle-aged woman who inhaled poisonous vapours while brushing the corals from live rock and compare it with the available literature. As the case revealed that the symptoms of PTX poisoning are not specific and neither is treatment, our aim was to give a brief tabulated review of the symptoms that may indicate such poisoning. Cases of palytoxin poisoning have been reported worldwide, and severe ones (mostly due to ingestion of contaminated sea food) can end in death. As it appears, most (if not all) poisonings result from unawareness of the risk and reckless handling by aquarists. This is one of the first articles which provides some practical advice about the use of personal protection equipment, including gloves, masks, eyewear, and other clothing during any coral manipulation to minimise the risk. We also draw attention to the lack of marketing/trading regulations for dangerous coral species and/or regulations or instructions dealing with their removal and health protection.


Pathfinding by peripheral pioneer neurons in grasshoppers.
Grasshopper neurons accurately project axons across long distances between peripheral structures and the central nervous system. Nerve-trunk pathways followed by these axons are established early in embryogenesis by pioneer neurons. Growth cones from the first pioneers navigate along a chain of cells to the CNS. The placement of these cells may constitute the initial guidance mechanism underlying long-distance pathfinding.


Structure and properties of single crystalline CaMg2Bi2, EuMg2Bi2, and YbMg2Bi2.
Single crystals of CaMg(2)Bi(2), EuMg(2)Bi(2), and YbMg(2)Bi(2) were obtained from a Mg-Bi flux cooled to 650 °C. These materials crystallize in the CaAl(2)Si(2) structure-type (P ̅3m1, No. 164), and crystal structures are reported from refinements of single crystal and powder X-ray diffraction data. EuMg(2)Bi(2) displays an antiferromagnetic transition near 7 K, which is observed via electrical resistivity, magnetization, and specific heat capacity measurements. Magnetization measurements on YbMg(2)Bi(2) reveal a weak diamagnetic moment consistent with divalent Yb. Despite charge-balanced empirical formulas, all three compounds are p-type conductors with Hall carrier concentrations of 2.0(3) × 10(19) cm(-3) for CaMg(2)Bi(2), 1.7(1) × 10(19) cm(-3) for EuMg(2)Bi(2), and 4.6(7) × 10(19) cm(-3) for YbMg(2)Bi(2), which are independent of temperature to 5 K. The electrical resistivity decreases with decreasing temperature and the resistivity ratios ρ(300 K)/ρ(10 K) ≤ 1.6 in all cases, indicating significant defect scattering.


Extranodal multicentric Castleman's disease with cutaneous involvement.
Castleman's disease (CAD) is an unusual lymphoid hyperplasia that might be induced by an immunologic reaction to a virus or another infectious organism or drug. CAD commonly occurs in the mediastinal lymph nodes and less frequently in the neck lymph nodes, but other lymph nodes might be involved. Extranodal CAD is rare. We present a 72-year-old woman with diffuse 2- to 5-cm cutaneous nodules that had appeared a few months earlier. The patient underwent a biopsy, and the lesions resolved within 4 months. The patient was asymptomatic except for mild joint pains. Histopathologic findings showed a circumscribed nodule composed of lymphoid follicles with contracted germinal centers and expanded mantle zones. The lymphoid follicles were separated by perivascular bands of sclerotic collagen. Aggregates of epithelioid and dendritic histiocytes were present between follicles, with a prominent lymphoplasmacytic infiltrate and vascular proliferation. Although extranodal CAD has been reported in the skin, we think that this is the first reported case of multicentric cutaneous CAD without evidence of lymph node involvement.


The effect of corticosteroid on collagen expression in injured rotator cuff tendon.
Subacromial corticosteroid injections are commonly used in the nonoperative management of rotator cuff disease. The effects of corticosteroid injection on injured rotator cuff tendons have not been studied. Our aims were to characterize the acute response of rotator cuff tendons to injury through the analysis of the type-III to type-I collagen expression ratio, a tendon injury marker, and to examine the effects of corticosteroid on this response. Sixty Sprague-Dawley rats were randomly assigned to four groups: control, tendon injury, steroid treatment, and tendon injury and steroid treatment. Six rats served as sham controls. Unilateral tendon injuries were created with full-thickness defects across 50% of the total width of the infraspinatus tendon, 5 mm from its humeral insertion. Steroid treatment with a single dose of methylprednisolone (0.6 mg/kg), equivalent to that given to humans, was injected into the subacromial space under direct visualization. Steroid treatment followed the creation of an injury in the rats in the injury and steroid treatment group. At one, three, and five weeks after the injury, the total RNA isolated from tendons was quantified with real-time polymerase chain reaction with use of primers for type-I and type-III collagen and ribosomal 18s RNA. The type-III to type-I collagen expression ratio remained at baseline at all time-points in the control and sham groups. At one week, the type-III to type-I collagen expression ratio increased more than fourfold above the control level in the tendon injury group (p = 0.017) and the tendon injury and steroid treatment group (p = 0.003). The ratio remained greater than twofold above the control at three weeks in both groups (p = 0.003 and p = 0.037) and returned to baseline at five weeks. Interestingly, the group that had steroid treatment only showed an increase of >4.5-fold (p = 0.001) in the type-III to type-I collagen expression ratio, without structural injury to the tendon. This ratio returned to baseline levels by three weeks. A single dose of corticosteroid does not alter the acute phase response of an injured rotator cuff tendon in the rat. However, the same steroid dose in uninjured tendons initiates a short-term response equivalent to that of structural injury.


Optical calorimetry in microfluidic droplets.
A novel microfluidic calorimeter that measures the enthalpy change of reactions occurring in 100 μm diameter aqueous droplets in fluoropolymer oil has been developed. The aqueous reactants flow into a microfluidic droplet generation chip in separate fluidic channels, limiting contact between the streams until immediately before they form the droplet. The diffusion-driven mixing of reactants is predominantly restricted to within the droplet. The temperature change in droplets due to the heat of reaction is measured optically by recording the reflectance spectra of encapsulated thermochromic liquid crystals (TLC) that are added to one of the reactant streams. As the droplets travel through the channel, the spectral characteristics of the TLC represent the internal temperature, allowing optical measurement with a precision of ≈6 mK. The microfluidic chip and all fluids are temperature controlled, and the reaction heat within droplets raises their temperature until thermal diffusion dissipates the heat into the surrounding oil and chip walls. Position resolved optical temperature measurement of the droplets allows calculation of the heat of reaction by analyzing the droplet temperature profile over time. Channel dimensions, droplet generation rate, droplet size, reactant stream flows and oil flow rate are carefully balanced to provide rapid diffusional mixing of reactants compared to thermal diffusion, while avoiding thermal "quenching" due to contact between the droplets and the chip walls. Compared to conventional microcalorimetry, which has been used in this work to provide reference measurements, this new continuous flow droplet calorimeter has the potential to perform titrations ≈1000-fold faster while using ≈400-fold less reactants per titration.


Options for management of municipal solid waste in New York City: a preliminary comparison of health risks and policy implications.
Landfill disposal and waste-to-energy (WTE) incineration remain the two principal options for managing municipal solid waste (MSW). One critical determinant of the acceptability of these options is the different health risks associated with each. In this analysis relying on published data and exposure modeling, we have performed health risk assessments for landfill disposal versus WTE treatment options for the management of New York City's MSW. These are based on the realistic scenario of using a waste transfer station (WTS) in Brooklyn and then transporting the untreated MSW by truck to a landfill in Pennsylvania or using a WTE facility in Brooklyn and then transporting the resultant ash by truck to a landfill in Pennsylvania. The overall results indicate that the individual cancer risks for both options would be considered generally acceptable, although the risk from landfilling is approximately 5 times greater than from WTE treatment; the individual non-cancer health risks for both options would be considered generally unacceptable, although once again the risk from landfilling is approximately 5 times greater than from WTE treatment. If one considers only the population in Brooklyn that would be directly affected by the siting of either a WTS or a WTE facility in their immediate neighborhood, individual cancer and non-cancer health risks for both options would be considered generally acceptable, but risks for the former remain considerably higher than for the latter. These results should be considered preliminary due to several limitations of this study such as: consideration of risks only from inhalation exposures; assumption that only volume and not composition of the waste stream is altered by WTE treatment; reliance on data from the literature rather than actual measurements of the sites considered, assuming comparability of the sites. However, the results of studies such as this, in conjunction with ecological, socioeconomic and equity considerations, should prove useful to environmental managers, regulators, policy makers, community representatives and other stakeholders in making sound and acceptable decisions regarding the optimal handling of MSW.


[Effect of empagliflosin on renal filtration in patients with coronary heart disease undergoing percutaneous coronary intervention].
Aim To evaluate the effect of empagliflozin on glycemia and renal filtration function in patients with stable ischemic heart disease (IHD) and type 2 diabetes mellitus (DM2) who underwent a percutaneous coronary intervention (PCI).Materials and methods This study included 40 patients with stable IHD and DM2 (age, 63 (58; 65) years; DM2 duration, 7 (4; 15) years) who had indications for an elective PCI. At baseline in the total sample, the level of glycated hemoglobin was 7.2 (6.5; 8.3)%; 48.7 % failed to achieve glycemic goals. A decrease in glomerular filtration rate (GFR) to below 60 ml/min/1.73 m2 was observed in 10.3 % of patients. All patients were divided into two group by simple randomization with successively assigned numbers. The main group consisted of 20 patients who received empagliflozin 10 mg/day in addition to their previous hypoglycemic therapy irrespective of their baseline glycemic control. Patients of the comparison group (n=20) continued on their previous hypoglycemic therapy as prescribed by their endocrinologist. The follow-up duration was 6 months. Statistical analysis was performed with the Statistica 10.0 software.Results The empagliflozin treatment improved the glycemic control; in the comparison group, no significant changes in glycemic control were observed. In both groups, GFR significantly decreased during the follow-up period; median decreases in GFR were -6.0 (-16.0; 4.0) and -8.4 (-26.5; 2.5) ml/min / 1.73 m2 in the main and comparison groups, respectively (p = 0.646). No significant changes in 24-h proteinuria were observed for patients taking empagliflozin. In the control group, the 24-h urinary protein excretion significantly progressed (p=0.011) during the follow-up period.Conclusion In patients with DM2 and stable IHD who underwent a PCI, addition of empagliflozin 10 mg/day to their current hypoglycemic therapy was associated with a significant improvement of glycemic control. The decrease in GFR during the empagliflozin treatment did not significantly differ from the value for patient receiving the other hypoglycemic therapy.


Neuromuscular function after a bout of low-load blood flow-restricted exercise.
This study compared endurance and neuromuscular function after bouts of low-load (LL), high-load (HL), and LL blood flow-restricted (LL(BFR)) resistance exercise. Eight recreationally active male subjects completed three sets of dynamic knee extensions to volitional failure under three conditions: HL (70% peak torque), LL (20% peak torque), and LL(BFR) (20% peak torque with an occlusive cuff inflated to 180 mm Hg wrapped around the thigh). Before and immediately after exercise, isometric torque, central activation, electrically evoked torque, and muscle activation via surface EMG were measured. Isometric torque and evoked torque decreased an average of 37% and 40%, respectively (P < 0.01) in all conditions after exercise. There were no differences in the toque decrements between the conditions (P > 0.05). Percent central activation did not change after any condition (P = 0.09). Rate of torque development declined an average of 26% after all three conditions (P = 0.003), and rate of half-relaxation time was depressed by 48% after the HL condition (P = 0.004) only. EMG amplitude was greater in the HL condition at the beginning and end of exercise compared with the LL and LL(BFR) conditions (P = 0.001). At the end of exercise, EMG amplitude rose 19% (P = 0.02) and was not different among conditions (P > 0.05). Subjects performed more repetitions during the LL and LL(BFR) conditions (P < 0.05). Although LL and LL(BFR) resistance exercise to volitional failure exhibit lower levels of muscle activation than HL exercise, similar torque decrements occur after all bouts of resistance exercise, and the muscle fatigue can be attributed to peripheral factors.


Multiscale Computational Model Predicts Mouse Skin Kinematics Under Tensile Loading.
Skin is a complex tissue whose biomechanical properties are generally understood in terms of an incompressible material whose microstructure undergoes affine deformations. A growing number of experiments, however, have demonstrated that skin has a high Poisson's ratio, substantially decreases in volume during uniaxial tensile loading, and demonstrates collagen fiber kinematics that are not affine with local deformation. In order to better understand the mechanical basis for these properties, we constructed multiscale mechanical models (MSM) of mouse skin based on microstructural multiphoton microscopy imaging of the dermal microstructure acquired during mechanical testing. Three models that spanned the cases of highly aligned, moderately aligned, and nearly random fiber networks were examined and compared to the data acquired from uniaxially stretched skin. Our results demonstrate that MSMs consisting of networks of matched fiber organization can predict the biomechanical behavior of mouse skin, including the large decrease in tissue volume and nonaffine fiber kinematics observed under uniaxial tension.


HLA molecules, bacteria and autoimmunity.
It has been well established that many diseases are linked to HLA antigens. Two of the most interesting HLA associations may provide some insight into the pathogenesis of rheumatic inflammatory conditions. In ankylosing spondylitis (AS), 96% of patients possess HLA-B27, whilst the frequency of this marker in the general population is c. 8%. In rheumatoid arthritis (RA), >90% of patients possess either HLA-DR1 or some subtypes of HLA-DR4, whilst the frequency of this marker in the general population is c. 35%. The association between HLA-B27 and reactive arthritis (ReA) has also been well established. Furthermore, it has been shown that ReA is triggered by infection via the gastrointestinal tract due to Yersinia, Salmonella or Campylobacter spp. and in the genitourinary tract due to chlamydia. In a similar way, microbiological and immunological studies have revealed an association between Klebsiella pneumoniae in AS and Proteus mirabilis in RA. This article reviews the possible pathological implications of the associations between HLA-B27, K. pneumoniae and AS, as well as HLA-DR1/DR4, P. mirabilis and RA.


[Acute fever without source in infants and children less than 36 months of age]
OBJECTIVES: The objective of this article is to review current information about fever without source in children less than 36 months of age and to discuss the practical aspects of diagnosis and management.METHODS: Articles from the specific area were reviewed and presented in a practical form with recommendations from the authors.RESULTS: Febrile children comprise a substantial proportion of ambulatory pediatric visits. The majority of children are less than 3 years old. At first examination, there may be considerable overlap in the clinical appearance of children with fever due to viral illness, occult bacteremia and serious bacterial infection. The authors present the recommendations and protocols cited in the literature to assist physicians in managing infants and children with fever without source.CONCLUSION: Although many approaches to the evaluation and management of the febrile infant exist, no diagnostic protocol or therapeutic scheme is optimal for all patients. Pediatricians may individualize their management of these patients, depending on their experience, or interpretation of the recommendations presented in the literature.


Relation of C-reactive protein, fibrinogen, and cardiorespiratory fitness to risk of systemic hypertension in men.
We investigated the relation between inflammation and incident hypertension, independent of obesity, and tested the associations of cardiorespiratory fitness (fitness) and indexes of inflammation for the development of hypertension in 2,475 normotensive men. Inflammatory markers were C-reactive protein (CRP) and fibrinogen. Fitness was directly measured by peak oxygen uptake during sign/symptom-limited treadmill exercise testing to volitional fatigue; 266 men (10.7%) developed hypertension during an average of 4 years follow-up. After adjusting for potential confounding variables, the relative risk (RR) and 95% confidence interval (CI) for incident hypertension in those in the upper tertile versus lower tertile were 1.55 (95% CI 1.15 to 2.09) for CRP and 1.51 (95% CI 1.10 to 2.06) for fibrinogen. Although the association between fibrinogen and incident hypertension persisted after adjusting for body mass index (p = 0.049), the relation between CRP and incident hypertension was no longer statistically significant (p = 0.08). Fit men had a 27% decreased (RR 0.73, 95% CI 0.56 to 0.94) risk of incident hypertension compared with unfit men in a multivariable adjusted model. In the joint analysis, unfit men with upper CRP had 1.81 times (95% CI 1.21 to 2.70) and unfit men with upper fibrinogen had 2.03 times (95% CI 1.33 to 3.12) greater risks of incident hypertension compared with fit men with low CRP and fibrinogen, respectively. However, these risks did not significantly increase in fit men with upper CRP (RR 1.12, 95% CI 0.76 to 1.63) and fibrinogen (RR 1.26, 95% CI 0.86 to 1.85) groups. In conclusion, these results suggest that heightened levels of fibrinogen, but not CRP, are associated with incident hypertension, independent of body weight, and that high fitness attenuates the risk of incident hypertension across upper levels of inflammatory markers in men.


Gene-respiratory disease interactions for rheumatoid arthritis risk.
We aimed to identify gene by respiratory tract disease interactions that increase RA risk. In this case-control study using the Mass General Brigham Biobank, we matched incident RA cases, confirmed by ACR/EULAR criteria, to four controls on age, sex, and electronic health record history. Genetic exposures included a validated overall genetic risk score (GRS) for RA, a Human Leukocyte Antigen (HLA) GRS for RA, and the MUC5B promoter variant, an established risk factor for RA-associated interstitial lung disease (ILD). Preceding respiratory tract diseases came from diagnosis codes (positive predictive value 86%). We estimated attributable proportions (AP) and multiplicative odds ratios (OR) with 95% confidence intervals (CI) for RA for each genetic and respiratory exposure using conditional logistic regression models, adjusting for potential confounders. We identified 653 incident RA cases and 2,607 matched controls (mean 54 years, 76% female). The highest tertile of the overall GRS and the HLA GRS were both associated with increased RA risk (OR 2.28, 95% CI 1.89,2.74; OR 2.02, 95% CI 1.67-2.45). ILD and the HLA GRS exhibited a synergistic relationship for RA risk (OR for both exposures 4.30, 95% CI 1.28,14.38; AP 0.51, 95% CI-0.16,1.18). Asthma and the MUC5B promoter variant also exhibited a synergistic interaction for seropositive RA (OR for both exposures 2.58, 95% CI 1.10,6.07; AP 0.62, 95% CI 0.24,1.00). ILD-HLA GRS and asthma-MUC5B promoter variant showed synergistic interactions for RA risk. Such interactions may prove useful for RA prevention and screening.


Stress, age, and immune function: toward a lifespan approach.
Both aging processes and psychological stress affect the immune system: Each can dysregulate immune function with a potentially substantial impact on physical health. Worse, the effects of stress and age are interactive. Psychological stress can both mimic and exacerbate the effects of aging, with older adults often showing greater immunological impairment to stress than younger adults. In addition, stressful experiences very early in life can alter the responsiveness of the nervous system and immune system. We review the unique impact of aging and stress on immune function, followed by evidence of interactions between age and stress. Further, we suggest that prenatal or early life stress may increase the likelihood of maladaptive immune responses to stress in late life. An understanding of the interactive effects of stress and age is critical to efforts to determine underlying mechanisms, clarify the directionality of effects, and develop effective interventions in early and late life.


Pediatric lung imaging features of COVID-19: A systematic review and meta-analysis.
Pediatric COVID-19 studies have been mostly restricted to case reports and small case series, which have prevented the identification of specific pediatric lung disease patterns in COVID-19. The overarching goal of this systematic review and meta-analysis is to provide the first comprehensive summary of the findings of published studies thus far describing COVID-19 lung imaging data in the pediatric population. A systematic literature search of PubMed was performed to identify studies assessing lung-imaging features of COVID-19 pediatric patients (0-18 years). A single-arm meta-analysis was conducted to obtain the pooled prevalence and 95% confidence interval (95% CI). A total of 29 articles (n = 1026 children) based on chest computerized tomography (CT) images were included. The main results of this comprehensive analysis are as follows: (1) Over a third of pediatric patients with COVID-19 (35.7%, 95% CI: 27.5%-44%) had normal chest CT scans and only 27.7% (95% CI: 19.9%-35.6%) had bilateral lesions. (2) The most typical pediatric chest CT findings of COVID-19 were ground-glass opacities (GGO) (37.2%, 95% CI: 29.3%-45%) and the presence of consolidations or pneumonic infiltrates (22.3%, 95% CI: 17.8%-26.9%). (3) The lung imaging findings in children with COVID-19 were overall less frequent and less severe than in adult patients. (4) Typical lung imaging features of viral respiratory infections in the pediatric population such as increased perihilar markings and hyperinflation were not reported in children with COVID-19. Chest CT manifestations in children with COVID-19 could potentially be used for early identification and prompt intervention in the pediatric population.


Non-lethal? Penetrating chest injury due to beanbag bullet.
We report a case of serious lung injury from beanbag bullet. A 46-year-old gentleman, shot with beanbag bullets was brought to the emergency department. Upon arrival he was in obvious respiratory distress and complained of severe pain in the right chest. A 3.0×3.5 cm entry wound on the right parasternal area was identified. Chest x-ray revealed a right haemopneumothorax, parenchymal changes at the right lung base and a radiopaque foreign body. A right-sided intercostal chest tube was inserted, draining air and 750 ml of blood. After stabilisation patient underwent a right thoracotomy. A beanbag bullet was found in the oblique fissure of the right lung, with extensive haematoma of the middle lobe. The bullet and skin fragments overlying the lung and along the bullet track were extracted. The pleural cavity was washed with normal saline and haemostasis was confirmed. The patient had an uneventful postoperative recovery.


Antioxidant Peptide Fractions Isolated from Wheat Germ Protein with Subcritical Water Extraction and Its Transport Across Caco-2 Cells.
Wheat germ protein (WGP) was extracted with subcritical water and then hydrolyzed with Alcalase 2.4 L to obtain antioxidant hydrolysates. Wheat germ peptides (WG-P, Mw < 1 kDa) were purified by using Sephadex G-15 column chromatography. The results showed that WG-P-4 possessed the strongest DPPH radical scavenging activity in comparison with other peptides fractions. In addition, free amino acids and LC-MS/MS analysis showed that Gly-Pro-Phe, Gly-Pro-Glu, and Phe-Gly-Glu were the major peptides of WG-P-4. Interestingly, the WG-P-4 fractions had good absorption characteristic. Moreover, the ratio of Papp both sides of apical compartment (AP) and basolateral compartment (BL) were between 0.5 and 1.0 on Caco-2 cell model, which indicated that transmembrane transportation was mainly passive transport. Therefore, WG-P could exert an effective antioxidant action by across the intestinal epithelium.


Anti-NMDA Receptor Autoantibody Is an Independent Predictor of Hospital Mortality but Not Brain Dysfunction in Septic Patients.
The presence of autoantibodies against neuronal cell surface or synaptic proteins and their relationship to autoimmune encephalitis have recently been characterized. These autoantibodies have been also reported in other pathologic conditions; however, their role during sepsis is not known. This study detected the presence of autoantibodies against neuronal cell surface or synaptic proteins in the serum of septic patients and determined their relationship to the occurrence of brain dysfunction and mortality. This prospective, observational cohort study was performed in four Brazilian intensive care units (ICUs). Sixty patients with community-acquired severe sepsis or septic shock admitted to the ICU were included. Blood samples were collected from patients within 24 h of ICU admission. Antibodies to six neuronal proteins were assessed, including glutamate receptors (types NMDA, AMPA1, and AMPA2); voltage-gated potassium channel complex (VGKC) proteins, leucine-rich glioma-inactivated protein 1 (LGI1), and contactin-associated protein-2 (Caspr2), as well as the GABAB1 receptor. There was no independent association between any of the measured autoantibodies and the occurrence of brain dysfunction (delirium or coma). However, there was an independent and significant relationship between anti-NMDAR fluorescence intensity and hospital mortality. In conclusion, anti-NMDAR was independently associated with hospital mortality but none of the measured antibodies were associated with brain dysfunction in septic patients.


Characterization of an extracellular metalloprotease with elastase activity from Staphylococcus epidermidis.
The gene sepA from Staphylococcus epidermidis TU3298-P, encoding the extracellular neutral metalloprotease SepP1, was cloned into pT181mcs. DNA sequencing revealed an open reading frame of 1,521 nucleotides encoding a 507-amino-acid protein with an M(r) of 55,819. The sepA-containing DNA fragment did not hybridize with Staphylococcus hyicus or Staphylococcus carnosus DNA. Expression of sepA in the protease-negative S. carnosus (pT181mcsP1) resulted in overproduction of a 33-kDa protease found in the culture medium. The first 15 N-terminal amino acids of the partially purified protease completely matched the deduced DNA sequence starting at GCA (Ala-208). This finding indicated that SepP1 is synthesized as a preproenzyme with a 28-amino-acid signal peptide, a 179-amino-acid hydrophilic pro region, and a 300-amino-acid extracellular mature form with a calculated M(r) of 32,739. In activity staining, the mature protease prepared from S. carnosus (pT181mcsP1) corresponded to the extracellular S. epidermidis Tü3298-P protease. The partially purified protease had a pH optimum between 5 and 7, and its activity could be inhibited by zinc- and metal-specific inhibitors such as EDTA and 1,10-phenanthroline, indicating that it is a neutral metalloprotease. The protease had a low substrate specificity. Glucagon was cleaved preferentially between aromatic (Phe) and hydrophobic (Val) amino acids. The protease hydrolyzed casein and elastin. The amino acid sequence of the mature form of SepP1 revealed pronounced similarities with the thermolabile and thermostable neutral proteases of various bacilli (44 to 55% identity) and a central part of the mature form of the Pseudomonas aeruginosa elastase (31% identity). From homology comparison with the Bacillus thermoproteolyticus thermolysin, we predict that mature SepP1 binds one zinc ion at a conserved zinc-binding site.


Diagnostic accuracy of fine needle aspiration cytology in thyroid lesions.
Evaluation of accuracy of fine needle aspiration cytology (FNAC) in the diagnosis of different thyroid lesions. This is a retrospective study of 296 diagnosed cases of thyroid nodules referred to cytology unit, pathology department, NCI, who underwent FNAC for diagnosis. The results were categorized according to the recent Bethesda classification into: insufficient for diagnosis, benign, atypical follicular lesion of undetermined significance, follicular neoplasm, suspicious for malignancy, and malignant sampling. The final histologic diagnosis and/or clinico-radiologic follow-up assessment for non-neoplastic lesions were considered the gold standard. The study included 296 cases presented with thyroid nodules who underwent diagnostic thyroid FNAC. Female to male ratio was 5.2:1, and the median age was 44years. Ninety-eight cases (33.1%) were diagnosed as benign, 40 cases (13.5%) as follicular lesion of undetermined significance, 49 cases (16.5%) as follicular neoplasm, 30 cases (10.1%) as suspicious for malignancy, 58 cases (19.5%) as malignant, and 21 cases (7.1%) as unsatisfactory. Nodular hyperplasia represented the majority of benign cases (89.8%), while papillary carcinoma was the most frequent malignant lesion (72.4%). Cytologic diagnoses were compared with their corresponding final histologic ones. FNAC achieved a sensitivity of 92.8, a specificity of 94.2%, a positive predictive value of 94.9%, a negative predictive value of 91.8%, a false positive rate of 7.2%, a false negative rate of 5.8%, and a total accuracy of 93.6%. FNA cytology is a sensitive, specific, and accurate initial diagnostic test for the evaluation of patients with thyroid swellings.


The Her7 node modulates the network topology of the zebrafish segmentation clock via sequestration of the Hes6 hub.
Using in vitro and in vivo assays, we define a network of Her/Hes dimers underlying transcriptional negative feedback within the zebrafish segmentation clock. Some of the dimers do not appear to be DNA-binding, whereas those dimers that do interact with DNA have distinct preferences for cis regulatory sequences. Dimerization is specific, with Hes6 serving as the hub of the network. Her1 binds DNA only as a homodimer but will also dimerize with Hes6. Her12 and Her15 bind DNA both as homodimers and as heterodimers with Hes6. Her7 dimerizes strongly with Hes6 and weakly with Her15. This network structure engenders specific network dynamics and imparts greater influence to the Her7 node. Computational analysis supports the hypothesis that Her7 disproportionately influences the availability of Hes6 to heterodimerize with other Her proteins. Genetic experiments suggest that this regulation is important for operation of the network. Her7 therefore has two functions within the zebrafish segmentation clock. Her7 acts directly within the delayed negative feedback as a DNA-binding heterodimer with Hes6. Her7 also has an emergent function, independent of DNA binding, in which it modulates network topology via sequestration of the network hub.


The comparative pathology of the glycosidase inhibitors swainsonine, castanospermine, and calystegines A3, B2, and C1 in mice.
To study various polyhydroxy-alkaloid glycosidase inhibitors, 16 groups of 3 mice were dosed using osmotic minipumps with swainsonine (0, 0.1, 1, and 10 mg/kg/day), castanospermine, and calystegines A(3), B(2), and C(1) (1, 10, and 100 mg/kg/day). After 28 days, the mice were euthanized, necropsied, and examined using light and electron microscopy. The high-dose swainsonine-treated mice developed neurologic disease with neuro-visceral vacuolation typical of locoweed poisoning. Castanospermine- and calystegines-treated mice were clinically normal; however, high-dose castanospermine-treated mice had thyroid, renal, hepatic, and skeletal myocyte vacuolation. Histochemically, swainsonine- and castanospermine-induced vacuoles contained mannose-rich oligosaccharides. High-dose calystegine A(3)-treated mice had increased numbers of granulated cells in the hepatic sinusoids. Electron microscopy, lectin histochemistry, and immunohistochemistry suggest these are pit cells (specialized NK cells). Histochemically, the granules contain glycoproteins or oligosaccharides with abundant terminal N-acetylglucosamine residues. Other calystegine-treated mice were histologically normal. These findings indicate that swainsonine produced lesions similar to locoweed, castanospermine caused vacuolar changes with minor changes in glycogen metabolism, and only calystegine A(3) produced minimal hepatic changes. These also suggest that in mice calystegines and castanospermine are less toxic than swainsonine, and as rodents are relatively resistant to disease, they are poor models to study such induced storage diseases.


The maize transposable element Ac induces recombination between the donor site and an homologous ectopic sequence.
The prominent repair mechanism of DNA double-strand breaks formed upon excision of the maize Ac transposable element is via nonhomologous end joining. In this work we have studied the role of homologous recombination as an additional repair pathway. To this end, we developed an assay whereby beta-Glucuronidase (GUS) activity is restored upon recombination between two homologous ectopic (nonallelic) sequences in transgenic tobacco plants. One of the recombination partners carried a deletion at the 5' end of GUS and an Ac or a Ds element inserted at the deletion site. The other partner carried an intact 5' end of the GUS open reading frame and had a deletion at the 3' end of the gene. Based on GUS reactivation data, we found that the excision of Ac induced recombination between ectopic sequences by at least two orders of magnitude. Recombination events, visualized by blue staining, were detected in seedlings, in pollen and in protoplasts. DNA fragments corresponding to recombination events were recovered exclusively in crosses with Ac-carrying plants, providing physical evidence for Ac-induced ectopic recombination. The occurrence of ectopic recombination following double-strand breaks is a potentially important factor in plant genome evolution.


Replication Fork Remodeling and Therapy Escape in DNA Damage Response-Deficient Cancers.
Most cancers have lost a critical DNA damage response (DDR) pathway during tumor evolution. These alterations provide a useful explanation for the initial sensitivity of tumors to DNA-targeting chemotherapy. A striking example is dysfunctional homology-directed repair (HDR), e.g., due to inactivating mutations in BRCA1 and BRCA2 genes. Extensive efforts are being made to develop novel targeted therapies exploiting such an HDR defect. Inhibitors of poly(ADP-ribose) polymerase (PARP) are an instructive example of this approach. Despite the success of PARP inhibitors, the presence of primary or acquired therapy resistance remains a major challenge in clinical oncology. To move the field of precision medicine forward, we need to understand the precise mechanisms causing therapy resistance. Using preclinical models, various mechanisms underlying chemotherapy resistance have been identified. Restoration of HDR seems to be a prevalent mechanism but this does not explain resistance in all cases. Interestingly, some factors involved in DNA damage response (DDR) have independent functions in replication fork (RF) biology and their loss causes RF instability and therapy sensitivity. However, in BRCA-deficient tumors, loss of these factors leads to restored stability of RFs and acquired drug resistance. In this review we discuss the recent advances in the field of RF biology and its potential implications for chemotherapy response in DDR-defective cancers. Additionally, we review the role of DNA damage tolerance (DDT) pathways in maintenance of genome integrity and their alterations in cancer. Furthermore, we refer to novel tools that, combined with a better understanding of drug resistance mechanisms, may constitute a great advance in personalized diagnosis and therapeutic strategies for patients with HDR-deficient tumors.


Robustness of holographic optical traps against phase scaling errors.
Complex three-dimensional patterns of multifunctional optical traps can be encoded in phase-only computer-generated holograms and projected with the holographic optical trapping technique. The trap-forming holograms, in turn, are implemented as diffractive optical elements whose phase transfer functions generally do not faithfully reproduce the design. We demonstrate that phase encoding errors reduce the overall intensities of the projected traps but, remarkably, do not affect their positions, relative intensities or mode structure. We exploit this robust performance to implement dual-color holographic optical tweezers with a single hologram.


Disrupting autophagy restores peroxisome function to an Arabidopsis lon2 mutant and reveals a role for the LON2 protease in peroxisomal matrix protein degradation.
Peroxisomes house critical metabolic reactions that are essential for seedling development. As seedlings mature, metabolic requirements change, and peroxisomal contents are remodeled. The resident peroxisomal protease LON2 is positioned to degrade obsolete or damaged peroxisomal proteins, but data supporting such a role in plants have remained elusive. Arabidopsis thaliana lon2 mutants display defects in peroxisomal metabolism and matrix protein import but appear to degrade matrix proteins normally. To elucidate LON2 functions, we executed a forward-genetic screen for lon2 suppressors, which revealed multiple mutations in key autophagy genes. Disabling core autophagy-related gene (ATG) products prevents autophagy, a process through which cytosolic constituents, including organelles, can be targeted for vacuolar degradation. We found that atg2, atg3, and atg7 mutations suppressed lon2 defects in auxin metabolism and matrix protein processing and rescued the abnormally large size and small number of lon2 peroxisomes. Moreover, analysis of lon2 atg mutants uncovered an apparent role for LON2 in matrix protein turnover. Our data suggest that LON2 facilitates matrix protein degradation during peroxisome content remodeling, provide evidence for the existence of pexophagy in plants, and indicate that peroxisome destruction via autophagy is enhanced when LON2 is absent.


Association of malocclusion and functional occlusion with subjective symptoms of TMD in adults: results of the Study of Health in Pomerania (SHIP).
An analysis of exclusively representative population-based studies on adults has shown that only few and inconsistent associations could be detected between malocclusions and clinical signs of temporomandibular disorders (TMD)--and none for functional occlusion factors (occlusal interferences, non-working side contacts, etc). The aim of this study was to analyze associations between morphologic occlusion as well as factors of functional occlusion and subjectively perceived symptoms of TMD--again on the basis of the population-based Study of Health in Pomerania (SHIP), providing a sample of 4310 subjects (out of 7008 subjects yielding a response rate of 68.8%) aged 20 to 81 years, and other international representative studies from the systematic review. Besides occlusal factors also parafunctions and socioeconomic status (SES) were taken into account (including age and sex). Multiple logistic regression analysis was used--adjusted for SES. In this study, none of the occlusal factors were significantly associated with the indication of more frequent subjective TMD symptoms. However, the parafunction "frequent clenching" was connected with subjective TMD symptoms (odds ratio = 3.4). Compared with other population-based studies few and (across studies) inconsistent associations between malocclusions and subjective TMD symptoms could be ascertained. No significant associations of factors of functional occlusion with TMD symptoms were identifiable.


Panic and suicidal ideation in primary care.
The objective of this article is to examine the relationship between panic attacks, panic disorder, and suicidal ideation among primary care patients. A probability sample of 1,007 primary care attenders from a large urban university practice was assessed for current mental disorders and suicidal ideation (past 2 week prevalence) with the PRIME-MD Patient Health Questionnaire. Controlling for major depression, substance use disorders, and sociodemographic variables simultaneously, patients with either panic attacks or panic disorder had significantly increased risks of suicidal ideation. Suicidal ideation was highly associated with major depression and comorbid panic disorder (OR = 15.4) or panic attacks (OR = 7.9). There is need for detection and possible treatment of patients with panic attacks or disorder in primary care, especially among those with co-occurring major depression.


Interobserver agreement in the evaluation of B-lines using bedside ultrasound.
We evaluated agreement among trained emergency physicians assessing the degree of B-line presence on bedside ultrasound in patients presenting to the emergency department (ED) with acute undifferentiated dyspnea. We also determined which thoracic zones offered the highest level of interobserver reliability for sonographic B-line assessment. We evaluated a prospective convenience sample of adult patients presenting with dyspnea to an academic ED. Two consecutive bedside lung ultrasounds were performed on 91 patients by a pair of physician-sonographers. The lung ultrasounds were structured 10-zone thoracic sonograms, documented as videos. Sonographer pairs were expert/expert (>100 lung ultrasounds performed) or expert/novice pairs (novices performed 5 supervised examinations after structured training) and blinded to clinical data. Sonographers reported B-line concentration with 3 assessment methods: (1) normal (<3 B-lines) or abnormal (≥3 B-lines); (2) ordinal (normal, mild, moderate, or severe), and (3) counting B-lines (0-10; >10) in each zone. All statistical analyses were performed using SPSS version 18.0 (Chicago, IL) and Stata 12.1 (College Station, TX). We evaluated interrater and intrarater agreement using Intraclass correlation coefficients (ICCs). The right and left anterior/superior lung zones showed substantial agreement in all assessment methods and demonstrated best overall agreement (ICC for right: counting, ordinal, and normal/abnormal, 0.811 [0.714-0.875], 0.875 [0.810-0.917], and 0.729 [0.590-0.821], respectively). Furthermore, both expert/expert pairs and expert/novice pairs showed substantial agreement in the right and left anterior/superior thoracic zones (expert/expert, 0.904 and 0.777, respectively; expert/novice, 0.862, and 0.834, respectively). Second best agreement was found for the lateral/superior lung zones (right: counting, ordinal, and normal/abnormal, 0.744 [0.612-0.831], 0.686 [0.524-0.792], and 0.639 [0.453-0.761], respectively; and ICC left: counting, ordinal, and normal/abnormal, 0.671 [0.501-0.782], 0.615 [0.417-0.746], and 0.720 [0.577-0.815], respectively). When comparing agreement to distinguish "normal vs abnormal" B-line findings, our results showed significant agreement in all zones with the exception of the right and left inferior/lateral lung fields and left posterior lung. Reinterpretation by 2 experts of all their own randomized video clips at a later date showed agreement of 0.697 (n=733 zones) and 0.647 (n=266) zones for ordinal assessment of B-line concentration. Interrater agreement was best in the anterior/superior thoracic zones followed by the lateral/superior zones for both expert/expert and expert/novice pairs. Agreement in the lateral/inferior lung zones was overall inferior. Intrarater agreement was highest at extreme high or low numbers of B-lines.


[Recurrent strongyloidiasis as an indicator of HTLV-1 infection].
A 53-year-old West African man presented two years after a travel to Guinea because of severe headache, neck stiffnes, fever and pruritus. The patient had been in orthopedical treatment for the last five months. Stool microscopy revealed a high number of Strongyloides stercoralis larvae. Hematology, biochemistry and all other parasitology results were normal. HIV-1/2 testing was negative and CD4+-lymphocyte count was normal. Concomitant infection by Human T Cell lymphotropic virus type 1 (HTLV-1) was confirmed by serology and PCR. The phylogenetic analysis confirmed African origin of the virus. The infection responded to a five-day course of albendazol at 400 mg/d but during the following five years repeat recrudescences were observed inspite of high-dosage and prolonged antiparasitic treatments. Eventually, eradication of the infection was achieved by a four day course of ivermectin 0.2 mg/kg/d. Although both strongyloidiasis and HTLV-1 infections occur most frequently in tropical areas, these may also be observed in temperate regions. Suppression of the immune system by HTLV-1 differs from that by HIV. CD4+-lymphocytes were rarely decreased. Prolonged treatment with ivermectin in a dosage exceeding the current recommendations may be required in HTLV-1 infected patients and was well tolerated. The unusual presentation of the infection with muscular symptoms contributed to the delay of the diagnosis. HTLV-1 positive patients must be monitored for years. They and their partners must be instructed how to prevent transmission of the virus.


The importance of artificial wetlands for birds: A case study from Cyprus.
The degradation of natural wetlands has significant effects on the ecosystem services they provide and the biodiversity they sustain. Under certain conditions, these negative effects can be mitigated by the presence of artificial wetlands. However, the conservation value of artificial wetlands needs to be explored further. In addition, it is unclear how certain anthropogenic variables, such as road networks and hunting reserves (i.e., areas where hunting of birds is prohibited) affect biodiversity in both artificial and natural wetlands. Here, we use data from thirteen artificial and six natural wetlands in Cyprus, to assess their similarities in bird species diversity and composition, and to quantify the relationship between species diversity and the density of road networks, hunting reserves, wetland size, and wetland depth. We found that while on average natural wetlands have more species and support higher abundances, certain artificial wetlands have the potential to support similarly diverse communities. Overall, regardless of the type, larger wetlands, with shallower waters tend to be more biodiverse. The same is true for wetlands surrounded by a higher percentage of hunting reserves and a lower density of road networks, albeit the effect of road networks was weaker. We conclude, from our results, that although the conservation value of natural wetlands is higher, artificial wetlands have the potential to play a complimentary role in the conservation of bird communities, assuming those wetlands have the right characteristics (e.g., in terms of size and depth) and assuming that the disturbances resulting from high-impact human-activities (e.g., hunting) are minimized.


Modeling the hepatitis A epidemiological transition in Brazil and Mexico.
Many low- to middle-income countries have completed or are in the process of transitioning from high or intermediate to low endemicity for hepatitis A virus (HAV). Because the risk of severe hepatitis A disease increases with age at infection, decreased incidence that leaves older children and adults susceptible to HAV infection may actually increase the population-level burden of disease from HAV. Mathematical models can be helpful for projecting future epidemiological profiles for HAV. An age-specific deterministic, dynamic compartmental transmission model with stratification by setting (rural versus urban) was calibrated with country-specific data on demography, urbanization, and seroprevalence of anti-HAV antibodies. HAV transmission was modeled as a function of setting-specific access to safe water. The model was then used to project various HAV-related epidemiological outcomes in Brazil and in Mexico from 1950 to 2050. The projected epidemiological outcomes were qualitatively similar in the 2 countries. The age at the midpoint of population immunity (AMPI) increased considerably and the mean age of symptomatic HAV cases shifted from childhood to early adulthood. The projected overall incidence rate of HAV infections decreased by about two thirds as safe water access improved. However, the incidence rate of symptomatic HAV infections remained roughly the same over the projection period. The incidence rates of HAV infections (all and symptomatic alone) were projected to become similar in rural and urban settings in the next decades. This model featuring population age structure, urbanization and access to safe water as key contributors to the epidemiological transition for HAV was previously validated with data from Thailand and fits equally well with data from Latin American countries. Assuming no introduction of a vaccination program over the projection period, both Brazil and Mexico were projected to experience a continued decrease in HAV incidence rates without any substantial decrease in the incidence rates of symptomatic HAV infections.


Evolution of gene regulation during transcription and translation.
Understanding how gene regulation evolves is a key area in the current evolutionary field. Gene regulation occurs at various levels. Previous work on the evolution of gene regulation has largely focused on gene transcription. In this study, we used a recently developed ribosomal footprint profiling method to investigate how gene regulation evolves at both the transcription (mRNA abundance) and translation (ribosomal density) levels. By constructing a hybrid between Saccharomyces cerevisiae (Scer) and Saccharomyces bayanus (Sbay), which diverged ∼20 Ma, and quantifying transcriptome and translatome in both parental strains and their hybrid, we showed that translation is much more conserved than transcription, mostly due to the buffering effect of translational regulation for the transcriptional divergence. More conservation in translation than transcription is also confirmed by the inheritance mode of transcription and translation between two species. Furthermore, cis and trans effects are widely involved in changes at both transcription and translation levels. Finally, our results showed that genes with certain functions and sequence features might employ specific modes for evolution at these two critical levels of gene regulation. Our results demonstrated that it is essential to investigate the evolution of gene regulation at various levels from different genetic backgrounds to obtain a complete picture of its evolutionary modes in nature.


Massive Distance Education: Barriers and Challenges in Shifting to a Complete Online Learning Environment.
The global pandemic has dramatically changed how the world functions and impacted all sectors of society including all educational institutions. Government and educators respond with immediate online teaching and learning for all students. Massive distance education has been drawn into the picture to provide non-stop learning in most countries worldwide. This study focuses on examining different orders of barriers educators have encountered during the Covid-19 pandemic. The barriers to massive online teaching and learning included the first-order barrier (technological or external barrier), the second-order barrier (internal barrier or teachers' and parents' beliefs), the third-order barrier (design thinking barrier), and the 2.5th order barrier (the classroom management barrier). Both teachers and students are suffering from unstable or limited internet connectivity and it directly hinders students' rights in the massive online education. Teachers are facing the need for sudden pedagogical redesign while parents are enduring the burden of providing all kinds of support for their children's online learning at home. Some learners are experiencing videoconferencing fatigue and struggling with overwhelming resources and an excessive amount of technology time. This study also identifies a group of forgotten learners, the videoconferencing refugees, who have limited access to the Internet and lost their learning opportunities. From a global perspective, shifting to massive online education may be possible with all four orders of barriers being overcome.


Indirect Lipid Transfer Protein Activity Measurements Using Quantification of Glycosphingolipid Production.
Here we summarize how glycosphingolipid production can be followed using metabolic labeling with radiolabeled lipid precursors. No assays are available yet that directly would address the lipid transfer protein activity in vivo. Therefore, these approaches can serve as tools to indirectly study the lipid transfer protein activity in cells, by monitoring their impact on the glycosphingolipid homeostasis.


Effects of 5-HT alone and its interaction with TRH on neurons in rat dorsal motor nucleus of the vagus.
The purpose of this study was to determine whether exposure to thyrotropin-releasing hormone (TRH) enhances the excitatory effect of 5-hydroxytryptamine (5-HT) on motoneurons of the dorsal motor nucleus of the vagus (DMV) as described in whole animal studies. For this purpose we used the patch-clamp technique applied to rat brain stem slices. Exposure of DMV motoneurons to concentrations of 5-HT (0.1-3 microM) resulted in a concentration-related increase in spontaneous firing rate. As previously described by Travagli et al. [Am. J. Physiol. 263 (Gastrointest. Liver Physiol. 26): G508-G517, 1992], TRH (1-30 microM) increased action potential firing rate. Indeed, when TRH perfusion increased the firing rate, addition of 5-HT to the perfusing solution exerted no further excitation of the DMV motoneuron, indicating that there was no summation of response. Studies using whole cell current recordings showed a common action of 5-HT and TRH in antagonizing the Ca(2+)-dependent afterhyperpolarizing current (IAHP). Again, interaction studies between TRH and 5-HT indicated no enhancing effect of TRH on 5-HT-induced antagonism of IAHP. In conclusion, our data indicated that the enhancement of 5-HT-induced excitation of DMV motoneurons by TRH described by in vivo rat experiments is not due to an interaction of TRH and 5-HT directly on the DMV motoneuron.


Endogenously synthesized peptide with an endoplasmic reticulum signal sequence sensitizes antigen processing mutant cells to class I-restricted cell-mediated lysis.
The HLA-A2-positive human mutant cell line T2 is not lysed by influenza virus-specific HLA-A2-restricted cytotoxic lymphocytes after virus infection. However, lysis does occur when cells are incubated with the antigenic influenza matrix protein-derived peptide M57-68. To examine the nature of this defect, T2 cells were transfected with two different plasmids. One plasmid encoded the peptide M57-68, and the other encoded the same peptide preceded by an endoplasmic reticulum translocation signal sequence. Mutant T2 cells expressing the M57-68 peptide without the signal sequence were not susceptible to lysis by M57-68-specific HLA-A2-restricted cytotoxic T lymphocytes, whereas T2 cells expressing the M57-68 peptide plus signal sequence were lysed effectively. Lysis of parental T1 cells with either plasmid was equally effective. These results suggest that the T2 mutant cells are defective in the transport of antigenic peptides from the cytosol into the secretory pathway.


Identification of a lens-specific regulatory region (LSR) of the murine alpha B-crystallin gene.
Previous studies have shown that the -661/+44 sequence of the murine alpha B-crystallin gene contains a muscle-preferred enhancer (-426/-257) and can drive the bacterial chloramphenicol acetyltransferase (CAT) gene in the lens, skeletal muscle and heart of transgenic mice. Here we show that transgenic mice carrying a truncated -164/+44 fragment of the alpha B-crystallin gene fused to the CAT gene expressed exclusively in the lens; by contrast mice carrying a -426/+44 fragment of the alpha B gene fused to CAT expressed highly in the lens, skeletal muscle and heart, and slightly in the lung, brain, kidney, spleen and liver. DNase I protection experiments indicated that the -147/-118 sequence is protected by nuclear proteins from alpha TN4-1 lens cell line, but not by nuclear proteins from myotubes of the C2C12 cell line. Site directed mutagenesis of this sequence decreased promoter activity in transiently-transfected lens cells, consistent with this sequence being a lens-specific regulatory region (LSR). We conclude that the -426/-257 enhancer is required for expression in skeletal muscle, heart and possibly other tissues, and that the -164/+44 sequence of the alpha B-crystallin gene is sufficient for expression in the lens of transgenic mice.