The Launch of the iCoDE-2 Standard Project: Integration of Connected Diabetes Device Data Into the Electronic Health Record.
Diabetes Technology Society virtually hosted the first meeting of the Integration of Connected Diabetes Device Data Into the Electronic Health Record #2 (iCoDE-2) Standards Project on May 31, 2023, via Zoom. Clinicians, patients, data aggregators, informaticists, manufacturers, attorneys, and cybersecurity experts discussed dosing data currently available from insulin delivery devices and data aggregators and the types of information that patients and clinicians want to see. This information along with technical and regulatory aspects of (1) data standards and (2) integration into the electronic health record (EHR) are the basis of iCoDE-2. Six sessions were hosted discussing themes including (1) What's Currently Available: The Continuous Glucose Monitor Experience; (2) What's Currently Available: The Aggregator Experience; (3) What's Currently Available: The Insulin Pump Experience; (4) What's Currently Available: The Insulin Pen Experience; (5) What Do Clinicians and Patients Want Out of iCoDE-2 Data?; and (6) Technical and Regulatory Aspects of Data Aggregation. Additional working group meetings to discuss data standard and clinical workflow will be held to create additional technical specifications and clinical workflows to aid in insulin dosing data integration into the EHR. A mid-project meeting will convene in the second half of 2023.


Ubiquitin binding site of the ubiquitin E2 variant (UEV) protein Mms2 is required for DNA damage tolerance in the yeast RAD6 pathway.
Different ubiquitin modifications to proliferating cell nuclear antigen (PCNA) signal distinct modes of lesion bypass in the RAD6 pathway of DNA damage tolerance. The modification of PCNA with monoubiquitin signals an error-prone bypass, whereas the extension of this modification into a Lys-63-linked polyubiquitin chain promotes error-free bypass. Chain formation is catalyzed by the Mms2/Ubc13 conjugating enzyme variant/conjugating enzyme (UEV.E2) complex together with the Rad5 ubiquitin ligase. In vitro studies of this UEV.E2 complex have identified a ubiquitin binding site that is mainly localized on Mms2. However, the role of this site in DNA damage tolerance and the molecular features of the ubiquitin/Mms2 interaction are poorly understood. Here we identify two molecular determinants, the side chains of Mms2-Ile-57 and ubiquitin-Ile-44, that are required for chain assembly in vitro and error-free lesion bypass in vivo. Mutating either of these side chains to alanine elicits a severe 10-20-fold inhibition of chain synthesis that is caused by compromised binding of the acceptor ubiquitin to Mms2. These results suggest that the ubiquitin binding site of Mms2 is necessary for error-free lesion bypass in the RAD6 pathway and provide new insights into ubiquitin recognition by UEV proteins.


Irritable bowel syndrome - less irritable, or better treatments?
Irritable bowel syndrome (IBS) is a chronic, often bothersome disorder of gut-brain interaction (DGBI) characterized by abdominal pain associated with a change in stool frequency and/or caliber. Recent advancements have improved our understanding of the underlying pathophysiology, thus opening new avenues for therapeutic intervention. The purpose of this review is to summarize the current literature regarding treatment modalities for IBS. Altering the gut microbiome via probiotic and antibiotic administration, avoiding dietary triggers, and modulating the gut-brain axis have all proven efficacious for the management of IBS symptoms. Several gut-specific pharmacotherapies are approved for the treatment of IBS, many of which primarily address either diarrhea or constipation, although many patients remain symptomatic despite appropriate use. Brain-gut behavioral therapies (BGBTs) are increasingly used to treat symptoms of IBS, particularly in those who do not respond to traditional therapies. Virtual reality represents an exciting new approach to treating DGBIs, like IBS, though data are limited. As our understanding of IBS continues to evolve, so should our therapeutic approach. Individualizing the therapeutic approach is of utmost importance.


Technique for the localization of intracranially implanted electrodes.
The anatomical localization of electrodes in the human brain is important for the interpretation of pathophysiological (epileptifom spikes, seizures) and functional data (stimulation mapping, evoked potentials). Electroencephalography and evoked potentials are volume-conducted field effects that are most easily interpreted with knowledge of the location and topology of adjacent structures, and brain stimulation techniques produce current fields whose effects are highly dependent on the geometry of electrode assemblies in relation to adjacent structures. In this paper, the authors describe a straightforward method for implanted electrode localization, and detail their experience to date with the technique. The described method is based on the coregistration of preoperative MR imaging studies with postimplant CT scans by using standard mutual information optimization of rigid body transformation of the CT to the MR image. Fused images of the MR and thresholded CT images are derived, and electrodes are visualized using various standard computer projections, renderings, and measurement tools. The authors have successfully used the described method over an extended period to localize electrode contacts in intracranial implants for seizure localization, and in long-term implants for movement disorders and seizure control. The accuracy of localization is very good, although it is dependent on image quality and possible brain shift between acquisition of the CT and MR images. This method is easily implemented and is useful for a wide variety of clinical and research applications. It is a straightforward process to extend it to additional image modalities that are emerging for surgical planning and image guidance.


Retest reliability of balance and mobility measurements in people with mild to moderate Alzheimer's disease.
To interpret changes of balance and mobility in people with Alzheimer's disease (AD), we require measures of balance and mobility that have demonstrated reliability in this population. The aim of the study was to determine the safety, feasibility and retest reliability of clinical and forceplate balance and mobility measurements in people with AD. Relative and absolute reliabilities were examined in 14 older people with mild to moderate AD. Relative reliability was calculated using the intraclass correlation coefficient, two-way mixed model (ICC3,1). Absolute reliability was calculated using the standard error of measurement (SEM), the minimum detectable change (MDC) and the coefficient of variation (CV). All measurements were clinically feasible and could be safely administered. ICC values were excellent and CVs were less than 11% in all clinical balance and mobility measures except the Timed Up & Go test with cognitive or manual task (ICC3,1 = 0.5 and 0.7, and CV = 14% and 10%, respectively). Most balance and mobility measures tested on the Neurocom™ forceplate (modified Clinical Test of Sensory Interaction on Balance, Walk Across (step width, step length parameters), and Sit to Stand (rising index parameter)) had excellent relative reliability (ICC3,1 ranging from 0.75 to 0.91). ICC values were fair to good for the other measures. Retest reliability of the balance and mobility measures used in this study ranged between fair to good, and good to excellent. Clinicians should consider retest reliability when deciding which balance and mobility measures are used to assess people with AD.


Bone markers and their prognostic value in metastatic bone disease: clinical evidence and future directions.
Bone metastases are prevalent among patients with advanced solid tumors. Metastatic bone disease alters bone homeostasis, resulting in reduced bone integrity and, consequently, increased skeletal complications. Biochemical markers of bone metabolism may meet an unmet need for useful, noninvasive, and sensitive surrogate information for following patients' skeletal health. Data for this review were identified by searches of PubMed, and references from relevant articles using the search terms "bone markers" or individual bone marker nomenclature, "cancer," and "metastases." Abstracts and reports from meetings were included only when they related directly to previously published work. Only papers published in English between 1990 and 2007 were included. Recent retrospective analyses with bisphosphonates, and particularly with zoledronic acid, have shown significant correlations between biochemical markers of bone metabolism levels and clinical outcomes, especially for bone resorption markers. Clinical results for biochemical markers of bone formation and resorption and other emerging markers of bone metabolism including bone sialoprotein, receptor-activator of nuclear factor-kappaB ligand, osteoprotegerin, and other markers are presented. However, biochemical markers of bone metabolism are not yet an established surrogate endpoint for treatment efficacy. Biochemical markers of bone metabolism may allow physicians to identify which patients with metastatic bone disease are at high risk for skeletal-related events or death and who may be responding to therapy. Prospective randomized clinical trials are underway to further assess the utility of markers of bone metabolism in patients with bone metastases.


Impact of a multicomponent integrated care delivery program on diabetes care goals achievement: a primary care quality improvement initiative.
To evaluate the impact of multicomponent integrated care (MIC) delivery program in a primary care real-life setting on diabetes care goals. Patients with T2D and usual primary care from the public health system in Mexico were invited to participate in a five-month ambulatory MIC quality initiative (DIAbetes Empowerment and Improvement of Care program, DIABEMPIC). 841 patients who finished the program and with complete data were analyzed. The patients had a mean decrease in hemoglobin A1c, systolic and diastolic pressure, and LDL-cholesterol of 2.4%, 9 mmHg, 3 mmHg, and 22.5 mg/dL, respectively (p < 0.001). The achievement of the optimal triple target goal increased from 1.8% to 26.7% (p < 0.001). In the adjusted analysis, the diabetes knowledge and global self-care behavior score post-intervention, as well as the increment of global self-care behavior score were associated with the optimal composite risk factor control achievement. The incorporation of diabetes therapeutic education interventions to improve self-care behaviors along with adequate treatment intensification in diabetes care are fundamental to attaining optimal risk factor control and attenuating disease burden.


Optimized T1-MPRAGE sequence for better visualization of spinal cord multiple sclerosis lesions at 3T.
Spinal cord lesions are highly prevalent in MS, and their visualization can help both in diagnosis and patient follow-up. However, the sensitivity of MR imaging to spinal cord lesions remains poor, primarily because of suboptimal contrast between lesions and a normal-appearing cord. Here, we propose an optimized 3D MPRAGE sequence for improved detection of MS lesions in the spinal cord at 3T. Images were acquired by use of T2 FSE, STIR, T1-gradient recalled-echo (for T1 mapping), and T1-MPRAGE in the sagittal plane, and T2*-weighted scans in the axial plane, on 40 patients with MS and 7 healthy volunteers. Two observers qualitatively evaluated the images for lesion conspicuity. Lesions seen between the C1 and C4 segments in 10 randomly selected patients with MS were further evaluated quantitatively for contrast-to-noise ratio between the lesion and normal-appearing cord, and for lesion burden. Spinal cord lesions were more conspicuous on the optimized T1-MPRAGE sequence than on any other sequence tested. Detailed analysis revealed that lesions were almost 3 times more conspicuous (P < .01), and the total lesion volume was 2 times greater (P < .05, n=10), in the T1-MPRAGE sequence compared with the standard STIR sequence. Correlation of clinical disability (Expanded Disability Status Score) with lesion load from each sequence also demonstrated the importance of the improved lesion conspicuity with T1-MPRAGE. The optimized T1-MPRAGE sequence described here improves the reliability of lesion visualization and estimation of lesion burden, especially when used in conjunction with other well-established clinical sequences.


[Therapeutic drug monitoring of 5-fluorouracil after its administration in high-dose protocols].
Therapeutic drug monitoring is currently investigated in patients receiving the drug by prolonged continuous infusion, either alone or associated with other chemotherapy agents. This arises from an increasing body of evidence that relates plasma fluorouracil concentrations to toxicity or effectiveness. Literature data indicate that threshold levels of exposure, as assessed by the area under the concentration-time curve, are associated with an increased risk of toxicity in patients treated for either a colorectal or a head and neck cancer. Kinetically guided dose adjustment may thus be proposed for the ongoing or the next cycle of chemotherapy. Historical comparisons and the results of a phase III multicenter randomized study in patients with metastatic colorectal cancer showed that dose-adapted fluorouracil resulted in significantly improved objective response rate, a trend to higher survival rate and fewer grade III-IV toxicities. Real-time fluorouracil monitoring may also help identifying patients with higher than expected exposure, thus providing a basis for intensifying supportive care in these patients.


Chronic exposure to tolbutamide and glibenclamide impairs insulin secretion but not transcription of K(ATP) channel components.
Clonal insulin-secreting BRIN-BD11 cells were used to examine effects of chronic 72-144 h exposure to the sulphonylureas tolbutamide and glibenclamide on insulin release, cellular insulin content, and mRNA levels of the Kir6.2 and SUR1 subunits of the beta-cell K(ATP) channel. Chronic exposure for 72-144 h to 5-100 microM tolbutamide and glibenclamide resulted in a time- and concentration-dependent irreversible decline in sulphonylurea-induced insulin secretion. In contrast, the decline in cellular insulin content induced by chronic exposure to high concentrations of sulphonylureas was readily reversible. Chronic exposure to tolbutamide or glibenclamide had no effect upon transcription of the Kir6.2 or SUR1 subunits of the pancreatic beta-cell K(ATP) channel. Whilst further studies are required to understand the precise nature of the chronic interactions of sulphonylurea with the insulin exocytotic mechanism, these observations may partially explain the well-known progressive failure of sulphonylurea therapy in type 2 diabetes.


Polymeric Self-Assembled Monolayers Anomalously Improve Thermal Transport across Graphene/Polymer Interfaces.
Ultralow thermal conductivities of bulk polymers greatly limit their applications in areas demanding fast heat dissipation, such as flexible electronics and microelectronics. Therefore, polymeric composites incorporating highly thermally conductive filler materials (e.g., graphene and carbon nanotubes) have been produced to address the issue. However, despite some enhancement, thermal conductivities of these materials are still far below theoretical predictions, mainly due to the inefficient thermal transport across material interfaces. Here, using molecular dynamics simulations, we demonstrate that polyethylene (PE) self-assembled monolayer (SAM) functionalized graphene surfaces at a high grafting density can drastically improve interfacial thermal conduction between graphene and the matrix of poly(methyl methacrylate) (PMMA). In contrast to abrupt temperature drop across pristine graphene/PMMA interfaces, temperature field in the vicinity of a PE-grafted graphene/PMMA interface is continuous with a smoother transition and higher thermal conductance. This anomalous improvement is attributed to three factors that closely relate to the grafting density of the SAM of PE. First, the SAM with high grafting densities features highly extended chains that enhance along-chain thermal conduction. Second, the strong covalent bonding between the SAM and the graphene facilitates heat transfer at their joints. Third, the SAM and the PMMA matrix are both organic materials, leading to enhanced interfacial vibrational coupling. Molecular mechanisms underpinning these phenomena are systematically elucidated by analyzing the temperature field, density distribution, Herman's orientation factor, the vibrational density of states, cumulative correlation factor, the integrated autocorrelation of interfacial heat power, and interfacial adhesion. All results suggest the incorporation of SAMs at a high grafting density or extremely extended polymer brushes for drastically improved interfacial thermal transport between hard and soft materials toward a wide range of applications.


Ovalbumin-induced airway inflammation and fibrosis in mice also exposed to ultrafine particles.
A murine model of allergen-induced airway inflammation was used to examine the effects of exposure to ultrafine particles (PM(2.5)) on airway inflammation and remodeling. Lung inflammation was measured by quantitative differential evaluation of lung lavage cells. Alterations in lung structure (airway remodeling and fibrosis) were evaluated by quantitative biochemical analysis of microdissected airways and by histological evaluation of stained lung sections. The same total number of cells was observed in lavage fluid from animals exposed for 4 wk to ovalbumin alone or to ovalbumin for 4 wk immediately before or after 6 exposures over a period of 2 wk to 235 ug/m(3) of PM(2.5). Mice exposed to ovalbumin for 6 wk with concurrent exposure to PM(2.5) during wk 5-6 had a significant decrease in the total number of cells recovered by lavage as compared with the group exposed to ovalbumin alone. There were no significant differences in the cell differential counts in the lavage fluid from mice exposed to ovalbumin alone as compared with values from mice exposed to ovalbumin and PM(2.5) under the protocols studied. Airway structural changes (remodeling) were examined by three different quantitative methods. None of the groups exposed to ovalbumin and PM had a significant increase in airway collagen content evaluated biochemically (i.e., total airway collagen) as compared to the matched groups of mice exposed to ovalbumin alone. Airway collagen content evaluated histologically by sirius red staining showed significant increases in all of the animals exposed to ovalbumin, with or without PM, and no apparent difference between the ovalbumin group and mice exposed to PM with ovalbumin. The findings were consistent with an additive, or less than additive, response of mice to exposure to PM and ovalbumin. Air or PM exposure alone for 2 wk did not result in observable goblet cells in the airways, while mice exposed to ovalbumin aerosol alone for 4 wk had about 20-25% goblet cells in their conducting airways. Sequential exposure to ovalbumin and PM (or vice versa) caused significant increases in goblet cells (to about 35% of total cells) in the conducting airways of the exposed mice. We conclude that when mice with allergen-induced airway inflammation induced by ovalbumin are also exposed to PM(2.5), the lung inflammatory response and airway remodeling may be modified, but that this altered response is dependent upon the sequence of exposure and the duration of exposure to ovalbumin aerosol. At the concentrations of PM tested, we did not see changes in airway fibrosis or airway reactivity for animals exposed to ovalbumin and PM(2.5) as compared with animals exposed only to ovalbumin aerosol. However, goblet-cell hyperplasia was significantly increased in mice exposed concurrently to ovalbumin and PM(2.5) as compared with mice exposed to ovalbumin alone.


Vascular endothelial growth factor in porcine coronary arteries following balloon angioplasty.
The present study was a preliminary inquiry into the presence of vascular endothelial growth factor (VEGF) in a model of coronary artery response to injury. We examined domestic pigs who had received a diet enriched in saturated fat and cholesterol and undergone balloon angioplasty of one or more coronary arteries. Immunohistochemical analysis of the coronary arteries 2 months after injury revealed the presence of VEGF distributed throughout the media and neointima of the angioplasty lesions and in association with blood vessels in the adventitia and those vessels growing into the base of the neointima. VEGF was also detected in areas of dietary-induced intimal proliferation. This study provided the first immunochemical demonstration of VEGF occuring naturally in a pig model of coronary response to injury.


Does the father's job matter? Parental occupation and preterm birth in Korea.
Limited evidence is available regarding the impact of paternal occupation and its combined effect with maternal occupation on preterm birth. Therefore, we assessed the association of maternal and paternal occupations with preterm birth. We used the national birth data of Korea between 2010 and 2020. Parental occupations were divided into 5 categories: (1) managers; (2) professionals, technicians, and related workers; (3) clerks and support workers; (4) service and sales workers; and (5) manual workers. A multinomial logistic regression model was used to calculate the adjusted odds ratios (aORs) of extremely, very, and moderate-to-late preterm births per occupational category considering individual risk factors. For the 4,004,976 singleton births, 40.2% of mothers and 95.5% of fathers were employed. Compared to non-employment, employment was associated with a lower risk of preterm birth. Among employed mothers, service and sales occupations were associated with a higher risk of preterm birth than managerial occupations (aOR, 1.06; 95% confidence interval [CI], 1.01 to 1.10 for moderate-to-late preterm births). The father's manual occupation was associated with a higher risk of preterm birth (aOR, 1.09; 95% CI, 1.05 to 1.13 for moderate-to-late preterm) than managerial occupations. When both parents had high-risk occupations, the risk of preterm birth was higher than in cases where only the mother or neither of the parents had a high-risk occupation. Paternal occupation was associated with preterm birth regardless of maternal employment and occupation and modified the effect of maternal occupation. Detailed occupational environment data are needed to identify the paternal exposures that increase the risk.


Correlates of longitudinal leukocyte telomere length in the Costa Rican Longevity Study of Healthy Aging (CRELES): On the importance of DNA collection and storage procedures.
The objective is to identify cofactors of leukocyte telomere length (LTL) in a Latin American population, specifically the association of LTL with 36 socio-demographic, early childhood, and health characteristics, as well as with DNA sample collection and storage procedures. The analysis is based on longitudinal information from a subsample of 1,261 individuals aged 60+ years at baseline from the Costa Rican Study of Longevity and Healthy Aging (CRELES): a nationally representative sample of elderly population. Random effects regression models for panel data were used to estimate the associations with LTL and its longitudinal changes. Sample collection procedures and DNA refrigerator storage time were strongly associated with LTL: telomeres are longer in blood collected in October-December, in DNA extracted from <1-year-old blood cells, and in DNA stored at 4°C for longer periods of time up to five years. The data confirmed that telomeres are shorter at older ages, as well as among males, and diabetic individuals, whereas telomeres are longer in the high-longevity Nicoya region. Most health, biomarkers, and early childhood indicators did not show significant associations with LTL. Longitudinal LTL variation over approximately two years was mainly associated with baseline LTL levels, as found in other studies. Our findings suggest that if there is unavoidable variability in season of sample collection and DNA storage time, these factors should be controlled for in all demographic and epidemiologic studies of LTL. However, due to unobserved components of measurement variation, statistical control may be inadequate as compared to standardization of data collection procedures.


Purification of antibody to galactosyl-protein conjugates.
A rapid procedure for purification of rabbit antibody to beta-D-galacto-sylphenylazo-bovine serum albumin is described. Antibody was precipitated with a heterologous antigen and was then dissociated from the precipitated complex with hapten. The antigen was partially removed by pH adjustment, and antibody was separated from residual antigen and from hapten by partition chromatography on Sephadex G-25. The antibody, obtained in 50 to 55 percent over-all yield, was 96 percent reactive with homologous antigen. It appeared to consist, of 95 percent of a single component, based upon physicochemical measurements.


[The participation of the genetic apparatus in the mechanisms of memory trace formation: the role of the calcium-regulatory system of the neurons in rats].
The dynamics of changes in some components of the calcium-regulated system of cortical and hippocampal neurons under the influence of long-term memory enhancers (ethylnorantifein and its analogues M1 and M2) was studied in rat brain. No change was found in the activity of transport Mg, Ca-ATPase and actomyosin-like Ca-ATPase in synaptic membranes 5, 15, 60, and 180 min after the injection of memory enhancers. The activation of the RNA transcription (60 min after the injection) was accompanied by an appreciable increase in activity of the chromatin Ca-ATPase. The amplification of synaptosomal protein synthesis (180 min) was accompanied by an increase in the activation of protein kinase C of synaptic membranes. The increase in Ca-ATPase activity of chromatin was also shown during the consolidation of the conditioned active avoidance in rats. The increase in the activity of protein kinase C seems to be connected with secondary rearrangement in synaptic membranes. The role in the long-term memory is discussed of direct activation of the genetic apparatus by neuroactive substances.


Fifth metacarpal stress fracture in a female softball pitcher.
This article describes a previously unreported fifth metacarpal stress fracture. A female softball pitcher presented with gradual development of dorsal pain in her pitching hand. A radioisotope scan showed increased uptake in the fifth metacarpal. This supported a clinical diagnosis of a stress fracture, and the patient was treated with a period of relative rest. At 6 weeks, the patient was able to return to pitching with technique modification. Seven cases of metacarpal stress fractures have been reported, yet none involved the fifth metacarpal. The causes of a fifth metacarpal stress fracture differ from those of other metacarpals and may be a combination of extrinsic forces from the ball and intrinsic forces from muscle pull. Although treatment involves the standard rest period, technique must be assessed and appropriately adjusted. Metacarpal stress fractures should be considered in athletes with persistent hand pain where repetitive grip function is used.


Adaptation and competition in deteriorating environments.
Evolution might rescue populations from extinction in changing environments. Using experimental evolution with microalgae, we investigated if competition influences adaptation to an abiotic stressor, and vice versa, if adaptation to abiotic change influences competition. In a first set of experiments, we propagated monocultures of five species with and without increasing salt stress for approximately 180 generations. When assayed in monoculture, two of the five species showed signatures of adaptation, that is, lines with a history of salt stress had higher population growth rates at high salt than lines without prior exposure to salt. When assayed in mixtures of species, however, only one of these two species had increased population size at high salt, indicating that competition can alter how adaptation to abiotic change influences population dynamics. In a second experiment, we cultivated two species in monocultures and in pairs, with and without increasing salt. While we found no effect of competition on adaptation to salt, our experiment revealed that evolutionary responses to salt can influence competition. Specifically, one of the two species had reduced competitive ability in the no-salt environment after long-term exposure to salt stress. Collectively, our results highlight the complex interplay of adaptation to abiotic change and competitive interactions.


[Molecular and cellular bases for the effects of interferon].
The data on the molecular mechanisms of interferons activity are analyzed. The analysis is based on the assumption that interferons are the substances of the hormonal nature. The initial step of the different interferons effects is attempted to be explained by the existence of the molecular and cellular reactions common for its realization. The dependence of the final results of interferons action on the metabolic processes in different cells and tissues determined by the specific functions and physiology of the latter is assumed.


Examining variation in treatment costs: a cost function for outpatient methadone treatment programs.
To estimate a hybrid cost function of the relationship between total annual cost for outpatient methadone treatment and output (annual patient days and selected services), input prices (wages and building space costs), and selected program and patient case-mix characteristics. Data are from a multistate study of 159 methadone treatment programs that participated in the Center for Substance Abuse Treatment's Evaluation of the Methadone/LAAM Treatment Program Accreditation Project between 1998 and 2000. Using least squares regression for weighted data, we estimate the relationship between total annual costs and selected output measures, wages, building space costs, and selected program and patient case-mix characteristics. Findings indicate that total annual cost is positively associated with program's annual patient days, with a 10 percent increase in patient days associated with an 8.2 percent increase in total cost. Total annual cost also increases with counselor wages (p<.01), but no significant association is found for nurse wages or monthly building costs. Surprisingly, program characteristics and patient case mix variables do not appear to explain variations in methadone treatment costs. Similar results are found for a model with services as outputs. This study provides important new insights into the determinants of methadone treatment costs. Our findings concur with economic theory in that total annual cost is positively related to counselor wages. However, among our factor inputs, counselor wages are the only significant driver of these costs. Furthermore, our findings suggest that methadone programs may realize economies of scale; however, other important factors, such as patient access, should be considered.


Mycobacterium tuberculosis complex and human coadaptation: a two-way street complicating host susceptibility to TB.
For centuries, the Mycobacterium tuberculosis complex (MTBC) has infected numerous populations, both human and non-human, causing symptomatic tuberculosis (TB) in some hosts. Research investigating the MTBC and how it has evolved with its host over time is sparse and has not resulted in many significant findings. There are even fewer studies investigating adaptation of the human host susceptibility to TB and these have largely focused on genome-wide association and candidate gene association studies. However, results emanating from these association studies are rarely replicated and appear to be population specific. It is, therefore, necessary to relook at the approach taken to investigate the relationship between the MTBC and the human host. Understanding that the evolution of the pathogen is coupled to the evolution of the host might be the missing link needed to effectively investigate their relationship. We hypothesize that this knowledge will bolster future efforts in combating the disease.


Preclinical anticancer effects and toxicologic assessment of hepatic artery infusion of fine-powder cisplatin with lipiodol in vivo.
We conducted an in vivo study to evaluate the anticancer effect and toxicity of fine-powder cisplatin suspended in lipiodol (fCDDP/LPD suspension) after a single administration of three different doses to rats via the intrahepatic artery after transplantation of rat ascites hepatoma cells. The toxicity of the fCDDP/LPD suspension was also assessed in the same protocol in noncancer-bearing rats and the observed toxicologic changes were compared among groups administered saline (Sal), an aqueous solution of fCDDP (fCDDP/Sal solution), and LPD alone. In parallel with the toxicity test, plasma CDDP concentrations were compared between the fCDDP/LPD suspension and fCDDP/Sal solution. The mean weight of the tumors in the fCDDP/LPD suspension groups was significantly less than in the LPD-alone group. The pathologic changes in the liver observed in the fCDDP/LPD suspension group increased with dose, were more marked compared with those in the fCDDP/Sal solution and LPD-alone groups, and were reversible. No other toxicologic effects were observed. The concentration of CDDP in the plasma in the fCDDP/LPD suspension group was slightly lower than that in the fCDDP/Sal solution group. In conclusion, the results indicate that the fCDDP/LPD suspension has sufficient anticancer efficacy and tolerability for use in the clinical treatment of hepatocellular carcinoma.


Tanshinone IIA promotes the apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis by up-regulating lncRNA GAS5.
Rheumatoid arthritis (RA) is a common chronic autoimmune joint disease characteristic of elevated proliferation and infiltration of fibroblast-like synoviocytes (FLS). Here, we aimed to explore the mechanisms of the Tanshinone IIA (Tan IIA)-induced apoptosis of FLS from patients with RA (termed RAFLS). Cell Counting Kit-8 (CCK-8) assay and Annexin V staining revealed that RAFLS viability decreased and apoptosis increased after Tan IIA treatment. Long non-coding RNA (lncRNA) GAS5 expression was significantly decreased in the synovial tissues and RAFLS, while Tan IIA treatment resulted in an up-regulation of GAS5. Consistently, knockdown of GAS5 using siRNA inhibited RAFLS apoptosis. Mechanistically, GAS5 knockdown down-regulated the expression of cleaved caspase-3 and caspase-9 in the RAFLS cells and activated the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. These data indicate that Tan IIA promotes RAFLS apoptosis by up-regulating lncRNA GAS5, with enhanced expression of cleaved caspase-3/caspase-9 and inhibited PI3K/AKT signaling.


Endobronchial argon plasma coagulation for treatment of Dieulafoy disease.
Dieulafoy disease of the bronchus is a rare condition. A 28-y-old male patient with recurrent exacerbations of hemoptysis (producing up to 300 mL) over the previous 4 y was admitted to our hospital following an acute episode. A 3-mm sessile lesion covered with somewhat ulcerated smooth mucosa was detected at the distal end of the left main bronchus, at the entrance to the upper lobe bronchus. Bleeding was controlled by argon plasma coagulation, and total coagulation of the lesion was also therapeutic. The patient has been followed for 2 y with no further bleeding. In the very few reported cases of Dieulafoy disease, surgical resection and selective embolization were the treatments used. Endobronchial treatment has not been described before as a therapeutic approach. We emphasize that argon plasma coagulation can be a treatment option in such cases.


Symptoms of food allergy.
Adverse reactions to foods can be due to many causes, but only those involving an immunological mechanism can be defined as food allergic disease. An increasing number of gastrointestinal and other diseases are being shown to involve food intolerances. Immediate reactions with symptoms within hours of eating a particular food are most readily shown to be due to food allergy and are often associated with the presence of food-specific IgE as shown by skin prick tests and RASTs. When reactions are delayed for 24 to 48 hours or more, underlying food intolerance is harder to recognize and much less often shown to be due to allergy. At present, diagnosis and management depends on dietary manipulation, showing that symptoms improve on food avoidance and are reproduced by food challenge (preferably double-blind). Further understanding of the mechanisms involved in food allergy, in Crohn's disease and irritable bowel syndrome may allow the development of simple tests to identify the foods concerned and perhaps, in the case of allergic disease, cure by the induction of tolerance.


Applications of Light Emitting Diodes in Health Care.
Light emitting diodes (LEDs) have become the main light sources for general lighting, due to their high lumen efficiency and long life time. Moreover, their high bandwidth and the availability of diverse wavelength contents ranging from ultraviolet to infrared empower them with great controllability in tuning brightness, pulse durations and spectra. These parameters are the essential ingredients of the applications in medical imaging and therapies. Despite the fast advances in both LED technologies and their applications, few reviews have been seen to link the controllable emission properties of LEDs to these applications. The objective of this paper is to bridge this gap by reviewing the main control techniques of LEDs that enable creating enhanced lighting patterns for imaging and generating effective photon doses for photobiomodulation. This paper also provides the basic mechanisms behind the effective LED therapies in treating cutaneous and neurological diseases. The emerging field of optogenetics is also discussed with a focus on the application of LEDs. The multidisciplinary topics reviewed in this paper can help the researchers in LEDs, imaging, light therapy and optogenetics better understand the basic principles in each other's field; and hence to stimulate the application of LEDs in health care.


A novel recognition site on laminin for the alpha 3 beta 1 integrin.
Synthetic peptide GD-2 is a sequence of amino acids derived from the carboxy-terminal long arm of the A chain of laminin. Previous studies have shown that peptide GD-2 promotes the adhesion of human squamous cell carcinoma (SCC) cells as well as a variety of other cell lines. In this study, we attempted to identify the receptor that SCC cells use to adhere to peptide GD-2. Monoclonal antibodies (mAbs) against a human SCC cell line were generated. One of these mAbs, ASC-1, bound to the surface of SCC cells as determined by flow cytometry. This mAb inhibited SCC cell adhesion to peptide GD-2 and laminin, but not fibronectin or type IV collagen, suggesting that mAb ASC-1 binds to the SCC receptor for the peptide GD-2 sequence of laminin. MAb ASC-1 immunoprecipitated a complex composed of two components of 135 and 116 kDa. Immunoadsorption of ASC-1-binding material from the SCC cell extract by incubation with mAb ASC-1 resulted in the removal of the alpha 3 beta 1 integrin from the extract. Immunohistochemical staining of tissue from a normal human tongue and from a patient with SCC of the tongue revealed that mAb ASC-1 stained the surface of epithelial cells that were in contact with the basement membrane, as well as those cells located two to three layers above the basement membrane. This mAb also stained blood vessels in the squamous tissue. This staining pattern was identical to that observed when the same tissues were stained by a mAb against the alpha 3 integrin subunit. In summary, by use of a new mAb, ASC-1, that recognizes the alpha 3 beta 1 integrin, we have determined that the alpha 3 beta 1 integrin mediates SCC cell adhesion to the peptide GD-2 sequence within laminin.


Photochemical Chemoselective Alkylation of Tryptophan-Containing Peptides.
We report a photochemical method for the chemoselective radical functionalization of tryptophan (Trp)-containing peptides. The method exploits the photoactivity of an electron donor-acceptor complex generated between the tryptophan unit and pyridinium salts. Irradiation with weak light (390 nm) generates radical intermediates right next to the targeted Trp amino acid, facilitating a proximity-driven radical functionalization. This protocol exhibits high chemoselectivity for Trp residues over other amino acids and tolerates biocompatible conditions.


Highlighting allelic variations at the interleukin-19 locus in term of preeclampsia predisposing factors and access to an accurate diagnostic/screening option.
Preeclampsia is the main cause of preterm parturition and maternal-fetal complications. T helper 1 and T helper 2 cytokines balance is a requirement in normal pregnancy and aberrant in this immunologic balance, play an important role in the pathology of preeclampsia. In previous studies single nucleotide polymorphisms have been associated with the alteration of serum cytokine levels. This study was aimed to discover association between interleukin-13 (rs20541, and rs56035208) and interleukin-19 (rs1028181 (T/C) and rs2243191(T/C)) polymorphisms with susceptibility to preeclampsia. In this case-control study 300 women with and without preeclampsia (n = 150/each) who referred to Zeynabieh Hospital- Shiraz, Iran, from February 2021 to April 2022 were enrolled. For genotyping the interleukin-13 and interleukin-19 polymorphisms, the Allele-specific polymerase chain reaction and direct sequencing method was carried out. Our statistical results revealed no significant differences in allele and genotype frequencies for interleukin-13 polymorphisms compared to controls. We found that the interleukin-13 polymorphisms are significantly associated with vulnerability to edema at rs20541 position and maternal drinking at rs56035208 position. But it was interesting to note that the differences of both the allele and genotype frequencies of interleukin-19 polymorphisms and their contribution to the risk of preeclampsia susceptibility were significant. No risk of preeclampsia was found in all comparisons for interleukin-13 polymorphisms. However, the interleukin-19 polymorphisms were found to confer the risk of preeclampsia in our population.


Multi-modality imaging of the leaking ureter: why does detection of traumatic and iatrogenic ureteral injuries remain a challenge?
Ureteral injuries are uncommon in trauma patients, accounting for fewer than 1% of all injuries to the urinary tract. These uncommon, yet problematic, injuries can often be overlooked in the standard search pattern on abdominal and pelvic multi-detector CT (MDCT) images, as radiologists focus on more immediate life-threatening injuries. However, early diagnosis and management are vital to reduce potential morbidity. If there is a high clinical index of suspicion for ureteral injuries with penetrating or blunt trauma, or if there is suspected iatrogenic ureteral injury, delayed-phase/urographic-phase MDCT images are essential for confirming the diagnosis. Moreover, making the distinction between partial and complete ureteral transection is critical, as it will guide management. The aim of this pictorial review is to overview the key imaging findings in blunt and penetrating traumatic and iatrogenic injuries of the ureter, as well as to discuss the advantages and disadvantages of different imaging modalities for accurately and rapidly establishing or excluding the diagnosis of ureteral injuries, with an emphasis on MDCT. The potential causes of missed ureteral injuries will also be discussed.


Patients highly value routine follow-up of skin cancer and cutaneous melanoma.
Skin cancer follow-up is a substantial burden to outpatient clinics. Few studies have investigated patients' views on skin cancer follow-up and cutaneous melanoma. The objective was to investigate patients' perceived benefits and the impact of follow-up. This study included an open sample of patients attending routine follow-up at the outpatient Departments of Plastic Surgery and Dermatology, Roskilde Hospital. A total of 218 follow-up patients diagnosed with cutaneous malignant melanoma (MM), non-melanoma skin cancer (NMSC) or actinic keratosis (AK) completed a structured interview. A total of 97% patients found follow-up useful. Continuity and consistency were important. One third of patients felt some degree of pre follow-up anxiety. The number of anxious MM patients was significantly greater than that of NMSC patients. No significant difference was found between the number of anxious MM and AK patients. Female gender, cohabitation and age younger than 50 years were associated with increased levels of anxiety. No relation was found between the number of anxious patients or the level of anxiety and the duration of the follow-up period. The majority of patients who attended found that the follow-up had been useful. Certain demographic characteristics were associated with higher levels of anxiety and may be addressed by supportive efforts targeting these groups.


Routine Angiographic Follow-Up versus Clinical Follow-Up after Percutaneous Coronary Intervention in Acute Myocardial Infarction.
Differences in the utility of routine angiographic follow-up (RAF) and clinical follow-up (CF) after percutaneous coronary intervention (PCI) in acute myocardial infarction (AMI) are not well understood. The present study aimed to compare the 3-year clinical outcomes of RAF and CF in AMI patients who underwent PCI with drug-eluting stents (DES). A total of 774 consecutive AMI patients who underwent PCI with DES were enrolled. RAF was performed at 6 to 9 months after index PCI (n=425). The remaining patients were medically managed and clinically followed (n=349); symptom-driven events were captured. To adjust for any potential confounders, a propensity score matched analysis was performed using a logistic regression model, and two propensity-matched groups (248 pairs, n=496, C-statistic=0.739) were generated. Cumulative clinical outcomes up to 3 years were compared between RAF and CF groups. During the 3-year follow-up period, the cumulative incidences of revascularization [target lesion revascularization: hazard ratio (HR), 2.40; 95% confidence interval (CI), 1.18-4.85; p=0.015, target vessel revascularization (TVR): HR, 3.33; 95% CI, 1.69-6.58; p=0.001, non-TVR: HR, 5.64; 95% CI, 1.90-16.6; p=0.002] and major adverse cardiac events (MACE; HR, 3.32; 95% CI, 1.92-5.73; p<0.001) were significantly higher in the RAF group than the CF group. However, the 3-year incidences of death and myocardial infarction were not different between the two groups. RAF following index PCI with DES in AMI patients was associated with increased incidences of revascularization and MACE. Therefore, CF seems warranted for asymptomatic patients after PCI for AMI.


[Size of the lesioned portion of the retina after laser exposure (electron microscopic study)].
Morphological pattern of the retina in the trauma focus and outside its borders has been studied electron microscopically after application of ruby laser monoimpulse radiation (wave length 0.69 mcm, energy 1.06-1.25 X 10(-4) J and impulse length 20 nc). The trauma focus is not limited only by the volcano-shaped inflation of the retina. The lesion of the photoreceptory ultrastructure and the disturbance of microcirculation in the vascular layer are defined in 3 zones investigated: at the distance 50, 100 and 1,000 mcm from the spot on the retina, visible in the light microscope. This phenomenon should be taken into consideration when choosing therapeutic doses of laser radiation, applied for treatment of various diseases of the eye.


A locally adaptive regularization based on anisotropic diffusion for deformable image registration of sliding organs.
We propose a deformable image registration algorithm that uses anisotropic smoothing for regularization to find correspondences between images of sliding organs. In particular, we apply the method for respiratory motion estimation in longitudinal thoracic and abdominal computed tomography scans. The algorithm uses locally adaptive diffusion tensors to determine the direction and magnitude with which to smooth the components of the displacement field that are normal and tangential to an expected sliding boundary. Validation was performed using synthetic, phantom, and 14 clinical datasets, including the publicly available DIR-Lab dataset. We show that motion discontinuities caused by sliding can be effectively recovered, unlike conventional regularizations that enforce globally smooth motion. In the clinical datasets, target registration error showed improved accuracy for lung landmarks compared to the diffusive regularization. We also present a generalization of our algorithm to other sliding geometries, including sliding tubes (e.g., needles sliding through tissue, or contrast agent flowing through a vessel). Potential clinical applications of this method include longitudinal change detection and radiotherapy for lung or abdominal tumours, especially those near the chest or abdominal wall.


Primary lipomatous tumors of the cardiac valves.
A tricuspid valve mass was identified on echocardiogram in a 69-year-old man with ischemic heart disease, chronic obstructive pulmonary disease, pulmonary embolism, and pneumonia; he died with progressive respiratory insufficiency. The abnormal mass was ascribed initially to infective endocarditis, and the diagnosis at autopsy was fibrolipoma, a benign tumor. Although rare, valve tumors should be included in the differential diagnosis of abnormal valve masses identified on echocardiogram.


SERS-based immunoassay on a plasmonic syringe filter for improved sampling and labeling efficiency of biomarkers.
Rapid, sensitive, and quantitative detection of biomarkers is needed for early diagnosis of disease and surveillance of infectious outbreaks. Here, we exploit a plasmonic syringe filter and surface-enhanced Raman spectroscopy (SERS) in the development of a rapid detection system, using human IgG as a model diagnostic biomarker. The novel assay design facilitates multiple passages of the sample and labeling solution through the detection zone enabling us to investigate and maximize sampling efficiency to the capture substrate. The vertical flow immunoassay process in this study involves the utilization of filter paper embedded with gold nanoparticles (AuNPs) to form a plasmonic substrate. Capture antibody (anti-human IgG) is then immobilized onto the prepared plasmonic paper and inserted into a vertical flow device (syringe filter holder). Sample solution is passed through the filter paper and the target antigen (human IgG) is selectively captured by the immobilized antibody to form an antibody-antigen complex. Next, functionalized AuNPs as extrinsic Raman labels (ERLs) are passed through the filter paper to label the captured biomarker molecules forming a layered structure. This sandwiched geometry enhances plasmonic coupling and SERS signal to provide highly sensitive detection of biomolecules. Systematic studies to investigate the impact of multiple infuse/withdraw cycles of the sample and labeling solutions reveal that antigen and ERL binding are maximized with 10 and 20 cycles, respectively. The optimized assay achieves a detection limit of ∼0.2 ng mL-1 for human IgG with a total assay time of less than 5 minutes, meeting the demands for rapid point of care diagnostics. Additionally, the optimized platform was implemented in the quantitative analysis of the SARS-CoV-2 nucleocapsid protein, the typical target in commercial, FDA-approved rapid antigen tests for COVID-19.


Detecting Differences of Fluorescent Markers Distribution in Single Cell Microscopy: Textural or Pointillist Feature Space?
We consider the detection of change in spatial distribution of fluorescent markers inside cells imaged by single cell microscopy. Such problems are important in bioimaging since the density of these markers can reflect the healthy or pathological state of cells, the spatial organization of DNA, or cell cycle stage. With the new super-resolved microscopes and associated microfluidic devices, bio-markers can be detected in single cells individually or collectively as a texture depending on the quality of the microscope impulse response. In this work, we propose, via numerical simulations, to address detection of changes in spatial density or in spatial clustering with an individual (pointillist) or collective (textural) approach by comparing their performances according to the size of the impulse response of the microscope. Pointillist approaches show good performances for small impulse response sizes only, while all textural approaches are found to overcome pointillist approaches with small as well as with large impulse response sizes. These results are validated with real fluorescence microscopy images with conventional resolution. This, a priori non-intuitive result in the perspective of the quest of super-resolution, demonstrates that, for difference detection tasks in single cell microscopy, super-resolved microscopes may not be mandatory and that lower cost, sub-resolved, microscopes can be sufficient.


A highly selective fluorescent sensor for glucosamine.
A new fluorescent chemical sensor for glucosamine is reported. The sensor is based on a boronic acid-containing coumarin aldehyde and shows excellent selectivity for glucosamine by forming a boronic ester with the sugar diol as well as an iminium ion with the amine group of glucosamine. The sensor successfully discriminates glucosamine over other similar biomolecules in terms of both fluorescence intensity and binding affinity. This method provides a new concept for the design and synthesis of very selective turn-on optical sensors for selective detection of multi-functional biomolecules.


Differentiated thyroid cancer in navarra (Spain): historic cohort results (1987-2003).
Introduction. Navarra has the highest incidence of differentiated thyroid cancer in Spain. The aim of this study was to review its management carried out by the Navarra's multidisciplinary Thyroid Disease Unit, from 1987 to 2003. Material and Methods. 325 patients were studied to find the incidence, prevalence, and prognostic factors. Statistical analysis comprised univariate and multivariate Cox proportional hazards regression models for survival and tumor recurrence. Results. The average annual incidence was 3.6 per 100,000 inhabitants, with a final prevalence of 82.4 per 100,000. Regarding survival and recurrence, statistical significance was observed for stage IV, follicular carcinoma, capsular and prethyroid muscles invasion, and T4 group. Only survival was related to tumour size larger than 40 mm. Only recurrence was related to lymph node metastases and radioiodine dose higher than 100 mCi. Conclusions. Attendance of patients in a functional unit setting has allowed us to classify them into three risk groups.


Allicin alleviates inflammation of trinitrobenzenesulfonic acid-induced rats and suppresses P38 and JNK pathways in Caco-2 cells.
Background. Allicin has anti-inflammatory, antioxidative and proapoptotic properties. Aims. To evaluate the effects and investigate the mechanism of allicin on trinitrobenzenesulfonic acid-induced colitis, specifically with mesalazine or sulfasalazine. Methods. 80 rats were divided equally into 8 groups: control; trinitrobenzenesulfonic acid; allicin prevention; allicin; mesalazine; sulfasalazine; allicin + sulfasalazine, and mesalazine + allicin. Systemic and colonic inflammation parameters were analysed. In addition, protein and culture medium of Caco-2 cells treated with various concentrations of IL-1β or allicin were collected for investigation of IL-8, NF-κB p65 P38, ERK, and JNK. One-way ANOVA and Kruskal-Wallis H test were used for parametric and nonparametric tests, respectively. Results. Allicin reduced the body weight loss of trinitrobenzenesulfonic acid-induced rats, histological score, serum TNF-α and IL-1β levels, and colon IL-1β mRNA level and induced serum IL-4 level, particularly in combination with mesalazine. In addition, 1 ng/mL IL-1β stimulated the P38, ERK, and JNK pathways, whereas pretreatment with allicin depressed this phenomenon, except for the ERK pathway. Conclusions. The inflammation induced by trinitrobenzenesulfonic acid is mitigated significantly by allicin treatment, particularly combined with mesalazine. Allicin inhibits the P38 and JNK pathways and the expression of NF-κB which explained the potential anti-inflammatory mechanisms of allicin.


Detection of First Marker Trait Associations for Resistance Against Sclerotinia sclerotiorum in Brassica juncea-Erucastrum cardaminoides Introgression Lines.
A set of 96 Brassica juncea-Erucastrum cardaminoides introgression lines (ILs) were developed with genomic regions associated with Sclerotinia stem rot (Sclerotinia sclerotiorum) resistance from a wild Brassicaceous species E. cardaminoides. ILs were assessed for their resistance responses to stem inoculation with S. sclerotiorum, over three crop seasons (season I, 2011/2012; II, 2014/2015; III, 2016-2017). Initially, ILs were genotyped with transferable SSR markers and subsequently through genotyping by sequencing. SSR based association mapping identified six marker loci associated to resistance in both A and B genomes. Subsequent genome-wide association analysis (GWAS) of 84 ILs recognized a large number of SNPs associated to resistance, in chromosomes A03, A06, and B03. Chromosomes A03 and A06 harbored the maximum number of resistance related SNPs. Annotation of linked genomic regions highlighted an array of resistance mechanisms in terms of signal transduction pathways, hypersensitive responses and production of anti-fungal proteins and metabolites. Of major importance was the clustering of SNPs, encoding multiple resistance genes on small regions spanning approximately 885 kb region on chromosome A03 and 74 kb on B03. Five SNPs on chromosome A03 (6,390,210-381) were associated with LRR-RLK (receptor like kinases) genes that encode LRR-protein kinase family proteins. Genetic factors associated with pathogen-associated molecular patterns (PAMPs) and effector-triggered immunity (ETI) were predicted on chromosome A03, exhibiting 11 SNPs (6,274,763-994). These belonged to three R-Genes encoding TIR-NBS-LRR proteins. Marker trait associations (MTAs) identified will facilitate marker assisted introgression of these critical resistances, into new cultivars of B. juncea initially and, subsequently, into other crop Brassica species.


Comparison between the polypeptide profile of halophilic bacteria and salt tolerant plants.
Changes in the polypeptide profile induced by salt stress in halotolerant and halophilic bacteria, isolated from the Atacama desert (northern Chile), were compared with those in the cotyledons of Prosopis chilensis (Leguminoseae) seedlings, a salt tolerant plant. SDS-PAGE analyses show the presence of four predominant polypeptides, with molecular weights around 78, 70, 60 and 44 kDa respectively, both in bacteria and in cotyledons from P. chilensis seedlings raised under salt stress conditions. Moreover, the 60 and 44 kDa polypeptides seem to be salt responsive, since their concentration increases with increasing NaCl in the growth medium. Our results suggest a common mechanism for salt tolerance in prokaryotes and in eukaryotes.


[Health economics analysis of diabetes is necessary. It facilitates decision-making and international comparison].
Cost-of-illness studies have shown diabetes to be associated with substantial direct and indirect costs, accounting for 5-6 percent of total health care expenditure. In a Swedish study, where total costs were divided into costs due to management of diabetes and costs due to complications, the total annual cost to the community was estimated to be SEK 5.7 billion in 1994, costs due to complications being the major item, accounting for over 75 per cent of the total. There have been few other Swedish studies of costs for diabetes or diabetes-related complications. The most widely studied category of complications is diabetes-related foot ulcers, with an estimated annual cost of SEK 1-2 billion. However, earlier studies were marred by shortcomings: costs estimated for the main diagnosis only, without breakdown into categories or distinction between type 1 and type 2 diabetes, sources of data other than official data-bases ignored, etc. Diabetes care in Sweden is of high quality, and substantial clinical, epidemiological and health economics research has been carried out. It is important that Sweden contributes to international research on health economics aspects of diabetes.


Effects of blood lead and cadmium levels on homocysteine level in plasma.
We studied the effect of non-occupational exposure to lead and cadmium on homocysteine level in plasma. Homocysteine is a marker for plasma folate folic acid metabolism in urban populations. 159 individuals from Beijing, Guangzhou, Shenzhen and Shanghai with no history of close exposure to heavy metals and no history of metabolic diseases were enrolled to participate in this study. Blood lead and cadmium levels were detected using ICP-MS method and the level of homocysteine was also measured using enzyme method. Our results showed that blood lead and cadmium levels in males were significantly higher than those in females. Also, blood lead and cadmium levels in smokers were higher than those in non-smokers; homocysteine level was significantly higher in smokers as well. According to blood lead and cadmium levels, cases were divided into four groups. Our results showed that a surge in blood lead and cadmium levels could result in an increase in homocysteine level. We concluded that in the Chinese population, smoking and gender might be the risk factors for elevated levels of lead and cadmium. Meanwhile, blood lead and cadmium levels may influence the homocysteine levels in the body. It is possible to speculate that non-occupational exposure to lead and cadmium, by increasing the homocysteine levels, negatively affect the cardiovascular and nervous system.


The Role of Endoplasmic Reticulum Stress in Differentiation of Cells of Mesenchymal Origin.
Endoplasmic reticulum (ER) is a multifunctional membrane-enclosed organelle. One of the major ER functions is cotranslational transport and processing of secretory, lysosomal, and transmembrane proteins. Impaired protein processing caused by disturbances in the ER homeostasis results in the ER stress. Restoration of normal ER functioning requires activation of an adaptive mechanism involving cell response to misfolded proteins, the so-called unfolded protein response (UPR). Besides controlling protein folding, UPR plays a key role in other physiological processes, in particular, differentiation of cells of connective, muscle, epithelial, and neural tissues. Cell differentiation is induced by the physiological levels of ER stress, while excessive ER stress suppresses differentiation and can result in cell death. So far, it remains unknown whether UPR activation induces cell differentiation or if UPR is initiated by the upregulated synthesis of secretory proteins during cell differentiation. Cell differentiation is an important stage in the development of multicellular organisms and is tightly controlled. Suppression or excessive activation of this process can lead to the development of various pathologies in an organism. In particular, impairments in the differentiation of connective tissue cells can result in the development of fibrosis, obesity, and osteoporosis. Recently, special attention has been paid to fibrosis as one of the major complications of COVID-19. Therefore, studying the role of UPR in the activation of cell differentiation is of both theoretical and practical interest, as it might result in the identification of molecular targets for selective regulation of cell differentiation stages and as well as the potential to modulate the mechanisms involved in the development of various pathological states.


Turning Cell Adhesions ON or OFF with High Spatiotemporal Precision Using the Green Light Responsive Protein CarH.
Spatiotemporal control of integrin-mediated cell adhesions to extracellular matrix regulates cell behavior with has numerous implications for biotechnological applications. In this work, two approaches for regulating cell adhesions in space and time with high precision are reported, both of which utilize green light. In the first design, CarH, which is a tetramer in the dark, is used to mask cRGD adhesion-peptides on a surface. Upon green light illumination, the CarH tetramer dissociates into its monomers, revealing the adhesion peptide so that cells can adhere. In the second design, the RGD motif is incorporated into the CarH protein tetramer such that cells can adhere to surfaces functionalized with this protein. The cell adhesions can be disrupted with green light, due to the disassembly of the CarH-RGD protein. Both designs allow for photoregulation with noninvasive visible light and open new possibilities to investigate the dynamical regulation of cell adhesions in cell biology.


Population structure of invasive and colonizing strains of Streptococcus agalactiae from neonates of six U.S. Academic Centers from 1995 to 1999.
The purpose of this study was to describe the population structure of group B streptococci (GBS) isolated from infected and colonized neonates during a prospective active-surveillance study of early-onset disease in six centers in the United States from July 1995 to June 1999 and to examine its relationship to bovine strains of GBS. The phylogenetic lineage of each GBS isolate was determined by multilocus sequence typing, and isolates were clustered into clonal complexes (CCs) using the eBURST software program. A total of 899 neonatal GBS isolates were studied, of which 129 were associated with invasive disease. Serotype Ia, Ib, and V isolates were highly clonal, with 92% to 96% of serotype Ia, Ib, and V isolates being confined to single clonal clusters. In contrast, serotype II and III isolates were each comprised of two major clones, with 39% of serotype II and 41% of serotype III isolates in CC 17 and 41% of serotype II and 54% of serotype III isolates in CC 19. Further analysis demonstrates that the CC 17 serotype II and III GBS are closely related to a previously described "ancestral" lineage of bovine GBS. While 120 (93%) of invasive GBS were confined to the same lineages that colonized neonates, 9 (7%) of the invasive GBS isolates were from rare lineages that comprised only 2.7% of colonizing lineages. These results are consistent with those for other geographic regions that demonstrate the highly clonal nature of GBS infecting and colonizing human neonates.


Enantioselective nitrone cycloadditions of alpha,beta-unsaturated 2-acyl imidazoles catalyzed by bis(oxazolinyl)pyridine-cerium(IV) triflate complexes.
[Structure: see text] Enantioselective nitrone cycloadditions with beta-substituted alpha,beta-unsaturated 2-acyl imidazoles catalyzed by bis(oxazolinyl)pyridine-cerium(IV) triflate complexes 1 have been reported. The isoxazolidine products were efficiently transformed into densely functionalized beta'-hydroxy-beta-amino acid derivatives.


hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization.
Telomerase, the ribonucleoprotein enzyme that elongates telomeres, is repressed in normal human somatic cells but is reactivated during tumor progression. We report the cloning of a human gene, hEST2, that shares significant sequence similarity with the telomerase catalytic subunit genes of lower eukaryotes. hEST2 is expressed at high levels in primary tumors, cancer cell lines, and telomerase-positive tissues but is undetectable in telomerase-negative cell lines and differentiated telomerase-negative tissues. Moreover, the message is up-regulated concomitant with the activation of telomerase during the immortalization of cultured cells and down-regulated during in vitro cellular differentiation. Taken together, these observations suggest that the induction of hEST2 mRNA expression is required for the telomerase activation that occurs during cellular immortalization and tumor progression.


Historical Alternans in the Emergency Department (H.A.t.E.D.) for Pain: An Analysis of Patient Pain Descriptors to Attending and Trainee Clinicians.
The concept of "historical alternans" (HA), a teaching folklore term referring to different descriptions of patient histories to trainees versus attending clinicians, can cause disjointed care and be a source of frustration for the trainee. Increased focus on the proper evaluation and treatment of pain in the emergency department (ED) setting prompts an evaluation of the HA concept as it relates to pain. We conducted a prospective observational pilot study comparing pain descriptions given to attending and trainee clinicians in the ED using a five-question pain description survey. Trainees included emergency medicine physician residents, emergency medicine physician assistant residents, off-service residents, and students. Trainees completed the first survey and attending clinicians repeated survey questions after at least a 10-minute washout. Surveys include descriptions of pain as part of patients' primary concern, severity indicated by a verbal numerical rating score (VNRS), and pain location, quality, and duration. During a 10-day period, surveys were completed for 97 patient encounters. Most trainee clinicians were emergency medicine physician residents (53%), followed by emergency medicine physician assistants (32%), students (13%), and off-service residents (2%). Pain complaints centered on the abdomen (18.5%), chest (12%), and knee (6%). Differences in pain description were found in the majority of cases (55%), with most having one categorical difference. The majority of categorical differences were VNRS (38%), although the difference in scores was not statistically significant (P = 0.20). Medical students had the highest variance in VNRS difference compared with attending clinicians. There was no significant difference in described duration (P = 0.99) or quality of pain (P = 0.99) between trainee and attending clinicians. Most patient encounters had at least one difference in categorical pain descriptors between trainee and attending clinicians. Although differences in severity of pain were present, they were not significant. HA does occur in the ED setting, but the magnitude of difference may be minimal.


Motivational conditions influence tongue motor performance.
Motivation plays an important role in the outcome of motor learning but has not received attention in tongue-training-induced plasticity of the corticomotor pathways. The present study investigated the influence of two different motivational conditions and gender on performance during a complex tongue-training paradigm using the tongue drive system (TDS). In addition, subject-based reports of motivation, fun, pain, and fatigue were compared between groups and genders. Sixteen subjects were randomized into three groups and were asked to use the TDS for 40 min. A motivational condition (monetary reward or self-controlled practice) was introduced in two groups and the third group served as the control. The subjects were instructed to play a computer game using the TDS, having control of the computer cursor through a magnet attached to the tongue, and performance was compared among groups. Performance improved in all groups and in both genders. The monetary reward group tended towards higher performance scores compared with the control group, whereas the self-controlled practice group performed significantly better compared with the control group. There was no significant difference between groups and genders in the subject-based report for level of motivation, fun, pain, or fatigue. In conclusion, introduction of motivational conditions influenced tongue motor performance.


Color constancy using 3D scene geometry derived from a single image.
The aim of color constancy is to remove the effect of the color of the light source. As color constancy is inherently an ill-posed problem, most of the existing color constancy algorithms are based on specific imaging assumptions (e.g., gray-world and white patch assumption). In this paper, 3D geometry models are used to determine which color constancy method to use for the different geometrical regions (depth/layer) found in images. The aim is to classify images into stages (rough 3D geometry models). According to stage models, images are divided into stage regions using hard and soft segmentation. After that, the best color constancy methods are selected for each geometry depth. To this end, we propose a method to combine color constancy algorithms by investigating the relation between depth, local image statistics, and color constancy. Image statistics are then exploited per depth to select the proper color constancy method. Our approach opens the possibility to estimate multiple illuminations by distinguishing nearby light source from distant illuminations. Experiments on state-of-the-art data sets show that the proposed algorithm outperforms state-of-the-art single color constancy algorithms with an improvement of almost 50% of median angular error. When using a perfect classifier (i.e, all of the test images are correctly classified into stages); the performance of the proposed method achieves an improvement of 52% of the median angular error compared with the best-performing single color constancy algorithm.


Modelling cultural systems and selective filters.
A specific goal of the field of cultural evolution is to understand how processes of transmission and selection at the individual level lead to population-wide patterns of cultural diversity and change. Models of cultural evolution have typically assumed that traits are independent of one another and essentially exchangeable. But culture has a structure: traits bear relationships to one another that affect the transmission and selection process itself. Here, we introduce a modelling framework to explore the effect of interdependencies on the process of learning. Through simulations, we find that introducing a simple structure changes the cultural dynamics. Based on a basic filtering mechanism for parsing trait relationships, more elaborate cultural filters emerge. In a mostly incompatible cultural domain of traits, these filters organize culture into mostly (but not fully) consistent and stable systems. Incompatible domains produce small homogeneous cultures, while more compatibility increases size, diversity and group divergence. When individuals copy based on a trait's features (here, its compatibility relationships), they produce more homogeneous cultures than when they copy based on the agent carrying the cultural trait. We discuss the implications of considering cultural systems and filters in the dynamics of cultural change. This article is part of the theme issue 'Foundations of cultural evolution'.


Replication of Coxiella burnetii in a Lysosome-Like Vacuole Does Not Require Lysosomal Hydrolases.
Coxiella burnetii is an intracellular bacterium that causes query, or Q fever, a disease that typically manifests as a severe flu-like illness. The initial target of C. burnetii is the alveolar macrophage. Here, it regulates vesicle trafficking pathways and fusion events to establish a large replication vacuole called the Coxiella-containing vacuole (CCV). Similar to a phagolysosome, the CCV has an acidic pH and contains lysosomal hydrolases obtained via fusion with late endocytic vesicles. Lysosomal hydrolases break down various lipids, carbohydrates, and proteins; thus, it is assumed C. burnetii derives nutrients for growth from these degradation products. To investigate this possibility, we utilized a GNPTAB-/- HeLa cell line that lacks lysosomal hydrolases in endocytic compartments. Unexpectedly, examination of C. burnetii growth in GNPTAB-/- HeLa cells revealed replication and viability are not impaired, indicating C. burnetii does not require by-products of hydrolase degradation to survive and grow in the CCV. However, although bacterial growth was normal, CCVs were abnormal, appearing dark and condensed rather than clear and spacious. Lack of degradation within CCVs allowed waste products to accumulate, including intraluminal vesicles, autophagy protein LC3, and cholesterol. The build-up of waste products coincided with an altered CCV membrane, where LAMP1 was decreased and CD63 and LAMP1 redistributed from a punctate to uniform localization. This disruption of CCV membrane organization may account for the decreased CCV size due to impaired fusion with late endocytic vesicles. Collectively, these results demonstrate lysosomal hydrolases are not required for C. burnetii survival and growth but are needed for normal CCV development. These data provide insight into mechanisms of CCV biogenesis while raising the important question of how C. burnetii obtains essential nutrients from its host.


Predictability rather than amplitude of temperature fluctuations determines stress resistance in a natural population of Drosophila simulans.
The adaptability of organisms to novel environmental conditions depends on the amount of genetic variance present in the population as well as on the ability of individuals to adjust their phenotype through phenotypic plasticity. Here, we investigated the phenotypic plasticity induced by a single generation's exposure to three different temperature regimes with respect to several life-history and stress-resistance traits in a natural population of Drosophila simulans. We studied a constant as well as a predictably and an unpredictably fluctuating temperature regime. We found high levels of phenotypic plasticity among all temperature regimes, suggesting a strong influence of both temperature fluctuations and their predictability. Increased heat tolerance was observed for flies developed in both types of fluctuating thermal environments compared with flies developed in a constant environment. We suggest that this was due to beneficial hardening when developing in either fluctuating temperature environment. To our surprise, flies that developed in constant and predictably changing environments were similar to each other in most traits when compared to flies from the unpredictably fluctuating environment. The unpredictably changing thermal environment imposed the most stressful condition, resulting in the lowest performance for stress-related traits, even though the absolute temperature changes never exceeded that of the predictably fluctuating environment. The overall decreased stress resistance of flies in the unpredictably fluctuating environment may be the consequence of maladaptive phenotypic plasticity in this setting, indicating that the adaptive value of plasticity depends on the predictability of the environment.


Structure and expression of the Drosophila melanogaster gene for the U1 small nuclear ribonucleoprotein particle 70K protein.
A genomic clone encoding the Drosophila U1 small nuclear ribonucleoprotein particle 70K protein was isolated by hybridization with a human U1 small nuclear ribonucleoprotein particle 70K protein cDNA. Southern blot and in situ hybridizations showed that this U1 70K gene is unique in the Drosophila genome, residing at cytological position 27D1,2. Polyadenylated transcripts of 1.9 and 3.1 kilobases were observed. While the 1.9-kilobase mRNA is always more abundant, the ratio of these two transcripts is developmentally regulated. Analysis of cDNA and genomic sequences indicated that these two RNAs encode an identical protein with a predicted molecular weight of 52,879. Comparison of the U1 70K proteins predicted from Drosophila, human, and Xenopus cDNAs revealed 68% amino acid identity in the most amino-terminal 214 amino acids, which include a sequence motif common to many proteins which bind RNA. The carboxy-terminal half is less well conserved but is highly charged and contains distinctive arginine-rich regions in all three species. These arginine-rich regions contain stretches of arginine-serine dipeptides like those found in transformer, transformer-2, and suppressor-of-white-apricot proteins, all of which have been identified as regulators of mRNA splicing in Drosophila melanogaster.


Protection against Escherichia coli-induced urinary tract infections with hybridoma antibodies directed against type 1 fimbriae or complementary D-mannose receptors.
Hybridoma antibodies directed against quaternary structural epitopes of the type 1 fimbrial adhesin of Escherichia coli or against D-mannose, the sugar determinant in the complementary host cell receptor, prevented the attachment of mannose-sensitive E. coli to various eucaryotic cells. Passive intraperitoneal administration of the fimbria-specific or D-mannose-specific antibodies protected mice against retrograde colonization with mannose-sensitive E. coli instilled into their urinary bladders. Monoclonal antibodies directed against fimbrial subunits rather than quaternary structural epitopes or against N-acetylgalactosamine rather than D-mannose residues lacked protective activity. These studies provide evidence that bacterial colonization can be blocked or interrupted by antibodies directed against either the adhesin or the complementary host cell receptor of pathogenic microorganisms.


Double-gradient denaturing gradient gel electrophoresis assay for identification of L-ferritin iron-responsive element mutations responsible for hereditary hyperferritinemia-cataract syndrome: identification of the new mutation C14G.
Hereditary hyperferritinemia-cataract syndrome is an autosomic dominant disorder caused by heterogeneous mutations on the iron-responsive element (IRE) of ferritin L-chain mRNA. The mutations described to date were identified by direct sequencing of DNA from probands with hyperferritinemia often associated to bilateral cataracts. A direct genetic approach on a large population is useful to recognize polymorphisms in the DNA region and the prevalence of mutations associated with minor increases in serum ferritin and subclinical cataracts. We developed a rapid DNA scanning technique to detect mutations in a single electrophoretic analysis. The double-gradient denaturing gradient gel electrophoresis (DG-DGGE) method consisted of PCR amplification of the target genomic DNA with GC-clamped oligonucleotides. The sequence encoded the 5' untranslated flanking region of ferritin L-chain mRNA, which includes an IRE stem-loop structure. The product was subjected to DG-DGGE (8.5-15% polyacrylamide and 50-95% denaturant) to separate the homo- and heteroduplexes. The method clearly identified all eight accessible mutations, including C-G transversions, which are the most difficult to detect. The method was applied to scan DNA samples from 50 healthy subjects and from 230 subjects with serum ferritin >400 microg/L. The new mutation G14C was identified. The DG-DGGE method detects all the mutations in the L-ferritin IRE sequence, is rapid and economical, and can be applied to scan large populations. The first population study indicated that the mutations are rare and may involve regions of the IRE structure not yet characterized.


Multiscale Imaging Approach for Studying the Central Nervous System: Methodology and Perspective.
Non-invasive imaging methods have become essential tools for understanding the central nervous system (CNS) in health and disease. In particular, magnetic resonance imaging (MRI) techniques provide information about the anatomy, microstructure, and function of the brain and spinal cord in vivo non-invasively. However, MRI is limited by its spatial resolution and signal specificity. In order to mitigate these shortcomings, it is crucial to validate MRI with an array of ancillary ex vivo imaging techniques. These techniques include histological methods, such as light and electron microscopy (EM), which can provide specific information on the tissue structure in healthy and diseased brain and spinal cord, at cellular and subcellular level. However, these conventional histological techniques are intrinsically two-dimensional (2D) and, as a result of sectioning, lack volumetric information of the tissue. This limitation can be overcome with genuine three-dimensional (3D) imaging approaches of the tissue. 3D highly resolved information of the CNS achievable by means of other imaging techniques can complement and improve the interpretation of MRI measurements. In this article, we provide an overview of different 3D imaging techniques that can be used to validate MRI. As an example, we introduce an approach of how to combine diffusion MRI and synchrotron X-ray phase contrast tomography (SXRPCT) data. Our approach paves the way for a new multiscale assessment of the CNS allowing to validate and to improve our understanding of in vivo imaging (such as MRI).


The Impact of 4-Nonylphenol on the Viability and Hormone Production of Mouse Leydig Cells.
Exogenous substances altering the function of the endocrine system and exhibiting adverse health effects on the organism are defined as endocrine disruptors. Nonylphenol is one of the most abundant alkylphenol ethoxylate derivatives, being detected in food products. Diverse studies have classified nonylphenol as hazardous to the health, especially to male reproduction. This in vitro study aimed to examine the effects of 4-nonylphenol on androstenedione and testosterone production as well as on the viability of Leydig cells of NMRI mice. The cells were cultured for 44 h with addition of 0.04; 0.2; 1.0; 2.5 and 5.0 μg/ml of 4-nonylphenol and compared to the control. Quantification of testosterone and androstenedione directly from aliquots of the medium was performed by enzyme-linked immunosorbent assay. Cell viability was measured by the metabolic activity assay for mitochondrial functional activity. Androstenedione production significantly (P < 0.001) increased with 1.0; 2.5 and 5.0 μg/ml 4-nonylphenol. Although cAMP-stimulated testosterone production was not significantly affected by 4-nonylphenol, a tendency to attenuate the level of testosterone in the Leydig cells treated with 2.5 and 5.0 μg/ml 4-nonylphenol was observed. The viability of mouse Leydig cells was slightly increased at the lowest doses of 4-nonylphenol (0.04 and 0.2 μg/ml). We also observed an increase at higher concentrations of the substance (1.0; 2.5 and 5.0 μg/ml), but this increase was not significant. Further investigations are required to establish the biological significance and possible reproductive implications.


Spectrum of asthma in children. II. Allergic components.
The incidence of a variety of clinical and immunological features of an allergic state was studied at 7, 10, and 14 years of age in a group of children suffering from one of four grades of asthma, ranging from mild subclinical to severe unremitting, and compared with the incidence in a control group of non-asthmatic children. The incidence of all features of allergy was significantly higher in the asthmatics but no one feature unequivocally distinguished the asthmatics from the controls. Almost all the asthmatics showed several features of the allergic state at 14 years of age. A cluster of allergic features was a differentiating characteristic of the asthmatics, and the children with the most allergic manifestations were usually the children with the most severe and persistent asthma. The first appearance of and subsequent variation in some of the allergic manifestations often did not correspond to the clinical course of the asthma.Though many manifestations of asthma can be understood on an allergic basis the mechanism by which emotional disturbance, exercise, viral infections, and non-allergenic stimuli precipitate attacks of asthma and the relation of these factors to allergy are unknown.


Proximal nail fold flap for digital mucous cyst excision.
The skin covering a digital mucous cyst is often very thin and is often excised with the cyst. Thus, transfer of a skin flap is needed for the defect. We have developed a proximal nail fold flap technique by which the thin skin covering the cyst can be preserved. We conducted a retrospective study to assess the effectiveness and reliability of this technique for digital mucous cyst excision. The study group comprised 26 patients treated for 28 digital mucous cysts. The flap was elevated on the nail matrix to expose the distal interphalangeal joint capsule. To preserve the skin in cases in which the skin covering the cyst was exceptionally thin, we did not excise the upper part of the cyst wall. Excision of the cyst and stalk was successful in all cases. Additional excision of the joint capsule or osteophyte(s) was achieved in 20 cases and 5 cases, respectively. No flap necrosis, skin defect or nail deformity resulted. Three of the cysts recurred and were treated successfully by reoperation involving the same flap elevation technique. We conclude that the proximal nail fold flap is useful for excision and reliable for wound coverage after digital mucous cyst excision. J. Med. Invest. 63: 278-280, August, 2016.


Femtosecond laser-assisted keratopigmentation for the management of visual disabilities due to peripheral iridectomies.
To report the technique and result of keratopigmentation using a femtosecond laser for the treatment of functional visual disabilities caused by peripheral iridectomies. Case report. Two eyes of 2 patients underwent femtosecond laser-assisted keratopigmentation (FLAK) for moderate to severe visual dysfunction secondary to peripheral iridectomies. The main outcomes measures of the study were changes in visual-related symptoms, cosmesis, and intraoperative surgical complications. Following FLAK, the visual-related symptoms (ghosting, glare, and monocular diplopia) improved in both cases with significant improvement to total elimination of symptoms. No patient lost any lines of visual acuity, and no significant complications were observed during the follow-up period. The cosmetic appearance was reported as very good. FLAK is minimally invasive and results in a significant decrease in the subjective glare and photophobia and even in resolution of monocular diplopia. The cosmetic outcome was also favorable. This technique allows surgeons to correct visual disabilities associated with iris defects with a high success rate while avoiding more aggressive intraocular surgery.


Single amino acid residue changes in subsite -1 of inulosucrase from Lactobacillus reuteri 121 strongly influence the size of products synthesized.
Bacterial fructansucrase enzymes belong to glycoside hydrolase family 68 and catalyze transglycosylation reactions with sucrose, resulting in the synthesis of fructooligosaccharides and/or a fructan polymer. Significant differences in fructansucrase enzyme product specificities can be observed, i.e. in the type of polymer (levan or inulin) synthesized, and in the ratio of polymer versus fructooligosaccharide synthesis. The Lactobacillus reuteri 121 inulosucrase enzyme produces a diverse range of fructooligosaccharide molecules and a minor amount of inulin polymer [with beta(2-1) linkages]. The three-dimensional structure of levansucrase (SacB) of Bacillus subtilis revealed eight amino acid residues interacting with sucrose. Sequence alignments showed that six of these eight amino acid residues, including the catalytic triad (D272, E523 and D424, inulosucrase numbering), are completely conserved in glycoside hydrolase family 68. The other three completely conserved residues are located at the -1 subsite (W271, W340 and R423). Our aim was to investigate the roles of these conserved amino acid residues in inulosucrase mutant proteins with regard to activity and product profile. Inulosucrase mutants W340N and R423H were virtually inactive, confirming the essential role of these residues in the inulosucrase active site. Inulosucrase mutants R423K and W271N were less strongly affected in activity, and displayed an altered fructooligosaccharide product pattern from sucrose, synthesizing a much lower amount of oligosaccharide and significantly more polymer. Our data show that the -1 subsite is not only important for substrate recognition and catalysis, but also plays an important role in determining the size of the products synthesized.


A Personalized Glomerulus Chip Engineered from Stem Cell-Derived Epithelium and Vascular Endothelium.
Progress in understanding kidney disease mechanisms and the development of targeted therapeutics have been limited by the lack of functional in vitro models that can closely recapitulate human physiological responses. Organ Chip (or organ-on-a-chip) microfluidic devices provide unique opportunities to overcome some of these challenges given their ability to model the structure and function of tissues and organs in vitro. Previously established organ chip models typically consist of heterogenous cell populations sourced from multiple donors, limiting their applications in patient-specific disease modeling and personalized medicine. In this study, we engineered a personalized glomerulus chip system reconstituted from human induced pluripotent stem (iPS) cell-derived vascular endothelial cells (ECs) and podocytes from a single patient. Our stem cell-derived kidney glomerulus chip successfully mimics the structure and some essential functions of the glomerular filtration barrier. We further modeled glomerular injury in our tissue chips by administering a clinically relevant dose of the chemotherapy drug Adriamycin. The drug disrupts the structural integrity of the endothelium and the podocyte tissue layers, leading to significant albuminuria as observed in patients with glomerulopathies. We anticipate that the personalized glomerulus chip model established in this report could help advance future studies of kidney disease mechanisms and the discovery of personalized therapies. Given the remarkable ability of human iPS cells to differentiate into almost any cell type, this work also provides a blueprint for the establishment of more personalized organ chip and 'body-on-a-chip' models in the future.


Prophylactic platelet transfusion threshold during therapy for adult acute myeloid leukemia: 10,000/microL versus 20,000/microL.
The threshold for prophylactic platelet transfusions has been classically established at 20,000/microL. In 48 patients with de novo acute myeloblastic leukemia (AML) we analyzed the effect of reducing the threshold for prophylactic platelet transfusion from 20,000/microL (group A) to 10,000/microL (group B) after induction and consolidation chemotherapy. Forty-eight adult patients with de novo AML diagnosed in a single institution in a nine year period were enrolled in the study. Between January 1989 and December 1993 the patients received prophylactic platelet transfusions when their platelet count was below 20,000/microL (group A), and from January 1994 to March 1998 prophylactic platelet transfusions were indicated below 10,000/microL or between 10,000/microL and 20,000/microL if there was any consumption factor. The mean number (SD) of platelet transfusions during induction was 8.4 (5.3) in group A and 8.5 (5.5) in group B; and during consolidation 4.7 (3.4) in group A and 4.6 (3.8) in group B (p = n.s.). Excluding the cases with consumption factors from the analysis, group B patients required 34% fewer transfusions during induction and 15.5% fewer during consolidation (p = 0.04). There were no differences between groups regarding major bleeding episodes. Our data show that the threshold for prophylactic platelet transfusion can be safely set at 10,000 microL during induction and consolidation chemotherapy for adult patients with de novo AML.


Nuclear magnetic resonance imaging of syringomyelia.
Five patients with syringomyelia were examined with a 3.5 kG nuclear magnetic resonance (NMR) imager. Syrinx cavities were visualized in all five cases, and image quality compared favorably with metrizamide computed tomography (CT). Axial images were optimal for identifying syrinx cavities, and sagittal views were useful in providing an overview of cord morphology and in examining the craniocervical junction. Inversion-recovery images were less valuable than the spin-echo sequences. By varying spin-echo imaging parameters, tissue relaxation times could be determined and the fluid nature of the syrinx cavities confirmed. This limited study suggests that NMR may challenge the current role of CT in the diagnostic workup of syringomyelia.


UDP-N-methyl-D-glucosamine-phosphate--a possible intermediate of N-methyl-L-glucosamine moiety of streptomycin.
The formation of N-methyl-L-glucosamine moiety of streptomycin from D-glucosamine by Streptomyces griseus was studied. The addition of thymine to the culture medium stimulated the formation of streptomycin and the incorporation of D-glucosamine into N-methyl-L-glucosamine moiety. During a study of sugar nucleotides in the mycelia, a novel UDP-amino sugar was isolated. The compound was formed before the maximum production of streptomycin. It was UDP-N-methyl-D-glucosamine-phosphate.