Conservative oxygen therapy for mechanically ventilated adults with suspected hypoxic ischaemic encephalopathy.
Liberal use of oxygen may contribute to secondary brain injury in patients with hypoxic-ischaemic encephalopathy (HIE). However, there are limited data on the effect of different oxygen regimens on survival and neurological disability in HIE patients. We undertook a post-hoc analysis of the 166 patients with suspected HIE enrolled in a trial comparing conservative oxygen therapy with usual oxygen therapy in 1000 mechanically ventilated ICU patients. The primary endpoint for the current analysis was death or unfavourable neurological outcome at day 180. Key secondary outcomes were day 180 mortality, and cause-specific mortality. Patients with HIE allocated to conservative oxygen spent less time in the ICU with an SpO2 ≥ 97% (26 h [interquartile range (IQR) 13-45 vs. 35 h [IQR 19-70], absolute difference, 9 h; 95% CI - 21.4 to 3.4). A total of 43 of 78 patients (55.1%) assigned to conservative oxygen and 49 of 72 patients (68.1%) assigned to usual oxygen died or had an unfavourable neurological outcome at day 180; odds ratio 0.58; 95% CI 0.3-1.12; P = 0.1 adjusted odds ratio 0.54; 95% CI 0.23-1.26; P = 0.15. A total of 37 of 86 patients (43%) assigned to conservative oxygen and 46 of 78 (59%) assigned to usual oxygen had died by day 180; odds ratio 0.53; 95% CI 0.28-0.98; P = 0.04; adjusted odds ratio 0.56; 95% CI 0.25-1.23; P = 0.15. Cause-specific mortality was similar by treatment group. Conservative oxygen therapy was not associated with a statistically significant reduction in death or unfavourable neurological outcomes at day 180. The potential for important benefit or harm from conservative oxygen therapy in HIE patients is not excluded by these data.


Is vascular calcification associated with bone mineral density and osteoporotic fractures in ambulatory, elderly women?
We analyzed the relationship between aortic calcification and two osteoporotic parameters (bone mineral density (BMD) and incident osteoporotic fractures) in 667 ambulatory, elderly women from the Epidemiology of Osteoporosis (EPIDOS) cohort (mean age, 80 years; range, 72-94 years). We did not find any correlation between the aortic calcification score and BMD or osteoporotic fractures. The aging process is associated with osteoporosis and aortic calcification; conditions which may have similar disease mechanisms. However, the relationship between these two settings remains to be elucidated. We analyzed the relationship between aortic calcification and osteoporotic parameters (BMD and incident osteoporotic fractures) in a cohort of ambulatory, elderly women. The study included 667 ambulatory women from the EPIDOS cohort (mean age, 80 years; age range, 72-94 years). The baseline examination included bone investigations, a clinical and functional examination, and a comprehensive questionnaire on health status and lifestyle. Semiquantitative methods were used to determine the abdominal aortic calcification score on baseline radiographs. Incident fractures were recorded via postal questionnaires issued every 4 months for about 4 years. Five hundred three women (75%) had aortic calcification. The mean aortic calcification score was 5.5 (median, 4). During the follow-up period, 186 (28%) women reported one or more incident osteoporotic fractures. We did not find any correlation between the aortic calcification score on one hand and the BMD or the occurrence of incident osteoporotic fractures on the other. Only age and systolic blood pressure were found to be independently associated with the aortic calcification score. Osteoporotic fractures were independently associated with age and BMD. Osteoporosis and aortic calcification appear to be independent processes in a cohort of ambulatory, elderly women. However, potential confounding factors may be present and prospective studies are needed to investigate this situation further.


Dynamics of sialyl Lewis(a) in aqueous solution and prediction of the structure of the sialyl Lewis(a)-SelectinE complex.
In this article we investigate all possible three-dimensional structures for sialyl Lewis(a) (SLe(a)) in aqueous solution and we predict without a priori experimental information its conformation when bound to SelectinE by using a combination of long molecular dynamics (MD) simulations. Based on 10ns MD studies, three structures differing in glycosidic conformations are proposed for SLe(a) in aqueous solution. Based on a 4ns MD study of the SLe(a)-SelectinE complex with initial structures derived from our prediction tools, we find that, fucose and N-acetyl neuraminic acid are in close contact with SelectinE and therefore expect interactions of the protein with these two sugar rings to be significantly more important than in the case of galactose and N-acetyl glucosamine. Our predictions indicate that the N-acetyl glucosamine of SLe(a) is positioned primarily in the aqueous phase. In order to be able to interact with SLe(a) the side chains of amino acid residues Lys99 and Lys111 in SelectinE appear to undergo large conformational changes when contrasted with the positions of these residues in the X-ray crystal structure. Furthermore, amino acid residues Arg97, Glu98 and Lys99 are acting as a holding arm to position the NeuNAc of SLe(a) in the binding pocket.


Movement timing and reach to reach variability during a repetitive reaching task in persons with chronic neck/shoulder pain and healthy subjects.
Neck/shoulder pain is linked to movement repetition, awkward postures, and muscular fatigue. Studies have examined the influence of pain and fatigue on movement characteristics, but few report spatial and temporal characteristics within- and between-dynamic movements. The combined influences of fatigue and pain on these parameters are examined here. A shoulder-injured group (PAIN) (intensity >3/10, duration >3 consecutive months) and an age-sex-matched control group (CTRL) (n = 16 in each) performed a repetitive reaching task to voluntary termination. Kinematics, heart rate, and muscle activity were recorded. Group comparisons were made at the beginning and end of task. Both pain and fatigue changed movement parameters with CTRL subjects adapting to fatigue with increased arm movement, while the PAIN group increased center of mass (CoM) and minimized shoulder movement. Fatigue was associated with decreased arm relative variability, while pain caused increased arm and decreased CoM relative variability. Time to peak velocity tended to indicate increased joint coupling in the CTRL group only. Together, this suggests that despite initial differences in movement strategies, both groups tend to move toward more fixed movement strategies at the end of the task. Those of CTRL are more fixed temporally and spatially (in the arm), whereas the PAIN group adaptations are primarily spatial in nature and appear to focus on controlling CoM.


Characteristics of hypertension and arterial stiffness in obstructive sleep apnea: A Scandinavian experience from a prospective study of 6408 normotensive and hypertensive patients.
The impact of obstructive sleep apnea (OSA) on arterial stiffness is less studied. We aimed to investigate the prevalence and covariates of increased pulse pressure (PP), a surrogate marker of arterial stiffness, in the entire study population as well as in separate analyses in normotensive and hypertensive patients. Further, we also explored the impact of smoking on brachial BP in hypertensive patients. Between 2012 and 2019, a total of 6408 participants with suspected OSA underwent a standard out-of-center respiratory polygraphy. OSA was defined by an apnea-hypopnea index (AHI) ≥15/h regardless of symptoms. PP ≥60 mmHg was used as a surrogate marker of increased arterial stiffness. Mean age was 49.3±13.7 years, 69.4% were male, and 34.5% had OSA. The prevalence of hypertension was 70.8% in OSA and 46.7% in No-OSA (AHI < 15/h) controls (P < .0001). Hypertension was controlled (clinic BP < 140/90 mmHg) in 45.5% and uncontrolled in 54.5% (P < .001). Mean PP was 50±12 mmHg in smokers and 52±12 mmHg in non-smokers (P = .001). Increased PP was found in 24.2% of the entire study population and was higher in patients with OSA compared to No-OSA group (27.5% vs 22.4%, P < .0001). In an unadjusted logistic regression model, OSA was associated with a 1.3-fold higher risk of having increased PP (95% CI 1.16-1.48, P < .001). In a multivariable-adjusted model, higher age, male sex, and history of hypertension, but not OSA (OR 0.89; 95% CI 0.77-1.02, P = .104) were associated with increased PP. In this large study of nearly 6500 participants who were referred with suspected OSA, one-third were diagnosed with OSA and a quarter had increased arterial stiffness by elevated brachial PP. Hypertension but not OSA per se was associated with increased arterial stiffness. Hypertension was highly prevalent and poorly controlled.


Sural nerve effects on medial gastrocnemius motoneurones in the cat.
1. Excitability cycles for medial gastrocnemius motoneurones were recorded following sural nerve stimuli with amplitudes of 1.5, 3, 5, 10 and 40 times the nerve threshold. The monosynaptic reflex was time-integrated to quantify the motoneuronal output and some implications of this technique are discussed. In particular it is calculated that this method specifically examines the 15-25% most excitable motoneurones of the medial gastrocnemius pool.2. In most cases high amplitude stimulation of the sural nerve caused a triphasic (excitatory-inhibitory-excitatory) change in excitability. Arguments are given to support the conclusion that this represents a corresponding post-synaptic alteration in medial gastrocnemius motoneurone potential. It is concluded that this pattern illustrates the nature of the sural nerve projections to this portion of the medial gastrocnemius pool.3. The first period of excitation began after a latency, corrected to be compatible with intracellular recordings, of 2.8 ms and had a mean duration of 4.0 ms. The minimum stimulus level necessary for this effect lay between 1.5 and 3 times the nerve threshold. The maximum amplitude of this facilitation occurred with stimuli of 3-5 times threshold and was 60-90% of the amplitude of the monosynaptic facilitation which followed stimulation of the lateral gastrocnemius-soleus nerve.4. A period of inhibition followed immediately after the period of excitation and had a mean duration of 16 ms. The minimum stimulus level necessary for this effect lay between 1.5 and 3 times the nerve threshold and the degree of inhibition increased with stimuli up to 40 times threshold.5. The second period of excitation lasted a mean period of 50 ms and was due to activity in high threshold fibres. On average its amplitude was 50% that of the initial excitation.6. Exceptions to this triphasic variation of excitability were found. This result is interpreted as indicating the presence of projections with opposing actions which were simultaneously activated by sural stimuli.


Staphylococcal septicemia and disseminated intravascular coagulation. Staphylococcus aureus endocarditis mimicking meningococcemia.
Two patients with acute endocarditis attributable to Staphylococcus aureus had a clinical syndrome similar to meningococcemia with meningitis. That fulminant S aureus septicemia may on occasion be associated with hemorrhagic skin lesions, thrombocytopenia, coagulation abnormalities, and meningitis is emphasized.


Thermodynamic assessment of Hg(II)-gibbsite interactions.
As discrete particles and/or as surface coatings on other minerals in natural systems, aluminum hydroxides are efficient sinks for Hg(II). The Hg(II) adsorption on gibbsite was determined as a function of temperature (T), pH, and the type of background electrolytes, i.e., NaNO(3), NaClO(4), and NaCl. When the equilibration time t(E) approximately 2 h, the Hg(II) retention on gibbsite was found to be a reversible process, which was ascribed to adsorption. The Hg(II) adsorption capacity, i.e., Gamma(Hg(II)), varied with the type of electrolyte used in accordance with the following order: Gamma(NO(3))(Hg(II)) > or = Gamma(ClO(4))(Hg(II)) > or = Gamma(Cl)(Hg(II)). In all cases, the estimated thermodynamic parameters showed that the Hg(II) adsorption on gibbsite was endothermic and spontaneous. The Hg(II) adsorption data were quantified with the Langmuir or Hill, and Dublin-Radushkevick (DR), isotherms at all temperatures and acidity levels examined. Always, the Hg(II) adsorption data were in compliance with the DR model. However, the Hg(II) adsorption in NaNO(3) or NaClO(4) was interpreted in terms of the Langmuir model. When NaCl was used as electrolyte, the Hg(II) adsorption was modeled well with the Hill equation. The mean free energy values calculated from DR plots concluded that Hg(II)-gibbsite interactions are a result of chemical bonding.


Light and electron microscopic study of vacuolated cells in immunoblastic lymphadenopathy-like T-cell lymphoma.
Three cases of immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma were analyzed immunologically, and the ultrastructure of mononuclear cells in the lymph nodes and peripheral blood was examined. In the peripheral blood, light microscopic examination revealed vacuolated lymphocytes. These vacuolated lymphocytes formed rosettes with sheep erythrocytes, and they were CD3- and CD4-positive using the avidin-biotin method in cases 1 and 2. Electron microscopic examination revealed two kinds of abnormal lymphocytes. One kind was of B-cell nature with rich lamellated rough-surfaced endoplasmic reticulum and mitochondria. The other kind had a large cytoplasm, in which Golgi apparatus, some endoplasmic reticulum, some mitochondria and a few vacuoles were seen. Some of these vacuoles had remnants of mitochondrial cristae or were enlarged endoplasmic reticulum. The vacuolated T lymphocytes and activated lymphocytes of B-cell nature disappeared with chemotherapy but reappeared with relapse of the disease. These observations suggest that vacuolated cells are related to pale cells in lymph node sections. In other words when these vacuolated cells are found in the peripheral blood of patients with lymphoid malignancies, IBL-like T-cell lymphoma can be suspected.


Idiopathic Pulmonary Fibrosis Mortality Risk Prediction Based on Artificial Intelligence: The CTPF Model.
Background: Idiopathic pulmonary fibrosis (IPF) needs a precise prediction method for its prognosis. This study took advantage of artificial intelligence (AI) deep learning to develop a new mortality risk prediction model for IPF patients. Methods: We established an artificial intelligence honeycomb segmentation system that segmented the honeycomb tissue area automatically from 102 manually labeled (by radiologists) cases of IPF patients' CT images. The percentage of honeycomb in the lung was calculated as the CT fibrosis score (CTS). The severity of the patients was evaluated by pulmonary function and physiological feature (PF) parameters (including FVC%pred, DLco%pred, SpO2%, age, and gender). Another 206 IPF cases were randomly divided into a training set (n = 165) and a verification set (n = 41) to calculate the fibrosis percentage in each case by the AI system mentioned previously. Then, using a competing risk (Fine-Gray) proportional hazards model, a risk score model was created according to the training set's patient data and used the validation data set to validate this model. Result: The final risk prediction model (CTPF) was established, and it included the CT stages and the PF (pulmonary function and physiological features) grades. The CT stages were defined into three stages: stage I (CTS≤5), stage II (5 < CTS<25), and stage III (≥25). The PF grades were classified into mild (a, 0-3 points), moderate (b, 4-6 points), and severe (c, 7-10 points). The AUC index and Briers scores at 1, 2, and 3 years in the training set were as follows: 74.3 [63.2,85.4], 8.6 [2.4,14.8]; 78 [70.2,85.9], 16.0 [10.1,22.0]; and 72.8 [58.3,87.3], 18.2 [11.9,24.6]. The results of the validation sets were similar and suggested that high-risk patients had significantly higher mortality rates. Conclusion: This CTPF model with AI technology can predict mortality risk in IPF precisely.


Randomized controlled trial of daily interruption of sedatives in critically ill children.
To study the feasibility of daily interruption of sedatives in critically ill children. Prospective randomized controlled open-label trial, performed in a pediatric intensive care unit of a tertiary care teaching and referring hospital. 30 children (0-12 years) receiving mechanically ventilation for >24 h were included. In the intervention group, all sedatives were stopped daily and restarted when COMFORT-behavior score ≥17. The control group received standard care. Primary end points were amounts of sedatives and number of bolus medications in the first 3 days after enrollment and number of (near) incidents. Secondary end points were duration of mechanical ventilation, length of stay in pediatric intensive care, and changes in COMFORT-behavior score. Midazolam and morphine use were lower in the intervention group compared with the control group (P = 0.007 and P = 0.02, respectively), whereas the number of bolus medications did not differ between groups. Two complications were recorded: one patient (intervention group) lost his intravenous line, and one patient (control group) had an unplanned extubation. Duration of mechanical ventilation was significantly shorter in the intervention group compared with the control group (median [interquartile range] of 4 [3-8] and 9 [4-10] days, respectively, P = 0.03). Length of stay in the PICU in the intervention group was significantly shorter than in the control group (median [interquartile range] of 6 [4-9] and 10 [7-15] days, respectively, P = 0.01). Daily interruption of sedatives in critically ill children is feasible, results in decreased use of sedation, earlier extubation, and shorter length of stay.


Convergent Evolution in a Murine Intestinal Parasite Rapidly Created the TGM Family of Molecular Mimics to Suppress the Host Immune Response.
The Transforming Growth Factor-β mimic (TGM) multigene family was recently discovered in the murine intestinal parasite Heligmosomoides polygyrus. This family was shaped by an atypical set of organismal and molecular evolutionary mechanisms along its path through the adaptive landscape. The relevant mechanisms are mimicry, convergence, exon modularity, new gene origination, and gene family neofunctionalization. We begin this review with a description of the TGM family and then address two evolutionary questions: "Why were TGM proteins needed for parasite survival" and "when did the TGM family originate"? For the former, we provide a likely answer, and for the latter, we identify multiple TGM building blocks in the ruminant intestinal parasite Haemonchus contortus. We close by identifying avenues for future investigation: new biochemical data to assign functions to more family members as well as new sequenced genomes in the Trichostrongyloidea superfamily and the Heligmosomoides genus to clarify TGM origins and expansion. Continued study of TGM proteins will generate increased knowledge of Transforming Growth Factor-β signaling, host-parasite interactions, and metazoan evolutionary mechanisms.


Does a prior hysterectomy complicate transvaginal/transumbilical hybrid NOTES cholecystectomy?-a comparative analysis of prospectively collected data.
Hysterectomy alters the anatomy of the posterior vaginal vault used as access for transvaginal/transumbilical hybrid NOTES cholecystectomy (NC), creating potential consequences for the feasibility and complication rate of the procedure. Therefore, the aim of our retrospective analysis of prospectively collected data was to analyze the postoperative course after NC in previously hysterectomized (PH) patients compared with patients who had not undergone hysterectomy (NH). A total of 126 NH patients and 50 PH patients aged over 42 who had an NC from 12/2008 to 04/2021 were compared regarding age, body mass index (BMI), ASA classification, number of percutaneous trocars, need for intraoperative urinary bladder catheterization, length of procedure, conversion rate, and intraoperative and postoperative complication rate according to the Clavien/Dindo classification, Comprehensive Complication Index (CCI), mortality, and hospital length of stay. PH patients were older than NH patients (63.0 vs 51.5 years; P < 0.001) but did not differ significantly in ASA classification (P = 0.595) and BMI (26.8 vs 27.9 kg/m2; P = 0.480). They required more percutaneous trocars (P = 0.047) and longer procedure time (66.0 vs. 58.5 min; P = 0.039). Out of all 287 scheduled NC only one had to be "converted" to traditional laparoscopic cholecystectomy. Intraoperative and postoperative complication rates, Clavien/Dindo classification, CCI, need for intraoperative urinary bladder catheterization, and length of stay did not differ significantly. Our results indicate an increased degree of difficulty of NC in PH patients, although there is no major impact on intraoperative and postoperative complication rates. Urinary bladder perforation is a specific access-related complication in PH patients.


Tumor angiogenesis in lymph node-negative rectal cancer: correlation with clinicopathological parameters and prognosis.
Intratumoral microvessel density (MVD) could be used as a prognostic factor in colorectal cancer. We retrospectively analyzed the value of microvessel count in predicting the clinical outcome of stage I and II (Dukes A and B) rectal cancer patients. Eighty-four patients who had undergone curative resection of lymph node-negative rectal cancer were included. Tumor type and differentiation, the depth of local invasion, venous invasion, the character of the invasive margin, and the degree of lymphocytic infiltration were evaluated for each tumor specimen. Immunohistochemical staining for the CD31 endothelial antigen was performed to highlight the microvessels. The median value of MVD was 45 microvessels. Low MVD (microvessels < or = 45) was observed in 41 patients (48.8%), and high MVD (>45) was found in 43 (51.2%). The presence of conspicuous lymphocytic infiltration was significantly associated with increased vessel density. With uni- and multivariate survival analysis MVD did not show any prognostic significance. The character of the invasive margin was the only parameter with independent prognostic value. MVD does not seem to provide any additional prognostic information when compared with standard histopathological parameters in lymph node-negative rectal cancer. It is likely that the strong association between MVD and the presence of conspicuous lymphocytic infiltration may interfere with its predictive value.


Platelet fibrinogen. Identity and initial observations on the mode of its degradation by plasmin.
Fibrinogen has been purified from human platelets. Platelet fibrinogen exhibits a characteristic pattern in agar gel immunoelectrophoresis different from that of plasma fibrinogen. Stepwise plasmin degradation has been used in further elucidation of the molecular properties of the platelet protein. Examination of comparative digests by immunologic and gel electrophoretic methods has revealed that (1) the platelet protein is more resistant to plasmin degradation, (2) the plasmin-produced fragments of platelet fibrinogen differ consistently from those of its plasma counterpart, and (3) platelet fibrinogen is different from fragment X of plasma fibrinogen. It is suggested that platelet fibrinogen may contribute to the stability of the thrombus.


Serum procalcitonin and C-reactive protein levels as diagnostic markers for distinguishing bacterial infections from lupus flares in systemic lupus erythematosus: A systematic review and meta‑analysis.
To systematically evaluate the diagnostic performance of procalcitonin (PCT) and C-reactive protein (CRP) for distinguishing bacterial infections from lupus flares in systemic lupus erythematosus (SLE) via meta-analysis. Electronic databases were comprehensively searched. The pooled standard mean difference (SMD) and 95% confidence interval (CI) were calculated to estimate the differences of serum PCT and CRP levels between bacterial infections and flares in SLE. Sensitivity, specificity and summary receiver operating characteristics (SROC) curve were used to assess the diagnostic values of PCT and CRP. The use of fixed or random effects model depended on heterogeneity. Fifteen studies were included in the analysis. Serum PCT and CRP levels were significantly higher in SLE patients with bacterial infections compared to SLE patients with flares (PCT: SMD = 1.035, 95 %CI = 0.708 to 1.362; CRP: SMD = 1.000, 95 %CI = 0.758 to 1.242). The overall sensitivity, specificity, area under the SROC curve, positive likelihood ratios (PLR) and negative likelihood ratios (NLR) of PCT were 0.62, 0.88, 0.862, 6.63 and 0.36, respectively, while the same indicators for CRP were 0.72, 0.70, 0.784, 2.45 and 0.38, respectively. Serum PCT and CRP levels were significantly increased in SLE with bacterial infections. PCT had a better diagnostic performance than CRP. PCT had a high value of PLR and could serve as a rule-in marker, while CRP testing may result in a high false-positive rate due to low PLR; both markers had a suboptimal value of NLR and are not appropriate for ruling out bacterial infections.


Hsp70: anti-apoptotic and tumorigenic protein.
Heat shock protein 70 (Hsp70) is a powerful chaperone whose expression is induced in response to a wide variety of physiological and environmental insults, including anticancer chemotherapy, thus allowing the cell to survive to lethal conditions. Hsp70 cytoprotective properties may be explained by its anti-apoptotic function. Indeed, this protein can inhibit key effectors of the apoptotic machinery at the pre- and postmitochondrial level. In cancer cells, the expression of Hsp70 is abnormally high, and Hsp70 may participate in oncogenesis and in resistance to chemotherapy. In rodent models, Hsp70 overexpression increases tumor growth and metastatic potential. Depletion or inhibition of Hsp70 frequently reduces the size of the tumors and even can cause their complete involution. But Hsp70 can also be found in the extracellular medium. Its role is then immunogenic and the term chaperokine to define the extracellular chaperones has been advanced. Hsp70 tumorigenic functions as well as the strategies that are being developed in cancer therapy in order to inhibit Hsp70 are commented in this chapter.


Local delivery of FTY720 induces neutrophil activation through chemokine signaling in an oronasal fistula model.
Cleft palate repair surgeries lack a regenerative reconstructive option and, in many cases, develop complications including oronasal fistula (ONF). Our group has developed a novel murine phenocopy of ONF to study the oral cavity wound healing program. Using this model, our team previously identified that delivery of FTY720 on a nanofiber scaffold had a unique immunomodulatory effect directing macrophages and monocytes into a pro-regenerative state during ONF healing. Here, the objective of this study was to determine the effects of local biomaterial-based FTY720 delivery in the ONF model on the early bulk gene expression and neutrophil phenotypic response within the regenerating tissue. Using a mouse model of ONF formation, a palate defect was created and was treated with FTY720 nanofiber scaffolds or (blank) vehicle control nanofibers. At 1 and 3 days post-implantation, ONF oral mucosal tissue from the defect region was collected for RNA sequencing analysis or flow cytometry. For the RNA-seq expression profiling, intracellular pathways were assessed using the KEGG Pathway database and Gene Ontology (GO) Terms enrichment interactive graph. To assess the effects of FTY720 on different neutrophil subpopulations, flow cytometry data was analyzed using pseudotime analysis based on Spanning-tree Progression Analysis of Density-normalized Events (SPADE). RNA sequencing analysis of palate mucosa injured tissue identified 669 genes that were differentially expressed (DE) during the first 3 days of ONF wound healing after local delivery of FTY720, including multiple genes in the sphingolipid signaling pathway. Evaluation of the DE genes at the KEGG Pathway database also identified the inflammatory immune response pathways (chemokine signaling, cytokine-cytokine receptor interaction, and leukocyte transendothelial migration), and the Gene Ontology enrichment analysis identified neutrophil chemotaxis and migration terms. SPADE dendrograms of CD11b+Ly6G+ neutrophils at both day 1 and day 3 post-injury showed significantly distinct subpopulations of neutrophils in oral mucosal defect tissue from the FTY720 scaffold treatment group compared to the vehicle control group (blank). Increased expression of CD88 and Vav1, among other genes, were found and staining of the ONF area demonstrated increased VAV1 staining in FTY720-treated healing oral mucosa. Treatment of oral mucosal defects using FTY720 scaffolds is a promising new immunotherapy to improve healing outcomes and reducing ONF formation during cleft palate surgical repair. Local delivery of FTY720 nanofiber scaffolds during ONF healing significantly shifted early gene transcription associated with immune cell recruitment and modulation of the immune microenvironment results in distinct neutrophil subpopulations in the oral mucosal defect tissue that provides a critical shift toward pro-regenerative immune signaling. The online version contains supplementary material available at 10.1007/s40883-021-00208-z.


A preparation of bacterial outer membrane with osmium tetroxide and uranyl acetate co-stain enables improved structural determination by transmission electron microscopy.
Biological nanoparticles, such as bacterial outer membrane vesicles (OMVs), are routinely characterized through transmission electron microscopy (TEM). In this study, we report a novel method to prepare OMVs for TEM imaging. To preserve vesicular shape and structure, we developed a dual fixation protocol involving osmium tetroxide incubation prior to negative staining with uranyl acetate. Combining osmium tetroxide with uranyl acetate resulted in preservation of sub-50 nm vesicles and improved morphological stability, enhancing characterization of lipid-based nanoparticles by TEM.


Chloride-depletion of photosynthetic water oxidase. No proton release during the second oxidation step, S2*==>S3*, and a transmembrane radical pair recombination from the third on.
Chloride depletion blocks the normal four-step progress of photosynthetic water oxidation. We studied proton release in chloride-depleted thylakoids which were dark-adapted and excited by flashing light. Proton release was blocked from the second flash on, possibly leaving an uncompensated positive charge in the catalytic centre. The reduction of P+680 by Tyrz was still very rapid (<< 10 microseconds). From the third flash on, P+680 was reduced more slowly (70 microseconds/200 microseconds), and by an electrogenic back-reaction. The uncompensated positive charge may be the reason why the rapid reduction of P+680 by Tyrz is prevented and the transmembrane charge-pair recombination is facilitated.


Mapping the short arm of human chromosome 16.
Physical mapping of 13 different breakpoints on the short arm of chromosome 16 using previously mapped probes and the subsequent mapping of additional probes enabled the division of this portion of the chromosome into six different intervals. D16S94 was mapped between HBA and D16S80 and is closer to PKD1 than either HBA or D16S80. A tight linkage group which includes FRA16A, D16S8, and D16S79 was identified. Seven breakpoints, including FRA16A, could not be separated by probe localizations. This study provides the basis for the development of detailed maps of the short arm of chromosome 16.


Role of Intracystic Cytokines and Nitric Oxide in Ovarian Neoplasms.
The development of new biomarkers for the diagnosis and prognosis of ovarian cancer may provide an opportunity for new therapies. In this study, we aimed to compare cytokines (interleukin [IL]-2, IL-5, IL-6, IL-8, IL-10 and tumour necrosis factor [TNF]-α) and nitric oxide (NO) metabolite levels in non-neoplastic tumours, benign primary ovarian tumours and malignant primary ovarian neoplasms. The secondary aim was to relate cytokine and intracystic NO metabolite levels to clinical, laboratory and pathologic characteristics for patients with primary ovarian malignancies. We evaluated 110 patients with adnexal masses. Cytokine concentrations were quantified by enzyme-linked immunosorbent assay and nitrate concentrations by enzymatic reduction of nitrite by nitrate reductase. Patients with malignant neoplasms had higher IL-6, IL-8 and NO levels compared to patients with benign neoplasms. Histologic grade 1 tumours were associated with elevated IL-2 levels, whereas anaemia was associated with elevated IL-6 levels. On average, those patients with elevated IL-8 levels also had a neutrophil/lymphocyte ratio (NLR) greater than 2.6 and less than 36 months of disease-free survival (DFS). Patients with normal CA 19-9 levels had elevated IL-10 levels. TNF-α was elevated in patients with two carcinogenesis and those with a platelet/lymphocyte ratio (PLR) less than 300. NO levels were higher in patients with an NLR less than 2.6 and CA 19-9 greater than 35 U/ml. Elevated intracystic cytokine levels, especially IL-6 and IL-8, are associated with worse prognosis in ovarian cancer.


Pre-existing, Concurrent/Early-Onset, and Late-Onset Diabetes in Chronic Pancreatitis: Do Outcomes Differ?
Diabetes secondary to endocrine insufficiency in chronic pancreatitis (CP) may develop at any time during the disease course. We sought to evaluate the differences in clinical characteristics and outcomes in CP patients with pre-existing, early-onset, and late-onset diabetes. We reviewed CP patients seen at our Pancreas Center during 2016-2021. We divided them into four groups: those without diabetes, with pre-existing diabetes, with early-onset diabetes, and with late-onset diabetes. We then compared clinical characteristics and outcomes. We identified 450 patients with CP: 271 without diabetes, 99 with pre-existing diabetes, 51 with early-onset diabetes, and 29 with late-onset diabetes. Early-onset diabetics were younger (54.1 vs 57.3 vs 62.5 vs 61.9 years), had more alcohol-related CP (45.1% vs 31.7% vs 32.3% vs 31%), had higher HbA1C levels (8.02% vs 5.11% vs 7.71% vs 7.66%), were more likely to be on insulin (78.4% vs 0% vs 48.4% vs 65.5%), and used more opioids (64.7% vs 43.9% vs 55.1% vs 44.8%) and gabapentinoids (66.7% vs 43.5% vs 48% vs 60.7%) compared to other groups (p < 0.05). Patients who developed diabetes after CP diagnosis had more exocrine insufficiency (72.4% vs 70.6% vs 65.7% vs 53.1%), anatomical complications, and interventions for pain control (p < 0.05). There was no difference in pancreatic cancer in the four groups. CP patients who are younger and use alcohol are at higher risk of having early-onset diabetes and have poorer glucose control compared other CP patients. Patients who develop diabetes after CP diagnosis have worse outcomes and use more resources.


Effect of late HIV diagnosis on HIV-related mortality among adults in general hospitals of Central Zone Tigray, northern Ethiopia: a retrospective cohort study.
The global incidence of HIV infection is not significantly decreasing, especially in sub-Saharan African countries, including Ethiopia. Though there is availability and accessibility of free HIV services, people are not being diagnosed early for HIV, and hence patients are still dying of HIV-related causes. This research is aimed at verifying the effect of late diagnosis of HIV on HIV-related mortality in Central Zone Tigray, Ethiopia. A retrospective cohort study among adult (≥15 years old) HIV patients in three general hospitals of Tigray was conducted. Record reviews were carried out retrospectively from 2010 to 2015. Sample size was determined using stpower Cox in Stata software. Data were entered into EpiData version 3.1 software and transferred to Stata version 12 for analysis. Both bivariable and multivariable analyses were performed using Cox regression model to compare the HIV-related mortality of exposed (cluster of differentiation 4 cells count <350 cells/mm3) and nonexposed (≥350 cells/mm3) patients using adjusted hazard ratio (AHR) at 95% confidence interval (CI). In all, 638 HIV patients were analyzed, contributing 2,105.6 person-years. Forty-eight (7.5%) patients died of HIV-related causes with a mortality rate of 2.28 per 100 person-years. In the multivariable Cox regression model, patients with late diagnosis of HIV had a higher risk of mortality (AHR =3.22, 95% CI: 1.17-8.82) than patients with early diagnosis of HIV. Rural residence (AHR =1.96, 95% CI: 1.05-3.68), unemployment (AHR =2.70, 95% CI: 1.03-7.08), bedridden patients (AHR =2.98, 95% CI: 1.45-6.13), ambulatory patients (AHR =2.54, 95% CI: 1.05-6.15), and baseline hemoglobin level of <11 mg/dL (AHR =3.06, 95% CI: 1.51-6.23) were other independent predictors of mortality. Late diagnosis of HIV increased HIV-related mortality. Rural residence, unemployment, bedridden and ambulatory patients, and baseline hemoglobin level <11 mg/dL were also independent predictors of HIV-related mortality.


A novel missense variant expands the phenotype and genotype of PAX6-associated foveal hypoplasia accompanied by various manifestations of anterior segment dysgenesis.
According to previous reports, PAX6-associated foveal hypoplasia (FH) could usually be accompanied by various anterior segment anomalies including variable iris changes. This study aims to exhibit unusual phenotypes of a novel missense variant of PAX6 from a Chinese pedigree. Ophthalmic examinations including slit-lamp biomicroscopy, gonioscopy, ophthalmic ultrasound, ultrasonic biomicroscopy, optical coherence tomography, wide-field fundus imaging, and visual field test were performed to evaluate the clinical manifestations. Whole-exome sequencing (WES) and bioinformatics analysis were conducted in eight members from this pedigree to identify the causative mutation. WES revealed a novel heterozygous substitution of PAX6 (NM_000280.5:c.157G > A, p.(Val53Met) (chr11:31823309 C > T, hg19)), which cosegregated with the phenotype of this pedigree. All the three patients (a pair of fraternal twins and their mother) exhibited bilateral FH and anterior segment dysgenesis (ASD) including microcornea, sclerocornea, obvious symmetrical corectopia, iris stromal dysplasia, goniodysgenesis, and abnormal distribution of fundus blood vessels. The girl of the fraternal twins also demonstrated bilateral temporal deviation of lenses and abnormal tissue membrane connecting anterior chamber angle and lens anterior capsule in the right eye. The mother additionally showed apparent cataract bilaterally and cupping of the optic disc in her left eye. A novel missense variant in PAX6 gene was detected in a Chinese pedigree demonstrating bilateral FH and ASD. It is really distinctive that the ASD involves almost all parts of the anterior segment, and bilateral symmetrical corectopia is the most perceptible sign. This study expands the phenotypic and genotypic spectrum of PAX6-associated ocular diseases, and facilitates the understanding of the crucial role that PAX6 plays in the development of the eye. Meanwhile, PAX6 could be considered as a candidate pathogenic gene of bilateral symmetrical corectopia.


Polyphosphonate induced coacervation of chitosan: encapsulation of proteins/enzymes and their biosensing.
Based on the polyphosphonate-assisted coacervation of chitosan, a simple and versatile procedure for the encapsulation of proteins/enzymes in chitosan-carbon nanotubes (CNTs) composites matrix was developed. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), energy dispersive spectrum (EDS) mapping demonstrated the hemoglobin (Hb) uniformly distributed into chitosan-CNTs composites matrix. Raman measurements indicated the CNTs in composites matrix retained the electronic and structural integrities of the pristine CNTs. Fourier transform infrared (FT-IR), ultraviolet-visible (UV-vis) and circular dichroism (CD) spectroscopy displayed the encapsulated Hb preserved their near-native structure, indicating the polyphosphonate-chitosan-CNTs composites possessed excellent biocompatibility for the encapsulation of proteins/enzymes. Electrochemical measurements indicated the encapsulated Hb could directly exchange electron with the substrate electrode. Moreover, the modified electrode showed excellent bioelectrocatalytic activity for the reduction of hydrogen peroxide. Under optimum experimental conditions, the fabricated electrochemical sensor displayed the fast response (less than 3s), wide linear range (7.0×10(-7) to 2.0×10(-3)M) and low detection limit (4.0×10(-7)M) for the determination of hydrogen peroxide. This newly developed protocol was simple and mild and would certainly find extensive applications in biocatalysis, biosensors, bioelectronics and biofuel cells.


Nordic Walking as an Exercise Intervention to Reduce Pain in Women With Aromatase Inhibitor-Associated Arthralgia: A Feasibility Study.
Women taking aromatase inhibitors as treatment for breast cancer commonly experience joint pain and stiffness (aromatase inhibitor-associated arthralgia [AIAA]), which can cause problems with adherence. There is evidence that exercise might be helpful, and Nordic walking could reduce joint pain compared to normal walking. To determine the feasibility of a trial of Nordic walking as an exercise intervention for women with AIAA. A feasibility study was carried out in a sample of women with AIAA using a randomized control design. Women were randomized to exercise (six-week supervised group Nordic walking training once per week with an increasing independent element, followed by six weeks 4 × 30 minutes/week independent Nordic walking); or enhanced usual care. Data were collected on recruitment, retention, exercise adherence, safety, and acceptability. The Brief Pain Inventory, GP Physical Activity Questionnaire, and biopsychosocial measures were completed at baseline, six and 12 weeks. Forty of 159 eligible women were recruited and attrition was 10%. There was no increased lymphedema and no long-term or serious injury. Adherence was >90% for weekly supervised group Nordic walking, and during independent Nordic walking, >80% women managed one to two Nordic walking sessions per week. From baseline to study end point, overall activity levels increased and pain reduced in both the intervention and control groups. Our findings indicate that women with AIAA are prepared to take up Nordic walking, complete a six-week supervised course and maintain increased activity levels over a 12-week period with no adverse effects.


Electron microscopic study of hemadsorption on vaccinia virus infected cells.
Hemadsorption (HAD) induced in HEp-2 cells infected with vaccinia virus was observed. In ultrathin sections, binding of 36 red blood cells (RBCs) was examined in detail and 3 types of HAD were observed: (1) direct and close binding of RBCs to infected HEp-2 cells (cyto-HAD) was observed in cross sections of 27 RBCs (2) binding of RBCs through microvilli of infected cells was found in 11 RBCs, and (3) five RBCs were distorted to form tentacle-like projections by which they were bound to the HEp-2 cell surface. Scanning electron microscopy revealed that more than 30% of the RBCs were bound to microvilli of vaccinia virus-infected HEp-2 cells, and that the number of microvilli twined round each RBC was over ten, RBCs were attached to certain microvilli through swollen sucker-like tips which were not observable in non-infected HEp-2 cells. RBCs sometimes revealed a polygonal shape at regions of binding to microvilli. Virion-mediated RBC HEp-2 cell binding could not be observed.


Attenuating regulatory T cell induction by TLR agonists through inhibition of p38 MAPK signaling in dendritic cells enhances their efficacy as vaccine adjuvants and cancer immunotherapeutics.
TLR ligands are potent adjuvants and promote Th1 responses to coadministered Ags by inducing innate IL-12 production. We found that TLR ligands also promote the induction of IL-10-secreting regulatory T (Treg) cells through p38 MAPK-induced IL-10 production by dendritic cells (DC). Inhibition of p38 suppressed TLR-induced IL-10 and PGE(2) and enhanced IL-12 production in DC. Incubation of Ag-pulsed CpG-stimulated DC with a p38 inhibitor suppressed their ability to generate Treg cells, while enhancing induction of Th1 cells. In addition, inhibition of p38 enhanced the antitumor therapeutic efficacy of DC pulsed with Ag and CpG and this was associated with an enhanced frequency of IFN-gamma-secreting T cells and a reduction of Foxp3(+) Treg cells infiltrating the tumors. Furthermore, addition of a p38 inhibitor to a pertussis vaccine formulated with CpG enhanced its protective efficacy in a murine respiratory challenge model. These data demonstrate that the adjuvant activity of TLR agonists is compromised by coinduction of Treg cells, but this can be overcome by inhibiting p38 signaling in DC. Our findings suggest that p38 is an important therapeutic target and provides a mechanism to enhance the efficacy of TLR agonists as vaccine adjuvants and cancer immunotherapeutics.


The autotransporter secretion system.
The type V secretion system includes the autotransporter family, the two-partner system and the Oca family. The autotransporter secretion process involving first the translocation of the precursor through the inner membrane and then its translocation through the outer membrane via a pore formed by a beta-barrel is reviewed.


Language impairment in Swedish bilingual children: a comparison between bilingual and monolingual children in Malmö.
In this study two groups of children were compared, 192 bilingual and 246 monolingual, referred over a 12-mo period to University Hospital in Malmö for suspected language impairment. Compared with monolingual children, bilingual children ran a significantly lower risk (RR = 0.76, p < 0.0001) of being referred by a child health centre, and a significantly higher risk (RR= 1.54, p < 0.0103) of being referred after 5 y of age. In the bilingual group there was also a significantly higher risk of parental refusal of assessment (RR = 2.35, p < 0.0016) and of the children being diagnosed with a severe language impairment (RR = 1.87, p < 0.0009). The risk for bilingual children with severe language impairment being discharged owing to non-attendance was high (RR = 6.20, p < 0.0002) compared to monolingual children. The risk increased with severity of language impairment. Bilingual children displayed a higher risk of a diagnosis of severe language impairment and of being discharged for non-attendance. The problem of low compliance among bilingual parents, despite strong indications of parental distress, must be addressed.


A systematic review of complementary and alternative medicine for asthma self-management.
This article is a systematic review of complementary and alternative medicine use for pediatric and adult asthma self-management. The aim of the review was to summarize the existing body of research regarding the types and patterns of, adverse events and risky behaviors associated with, and patient-provider communication about complementary therapies in asthma. This evidence serves as the basis for a series of recommendations in support of patient-centered care, which addresses both patient preferences for integrated treatment and patient safety.


A Case of Lung Adenocarcinoma with Pulmonary Hypertrophic Osteoarthropathy Showing Pathological Complete Response to a Pembrolizumab-Containing Chemoimmunotherapy.
A 65-year-old woman with prolonged cough and presumed pulmonary hypertrophic osteoarthropathy was referred to our hospital. Computed tomography showed 2 tumors larger than 3 cm in size and massive hilar lymph node enlargement in the right lung. Pathological examination of the transbronchial lung biopsy specimen showed atypical malignant cells, presumed adenocarcinoma, with 1% positivity of programmed cell death 1 ligand (PD-L1). Three courses of chemoimmunotherapy with pembrolizumab (400 mg q3w), carboplatin (AUC 5 mg/mL · min q3w), and pemetrexed (500 mg/m2 q3w) were well tolerated and brought about a quasi-complete response both of the lung tumors and lymph nodes and complete symptom relief of the pulmonary hypertrophic osteoarthropathy, finally leading to the surgical intervention, that is, lobectomy and lymph node dissection. Postoperative pathological examination showed no viable cancer foci both in the lung tumors and lymph nodes. The patient recovered uneventfully. Physicians should note the combination chemoimmunotherapy including pembrolizumab, with curative intent, to optimally treat patients with locally advanced non-small cell lung cancer (NSCLC) even if the NSCLC bears a small amount of PD-L1.


Comparison of radionuclide and computed tomography scanning in non-neoplastic intracranial disease.
In this comparison of computed tomography (CT) and radionuclide scanning (RN) in non-neoplastic disease, the CT and RN patterns in vascular occlusive disease, vascular anomalies, cerebral hemorrhage, cerebral infections, and trauma are described in detail. A retrospective evaluation of CT and RN findings in 514 patients provides a basis for determining which modality is more sensitive in a given disease entity. Illustrative cases demonstrate pertinent findings in both CT and RN scanning in the aforementioned disease categories.


Quantifying OH radical generation in hydrodynamic cavitation via coumarin dosimetry: Influence of operating parameters and cavitation devices.
Hydrodynamic cavitation (HC) has been extensively investigated for effluent treatment applications. Performance of HC devices or processes is often reported in terms of degradation of organic pollutants rather than quantification of hydroxyl (OH) radicals. In this study, generation of OH radicals in vortex based cavitation device using coumarin dosimetry was quantified. Coumarin was used as the chemical probe with an initial concentration of 100 µM (15 ppm). Generation of OH radicals was quantified by analysing generated single hydroxylated products. The influence of operating parameters such as pH and type of acid used to adjust pH, dissolved oxygen, and inlet and outlet pressures was investigated. Acidic pH was found to be more conducive for generating OH radicals and therefore subsequent experiments were performed at pH of 3. Sulphuric acid was found to be more than three times effective than hydrochloric acid in generating OH radicals. Effect of initial levels of dissolved oxygen was found to influence OH radical generation. Performance of vortex based cavitation device was then compared with other commonly used cavitation devices based on orifice and venturi. The vortex based cavitation device was found to outperform the orifice and venturi based devices in terms of initial per-pass factor. Influence of device scale (nominal flow rate through the device) on performance was then evaluated. The results presented for these devices unambiguously quantifies their cavitational performance. The presented results will be useful for evaluating computational models and stimulate further development of predictive computational models in this challenging area.


Response surface optimization of corn stover pretreatment using dilute phosphoric acid for enzymatic hydrolysis and ethanol production.
Dilute H(3)PO(4) (0.0-2.0%, v/v) was used to pretreat corn stover (10%, w/w) for conversion to ethanol. Pretreatment conditions were optimized for temperature, acid loading, and time using central composite design. Optimal pretreatment conditions were chosen to promote sugar yields following enzymatic digestion while minimizing formation of furans, which are potent inhibitors of fermentation. The maximum glucose yield (85%) was obtained after enzymatic hydrolysis of corn stover pretreated with 0.5% (v/v) acid at 180°C for 15min while highest yield for xylose (91.4%) was observed from corn stover pretreated with 1% (v/v) acid at 160°C for 10min. About 26.4±0.1g ethanol was produced per L by recombinant Escherichia coli strain FBR5 from 55.1±1.0g sugars generated from enzymatically hydrolyzed corn stover (10%, w/w) pretreated under a balanced optimized condition (161.81°C, 0.78% acid, 9.78min) where only 0.4±0.0g furfural and 0.1±0.0 hydroxylmethyl furfural were produced.


Comparative assessment of groundwater vulnerability using GIS-based DRASTIC and DRASTIC-AHP for Thoothukudi District, Tamil Nadu India.
The groundwater is very precious in the world. Rapid urbanization and industrialization create tremendous stress on groundwater quality and quantity. Unscientific groundwater extraction and waste disposal methods impact the groundwater aquifer's susceptibility in the coastal area. This research examines how industrial waste, seawater intrusion, and solid waste dumping affect the Thoothukudi District, located on the southwest coast of Tamil Nadu, India. The groundwater vulnerability potential is determined using the DRASTIC and analytical hierarchy process (AHP)-based DRASTIC model. DRASTIC-AHP method's weights and ranks are determined using multi-criteria decision analysis (MCDA)-based pairwise comparison method. Remote sensing (RS) and geographical information system (GIS) are implemented to prepare the input layers for DRASTIC and DRASTIC-AHP. The findings reveal a very high category of vulnerability along the coastline that is covered in sand and loose sediments from an aquifer. Similar conditions exist on the southeast side, which is covered with gravel, sand, and sandstone with shale and has relatively low-slope topography. This enables higher contaminant percolation into the groundwater and raises the possibility for pollution. The DRASTIC-AHP method's results reveal that the southeast side has a significant possibility of contamination. The water table, net recharge, vadose zone, and conductivity greatly impacted the DRASTIC vulnerability assessment due to their stronger weight than theoretical weight. It may be stated that the DRASTIC technique is more cost-effective and time-efficient in analyzing a wide range of regional groundwater risks while avoiding sloppy, uncontrolled land development and other unwanted activities.


Comparison of Nocturnal Glucose After Exercise Among Dual-Hormone, Single-Hormone Algorithm-Assisted Insulin Delivery System and Usual Care in Adults and Adolescents Living with Type 1 Diabetes: A Pooled Analysis.
Background: Available studies comparing the efficacy of dual-hormone (DH)-algorithm-assisted insulin delivery (AID), single-hormone (SH)-AID and usual care on postexercise overnight glucose in people with type 1 diabetes (T1D) have had different outcomes. By pooling data from all available studies, we aim to draw stronger conclusions. Methods: Data were pooled from two three-arm, open-label, randomized, controlled, crossover studies. Forty-one adults [median (Q1, Q3) age: 34.0 years (29.5, 51.0), mean HbA1c: 7.5% ± 1.0%] and 17 adolescents with T1D [age: 14.0 (13.0, 16.0), HbA1c: 7.8% ± 0.8%] underwent DH-AID, SH-AID, and usual care. Each intervention involved evening aerobic exercise (60-min). The primary outcome, time in range% (TIR%) overnight (00:00-06:00) postexercise based on continuous glucose monitoring, was compared among treatments using linear mixed effect model or generalized linear mixed model. Results: Among adults, mean TIR% was 94.0% ± 11.9%, 83.1% ± 20.5%, and 65.1% ± 37.0% during DH-AID, SH-AID, and usual care intervention, respectively (P < 0.05 for all between-group comparisons). DH-AID was superior to SH-AID and usual care, and SH-AID was superior to usual care regarding hypoglycemia and hyperglycemia prevention, but not glycemic variability. Among adolescents, DH-AID and SH-AID reduced dysglycemia, but not glycemic variability, better than usual care. Glycemic outcomes were similar between DH-AID and SH-AID. Conclusion: AID systems allow improved postexercise nocturnal glycemic management than usual care for both adults and adolescents. DH-AID was better than SH-AID among adults, but not adolescents.


Estimated Costs for Delivery of HIV Antiretroviral Therapy to Individuals with CD4+ T-Cell Counts >350 cells/uL in Rural Uganda.
Evidence favoring earlier HIV ART initiation at high CD4+ T-cell counts (CD4>350/uL) has grown, and guidelines now recommend earlier HIV treatment. However, the cost of providing ART to individuals with CD4>350 in Sub-Saharan Africa has not been well estimated. This remains a major barrier to optimal global cost projections for accelerating the scale-up of ART. Our objective was to compute costs of ART delivery to high CD4+count individuals in a typical rural Ugandan health center-based HIV clinic, and use these data to construct scenarios of efficient ART scale-up. Within a clinical study evaluating streamlined ART delivery to 197 individuals with CD4+ cell counts >350 cells/uL (EARLI Study: NCT01479634) in Mbarara, Uganda, we performed a micro-costing analysis of administrative records, ART prices, and time-and-motion analysis of staff work patterns. We computed observed per-person-per-year (ppy) costs, and constructed models estimating costs under several increasingly efficient ART scale-up scenarios using local salaries, lowest drug prices, optimized patient loads, and inclusion of viral load (VL) testing. Among 197 individuals enrolled in the EARLI Study, median pre-ART CD4+ cell count was 569/uL (IQR 451-716). Observed ART delivery cost was $628 ppy at steady state. Models using local salaries and only core laboratory tests estimated costs of $529/$445 ppy (+/-VL testing, respectively). Models with lower salaries, lowest ART prices, and optimized healthcare worker schedules reduced costs by $100-200 ppy. Costs in a maximally efficient scale-up model were $320/$236 ppy (+/- VL testing). This included $39 for personnel, $106 for ART, $130/$46 for laboratory tests, and $46 for administrative/other costs. A key limitation of this study is its derivation and extrapolation of costs from one large rural treatment program of high CD4+ count individuals. In a Ugandan HIV clinic, ART delivery costs--including VL testing--for individuals with CD4>350 were similar to estimates from high-efficiency programs. In higher efficiency scale-up models, costs were substantially lower. These favorable costs may be achieved because high CD4+ count patients are often asymptomatic, facilitating more efficient streamlined ART delivery. Our work provides a framework for calculating costs of efficient ART scale-up models using accessible data from specific programs and regions.


Intergenerational continuity of parent-child separation among mother-offspring dyads: Implication for child cognitive development in rural China.
The labor migration in China often leads to parent-child separation. Research to date has primarily focused on understanding mental and cognitive outcomes for children exposed to parent-child separation, with little consideration for the cumulative effects of intergenerational continuity of parent-child separation. A total of 2729 children between the ages of 4 and 6, along with one parent (primarily mothers, 86.2%) and/or one primary caregiver (if the child is separated from both parents), were recruited in the rural area of Anhui Province, China. A unique subsample of children (n = 249) with persistent experience of parent-child separation or whose mother reported being left behind by her parents during early childhood were enrolled for cognitive assessment. A total of 239 age-, gender- and residence-matched children without any parent-child separation experience were selected as the control group. Child cognitive performance was examined with the Chinese version of the Wechsler Preschool and Primary Scale of Intelligence, Fourth edition (WPPSI-IV). The association between intergenerational continuity of parent-child separation and child cognition was described using multivariate OLS regression models. Compared to mother-offspring dyads without any history of parent-child separation, girls (n = 236/450, 52.4%) who experienced the intergenerational continuity of parent-child separation showed a significant decrease of 5.73 points (95% CI: -9.83, -1.62; p = 0.006) on full-scale intelligence quotient (FSIQ) and a decrease of 5.71 points (95% CI: -9.80, 1.63; p = 0.006) on verbal comprehension index. No similar result was observed in boys. The cumulative effects of parent-child separation among mother-offspring dyads on child cognitive development highlight the need for effective early intervention to break the intergenerational cycle of disadvantage. Sex differences and possible epigenetic mechanisms underlying the intergenerational effects of parent-child separation warrant further investigation.


Survival status and predictors of mortality among traumatic brain injury patients in an Ethiopian hospital: A retrospective cohort study.
Traumatic brain injury is a major global public health problem causing substantial mortality among the adult population. Hence, this study aimed to determine the predictors of mortality among adult traumatic brain injury patients in Felegehiwot Comprehensive Specialized Hospital in Northwest Ethiopia during 2020. A retrospective cohort study was conducted at Felegehiwot Comprehensive Specialized Hospital using anonymized patient data obtained from chart review. Descriptive statistics were used to summarise the patient characteristics. The Kaplan-Meier survival curve and log-rank test were used to test for differences in survival status among groups. The Cox proportional hazards regression model was used at the 5% level of significance to determine the net effect of each explanatory variable on time to death. In total, 338 patients aged ≥15 years and diagnosed with traumatic brain injury were included in the analysis. Among these patients, 103 (30.45%) died, giving a crude death rate of 25.53 per 1000 (95% CI: 21.05-30.98) person-days of follow-up. The overall median survival time was 44 days. The independent predictors of mortality after diagnosis of traumatic brain injury were admission Glasgow coma scale score ≤ 8 (adjusted hazard ratio (AHR): 4.85; 95% confidence interval (CI): 1.73-13.62), bilateral non-reactive pupils at admission (AHR: 2.00 (95% CI: 1.10-3.71), elevated systolic blood pressure at admission (AHR: 0.31; 95% CI:0.11-0.86), elevated diastolic blood pressure at admission (AHR: 3.54; 95% CI: 1.33-9.43), and haematoma evacuation (AHR: 0.42; 95% CI: 0.16-0.90). The Survival status of traumatic brain injury patients was relatively low in this study. Glasgow coma scale score, bilateral non-reactive pupils, and elevated blood pressure were significant predictors of mortality. Further prospective follow-up studies that include residence and occupation are recommended.


Heavy metal(loid)s in agricultural soil from main grain production regions of China: Bioaccessibility and health risks to humans.
Unintentional ingestion of metal-contaminated soils may pose a great threat to human health. To accurately evaluate the health risks of heavy metal(loid)s in soils, their bioaccessibility has been widely determined by in vitro assays and increasingly employed to optimize the assessment parameters. Given that, using meta-analysis, we analyzed the literature on farmland heavy metal(loid)s (As, Cd, Cr, Cu, Hg, Pb, Ni, and Zn) in Chinese main grain production regions, and collected their total and bioaccessibility data to accurately assess their human health risks. Monte Carlo simulation was used to reduce the uncertainty in metal concentration, intake rate, toxicity coefficient, and body weight. We found that the mean concentration (0.47 mg/kg) and geological accumulation index (Igeo, 0-5.24) of Cd were the priority position of controlling metals. Moreover, children are more vulnerable to carcinogenic risks than adults. Soil mineralogy, physicochemical properties, Fe, and the types of in vitro assays are the influencing factors of bioaccessibility discrepancy. Furthermore, appropriate bioaccessibility determination methods can be adapted according to the differences in ecological receptors for the risk assessment, like developing a "personalized assessment" scheme for polluted farmland soil management. Collectively, bioaccessibility-based models may provide an accurate and effective approach to human health risk assessment.


A genome-wide landscape of mRNAs, miRNAs, lncRNAs, and circRNAs of skeletal muscles during dietary restriction in Mongolian horses.
The proportion of different muscle fibers is essential for the horse breed's aptitude for athletic activities. Adaptation of locomotor muscle is correlated with altered physiologic conditions. To investigate the adaptive changes of muscle fiber phenotype and transcriptome in horse skeletal muscle during dietary restriction (DR). The muscle fiber type distribution and deep RNA-seq analysis of detecting differentially expressed mRNAs (DEGs), miRNA (DEMIRs), lncRNAs (DELs), circRNAs (DECs), and their function analysis were investigated in gluteus medius muscle of Mongolian horses during DR. A total of 1433 DEGs, 5 DEMIRs, 329 DELs, and 53 DECs were identified. Differing from non-uniform muscle fiber type changing, functional enrichment analysis showed that most downregulated DEGs were associated in muscle contraction, fuel energy metabolism, and protein balance. Linkages between non-coding RNA and mRNA landscape were detected from their functional changes. Our study provides new insights into the expressional changes of mRNA and non-coding RNA in horse skeletal muscles during DR, which might improve our understanding of the molecular mechanisms regulating muscle adaption during DR for racing horses.


Aloin promotes osteogenesis of bone-marrow-derived mesenchymal stem cells via the ERK1/2-dependent Runx2 signaling pathway.
Osteoporosis is characterized by low bone mass and the degeneration of bone structure, conditions which increase the risk of fracture. Aloin has been shown to affect bone metabolism, but its role in osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) remains unclear. The aim of our study was to determine whether aloin promotes the proliferation and osteogenic differentiation of BMSCs and, if so, whether it acts via activation of the ERK1/2-Runx2 signaling pathway. We found that the different concentrations of aloin tested had no obvious cytotoxic effects on the viability of BMSCs. Under osteogenic induction conditions, aloin increased cellular alkaline phosphatase activity, promoted BMSC mineralization, and increased osteogenic-related gene expression. In addition, treating the BMSCs with the signal transduction inhibitor PD98059 (ERK1/2) effectively attenuated Runx2 activation in these cells and also suppressed osteoblastic differentiation. Overall, our study demonstrates that aloin promotes osteogenic differentiation of BMSCs through activation of the ERK1/2-Runx2 signaling pathway.


Reactivation of Kaposi's sarcoma-associated herpesvirus from latency requires MEK/ERK, JNK and p38 multiple mitogen-activated protein kinase pathways.
Lytic replication of Kaposi's sarcoma-associated herpesvirus (KSHV) promotes the progression of Kaposi's sarcoma (KS), a dominant malignancy in patients with AIDS. While 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced KSHV reactivation from latency is mediated by the protein kinase C delta and MEK/ERK mitogen-activated protein kinase (MAPK) pathways, we have recently shown that the MEK/ERK, JNK and p38 MAPK pathways modulate KSHV lytic replication during productive primary infection of human umbilical vein endothelial cells [Pan, H., Xie, J., Ye, F., Gao, S.J., 2006. Modulation of Kaposi's sarcoma-associated herpesvirus infection and replication by MEK/ERK, JNK, and p38 multiple mitogen-activated protein kinase pathways during primary infection. J. Virol. 80 (11), 5371-5382]. Here, we report that, besides the MEK/ERK pathway, the JNK and p38 MAPK pathways also mediate TPA-induced KSHV reactivation from latency. The MEK/ERK, JNK and p38 MAPK pathways were constitutively activated in latent KSHV-infected BCBL-1 cells. TPA treatment enhanced the levels of activated ERK and p38 but not those of activated JNK. Inhibitors of all three MAPK pathways reduced TPA-induced production of KSHV infectious virions in BCBL-1 cells in a dose-dependent fashion. The inhibitors blocked KSHV lytic replication at the early stage(s) of reactivation, and reduced the expression of viral lytic genes including RTA, a key immediate-early transactivator of viral lytic replication. Activation of MAPK pathways was necessary and sufficient for activating the promoter of RTA. Furthermore, we showed that the activation of RTA promoter by MAPK pathways was mediated by their downstream target AP-1. Together, these findings suggest that MAPK pathways might have general roles in regulating the life cycle of KSHV by mediating both viral infection and switch from viral latency to lytic replication.


Triclosan Disrupts SKN-1/Nrf2-Mediated Oxidative Stress Response in C. elegans and Human Mesenchymal Stem Cells.
Triclosan (TCS), an antimicrobial chemical with potential endocrine-disrupting properties, may pose a risk to early embryonic development and cellular homeostasis during adulthood. Here, we show that TCS induces toxicity in both the nematode C. elegans and human mesenchymal stem cells (hMSCs) by disrupting the SKN-1/Nrf2-mediated oxidative stress response. Specifically, TCS exposure affected C. elegans survival and hMSC proliferation in a dose-dependent manner. Cellular analysis showed that TCS inhibited the nuclear localization of SKN-1/Nrf2 and the expression of its target genes, which were associated with oxidative stress response. Notably, TCS-induced toxicity was significantly reduced by either antioxidant treatment or constitutive SKN-1/Nrf2 activation. As Nrf2 is strongly associated with aging and chemoresistance, these findings will provide a novel approach to the identification of therapeutic targets and disease treatment.


Population-scale study of eRNA transcription reveals bipartite functional enhancer architecture.
Enhancer RNAs (eRNA) are unstable non-coding RNAs, transcribed bidirectionally from active regulatory sequences, whose expression levels correlate with enhancer activity. We use capped-nascent-RNA sequencing to efficiently capture bidirectional transcription initiation across several human lymphoblastoid cell lines (Yoruba population) and detect ~75,000 eRNA transcription sites with high sensitivity and specificity. The use of nascent-RNA sequencing sidesteps the confounding effect of eRNA instability. We identify quantitative trait loci (QTLs) associated with the level and directionality of eRNA expression. High-resolution analyses of these two types of QTLs reveal distinct positions of enrichment at the central transcription factor (TF) binding regions and at the flanking eRNA initiation regions, both of which are associated with mRNA expression QTLs. These two regions-the central TF-binding footprint and the eRNA initiation cores-define a bipartite architecture of enhancers, inform enhancer function, and can be used as an indicator of the significance of non-coding regulatory variants.


An economic evaluation of diabetes mellitus in India: A systematic review.
Approximately 88 million people throughout Southeast Asia have diabetes, of which 77 million reside in India. India had an annual estimated diabetes treatment cost of Rs.10,000 to 12,000 crore in 2003, which is likely to rise to as high as Rs. 1,26,000 crores by 2025. To assess the annual mean expenditure for an individual with diabetes mellitus. We used search terms and Boolean operators to identify studies on the economic burden of diabetes mellitus patients in India between 2015 and 2020. Thereafter, we used the checklists of STROBE, as well as the PRISMA, Drummonds and down, and Black criteria, to assess the quality of included studies. Then, Meta xl was used to calculate the weighted means and weighted proportion based on the quality assessment findings. Mean expenditure with a maximum weightage of 100% was found in a community-based study, whereas the lowest weightage obtained was 20%. The mean expenditure on diabetes and its complications was calculated as INR 15,535/-(USD 209.3) per year, with a pooled mean of INR 17, 080(USD 230.1)per year. On average, the OPD charges were 3%-5% of the total annual income of the individual. However, when there were complications and hospitalization, the average expenditure was higher (21%) which averages to around 11,000 INR. While patients spent 3% of their annual income on only OPD charges on average, complications substantially increased the total cost by more than 10%, which amounts to catastrophic health expenditure.


Effect of temporal resolution on the estimation of left ventricular function by cardiac MR imaging.
The aim of this study was to investigate the effect of temporal resolution on the estimation of left ventricular (LV) function by cardiac magnetic resonance (MR) imaging using a steady-state free precession (SSFP) sequence. Left ventricular function was assessed by cine MR imaging using a segmented SSFP sequence in 10 healthy volunteers. Views per segment (VPS) were set at 8 and 20, resulting in high and low true temporal resolution, respectively. Irrespective of VPS, images were reconstructed at 40 cardiac phases, providing high apparent temporal resolution. Data were analyzed using 40, 20 and 10 phases to simulate different apparent temporal resolutions. Increasing the cardiac phases used for analysis slightly decreased mean end-systolic volume (ESV) and slightly increased mean ejection fraction (EF). No substantial difference in estimates of end-diastolic volume (EDV) was found between VPSs of 8 and 20. Imaging with a VPS of 20 yielded a larger ESV and smaller EF than imaging with a VPS of 8 when 40 phases were used. In conclusion, low true temporal resolution causes overestimation of ESV and underestimation of EF. Improvement of apparent temporal resolution mildly reduces but does not eliminate the errors caused by low true temporal resolution.


Outcomes after complete dissolution of everolimus-eluting bioresorbable scaffolds implanted during routine practice.
Long-term outcomes of unselected patients treated with bioresorbable vascular scaffold (BVS) implantation are lacking, especially for the period after complete dissolution of the BVS. This study sought to evaluate 5-year outcomes in patients treated with BVS in routine practice. Consecutive patients who underwent implantation of everolimus-eluting BVS during routine clinical practice at 2 high-volume centres in Germany were studied. The patients were followed-up for up to 5 years. The primary endpoints of interest were the composite of death, myocardial infarction and target lesion revascularization, as well as definite scaffold thrombosis. A total of 419 patients (mean age 66.6 ± 10.9 years; 31.5% had diabetes) were included, of whom 38.9% presented with an acute coronary syndrome. Of the 527 lesions treated, 49.0% were classified as complex and 13.1% were bifurcation lesions. At 5 years, the composite clinical endpoint occurred in 33.1% of patients and definite scaffold thrombosis occurred in 4.7%. Most definite scaffold thrombosis occurred within 2 years after BVS implantation. In patients treated with BVS implantation in routine clinical practice the rates of adverse clinical events at 5 years were high, including a considerable incidence of scaffold thrombosis.


Nicotine releases stereoselectively and Ca2(+)-dependently endogenous 3,4-dihydroxyphenylalanine from rat striatal slices.
In superfused slices of rat striatum, nicotine-evoked release of endogenous 3,4-dihydroxyphenylalanine (DOPA) was studied in comparison with that of dopamine (DA). (+/-)-Nicotine (0.1-10 microM) constantly and repetitively released DOPA and DA over a similar time course in a concentration-dependent manner. The ratio of DOPA and DA evoked was approximately 1:2-3. The turnover rate of DOPA was about 300 times higher compared to DA. (+/-)-Nicotine (10 microM)-induced DOPA release was mecamylamine (20 microM)-sensitive, Ca2(+)-dependent and tetrodotoxin (0.3 microM)-insensitive. The (+)-isomer induced no DOPA release. These characteristics of DOPA release were almost the same as those of DA. Nicotine evokes endogenous DOPA via nicotinic cholinergic receptors in a manner similar to the transmitter DA. These findings further support a probable role of DOPA as a neuroactive substance in the rat central nervous system.


Screening of plant growth promoting attributes and arsenic remediation efficacy of bacteria isolated from agricultural soils of Chhattisgarh.
Arsenic (As) resistant indigenous bacteria with discrete minimum inhibitory concentration values for arsenate [As(V)] and arsenite [As(III)] were isolated from the paddy fields of different regions of Chhattisgarh, India, following enrichment culture technique. Evaluation of the plant growth promoting (PGP) properties of the isolates revealed that two rod-shaped Gram-positive bacteria viz., ARP2 and ART2 acquired various PGP traits, including phosphate solubilization, production of siderophore, indole acetic acid, ammonia, and exopolysaccharide. Both the isolates significantly increased (40-80%) the root length of Oryza sativa L. even under As-exposure. Sequencing of 16S rRNA gene identified these isolates as Bacillus nealsonii strain ARP2 and Bacillus tequilensis strain ART2, respectively. Isolate ARP2 exhibited arsenate reductase activity thereby rapidly reduced As(V) into As(III), achieving a reduction rate of 37.5 μM min-1. Alike, strain ART2 was capable of oxidizing As(III) into As(V) via arsenite oxidase enzyme, and revealed the oxidation rate of 21.8 μM min-1. Quantitative estimation of As through atomic absorption spectrophotometer revealed that the isolates ARP2 and ART2 removed 93 ± 0.2% and 77 ± 0.14% of As(V) and As(III), respectively, from As-containing culture media. The FTIR analysis showed the interaction of As with the cell membrane and was further confirmed by SEM and TEM techniques, which marked the increase in cell volume owing to successive accumulation of As. The As-resistant and PGP properties of above two isolates demonstrates their potentiality for sustainable bioremediation of As, and establishment of flora in As-rich environment.


Reducing the effect of non-volatile fatty acids (non-VFAs) on polyhydroxyalkanoates (PHA) production from fermented thermal-hydrolyzed sludge.
Thermal-hydrolyzed sludge (THS) can be fermented to produce volatile fatty acids (VFAs) rich liquids. These fermentative liquids are considered as a potential feedstock for polyhydroxyalkanoates (PHA) production. However, the presence of high levels of non-VFAs organics supporting the growth response instead of PHA accumulation hindered an efficient culture selection in a feast and famine regime. Lowering the non-VFAs content can compromise activities of microorganisms to take up these external carbon sources and improve the selective pressure; thus two enhanced strategies were tested to optimize the selection process: 1) increasing the proportion of VFAs in the original substrate or 2) removing most of the non-VFAs at the end of the feast phase. Results showed that the strategies resulted in PHA yields on VFAs respectively of 0.62 and 0.54 Cmol/Cmol, significantly higher than that in original SBR (0.16-0.35 Cmol/Cmol), confirming that reducing the adverse effect of non-VFAs can impose effective internal limitation and induce high PHA storage responses. In PHA accumulation tests, cultures selected with synthetic substrates accumulated a maximum PHA content of 61.4 wt%, which is the highest as ever reported among PHA production from THS. In summary, the study provided valuable references for improving PHA production from complex substrates.


Echo planar imaging of perfluorocarbons.
Emulsions of perfluorotributylamine (FTBA) and perflubron were evaluated for their utility in 19F echo planar imaging. Fluorine images of the emulsions were obtained in a phantom and two mice that had been predosed. Both agents, but particularly perflubron, show potential for fluorine echo planar studies because of the long spin-spin relaxation times of the CF3 resonances. High resolution thin slice images obtained in as little as 26.6 ms are presented.


Fifteen years of NaV 1.7 channels as an analgesic target: Why has excellent in vitro pharmacology not translated into in vivo analgesic efficacy?
In 2006, humans with a congenital insensitivity to pain (CIP) were found to lack functional NaV 1.7 channels. In the subsequent 15 years there was a rush to develop selective inhibitors of NaV 1.7 channels with the goal of producing broadly effective analgesics without the problems of addiction and tolerance associated with opioids. Pharmacologically, this mission has been highly successful, leading to a number of highly potent and selective inhibitors of NaV 1.7 channels. However, there are very few examples where these inhibitors have yielded effective analgesia in preclinical pain models or human clinical trials. In this review, we summarise the role of the NaV 1.7 channel in nociception, its history as a therapeutic target and the quest to develop potent inhibitors of this channel. Finally, we discuss possible reasons why the pain-free state seen in humans with CIP has been so difficult to replicate pharmacologically. LINKED ARTICLES: This article is part of a themed issue on Structure Guided Pharmacology of Membrane Proteins (BJP 75th Anniversary). To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v179.14/issuetoc.


Endotracheobronchial neurofibroma treated by right carinal pneumonectomy.
Neurofibromas arising from the trachea and bronchus are relatively rare. We report the case of a 42-year-old woman who came to our hospital with severe shortness of breath. Right sleeve pneumonectomy was performed, and neurofibroma was confirmed by pathologic examination. In a search of the world literature, 28 cases of endotracheobronchial neurofibromas were found. The literature is reviewed.


A mechanistic insight for the biosynthesis of N,N-dimethyltryptamine: An ONIOM theoretical approach.
Psychoactive natural products are potent serotonergic agonists capable of modulating brain functions such as memory and cognition. These substances have shown therapeutic potential for treating various mental disorders. The fact that N,N-dimethyltryptamine (DMT) is produced endogenously in several plants and animals, including humans, makes it particularly attractive. As an amino acid-derived alkaloid, the DMT biosynthetic pathway is part of the L-tryptophan biochemical cascade and can be divided into the decarboxylation by an aromatic L-amino acid decarboxylase (AADC) for tryptamine formation and the subsequent double-methylation by the indolethylamine-N-methyltransferase (INMT) through the cofactor S-adenosyl-L-methionine (SAM), a methyl donor. Unlike the decarboxylation mechanism of L-tryptophan, the molecular details of the double methylation of tryptamine have not been elucidated. Therefore, we propose an in silico model using molecular dynamics (MD), non-covalent interaction index (NCI) and density functional theory (DFT) calculations with the ONIOM QM:MM B3LYP/6-31+G(d,p):MM/UFF level of theory. Based on the obtained energetic data, the potential energy surface (PES) indicates an SN2 mechanism profile, with the second methylation energy barrier being the rate-limiting step with δG‡=60kJ∙mol-1 larger than the previous methylation, following the NCI analysis showing more repulsive interactions for the second transition state. In addition, the hybridization information of each reaction step provides geometric details about the double-methylation.


The effect of streptozotocin diabetes on insulin binding by isolated rat kidney tubules.
Preparations of kidney tubules were isolated from rat kidney cortex and were demonstrated to possess specific binding sites for insulin. The binding was time-and temperature-dependent and the label was displaced by bovine insulin, A1-B29 dodecoyl insulin, proinsulin and insulin A- and B-chains in proportion to their relative activity. Cell-associated degradation was studied by incubating tubules in the presence of fatty-acid-free albumin. The tubules showed high insulin-degrading activity, which was dependent on temperature, time and cell concentration. The number and affinity of insulin receptors on tubules isolated from kidneys taken from streptozotocin-diabetic rats was not significantly different from tubules isolated from untreated control or insulin-treated diabetic rats. Diabetes did not alter the kinetics of insulin degradation by the tubules. This lack of response by the tubules to changes in the concentration of circulating insulin supports the hypothesis that the kidneys do not play an active role in modulating the rate of insulin removal from the circulation.


Rabbit erythrocyte survival following diminished sialic acid and ATP depletion.
The limited deformability and ATP depletion was considered by some authors to be the factor limiting the life span of old red blood cells (RBC) in circulation. Others believed that sialic acid on the RBC surface determines their life span. We compared the life span of 51Cr labelled, neuraminidase treated rabbit RBCs with ATP depleted by incubation at 37 degree C rabbit RBCs. Osmotic fragility, agglutinability, glucose-6-phosphate dehydrogenase (G6PD) and hexokinase activity and ATP levels of these cells were determined. Desyalated RBCs were removed from the circulation within 24 hours. ATP levels, G6d and hexokinase activity and osmotic fragility were normal in these cells. The agglutination by poly(L-lysine) was affected by the loss of surface charge on these cells. Half the ATP depleted RBCs were out of the circulation within three days. Reconstitution of ATP by reincubation with adenosine, elevated the ATP levels to about 80% of their original level, but survival of these cells did not improve. Analysis of sialic acid showed tha 50% of it was removed during the incubation for ATP depletion. The low ATP level and loss of sialic acid fromt he RBC membrane appeared to be conincidental rather than dependent on each other. The latter appears to be a primary factor in red cell survival.


Response of limbic neurotensin systems to methamphetamine self-administration.
Methamphetamine (METH) abuse is personally and socially devastating. Although effects of METH on dopamine (DA) systems likely contribute to its highly addictive nature, no medications are approved to treat METH dependence. Thus, we and others have studied the METH-induced responses of neurotensin (NT) systems. NT is associated with inhibitory feedback action on DA projections, and NT levels are elevated in both the nucleus accumbens and dorsal striatum after noncontingent treatment with high doses of METH. In the present study, we used a METH self-administration (SA) model (linked to lever pressing) to demonstrate that substitution of an NT agonist for METH, while not significantly affecting motor activity, dramatically reduced lever pressing but was not self-administered per se. We also found that nucleus accumbens NT levels were elevated via a D1 mechanism after five sessions in rats self-administering METH (SAM), with a lesser effect in corresponding yoked rats. Extended (15 daily sessions) exposure to METH SA manifested similar NT responses; however, more detailed analyses revealed (i) 15 days of METH SA significantly elevated NT levels in the nucleus accumbens shell and dorsal striatum, but not the nucleus accumbens core, with a lesser effect in the corresponding yoked METH rats; (ii) the elevation of NT in both the nucleus accumbens shell and dorsal striatum significantly correlated with the total amount of METH received in the self-administering, but not the corresponding yoked METH rats; and (iii) an NT agonist blocked, but an NT antagonist did not alter, lever-pressing behavior on day 15 in SAM rats. After 5 days in SAM animals, NT levels were also elevated in the ventral tegmental area, but not frontal cortex of rats self-administering METH.


Induction of hyperthermia using an intracavitary multielement ultrasonic applicator.
In this paper, the possibility of inducing controlled hyperthermia in rectal or vaginal wall tumors using an intracavitary ultrasonic applicator was investigated. A computer model that took into account the thermal and ultrasonic properties of tissues and surface cooling was used to optimize the transducer parameters to obtain desirable temperature distributions for different perfusion situations in the tumor. Also, an applicator that consisted of a cylindrical array of five independently controllable ultrasonic transducers was developed. This array was then tested in degassed water to determine the functional characteristics. This same applicator, modified to include water cooling of the tissue surface, was tested in vivo in dogs. The temperature distributions were found to be promising and with modifications this approach will be used in clinical treatments of suitable tumors.


Spinal subarchnoid metastatic spread from non-neuraxial primary neoplasms. Case report.
A rare case of metastasis to the spinal subarachnoid space from a non-neuraxial primary tumor is presented. Dissemination was shown by computerized tomography to be via the cerebrospinal fluid from secondary deposits in the central nervous system and meninges. This route would seem to be the most common mode of spread to the spinal subarachnoid space.


Origin of deoxycorticosterone and deoxycorticosterone sulfate in human pregnancy: absence of steroid 21-sulfatase activity in sulfatase-deficient placenta.
The activity of steroid 21-sulfatase, the enzyme that catalyzes the hydrolysis of deoxycorticosterone sulfate (DOC-SO4) is demonstrable in human placenta. Thus, it is possible that this placental enzyme, by way of the hydrolysis of either DOC-SO4 or 21-hydroxypregnenolone mono- or di-sulfate of fetal origin, may be important in the biosynthesis of DOC, which is present in the plasma of pregnant women in high concentration. To investigate this issue further, we evaluated steroid 21-sulfatase activity in microsomal preparations of a sulfatase-deficient placenta. Immediately after delivery, at term, of a living male fetus with sulfatase deficiency, a microsome-enriched fraction of placental tissue was prepared; sulfatase activity was evaluated by use of three substrates, viz. dehydroisoandrosterone sulfate (DS), estrone sulfate (E1-SO4), and DOC-SO4, in various concentrations. Similar incubations were conducted with aliquots of a microsome-enriched fraction prepared from placental tissue of a normal fetus that was delivered, at term, within minutes of the time of delivery of the infant with sulfatase deficiency. In microsomal fractions from the normal placenta, each of the steroid sulfates was hydrolyzed. In the absence of microsomes, and in the presence of microsomal fractions from the sulfatase-deficient placenta, the hydrolysis of DOC-SO4 and DS was not detected. Moreover, in microsomes prepared from the sulfatase-deficient placenta, E1-SO4 was hydrolyzed at a rate that was only 10% of that in incubations with microsomal preparations of the normal placenta. We conclude that with sulfatase deficiency, the placenta is deficient not only in sulfatase activity for steroid-3-sulfates but for steroid 21-sulfates, e.g. DOC-SO4, as well.


Time changes in predictive power of established and recently proposed clinical, cytogenetical and comorbidity scores for Myelodysplastic Syndromes.
Recent improvements in the treatment of Myelodysplastic Syndromes have fostered further interest in the development of prognostic scores. Prognostic indices such as the IPSS were developed and later validated assuming their predictive values to be unchanged over time. A systematic analysis of the possible variability of predictive power over time in different scores is still lacking and was the aim of this study. For 243 primary MDS patients from a single institution treated with supportive care, 19 established or modified scoring systems based on different prognostic factors (clinical, cytogenetical and/or comorbidity) were analysed for their variability over time by statistical methods that quantify time variations in the risk relations (specifically the risk ratios of Cox models) between prognostic subgroups. Established scores based mainly on clinical parameters showed strong to moderate loss of predictive power over time whereas cytogenetic scores maintained their predictive power. Scores including comorbidity data showed gain of predictive power over time. The development and comparison of prognostic systems have to take into account their stability versus the possibility or need for re-evaluation. Possibly not only re-evaluation after time is of importance, but also different weighting of items constituting scores.


Pheochromocytoma-induced cardiogenic shock: A multicentre analysis of clinical profiles, management and outcomes.
There is still uncertainty about the management of patients with pheochromocytoma-induced cardiogenic shock (PICS). This study aims to investigate the clinical presentation, management, and outcome of patients with PICS. We collected, retrospectively, the data of 18 patients without previously known pheochromocytoma admitted to 8 European hospitals with a diagnosis of PICS. Among the 18 patients with a median age of 50 years (Q1-Q3: 40-61), 50% were men. The main clinical features at presentation were pulmonary congestion (83%) and cyclic fluctuation of hypertension peaks and hypotension (72%). Echocardiography showed a median left ventricular ejection fraction (LVEF) of 25% (Q1-Q3: 15-33.5) with an atypical- Takotsubo (TTS) pattern in 50%. Inotropes/vasopressors were started in all patients and temporary mechanical circulatory support (t-MCS) was required in 11 (61%) patients. All patients underwent surgical removal of the pheochromocytoma; 4 patients (22%) were operated on while under t-MCS. The median LVEF was estimated at 55% at discharge. Only one patient required heart transplantation (5.5%), and all patients were alive at a median follow-up of 679 days. PICS should be suspected in case of a CS with severe cyclic blood pressure fluctuation and rapid hemodynamic deterioration, associated with increased inflammatory markers or in case of TTS progressing to CS, particularly if an atypical TTS echocardiographic pattern is revealed. T-MCS should be considered in the most severe cases. The main challenge is to stabilize the patient, with medical therapy or with t-MCS, since it remains a reversible cause of CS with a low mortality rate.


Accuracy of heparin-induced platelet aggregation test for the diagnosis of heparin-induced thrombocytopenia.
Whereas the utility of washed platelet assays such as the heparin-induced platelet activation test (HIPA) for the diagnosis of heparin-induced thrombocytopenia (HIT) is regarded as high, the performance of simpler assays such as the heparin-induced platelet aggregation test (PAT) is still elusive. Using well-characterized samples of a large cohort study, we aimed to assess the diagnostic accuracy of PAT for the diagnosis of HIT. One-hundred twenty-two immunoassay-positive serum samples of a previous, prospective single-center cohort study including consecutive patients with suspected HIT (n = 1291) were used. HIPA was determined as reference gold standard; samples were previously analyzed using PAT as well as polyspecific platelet factor 4/heparin enzyme-linked immunosorbent assay (ELISA). 4Ts score was calculated using the patient documentation. Diagnosis of HIT was defined as a positive HIPA, which is a positive reaction in 2 out of 4 donor platelets within 30 min. HIT was diagnosed in 39 out of 122 patients corresponding to a prevalence of 32%. Median optical density (ELISA) was 2.8 (inter-quartile range 2.3, 3.0) in patients with HIT and 0.7 (0.5, 1.3) in patients without HIT. PAT was positive in 27 out of 39 HIT patients and it was negative in 83 out of 83 HIT-negative patients. Thus, the sensitivity of PAT for the diagnosis of HIT was 69% (95% confidence interval 52%, 83%) and the specificity 100% (96%, 100%). Our results demonstrate that PAT is a valuable test to confirm HIT but cannot be applied to rule-out HIT in clinical practice.


Temporal characteristics of potassium-stimulated acetylcholine release and inactivation of calcium influx in rat brain synaptosomes.
The time course of Ca2+-dependent [3H]acetylcholine [( 3H]ACh) release and inactivation of 45Ca2+ entry were examined in rat brain synaptosomes depolarized by 45 mM [K+]0. Under conditions where the intrasynaptosomal stores of releasable [3H]ACh were neither exhausted nor replenished in the course of stimulation, the K+-evoked release consisted of a major (40% of the releasable [3H]ACh pool), rapidly terminating phase (t1/2 = 17.8 s), and a subsequent, slow efflux that could be detected only during a prolonged, maintained depolarization. The time course of inactivation of K+-stimulated Ca2+ entry suggests the presence of fast-inactivating, slow-inactivating, and noninactivating, or very slowly inactivating, components. The fast-inactivating component of the K+-stimulated Ca2+ entry into synaptosomes appears to be responsible for the rapidly terminating phase of transmitter release during the first 60 s of K+ stimulus. The noninactivating Ca2+ entry may account for the slow phase of transmitter release. These results indicate that under conditions of maintained depolarization of synaptosomes by high [K+]0 the time course and the amount of transmitter released may be a function of the kinetics of inactivation of the voltage-dependent Ca channels.


A new microcontroller supervised thermoelectric renal hypothermia system.
In the present study, a thermoelectric system controlled by a microcontroller is developed to induce renal hypothermia. Temperature value was managed by 8-byte microcontroller, PIC16F877, and was programmed using microcontroller MPASM package. In order to ensure hypothermia in the kidney 1-4 modules and sensors perceiving temperature of the area can be selected. Temperature values are arranged proportionately for the selected area and the determined temperature values can be monitored from an Liquid Crystal Display (LCD) screen. The temperature range of the system is between -50 and +50 degrees C. Renal hypothermia system was tried under in vivo conditions on the kidney of a dog.