Antigen engineering can play a critical role in the protective immunity elicited by Yersinia pestis DNA vaccines.
The use of a DNA immunization approach to deliver protective antigens against Yersinia pestis (Y. pestis) has been successful in previously reported studies. In the current study, the gene designs for V and F1, two well-studied virulent factors serving as main targets for vaccine development, were altered to explore additional options in hopes of improving the protective immunity of DNA vaccines expressing these two antigens. Compared to the wild type V gene DNA vaccines, the use of codon optimized V gene sequences was effective in improving the antigen expression, titers of anti-V antibody responses, and survival against a mucosal lethal challenge. For the F1 DNA vaccine, removal of the N-terminal hydrophobic region was able to improve protective immunity. However, adding a mammalian signal peptide sequence to F1 actually led to reduced protection despite it inducing slightly higher anti-F1 antibody responses. The F1 gene can be fused with a gene coding for YscF, a newly confirmed partial protective antigen for Y. pestis, to produce DNA vaccines that express fused F1 and YscF antigens. One design, in particular, that had YscF fused to the downstream sequence of F1, produced better protection than separate F1 or YscF DNA vaccines, suggesting a potential synergistic effect between these two antigens. Findings from the above studies indicated that there are multiple approaches to optimize the protective immunity for plague DNA vaccines. Most importantly, proper antigen engineering to produce optimal antigen gene inserts in DNA vaccines can clearly play a major role in the future designs of a wide range of DNA vaccines.


Biosynthesis of Haloterpenoids in Red Algae via Microbial-like Type I Terpene Synthases.
Red algae or seaweeds produce highly distinctive halogenated terpenoid compounds, including the pentabromochlorinated monoterpene halomon that was once heralded as a promising anticancer agent. The first dedicated step in the biosynthesis of these natural product molecules is expected to be catalyzed by terpene synthase (TS) enzymes. Recent work has demonstrated an emerging class of type I TSs in red algal terpene biosynthesis. However, only one such enzyme from a notoriously haloterpenoid-producing red alga (Laurencia pacifica) has been functionally characterized and the product structure is not related to halogenated terpenoids. Herein, we report 10 new type I TSs from the red algae Portieria hornemannii, Plocamium pacificum, L. pacifica, and Laurencia subopposita that produce a diversity of halogenated mono- and sesquiterpenes. We used a combination of genome sequencing, terpenoid metabolomics, in vitro biochemistry, and bioinformatics to establish red algal TSs in all four species, including those associated with the selective production of key halogenated terpene precursors myrcene, trans-β-ocimene, and germacrene D-4-ol. These results expand on a small but growing number of characterized red algal TSs and offer insight into the biosynthesis of iconic halogenated algal compounds that are not without precedence elsewhere in biology.


Chemotherapy-induced infiltration of neutrophils promotes pancreatic cancer metastasis via Gas6/AXL signalling axis.
Pancreatic ductal adenocarcinoma (PDAC) is a highly metastatic disease and cytotoxic chemotherapy is the standard of care treatment for patients with advanced disease. Here, we investigate how the microenvironment in PDAC liver metastases reacts to chemotherapy and its role in metastatic disease progression post-treatment, an area which is poorly understood. The impact of chemotherapy on metastatic disease progression and immune cell infiltrates was characterised using flow and mass cytometry combined with transcriptional and histopathological analysis in experimental PDAC liver metastases mouse models. Findings were validated in patient derived liver metastases and in an autochthonous PDAC mouse model. Human and murine primary cell cocultures and ex vivo patient-derived liver explants were deployed to gain mechanistical insights on whether and how chemotherapy affects the metastatic tumour microenvironment. We show that in vivo, chemotherapy induces an initial infiltration of proinflammatory macrophages into the liver and activates cytotoxic T cells, leading only to a temporary restraining of metastatic disease progression. However, after stopping treatment, neutrophils are recruited to the metastatic liver via CXCL1 and 2 secretion by metastatic tumour cells. These neutrophils express growth arrest specific 6 (Gas6) which leads to AXL receptor activation on tumour cells enabling their regrowth. Disruption of neutrophil infiltration or inhibition of the Gas6/AXL signalling axis in combination with chemotherapy inhibits metastatic growth. Chemotherapy increases Gas6 expression in circulating neutrophils from patients with metastatic pancreatic cancer and recombinant Gas6 is sufficient to promote tumour cell proliferation ex vivo, in patient-derived metastatic liver explants. Combining chemotherapy with Gas6/AXL or neutrophil targeted therapy could provide a therapeutic benefit for patients with metastatic pancreatic cancer.


Gastrointestinal Kaposi's sarcoma as the first manifestation of acquired immunodeficiency syndrome.
Gastrointestinal Kaposi's sarcoma can occur in HIV-infected patients without previously diagnosed AIDS. Gastrointestinal symptoms in such patients should be thoroughly investigated because of the possibility of gastrointestinal Kaposi's sarcoma, despite the absence of the cutaneous form. The discovery of gastrointestinal Kaposi's sarcoma establishes the diagnosis of AIDS, as it did in our two patients.


Erratum: Reproducibility and Harmonization in Research using Biological Standards: The Example of Platelet Agonist Collagen-Related Peptide.
An erratum was issued for: Reproducibility and Harmonization in Research using Biological Standards: The Example of Platelet Agonist Collagen-Related Peptide. The Authors section was updated from: Carmen H. Coxon1 Peter Rigsby1 1National Institute for Biological Standards and Control to: Carmen H. Coxon1 Peter Rigsby1 1Medicines and Healthcare products Regulatory Agency.


Evolutionary conservation and expression patterns of maize starch branching enzyme I and IIb genes suggests isoform specialization.
Expression of the maize (Zea mays L.) starch branching enzyme (SBE) genes Sbe1 and Sbe2 were characterized during kernel development and in vegetative tissues. The onset of Sbe1 and Sbe2 expression during endosperm development was similar to that of other genes involved in starch biosynthesis (Wx, Sh2 and Bt2). However, the expression of Sbe2 peaked earlier than that of Sbe1 in developing endosperm and embryos resulting in a shift in the ratio of Sbe1 to Sbe2 relative message levels during kernel and embryo development. Transcripts hybridizing to the Sbe2 probe were not detectable in leaves or roots which nonetheless have SBEII enzymatic activity, suggesting that there may be another divergent SBEII-like gene(s) in maize. A similar expression pattern is shared between the maize genes and related genes in pea, which together with their evolutionary conservation, suggests that the SBE isoforms may play unique roles in starch biosynthesis during plant development.


Interaction of beta-adrenergic receptors with the inhibitory guanine nucleotide-binding protein of adenylate cyclase in membranes prepared from cyc- S49 lymphoma cells.
beta-Adrenergic receptors on membranes prepared from L6 myoblasts, wild-type S49 lymphoma cells, and an adenylate cyclase-deficient variant (cyc-) of S49 lymphoma cells bind the agonist [3H]hydroxybenzylisoproterenol ([3H]HBI) with high affinity. In each case the agonist [3H]HBI is associated with a larger complex than is the antagonist [125I]iodopindolol, and the binding of [3H]HBI can be inhibited by GTP. These observations suggest that there is an agonist-dependent association of the receptor with a guanine nucleotide-binding protein. The goal of the present experiments was to investigate the possibility that an interaction of beta-adrenergic receptors with the inhibitory guanine nucleotide-binding protein of adenylate cyclase was responsible for these observations. Treatment of S49 cells with pertussis toxin decreased the extent of pertussis toxin-catalyzed [32P]ADP-ribosylation of a 41,000-dalton protein, measured in vitro, and decreased the inhibition of adenylate cyclase activity observed in the presence of somatostatin or analogues of GTP. Isoproterenol-stimulated adenylate cyclase activity was potentiated following treatment of wild-type S49 cells and L6 myoblasts with pertussis toxin. Although the ability of receptors on membranes prepared from L6 myoblasts to bind the agonist [3H]HBI was not affected by treatment of cells with pertussis toxin, treatment of cyc- S49 cells with pertussis toxin markedly decreased the ability of receptors to bind [3H]HBI. The observed inhibition of the binding of the agonist [3H]HBI to beta-adrenergic receptors on membranes prepared from cyc- S49 cells after treatment with pertussis toxin could be explained by an interaction between beta-adrenergic receptors and the inhibitory guanine nucleotide-binding protein. Such an interaction may represent a mechanism through which stimulation of the activity of adenylate cyclase by beta-adrenergic receptors can be regulated or through which beta-adrenergic receptors can affect the activity of cyclic AMP-independent cellular processes.


HetEnc: a deep learning predictive model for multi-type biological dataset.
Researchers today are generating unprecedented amounts of biological data. One trend in current biological research is integrated analysis with multi-platform data. Effective integration of multi-platform data into the solution of a single or multi-task classification problem; however, is critical and challenging. In this study, we proposed HetEnc, a novel deep learning-based approach, for information domain separation. HetEnc includes both an unsupervised feature representation module and a supervised neural network module to handle multi-platform gene expression datasets. It first constructs three different encoding networks to represent the original gene expression data using high-level abstracted features. A six-layer fully-connected feed-forward neural network is then trained using these abstracted features for each targeted endpoint. We applied HetEnc to the SEQC neuroblastoma dataset to demonstrate that it outperforms other machine learning approaches. Although we used multi-platform data in feature abstraction and model training, HetEnc does not need multi-platform data for prediction, enabling a broader application of the trained model by reducing the cost of gene expression profiling for new samples to a single platform. Thus, HetEnc provides a new solution to integrated gene expression analysis, accelerating modern biological research.


A diagnostic microdosing approach to investigate platinum sensitivity in non-small cell lung cancer.
The platinum-based drugs cisplatin, carboplatin and oxaliplatin are often used for chemotherapy, but drug resistance is common. The prediction of resistance to these drugs via genomics is a challenging problem since hundreds of genes are involved. A possible alternative is to use mass spectrometry to determine the propensity for cells to form drug-DNA adducts-the pharmacodynamic drug-target complex for this class of drugs. The feasibility of predictive diagnostic microdosing was assessed in non-small cell lung cancer (NSCLC) cell culture and a pilot clinical trial. Accelerator mass spectrometry (AMS) was used to quantify [14 C]carboplatin-DNA monoadduct levels in the cell lines induced by microdoses and therapeutic doses of carboplatin, followed by correlation with carboplatin IC50 values for each cell line. The adduct levels in cell culture experiments were linearly proportional to dose (R2 = 0.95, p < 0.0001) and correlated with IC50 across all cell lines for microdose and therapeutically relevant carboplatin concentrations (p = 0.02 and p = 0.01, respectively). A pilot microdosing clinical trial was conducted to define protocols and gather preliminary data. Plasma pharmacokinetics (PK) and [14 C]carboplatin-DNA adducts in white blood cells and tumor tissues from six NSCLC patients were quantified via AMS. The blood plasma half-life of [14 C]carboplatin administered as a microdose was consistent with the known PK of therapeutic dosing. The optimal [14 C]carboplatin formulation for the microdose was 107 dpm/kg of body weight and 1% of the therapeutic dose for the total mass of carboplatin. No microdose-associated toxicity was observed in the patients. Additional accruals are required to significantly correlate adduct levels with response.


GHK and DNA: resetting the human genome to health.
During human aging there is an increase in the activity of inflammatory, cancer promoting, and tissue destructive genes plus a decrease in the activity of regenerative and reparative genes. The human blood tripeptide GHK possesses many positive effects but declines with age. It improves wound healing and tissue regeneration (skin, hair follicles, stomach and intestinal linings, and boney tissue), increases collagen and glycosaminoglycans, stimulates synthesis of decorin, increases angiogenesis, and nerve outgrowth, possesses antioxidant and anti-inflammatory effects, and increases cellular stemness and the secretion of trophic factors by mesenchymal stem cells. Recently, GHK has been found to reset genes of diseased cells from patients with cancer or COPD to a more healthy state. Cancer cells reset their programmed cell death system while COPD patients' cells shut down tissue destructive genes and stimulate repair and remodeling activities. In this paper, we discuss GHK's effect on genes that suppress fibrinogen synthesis, the insulin/insulin-like system, and cancer growth plus activation of genes that increase the ubiquitin-proteasome system, DNA repair, antioxidant systems, and healing by the TGF beta superfamily. A variety of methods and dosages to effectively use GHK to reset genes to a healthier state are also discussed.


A wearable walking monitoring system for gait analysis.
In this paper, both hardware and software design to develop a wearable walking monitoring system for gait analysis are presented. For hardware, the mechanism proposed is adaptive to different individuals to wear, and the portability of the design makes it easy to perform outdoor experiments. Four force sensors and two angle displacement sensors were used to measure plantar force distribution and the angles of hip and knee joints. For software design, a novel algorithm was developed to detect different gait phases and the four gait periods during the stance phase. Furthermore, the center of ground contact force was calculated based on the relationships of the force sensors. The results were compared with the VICON motion capture system and a force plate for validation. Experiments showed the behavior of the joint angles are similar to VICON system, and the average error in foot strike time is less than 90 ms.


Motor training and the combination of action observation and peripheral nerve stimulation reciprocally interfere with the plastic changes induced in primary motor cortex excitability.
AO-PNS is a stimulation protocol combining action observation (AO) and peripheral nerve stimulation (PNS) to induce plasticity in the primary motor cortex (M1) (increased excitability). Another method to increase M1 excitability is motor training. The combination of two protocols, which individually induce long-term potentiation (LTP)-like plasticity in overlapping neural circuits, results in a transitory occlusion or reverse of this phenomenon. This study aimed to understand the neurophysiological mechanisms underlying AO-PNS by testing whether AO-PNS and motor training induced LTP-like plasticity in, at least partially, overlapping neural networks. One group of participants practiced a motor training (finger opposition movements) followed by AO-PNS, whereas another group performed the two protocols in reverse order. Motor performance was evaluated by means of a sensor-engineered glove and transcranial magnetic stimulation was used to assess M1 excitability before and after each conditioning protocol. Motor training increased movement frequency, suggesting the occurrence of motor learning in both groups. When applied on first, both motor training and AO-PNS significantly increased the motor-evoked potential (MEP), but occluded the increase of cortical excitability expected after the following protocol, leading to a significant decrease of MEP amplitude. These results suggest that motor training and AO-PNS act on partially overlapping neuronal networks, which include M1, and that AO-PNS might be able to induce LTP-like plasticity in a similar way to overt movement execution. This candidates AO-PNS as methodology potentially useful when planning rehabilitative interventions on patients who cannot voluntarily move.


Inequality and Mobility using Income, Consumption, and Wealth for the Same Individuals.
Recent studies of economic inequality almost always separately examine income inequality, consumption inequality, and wealth inequality, and hence, these studies miss the important synergy between the three measures explicit in the life-cycle budget constraint. Using the Panel Study of Income Dynamics (PSID), we study inequality in three dimensions, focusing on the conjoint distributions of income, consumption, and wealth for the same individuals. We find that the trends in inequality in income, consumption, and wealth similarly increase between 1999 and 2013. We examine the pairwise distributions of our measures using the average propensity to consume and the wealth-income ratios. Using the longitudinal nature of the PSID, we follow people over this period and find mobility is similar using income, consumption and wealth. We conclude that while all three types of inequality are rising, wealth increasingly acts as a buffer to cushion income changes, which could reduce mobility - both intra- and inter-generational mobility.


Response Characteristics Study of Ethylene Sensor for Fruit Ripening under Temperature Control.
Post-ripening fruits need to be ripened to reach edible conditions, as they are not yet mature enough when picked. Ripening technology is based mainly on temperature control and gas regulation, with the proportion of ethylene being one of the key gas regulation parameters. A sensor's time domain response characteristic curve was obtained through the ethylene monitoring system. The first experiment showed that the sensor has good response speed (maximum of first derivative: 2.01714; minimum of first derivative: -2.01714), stability (xg: 2.42%; trec: 2.05%; Dres: 3.28%), and repeatability (xg: 20.6; trec: 52.4; Dres: 2.31). The second experiment showed that optimal ripening parameters include color, hardness (Change Ⅰ: 88.53%, Change Ⅱ: 75.28%), adhesiveness (Change Ⅰ: 95.29%, Change Ⅱ: 74.72%), and chewiness (Change Ⅰ: 95.18%, Change Ⅱ: 74.25%), verifying the response characteristics of the sensor. This paper proves that the sensor was able to accurately monitor changes in concentration which reflect changes in fruit ripeness, and that the optimal parameters were the ethylene response parameter (Change Ⅰ: 27.78%, Change Ⅱ: 32.53%) and the first derivative parameter (Change Ⅰ: 202.38%, Change Ⅱ: -293.28%). Developing a gas-sensing technology suitable for fruit ripening is of great significance.


The Effect of Shoulder Plyometric Training on Amortization Time and Upper-Extremity Kinematics.
Plyometric training is credited with providing benefits in performance and dynamic restraint. However, limited prospective data exist quantifying kinematic adaptations such as amortization time, glenohumeral rotation, and scapulothoracic position, which may underlie the efficacy of plyometric training for upper-extremity rehabilitation or performance enhancement. To measure upper-extremity kinematics and plyometric phase times before and after an 8-wk upper-extremity strength- and plyometric-training program. Randomized pretest-posttest design. Research laboratory. 40 recreationally active men (plyometric group, age 20.43 ± 1.40 y, height 180.00 ± 8.80 cm, weight 73.07 ± 7.21 kg; strength group, age 21.95 ± 3.40 y, height 173.98 ± 11.91 cm, weight 74.79 ± 13.55 kg). Participants were randomly assigned to either a strength-training group or a strength- and plyometric-training group. Each participant performed the assigned training for 8 wk. Dynamic and static glenohumeral and scapular-rotation measurements were taken before and after the training programs. Dynamic measurement of scapular rotation and time spent in each plyometric phase (concentric, eccentric, and amortization) during a ball-toss exercise were recorded while the subjects were fitted with an electromagnetic tracking system. Static measures included scapular upward rotation at 3 different glenohumeral-abduction angles, glenohumeral internal rotation, and glenohumeral external rotation. Posttesting showed that both groups significantly decreased the time spent in the amortization, concentric, and eccentric phases of a ball-toss exercise (P < .01). Both groups also exhibited significantly decreased static external rotation and increased dynamic scapular upward rotation after the training period (P < .01). The only difference between the training protocols was that the plyometric-training group exhibited an increase in internal rotation that was not present in the strength-training group (P < .01). These findings support the use of both upper-extremity plyometrics and strength training for reducing commonly identified upper-extremity-injury risk factors and improving upper-extremity performance.


Aspirin promotes low density lipoprotein susceptibility to oxidative modification in healthy volunteers.
Low density lipoprotein oxidation is a major pathogenic pathway in atherosclerosis. Previous studies suggested that aspirin, a commonly prescribed drug in patients with atherosclerosis, when given in a dose of 300 mg/day may decrease LDL susceptibility to oxidative modification. However, the effect of the more common lower dose aspirin on LDL oxidation is not known. To examine the effect of aspirin administration (low dosage) on the susceptibility of LDL to oxidative modification in healthy volunteers. Aspirin 75 mg was administered daily for 2 weeks to 10 healthy volunteers selected from the medical staff and students at the faculty of medicine. The main outcome measure was ex vivo oxidation of LDL by ultraviolet C irradiation or by peroxyl free radicals generated by AAPH (2,2'-azobis 2-amidinopropane hydrochloride). The extent of LDL oxidation was determined by measuring the formed amounts of thiobarbituric acid-reactive substances, lipid peroxides and conjugated dienes. Following exposure to UVc irradiation there was a significant (P 0.01) increase (10.8%) in TBARS concentrations and a significant (P < 0.05) increase (5.4%) in PD concentrations in LDL withdrawn after aspirin treatment as compared to LDL withdrawn before aspirin treatment. Following incubation with AAPH there was a significant (P < 0.05) increase (15%) in PD concentrations and a significant (P < 0.05) reduction (10%) of the LDL oxidation lag time in LDL withdrawn after aspirin intake as compared to LDL withdrawn before aspirin treatment. Aspirin treatment given to healthy volunteers at a dose of 75 mg/day increased the susceptibility of their plasma LDL to oxidative modification ex vivo. Our study provides, for the first time, in vivo evidence of pro-oxidative properties of aspirin already suggested by previous in vitro trials.


[Immunochemical tests in the diagnosis of diseases of the male reproductive system].
Immunochemical tests were employed to measure proteins (acid phosphatase, prostatic beta-globulin, endometrial alpha-2-globulin, lactoferrin, carcinoembryonal antigen) in spermatic plasma and prostatic fluid from healthy subjects and patients with prostatic adenoma, cancer, chronic inflammation, defects of spermatogenesis. It was found that the overall concentration of acid phosphatase and prostatic beta-globulin may serve a diagnostic criteria to differentiate prostatic adenoma from cancer as in 93% of prostatic cancer this parameter did not exceed 400 micrograms/ml whereas in 75% of adenomas it was above 1200 micrograms/ml. Activity of chronic prostatitis can be assessed from lactoferrin test. The level of the organ-specific antigens (acid phosphatase and prostatic beta-globulin) and lactoferrin correlated with the severity of spermatogenesis disorders.


New patterns of cocaine use: changing doses and routes.
The history of coca and cocaine use is reviewed in terms of medical and nonmedical patterns of use. Use of coca leaves and coca extract products involved daily use of no more than 200 mg to 500 mg of cocaine. When cocaine became available after 1860, daily doses increased to as much as 1,620 mg and the oral route of administration became supplemented by intranasal, injection, topical, and smoking administration. Contemporary patterns of use between 1970 and 1978 involved social-recreational intranasal doses of 1 to 4 grams per month. From 1978 to 1982, doses increased to 1 to 3 grams per week with increased use of smoking cocaine freebase. Between 1982 and 1984, episodes of concentrated binding became more common, as did the development of experimental practices including intranasal cocaine freebase and the smoking of coca paste. These patterns are discussed in terms of several variables, including purity, dosages, dose regimes, routes of administration, paraphernalia, and changing perceptions of cocaine.


[Stress fractures in children: clinico-radiologic diagnosis].
The authors present 4 children with stress bone fractures and review this entity. The most significant aspects in the mechanism of production of stress fractures are considered along with the appropriate steps for the diagnosis after careful history and physical examination, with special emphasis to the imaging diagnosis.


Impact of therapeutic hypothermia onset and duration on survival, neurologic function, and neurodegeneration after cardiac arrest.
Post-cardiac-arrest therapeutic hypothermia improves outcomes in comatose cardiac arrest survivors. This study tests the hypothesis that the efficacy of post-cardiac-arrest therapeutic hypothermia is dependent on the onset and duration of therapy. Prospective randomized laboratory investigation. University research laboratory. A total of 268 male Long Evans rats. Post-cardiac-arrest therapeutic hypothermia. Adult male Long Evans rats that achieved return of spontaneous circulation after a 10-min asphyxial cardiac arrest were block randomized to normothermia (37°C ± 1°C) or therapeutic hypothermia (33°C ± 1°C) initiated 0, 1, 4, or 8 hrs after return of spontaneous circulation and maintained for 24 or 48 hrs. Therapeutic hypothermia initiated 0, 1, 4, and 8 hrs after return of spontaneous circulation resulted in 7-day survival rates of 45%*, 36%*, 36%*, and 14%, respectively, compared to 17% for normothermic controls and survival with good neurologic function rates of 24%*, 24%*, 19%*, and 0%, respectively, compared to 2% for normothermic controls (*p < .05 vs. normothermia). These outcomes were not different when therapeutic hypothermia was maintained for 24 vs. 48 hrs. In contrast, hippocampal CA1 pyramidal neuron counts were 53% ± 27%*, 53% ± 19%*, 51% ± 24%*, and 65% ± 16%* of normal, respectively, when therapeutic hypothermia was initiated 0, 1, 4, or 8 hrs after return of spontaneous circulation compared to 9% in normothermic controls (*p < .01 vs. normothermia). Furthermore, surviving neuron counts were greater when therapeutic hypothermia was maintained for 48 hrs compared to 24 hrs (68% ± 15%* vs. 42% ± 22%, *p < .0001). In this study, post-cardiac-arrest therapeutic hypothermia resulted in comparable improvement of survival and survival with good neurologic function when initiated within 4 hrs after return of spontaneous circulation. However, histologic assessment of neuronal survival revealed a potentially broader therapeutic window and greater neuroprotection when therapeutic hypothermia was maintained for 48 vs. 24 hrs.


Blocking of collagenase secretion by estramustine during in vitro tumor cell invasion.
The effects of the antitumorigenic drug estramustine on tumor cell membrane penetration (invasion) were investigated in vitro by utilizing a synthetic basement membrane system (a modified Boyden chamber). Tumor cells were plated on a "partition barrier," consisting of a porous filter (8-micron pores) which was coated with a reconstituted basement membrane matrix (Matrigel), and induced to migrate across the barrier with conditioned medium obtained from 9DU 145 human prostatic tumor cells (passage 9). Quantitative radiolabeling studies demonstrated that specially isolated lines (isolated by several passages through the Matrigel) of DU 145 cells, A2058 melanoma, and B16-F10 melanoma cells were highly invasive such that 15 to 20% migrated across a 1-mm-thick Matrigel layer within 5 h at 37 degrees C. NIH-3T3 cells, mouse fibroblasts, and 20DU 145 cells (passage 20) exhibited little or no membrane invasive behavior. Micromolar concentrations of estramustine (30 to 120 microM) inhibited invasion by the invasive cell lines in a dosage-dependent fashion. Quantitative enzymatic assays and radioimmune assays demonstrated that estramustine inhibited membrane invasion by blocking type IV collagenase secretion. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blots confirmed that 30 to 60 microM estramustine blocked secretion of a Mr 105,000 collagenase protein. Indirect studies showed that a collagenase antibody raised against the Mr 105,000 protein and inhibitors of proteinase activity, including a metalloproteinase inhibitor, and 1,10-phenanthroline, blocked invasion. Because the antibodies inhibited type IV collagenase digestion of 3H-mouse type IV collagen, and invasion simultaneously, it is proposed that collagenolytic activity is involved in invasion. These data demonstrate that estramustine blocks proteinase secretion, and suggest that estramustine may be a useful therapeutic drug for the prevention of metastasis.


The benefits and costs of changing treatment technique in electroconvulsive therapy due to insufficient improvement of a major depressive episode.
Electroconvulsive therapy (ECT) technique is often changed after insufficient improvement, yet there has been little research on switching strategies. To document clinical outcome in ECT nonresponders who were received a second course using high dose, brief pulse, bifrontotemporal (HD BP BL) ECT, and compare relapse rates and cognitive effects relative to patients who received only one ECT course and as a function of the type of ECT first received. Patients were classified as receiving Weak, Strong, or HD BP BL ECT during three randomized trials at Columbia University. Nonresponders received HD BP BL ECT. In a separate multi-site trial, Optimization of ECT, patients were randomized to right unilateral or BL ECT and nonresponders also received further treatment with HD BP BL ECT. Remission rates with a second course of HD BP BL ECT were high in ECT nonresponders, approximately 60% and 40% in the Columbia University and Optimization of ECT studies, respectively. Clinical outcome was independent of the type of ECT first received. A second course with HD BP BL ECT resulted in greater retrograde amnesia immediately, two months, and six months following ECT. In the largest samples of ECT nonresponders studied to date, a second course of ECT had marked antidepressant effects. Since the therapeutic effects were independent of the technique first administered, it is possible that many patients may benefit simply from longer courses of ECT. Randomized trials are needed to determine whether, when, and how to change treatment technique in ECT.


Spontaneous repair of the atrophic contralateral ovary without ovariectomy in the case of a granulosa theca cell tumor (GTCT) affected mare.
A 21 year old thoroughbred mare with granulosa theca cell tumor (GTCT) in the right side and atrophic contralateral ovary was investigated in this study. After arrival at our laboratory on 10th December 1999, the clinical diagnosis of GTCT was examined by rectal palpation and ultrasonographic image of ovaries. Plasma from peripheral blood was collected in the breeding and non-breeding seasons for hormonal analysis. The results showed that the contralateral ovary regained normal activity without any treatment of the GTCT affected ovary and contained follicles showing different sizes 19 months later. However, the affected right ovary, which became smaller after 4 months, was totally inactive without any follicle. The observations clearly demonstrate that without any treatment of the GTCT affected ovary, a mare can return to her normal estrous cycle within a certain period in some GTCT cases.


TCF12 is mutated in anaplastic oligodendroglioma.
Anaplastic oligodendroglioma (AO) are rare primary brain tumours that are generally incurable, with heterogeneous prognosis and few treatment targets identified. Most oligodendrogliomas have chromosomes 1p/19q co-deletion and an IDH mutation. Here we analysed 51 AO by whole-exome sequencing, identifying previously reported frequent somatic mutations in CIC and FUBP1. We also identified recurrent mutations in TCF12 and in an additional series of 83 AO. Overall, 7.5% of AO are mutated for TCF12, which encodes an oligodendrocyte-related transcription factor. Eighty percent of TCF12 mutations identified were in either the bHLH domain, which is important for TCF12 function as a transcription factor, or were frameshift mutations leading to TCF12 truncated for this domain. We show that these mutations compromise TCF12 transcriptional activity and are associated with a more aggressive tumour type. Our analysis provides further insights into the unique and shared pathways driving AO.


Indocyanine green enhanced near-infrared fluorescence imaging for perfusion assessment of colonic conduit for esophageal replacement: Utility of a novel technique.
Esophagectomy, followed by esophageal replacement using gastric/colonic conduits, is a complex surgical procedure with significant perioperative morbidity. The most significant and potentially life-threatening complication associated with esophageal replacement is conduit ischaemia, resulting in anastomotic leak and conduit necrosis. Ensuring adequate perfusion of the conduit remains the key to preventing conduit ischaemia. Indocyanine green (ICG) enhanced near-infrared fluorescence imaging is a novel technique which has been used for assessing bowel perfusion. While numerous studies have focused on ICG fluorescence imaging for assessment of gastric conduit perfusion after esophagectomy, data regarding its use for colonic conduits is limited to case reports. ICG fluorescence imaging can help in resolving intraoperative issues by predicting the adequacy of colonic conduit perfusion, thereby preventing postoperative morbidity. To the best of our knowledge, this is the first report in Indian literature describing the utility of ICG fluorescence imaging for assessment of perfusion of colonic interposition.


Intravenous synthetic platelet (SynthoPlate) nanoconstructs reduce bleeding and improve 'golden hour' survival in a porcine model of traumatic arterial hemorrhage.
Traumatic non-compressible hemorrhage is a leading cause of civilian and military mortality and its treatment requires massive transfusion of blood components, especially platelets. However, in austere civilian and battlefield locations, access to platelets is highly challenging due to limited supply and portability, high risk of bacterial contamination and short shelf-life. To resolve this, we have developed an I.V.-administrable 'synthetic platelet' nanoconstruct (SynthoPlate), that can mimic and amplify body's natural hemostatic mechanisms specifically at the bleeding site while maintaining systemic safety. Previously we have reported the detailed biochemical and hemostatic characterization of SynthoPlate in a non-trauma tail-bleeding model in mice. Building on this, here we sought to evaluate the hemostatic ability of SynthoPlate in emergency administration within the 'golden hour' following traumatic hemorrhagic injury in the femoral artery, in a pig model. We first characterized the storage stability and post-sterilization biofunctionality of SynthoPlate in vitro. The nanoconstructs were then I.V.-administered to pigs and their systemic safety and biodistribution were characterized. Subsequently we demonstrated that, following femoral artery injury, bolus administration of SynthoPlate could reduce blood loss, stabilize blood pressure and significantly improve survival. Our results indicate substantial promise of SynthoPlate as a viable platelet surrogate for emergency management of traumatic bleeding.


Epidemiological transition of colorectal cancer in developing countries: environmental factors, molecular pathways, and opportunities for prevention.
Colorectal cancer (CRC) is one of the leading causes of cancer and cancer-related mortality worldwide. The disease has been traditionally a major health problem in industrial countries, however the CRC rates are increasing in the developing countries that are undergoing economic growth. Several environmental risk factors, mainly changes in diet and life style, have been suggested to underlie the rise of CRC in these populations. Diet and lifestyle impinge on nuclear receptors, on the intestinal microbiota and on crucial molecular pathways that are implicated in intestinal carcinogenesis. In this respect, the epidemiological transition in several regions of the world offers a unique opportunity to better understand CRC carcinogenesis by studying the disease phenotypes and their environmental and molecular associations in different populations. The data from these studies may have important implications for the global prevention and treatment of CRC.


[Nacissistic personality disorder].
The narcissistic personality disorder is the extreme form of the so-called narcissistic personality style, which is characterized by exaggerated self-importance coupled with an inordinate desire to be admired, ideas of grandiosity and a strong sense of entitlement, an absence of empathy and feelings of envy. In contact with others, the narcissist appears arrogant, readily feels injured, and takes advantage of other to achieve his own ends. A central psychodynamic role in the development of the condition is a labile sense of self-worth and its overcompensation. Psychotherapeutic treatment aims initially to develop an awareness of the problem in the patient, correction of cognitive dysfunction and a reduction in aggressive or disdainful behavior. Impulsive and aggressive behavior and attacks of depression may be reduced through the use of selective serotonin reuptake inhibitors.


[Heart rhythm in cardiovascular disorders of neurotic origin].
Cardiac rhythm (CR) and vegetative parameters at rest and after a series of short-term loads were compared in normal subjects and neurotic patients with the cardiologic syndrome (group 1) and those with vegetovascular crisis (Group 2). At rest, second-group patients demonstrated the best CR stabilization (by sigma R-R), with the CR oscillation pattern respiratory constituent (RC) diminished and its slow-wave (SW2) constituent prevailing as well as the highest arterial BP and intrasystemic dissociations (the Kërdo index). First-group patients showed the greatest sigma R-R, with the SW2 prevailing in the presence of a greater RC contribution, as compared to group 2 patients. Different patterns of post-exercise change were demonstrated: normal subjects showed increased heart rate, BP and sigma R-R, with doubled SW1 contribution, reduced SW2 and unchanged RC, an evidence of sympathetic mobilization in the presence of vagal pacemaker effects. In second-group patients, heart rate and BP were reduced, the contribution of SW2 increased, and that of SW1 and RC diminished, while the initial CR stabilization by the sigma R-R remained unchanged. Inter-systemic dissociations were demonstrated, using the Hildebrandt index; this combination signals overstrain of the controlling mechanisms, growing centralization of CR control and disadaptation. In first-group patients, BP rose, heart rate and sigma R-R declined, while the CR oscillation pattern remained basically unchanged, suggesting a rigidity of the central adaptation mechanisms.


Support groups for Alzheimer's caregivers: Creating our own space in uncertain times.
Older men are often excluded from family caregiving research despite the steady increase in the number of husbands assuming primary caregiving roles. We explored perceptions of older, male caregivers' experiences with caring for a wife with Alzheimer's Disease (AD) and examined what aspects of the support group were beneficial. Our qualitative research methods invited six caregivers ranging in age from 74 to 85 years to narratively construct their perspectives on caring for their wives with Alzheimer's Disease and benefits of participation in an all-male support group. Thematic analyses revealed care-givers faced several transitions. "Losses related to their personal relationships with their wife, family, and self," captured as loss of golden years. The second area, benefits and improvements of support groups, were captured in the following theme: "creating our own space," which included two sub-themes: "releasing our frustration" and "developing coping strategies." There was also "Gendered experience of caregiving." This study revealed that male caregivers benefited from the support and company of other men in similar caregiving situations. Results from this study have implications for health care professionals for the development of psychosocial educational groups aimed at providing support to male caregivers.


Chemotherapy in Combination With Radiotherapy for Definitive-Intent Treatment of Stage II-IVA Nasopharyngeal Carcinoma: CSCO and ASCO Guideline.
The aim of this joint guideline is to provide evidence-based recommendations to practicing physicians and other healthcare providers on definitive-intent chemoradiotherapy for patients with stage II-IVA nasopharyngeal carcinoma (NPC). The Chinese Society of Clinical Oncology (CSCO) and ASCO convened an expert panel of radiation oncology, medical oncology, surgery, and advocacy representatives. The literature search included systematic reviews, meta-analyses, and randomized controlled trials published from 1990 through 2020. Outcomes of interest included survival, distant and locoregional disease control, and quality of life. Expert panel members used this evidence and informal consensus to develop evidence-based guideline recommendations. The literature search identified 108 relevant studies to inform the evidence base for this guideline. Five overarching clinical questions were addressed, which included subquestions on radiotherapy (RT), chemotherapy sequence, and concurrent, induction, and adjuvant chemotherapy options. Evidence-based recommendations were developed to address aspects of care related to chemotherapy in combination with RT for the definitive-intent treatment of stage II to IVA NPC.Additional information is available at www.asco.org/head-neck-cancer-guidelines.


Comparison of fasting blood sugar and serum lipid profile changes after treatment with atypical antipsychotics olanzapine and risperidone.
This study aimed to compare the effects of the two most commonly prescribed atypical antipsychotics, olanzapine and risperidone, on fasting blood sugar and serum lipid profile of the recipients. A randomised, comparative, open clinical study was conducted on 60 schizophrenic patients. The patients were divided into two groups, one receiving olanzapine and the other receiving risperidone. The patients were assessed for changes in fasting blood sugar and serum lipid profile (triglycerides [TG], high-density lipoprotein [HDL], low-density lipoprotein [LDL], very-low-density lipoprotein [VLDL] and total cholesterol) eight weeks after starting treatment. The number of patients positive for fasting blood sugar and lipid profile criteria of metabolic syndrome was calculated by applying the modified National Cholesterol Education Programme Adult Treatment Panel III guidelines (NCEP ATP III) criteria at eight weeks. Patients treated with olanzapine showed a highly significant increase in the observed parameters, whereas those treated with risperidone showed a significant increase in fasting blood sugar, HDL and LDL levels, and a highly significant increase in other parameters. Intergroup comparison was insignificant except for TG, VLDL and total cholesterol levels. More men as compared to women fulfilled the NCEP ATP III criteria for metabolic syndrome in both groups. Olanzapine has a higher propensity to cause derangement of some parameters of lipid profile than risperidone. These parameters include TG, VLDL and total cholesterol levels.


Evaluating aeration and stirring effects to improve itaconic acid production from glucose using Aspergillus terreus.
The effects of the bioreactor conditions, in particular the mode and intensity of aeration and mixing were studied on itaconic acid (IA) fermentation efficiency by Aspergillus terreus strain from glucose substrate. IA was produced in batch system by systematically varying the oxygen content of the aeration gas (from 21 to 31.5 vol% O2) and the stirring rate (from 150 to 600 rpm). The data were analyzed kinetically to characterize the behavior of the process, and besides, the performances were evaluated comparatively with the literature. It turned out that the operation of the bioreactor with either the higher inlet O2 concentration (31.5 vol% O2) or faster stirring (600 rpm) could enhance biological IA generation the most, resulting in yield and volumetric productivity of 0.31 g IA/g glucose and 0.32 g IA/g glucose and 3.15 g IA/L day and 4.26 g IA/L day, respectively. Overall, the significance of fermentation settings was shown in this work regarding IA production catalyzed by A. terreus and notable advances could be realized by adjusting the aeration and stirring towards an optimal combination.


Downregulation of Cohesin Loading Factor Nipped-B-Like Protein (NIPBL) Induces Cell Cycle Arrest, Apoptosis, and Autophagy of Breast Cancer Cell Lines.
BACKGROUND The cohesin loading factor, nipped-B-like protein (NIPBL), is also known as the sister chromatid cohesion 2 (SCC2) human homolog. Recently, we have studied the role of expression levels of NIPBL in cell proliferation and chemotherapy resistance of non-small cell lung cancer (NSCLC) cells in vitro. The aim of this study was to investigate the effects of expression of the cohesin loading factor, NIPBL, on the cell cycle, apoptosis, and autophagy of breast cancer cell lines in vitro. MATERIAL AND METHODS Expression levels of the NIPBL in the breast cancer cell lines, MCF7, Bcap37, MDA-MB 453 and MDA-MB 231, were measured using Western blot and flow cytometry. Small interfering RNA (si-RNA) was used to study the biological functions of NIPBL. The cell counting kit-8 (CCK-8) assay and the colony formation assay were used to measure cell proliferation; the wound scratching assay and transwell chamber assay were used to investigate cell invasion and migration. RESULTS NIPBL gene and protein expression were upregulated in the MCF7 and Bcap37 cells; si-NIPBL transfection inhibited cell proliferation, invasion, and migration of breast cancer cells. Downregulation of NIPBL arrested cells in the G0/G1 phase of the cell cycle and induced apoptosis and autophagy of breast cancer cells through the caspase3 and mammalian target of rapamycin (mTOR) signaling pathways. CONCLUSIONS [color=black]Downregulation of cohesin loading factor NIPBL arrested breast cancer cells in vitro in the G0/G1 phase of the cell cycle and induced apoptosis and autophagy. [/color].


Improving the delivery of a fixed bridge.
Force loading of dental prosthesis is generally a guessing game played by the operator with the patient. Relying on a patient is comfort level to adjust occlusion is now being replaced by sound force information provided by the T Scan II. Proper use of the T Scan II when installing a segmental fixed prosthesis gives a more precise, comprehensive occlusion. Porcelain surfaces, solder joints, and the underlying abutment teeth can now be loaded with low-to-moderate forces because the operator can fine-tune the occlusal forces precisely. With proper force loading, greater material and prosthesis longevity is significantly enhanced.


Including uncertainty of the expected mortality rates in the prediction of loss in life expectancy.
This study introduces a novel method for estimating the variance of life expectancy since diagnosis (LEC) and loss in life expectancy (LLE) for cancer patients within a relative survival framework in situations where life tables based on the entire general population are not accessible. LEC and LLE are useful summary measures of survival in population-based cancer studies, but require information on the mortality in the general population. Our method addresses the challenge of incorporating the uncertainty of expected mortality rates when using a sample from the general population. To illustrate the approach, we estimated LEC and LLE for patients diagnosed with colon and breast cancer in Sweden. General population mortality rates were based on a random sample drawn from comparators of a matched cohort. Flexible parametric survival models were used to model the mortality among cancer patients and the mortality in the random sample from the general population. Based on the models, LEC and LLE together with their variances were estimated. The results were compared with those obtained using fixed expected mortality rates. By accounting for the uncertainty of expected mortality rates, the proposed method ensures more accurate estimates of variances and, therefore, confidence intervals of LEC and LLE for cancer patients. This is particularly valuable for older patients and some cancer types, where underestimation of the variance can be substantial when the entire general population data are not accessible. The method can be implemented using existing software, making it accessible for use in various cancer studies. The provided example of Stata code further facilitates its adoption.


Hyperuricemia in the hemolytic-uremic syndrome.
Elevated serum uric acid concentrations were found in 18 children with hemolytic-uremic syndrome. The highest recorded level in each patient ranged from 10 to 29 mg/100 ml, and nine patients had concentrations of greater than 20 mg/100 ml. During a 53-hour period of peritoneal dialysis, 5.5 gm of uric acid was removed from one infant. Although acute renal failure was probably the major factor, erythrocyte destruction and hyperlacticacidemia may also have been important in the pathogenesis of the hyperuricemia.


Differential gene expression analysis of a known hepatotoxin, N-acetyl-p-amino-phenol (APAP) as compared to its non-toxic analog, N-acetyl-m-amino-phenol (AMAP) in mouse liver.
Drug-induced hepatotoxicity is one of the most common adverse events associated with drug withdrawal from the market. Elucidating the molecular mechanism of hepatotoxicity is essential to predict the safety of a new molecule. To examine genes involved in hepatotoxicity, we have used oligonucleotide CodeLink Bioarrays and determined the transcriptional profile of mice liver treated with hepatotoxic drug N-acetyl-p-amino-phenol (APAP) as well as its non-toxic analog N-acetyl-m-amino-phenol (AMAP). Out of 20,000 genes analyzed, 896 showed differential expression of > or = 2-fold (648 upregulated and 248 downregulated) within the liver of APAP treated mice as compared to control. In comparison to AMAP treated mice, 62 genes were upregulated and 70 genes were downregulated in mice liver after APAP treatment. Functional classification of these differentially expressed genes identified genes associated with stress response, cell cycle, growth inhibition, cell death, structural components, cell signaling and inflammation. Gene expression profile was further correlated with biochemical analysis and histopathological lesions. These data show that gene expression profiling would help in better understanding the molecular basis of drug-induced hepatotoxicity that will lead to rational development of safer drugs, particularly in pre-clinical stages.


Black carbon and kerogen in soils and sediments. 2. Their roles in equilibrium sorption of less-polar organic pollutants.
The first paper of this series reported that soil/sediment organic matter (SOM) can be fractionated into four fractions with a combined wet chemical procedure and that kerogen and black carbon (BC) are major SOM components in soil/sediment samples collected from the industrialized suburban areas of Guangzhou, China. The goal of this study was to determine the sorptive properties forthe four SOM fractions for organic contaminants. Sorption isotherms were measured with a batch technique using phenanthrene and naphthalene as the sorbates and four original and four Soxhlet-extracted soil/sediment samples, 15 isolated SOM fractions, and a char as the sorbents. The results showed that the sorption isotherms measured for all the sorbents were variously nonlinear. The isolated humic acid (HA) exhibited significantly nonlinear sorption, but its contribution to the overall isotherm nonlinearity and sorption capacity of the original soil was insignificant because of its low content in the tested soils and sediments. The particulate kerogen and black carbon (KB) fractions exhibited more nonlinear sorption with much higher organic carbon-normalized capacities for both sorbates. They dominate the observed overall sorption by the tested soils and sediments and are expected to be the most important soil components affecting bioavailability and ultimate fate of hydrophobic organic contaminants (HOCs). The fact that the isolated KB fractions exhibited much higher sorption capacities than when they were associated with soil/sediment matrixes suggested that a large fraction of the particulate kerogen and BC was not accessible to sorbing HOCs. Encapsulation within soil aggregates and surface coverage by inorganic and organic coatings may have caused large variations in the accessibility of fine kerogen and BC particles to HOCs and hence lowered the sorption capacity of the soil. This variability posts an ultimate challenge for precisely predicting HOC sorption by soils from the contents of different types of SOM.


Selenium Protects ARPE-19 and ACBRI 181 Cells against High Glucose-Induced Oxidative Stress.
Diabetic retinopathy (DR), a complication of diabetes mellitus (DM), can cause severe visual loss. The retinal pigment epithelium (RPE) plays a crucial role in retinal physiology but is vulnerable to oxidative damage. We investigated the protective effects of selenium (Se) on retinal pigment epithelium (ARPE-19) and primary human retinal microvascular endothelial (ACBRI 181) cells against high glucose (HG)-induced oxidative stress and apoptotic cascade. To achieve this objective, we utilized varying concentrations of D-glucose (ranging from 5 to 80 mM) to induce the HG model. HG-induced oxidative stress in ARPE-19 and ACBRI 181 cells and the apoptotic cascade were evaluated by determining Ca2+ overload, mitochondrial membrane depolarization, caspase-3/-9 activation, intracellular reactive oxygen species (ROS), lipid peroxidation (LP), glutathione (GSH), glutathione peroxidase (GSH-Px), vascular endothelial growth factor (VEGF) and apoptosis levels. A cell viability assay utilizing MTT was conducted to ascertain the optimal concentration of Se to be employed. The quantification of MTT, ROS, VEGF levels, and caspase-3 and -9 activation was accomplished using a plate reader. To quantitatively assess LP and GSH levels, GSH-Px activities were utilized by spectrophotometer and apoptosis, mitochondrial membrane depolarization, and the release of Ca2+ from intracellular stores were evaluated by spectrofluorometer. Our investigation revealed a significant augmentation in oxidative stress induced by HG, leading to cellular damage through modulation of mitochondrial membrane potential, ROS levels, and intracellular Ca2+ release. Incubation with Se resulted in a notable reduction in ROS production induced by HG, as well as a reduction in apoptosis and the activation of caspase-3 and -9. Additionally, Se incubation led to decreased levels of VEGF and LP while concurrently increasing levels of GSH and GSH-Px. The findings from this study strongly suggest that Se exerts a protective effect on ARPE-19 and ACBRI 181 cells against HG-induced oxidative stress and apoptosis. This protective mechanism is partially mediated through the intracellular Ca2+ signaling pathway.


Dietary patterns and nutrient intakes of 7-year-old children taking part in an atherosclerosis prevention project in Finland.
To evaluate the dietary patterns of 7-year-old children participating in an atherosclerosis prevention project and the relationship of those dietary patterns to nutrient intakes and serum cholesterol values. In the randomized, prospective Special Turku Coronary Risk Factor Intervention Project (STRIP) 1,062 children were randomly assigned to an intervention group (n=540; low-saturated fat, low-cholesterol diet) or to a control group (n=522; unrestricted diet) at 7 months of age. The intervention families received, at 6-month intervals, individualized counseling that focused on the known environmental atherosclerosis risk factors and aimed at reducing children's saturated fat and cholesterol intake. Nutrition counseling was targeted at the child but, because of the young age of the children, was given to the parents. When children were 7 years old, food and nutrient intakes of 307 intervention and 323 control children were studied using 4-day food records. K-means cluster analysis was used to classify children into 4 groups on the basis of similarity of food intake. Differences in nutrient intakes and serum lipid concentrations between children in the 4 food intake clusters were evaluated using Tukey's multiple comparison test. Intervention children dominated the bread, skim milk, and margarine cluster and the cereals, rice, and pasta cluster whereas the 1.5%-fat milk and butter cluster included mainly control children. Saturated fat intake was nearest to the recommendations, that is 11.7% and 11.9% of energy, in the bread, skim milk, and margarine cluster and the cereals, rice, and pasta cluster, respectively. Children in the bread, skim milk, and margarine cluster had 20% to 27% higher fiber intakes (P<.001) whereas children in the sugar and sweets cluster had markedly higher sugar intakes than children in other clusters (P<.001). Serum cholesterol concentrations were lower in those clusters with high dietary ratios of polyunsaturated to saturated fat. Detailed and repeated dietary counseling of parents, starting when children are aged 7 months, that aims at decreasing children's exposure to known nutrition risk factors for coronary heart disease modifies children's food patterns and nutrient intakes toward expected values.


Modelling time-varying volatility using GARCH models: evidence from the Indian stock market.
Background: In this study, we examined the volatility of the Indian stock market from 2008 to 2021. Owing to the financial crisis, volatility forecasting of the Indian stock market has become crucial for economic and financial analysts. An empirical study of the returns of the NSE indices revealed an autoregressive conditional heteroskedastic trend in the Indian stock market. Methods: Using GARCH 1, 1 (generalized autoregressive conditional heteroskedasticity) and FIGARCH (fractionally integrated GARCH), we examine investor behaviour and the persistence of long-term volatility. Results: The empirical findings of the estimated models suggest that shocks persist for a long time in NSE returns. Furthermore, bad news has a greater impact on stock volatility than good news. The return on assets is stable but highly volatile, even though the Indian economy has experienced the global crash to some extent. Conclusions: Models of volatility derived from the GARCH equation provide accurate forecasts and are useful for portfolio allocation, performance measurement, and option valuation.


Crosstalk between Gut Sensory Ghrelin Signaling and Adipose Tissue Sympathetic Outflow Regulates Metabolic Homeostasis.
The stomach-derived orexigenic hormone ghrelin is a key regulator of energy homeostasis and metabolism in humans. The ghrelin receptor, growth hormone secretagogue receptor 1a (GHSR), is widely expressed in the brain and gastrointestinal vagal sensory neurons, and neuronal GHSR knockout results in a profoundly beneficial metabolic profile and protects against diet-induced obesity (DIO) and insulin resistance. Here we show that in addition to the well characterized vagal GHSR, GHSR is robustly expressed in gastrointestinal sensory neurons emanating from spinal dorsal root ganglia. Remarkably, sensory neuron GHSR deletion attenuates DIO through increased energy expenditure and sympathetic outflow to adipose tissue independent of food intake. In addition, neuronal viral tract tracing reveals prominent crosstalk between gut non-vagal sensory afferents and adipose sympathetic outflow. Hence, these findings demonstrate a novel gut sensory ghrelin signaling pathway critical for maintaining energy homeostasis.


Spermidine exodus and oxidation in the apoplast induced by abiotic stress is responsible for H2O2 signatures that direct tolerance responses in tobacco.
Polyamines (PAs) exert a protective effect against stress challenges, but their molecular role in this remains speculative. In order to detect the signaling role of apoplastic PA-derived hydrogen peroxide (H2O2) under abiotic stress, we developed a series of tobacco (Nicotiana tabacum cv Xanthi) transgenic plants overexpressing or downregulating apoplastic polyamine oxidase (PAO; S-pao and A-pao plants, respectively) or downregulating S-adenosyl-l-methionine decarboxylase (samdc plants). Upon salt stress, plants secreted spermidine (Spd) into the apoplast, where it was oxidized by the apoplastic PAO, generating H2O2. A-pao plants accumulated less H2O2 and exhibited less programmed cell death (PCD) than did wild-type plants, in contrast with S-pao and samdc downregulating plants. Induction of either stress-responsive genes or PCD was dependent on the level of Spd-derived apoplastic H2O2. Thus, in wild-type and A-pao plants, stress-responsive genes were efficiently induced, although in the latter at a lower rate, while S-pao plants, with higher H2O2 levels, failed to accumulate stress-responsive mRNAs, inducing PCD instead. Furthermore, decreasing intracellular PAs, while keeping normal apoplastic Spd oxidation, as in samdc downregulating transgenic plants, caused enhanced salinity-induced PCD. These results reveal that salinity induces the exodus of Spd into the apoplast, where it is catabolized by PAO, producing H2O2. The accumulated H2O2 results in the induction of either tolerance responses or PCD, depending also on the levels of intracellular PAs.


Analysis of particle-borne odorants emitted from concentrated animal feeding operations.
Airborne particles are known to serve as a carrier of odors emanating from concentrated animal feeding operations (CAFOs). However, limited quantitative data about particle-borne odorants preclude an accurate assessment of the role of particles in odor transport. This study collected total suspended particulates (TSP) and PM10 (particles with aerodynamic diameter smaller than 10 μm) at the air exhaust of eight types of CAFOs (swine: farrowing, gestation, weaning, and finishing; poultry: manure-belt layer hen, tom turkey, chicken broiler, and cage-free layer hen; in total 20 animal buildings) in multiple seasons, and examined the variability in particle odorant composition with animal operation type, season, and particle size. Fifty-seven non-sulfur-containing odorants were identified and quantitated, including carbonyls, alcohols, acids, phenols, and nitrogen-containing compounds. They in total accounted for 2.19±1.52% TSP and 4.97±3.25% PM10 mass. Acetic acid and ethanol were most abundant but less odor-contributing than phenylacetic acid, indole, dodecanoic acid, and (E,E)-2,4-decadienal, as determined by odor activity value. Particle odorant composition varied significantly with animal operation type, season, and particle size. The TSP and PM10 samples from swine gestation buildings, for example, showed distinctly different odorant compositions than those from tom turkey buildings. The summer TSP and PM10 samples contained in general lower concentrations of short-chain fatty acids but higher concentrations of long-chain fatty acids, aldehydes, and short-chain alcohols than the winter samples. Compared to TSP, PM10 samples from different types of CAFOs shared a more similar odorant composition, contained higher odorant concentrations per mass of particles, and accounted for on average 53.2% of the odor strength of their corresponding TSP samples.


Conditional expression of a truncated fragment of nonmuscle myosin II-A alters cell shape but not cytokinesis in HeLa cells.
A truncated fragment of the nonmuscle myosin II-A heavy chain (NMHC II-A) lacking amino acids 1-591, delta N592, was used to examine the cellular functions of this protein. Green fluorescent protein (GFP) was fused to the amino terminus of full-length human NMHC II-A, NMHC II-B, and delta N592 and the fusion proteins were stably expressed in HeLa cells by using a conditional expression system requiring absence of doxycycline. The HeLa cell line studied normally expressed only NMHC II-A and not NMHC II-B protein. Confocal microscopy indicated that the GFP fusion proteins of full-length NMHC II-A, II-B, and delta N592 were localized to stress fibers. However, in vitro assays showed that baculovirus-expressed delta N592 did not bind to actin, suggesting that delta N592 was localized to actin stress fibers through incorporation into endogenous myosin filaments. There was no evidence for the formation of heterodimers between the full-length endogenous nonmuscle myosin and truncated nonmuscle MHCs. Expression of delta N592, but not full-length NMHC II-A or NMHC II-B, induced cell rounding with rearrangement of actin filaments and disappearance of focal adhesions. These cells returned to their normal morphology when expression of delta N592 was repressed by addition of doxycycline. We also show that GFP-tagged full-length NMHC II-A or II-B, but not delta N592, were localized to the cytokinetic ring during mitosis, indicating that, in vertebrates, the amino-terminus part of mammalian nonmuscle myosin II may be necessary for localization to the cytokinetic ring.


Common Variants Associated With OSMR Expression Contribute to Carotid Plaque Vulnerability, but Not to Cardiovascular Disease in Humans.
Background and Aims: Oncostatin M (OSM) signaling is implicated in atherosclerosis, however the mechanism remains unclear. We investigated the impact of common genetic variants in OSM and its receptors, OSMR and LIFR, on overall plaque vulnerability, plaque phenotype, intraplaque OSMR and LIFR expression, coronary artery calcification burden and cardiovascular disease susceptibility. Methods and Results: We queried Genotype-Tissue Expression data and found that rs13168867 (C allele) was associated with decreased OSMR expression and that rs10491509 (A allele) was associated with increased LIFR expression in arterial tissues. No variant was significantly associated with OSM expression. We associated these two variants with plaque characteristics from 1,443 genotyped carotid endarterectomy patients in the Athero-Express Biobank Study. After correction for multiple testing, rs13168867 was significantly associated with an increased overall plaque vulnerability (β = 0.118 ± s.e. = 0.040, p = 3.00 × 10-3, C allele). Looking at individual plaque characteristics, rs13168867 showed strongest associations with intraplaque fat (β = 0.248 ± s.e. = 0.088, p = 4.66 × 10-3, C allele) and collagen content (β = -0.259 ± s.e. = 0.095, p = 6.22 × 10-3, C allele), but these associations were not significant after correction for multiple testing. rs13168867 was not associated with intraplaque OSMR expression. Neither was intraplaque OSMR expression associated with plaque vulnerability and no known OSMR eQTLs were associated with coronary artery calcification burden, or cardiovascular disease susceptibility. No associations were found for rs10491509 in the LIFR locus. Conclusions: Our study suggests that rs1316887 in the OSMR locus is associated with increased plaque vulnerability, but not with coronary calcification or cardiovascular disease risk. It remains unclear through which precise biological mechanisms OSM signaling exerts its effects on plaque morphology. However, the OSM-OSMR/LIFR pathway is unlikely to be causally involved in lifetime cardiovascular disease susceptibility.


[The mycoplasmas isolated from respiratory specimens].
Mycoplasma pneumoniae is the species that is considered pathogen for respiratory tract. Other species, M. hominis and U. urealyticum, have been isolated from respiratory specimens and its aetiological relation with respiratory infections is not clear. In this study, mycoplasma from respiratory specimens have been isolated from HIV infected patients, patients suffering from chronic bronchitis and patients with respiratory infections without any underlying disease, using SP-4 media and being identified with glucose fermentation test, guinea pig blood cells adsorption, tetrazolium reduction, urea test, arginine test, and antigenic study by an immunoassay on the colonies grown on SP-4 solid medium. M. hominis has been the species most frequent isolated, mainly from patients without any underlying disease and from HIV infected patients, and less frequently from patients suffering from chronic bronchitis, the differences between both groups being significative. Other Mycoplasma, no identified at he species level, have been isolated although without any significative differences. In only one case U. urealyticum has been isolated. No M. pneumoniae isolation was obtained.


Implantable polymeric microneedles with phototriggerable properties as a patient-controlled transdermal analgesia system.
Adequate pain control can be achieved using a patient-controlled drug delivery system that can provide analgesia to patients as needed. To achieve this objective, we developed a phototriggered microneedle (MN) system that enables the on-demand delivery of pain medications to the skin under external near-infrared (NIR) light stimulation. In this system, polymeric MNs, containing NIR absorbers and analgesics, are combined with a poly(l-lactide-co-d,l-lactide) supporting array. A "removable design" of the supporting array enables the quick implantation of the MNs into the skin to act as a drug depot, thus shortening the patch application time. Upon irradiation with NIR light, the NIR absorbers in the implanted MNs can absorb light energy and induce a phase transition in the MNs to activate drug release. We demonstrated that lidocaine release can be modulated or repeatedly triggered by varying the duration of irradiation and controlling the on and off status of the laser. Lidocaine delivered by the implanted MNs can be rapidly absorbed into the blood circulation within 10 min and has a bioavailability of at least 95% relative to the subcutaneous injection, showing that the proposed system has the potential to provide a rapid onset of pain relief. Such an implantable device may allow pain sufferers receiving the painkiller without the need for multiple needle injections, and may enable controlling pain more conveniently and comfortably.


Current concepts of integrated coronary physiology in the catheterization laboratory.
Over the last 15 years, the use of invasive coronary physiology in the catheterization laboratory has demonstrated favorable outcomes for decision making in patients with intermediate single-vessel stenoses, complex bifurcation and ostial branch stenoses, multivessel coronary artery disease, and left main stenoses. A recent large multicenter study (FAME [FFR versus Angiography for Multivessel Evaluation]) found that a physiologically-guided approach was superior to the standard angiographically-guided approach for percutaneous revascularization in patients with multivessel coronary artery disease. This review addresses selected pertinent concepts and studies supporting the integration of coronary physiology in the catheterization laboratory for optimal patient outcomes.


Isolation and characterization of a novel alkalitolerant thermophile, Anaerobranca horikoshii gen. nov., sp. nov.
Nine moderately alkalitolerant thermophilic bacteria with similar properties were isolated from water and soil samples obtained from Yellowstone National Park. These Gram-type-positive, rod-shaped bacteria produce cells with primary branches. The cells are peritrichous and exhibit only slight tumbling motility. At 60 degrees C the pH range for growth is 6.9 to 10.3, and the optimum pH is 8.5. At pH 8.5 the temperature range for growth is 34 to 66 degrees C, with an optimum temperature of 57 degrees C. The strains are mainly proteolytic. The fermentation products from yeast extract are acetate, CO2, and H2. Fumarate added to minimal medium containing yeast extract is stoichiometrically converted to succinate, indicating that it is used as an alternative electron acceptor. The DNA G + C content is 33 to 34 mol%. On the basis of its unique properties, such as branch formation, growth at alkaline pH values at elevated temperatures, and the relative distance of its 16S rRNA sequence from those of other known bacteria, we propose that strain JW/YL-138T (T = type strain) and eight similar strains represent a new genus and species, Anaerobranca horikoshii. Strain JW/YL-138 is designated the type strain of the type species, A. horikoshii, which was named in honor of Koki Horikoshi, a pioneer in the field of alkaliphilic bacteria.


Immunomorphological Pattern of Molecular Chaperones in Normal and Pathological Thyroid Tissues and Circulating Exosomes: Potential Use in Clinics.
The thyroid is a major component of the endocrine system and its pathology can cause serious diseases, e.g., papillary carcinoma (PC). However, the carcinogenic mechanisms are poorly understood and clinical useful biomarkers are scarce. Therefore, we determined if there are quantitative patterns of molecular chaperones in the tumor tissue and circulating exosomes that may be useful in diagnosis and provide clues on their participation in carcinogenesis. Hsp27, Hsp60, Hsp70, and Hsp90 were quantified by immunohistochemistry in PC, benign goiter (BG), and normal peritumoral tissue (PT). The same chaperones were assessed in plasma exosomes from PC and BG patients before and after ablative surgery, using Western blotting. Hsp27, Hsp60, and Hsp90 were increased in PC in comparison with PT and BG but no differences were found for Hsp70. Similarly, exosomal levels of Hsp27, Hsp60, and Hsp90 were higher in PC than in BG, and those in PC were higher before ablative surgery than after it. Hsp27, Hsp60, and Hsp90 show distinctive quantitative patterns in thyroid tissue and circulating exosomes in PC as compared with BG, suggesting some implication in the carcinogenesis of these chaperones and indicating their potential as biomarkers for clinical applications.


Haploidentical bone marrow and stem cell transplantation: experience with post-transplantation cyclophosphamide.
Allogeneic blood or bone marrow transplantation (BMT) is a potentially curative therapy for high-risk hematologic malignancies not curable by standard chemotherapy, but the procedure is limited by the availability of human leukocyte antigen-matched donors for many patients, as well as toxicities including graft-versus-host disease (GVHD). Our group has developed the use of high-dose post-transplantation cyclophosphamide (PTCy) to selectively remove alloreactive T cells without compromising engraftment. This protocol has allowed for successful transplantation of human leukocyte antigen (HLA)-haploidentical (haplo) grafts, thus expanding the donor pool for the many patients who would not otherwise be a candidate for this life-saving procedure. In this review we will summarize the data that led to the development of PTCy, then focus on the outcomes of haploBMT trials with PTCy across different transplant platforms for patients with malignant hematologic diseases, and finally we will discuss emerging evidence that suggests equivalency of haploBMT with PTCy compared with more traditional transplants.


Extent of cell differentiation and capacity for cartilage synthesis in human adult adipose-derived stem cells: comparison with fetal chondrocytes.
This study evaluated the extent of differentiation and cartilage biosynthetic capacity of human adult adipose-derived stem cells relative to human fetal chondrocytes. Both types of cell were seeded into nonwoven-mesh polyglycolic acid (PGA) scaffolds and cultured under dynamic conditions with and without addition of TGF-beta1 and insulin. Gene expression for aggrecan and collagen type II was upregulated in the stem cells in the presence of growth factors, and key components of articular cartilage such as glycosaminoglycan (GAG) and collagen type II were synthesized in cultured tissue constructs. However, on a per cell basis and in the presence of growth factors, accumulation of GAG and collagen type II were, respectively, 3.4- and 6.1-fold lower in the stem cell cultures than in the chondrocyte cultures. Although the stem cells synthesized significantly higher levels of total collagen than the chondrocytes, only about 2.4% of this collagen was collagen type II. Relative to cultures without added growth factors, treatment of the stem cells with TGF-beta1 and insulin resulted in a 59% increase in GAG synthesis, but there was no significant change in collagen production even though collagen type II gene expression was upregulated 530-fold. In contrast, in the chondrocyte cultures, synthesis of collagen type II and levels of collagen type II as a percentage of total collagen more than doubled after growth factors were applied. Although considerable progress has been achieved to develop differentiation strategies and scaffold-based culture techniques for adult mesenchymal stem cells, the extent of differentiation of human adipose-derived stem cells in this study and their capacity for cartilage synthesis fell considerably short of those of fetal chondrocytes.


High-dose chemotherapy and autologous stem cell transplantation for primary refractory or relapsed Hodgkin lymphoma: long-term outcome in the first 100 patients treated in Vancouver.
Beginning in 1985, patients in British Columbia with Hodgkin lymphoma (HL) that was not controlled by conventional chemotherapy routinely underwent high-dose chemotherapy and autologous stem cell transplantation (HD-ASCT). Long-term complications of HD-ASCT have become apparent as more patients survive without recurrence of HL. Data were obtained retrospectively on the first 100 patients that underwent HD-ASCT for HL in Vancouver, focusing on relapse, treatment-related complications, and the occurrence of late events. Fifty-three patients remain alive (median follow-up, 11.4 years [range, 10.0-17.4 years]) with an overall survival (OAS) of 54% at 15 years. OAS was significantly better in patients in first relapse (67%) than in patients with primary refractory-induction failure (39%) and advanced disease (29%) (P = .002). The major cause of death was progression of HL (32% at 15 years). Treatment-related mortality, including death from second malignancy, was 17% at 15 years. Cumulative risk of a second malignancy was 9% at 15 years. Karnofsky performance status was at least 90% in 47 patients although hypogonadism (20 patients), hypothyroidism (12 patients), unusual infections (10 patients), anxiety or depression (7 patients), and cardiac disease (5 patients) were not uncommon in survivors. HD-ASCT can lead to durable remissions in relapsed or refractory HL with acceptable but definite late toxicity. The occurrence of late events necessitates lifelong medical surveillance.


Delayed presentation of small bowel injury during suprapubic catheterisation.
We present a case of small intestine injury resulting from suprapubic catheter insertion. This case is of particular interest for three reasons. Firstly, the presentation of the injury was delayed by three months, until the time of the first catheter exchange. Secondly, the injury was managed conservatively, without surgical exploration. Finally, the injury occurred using a newer, Seldinger-type suprapubic catheter insertion kit.


Monitoring Response to Home Parenteral Nutrition in Adult Cancer Patients.
Current guidelines recommend home parenteral nutrition (HPN) for cancer patients with chronic deficiencies of dietary intake or absorption when enteral nutrition is not adequate or feasible in suitable patients. HPN has been shown to slow down progressive weight loss and improve nutritional status, but limited information is available on the monitoring practice of cancer patients on HPN. Clinical management of these patients based only on nutritional status is incomplete. Moreover, some commonly used clinical parameters to monitor patients (weight loss, body weight, body mass index, and oral food intake) do not accurately reflect patient's body composition, while bioelectrical impedance analysis (BIA) is a validated tool to properly assess nutritional status on a regular basis. Therefore, patient's monitoring should rely on other affordable indicators such as Karnofsky Performance Status (KPS) and modified Glasgow Prognostic Score (mGPS) to also assess patient's functional status and prognosis. Finally, catheter-related complications and quality of life represent crucial issues to be monitored over time. The purpose of this narrative review is to describe the role and relevance of monitoring cancer patients on HPN, regardless of whether they are receiving anticancer treatments. These practical tips may be clinically useful to better guide healthcare providers in the nutritional care of these patients.


Nucleotide sequence and expression in vitro of cDNA derived from mRNA of int-1, a provirally activated mouse mammary oncogene.
The mouse int-1 gene is a putative mammary oncogene discovered as a target for transcriptionally activating proviral insertion mutations in mammary carcinomas induced by the mouse mammary tumor virus in C3H mice. We have isolated molecular clones of full- or nearly full-length cDNA transcribed from int-1 RNA (2.6 kilobases) in a virus-induced mammary tumor. Comparison of the nucleotide sequence of the cDNA clones with that of the int-1 gene (A. van Ooyen and R. Nusse, Cell 39:233-240, 1984) shows the following. The coding region of the int-1 gene is composed of four exons. The splice donor and acceptor sites conform to consensus; however, at least two closely spaced polyadenylation sites are used, and the transcriptional initiation site remains ambiguous. The major open reading frame is preceded by an open frame 10 codons in length. The mRNA encodes a 41-kilodalton protein with several striking features--a strongly hydrophobic amino terminus, a cysteine-rich carboxy terminus, and four potential glycosylation sites. There are no differences in nucleotide sequence between the known exons of the normal and a provirally activated allele. The length of the deduced open reading frame was further confirmed by in vitro translation of RNA transcribed from the cDNA clones with SP6 RNA polymerase.


Accuracy of the gaussian point spread function model in 2D localization microscopy.
The gaussian function is simple and easy to implement as Point Spread Function (PSF) model for fitting the position of fluorescent emitters in localization microscopy. Despite its attractiveness the appropriateness of the gaussian is questionable as it is not based on the laws of optics. Here we study the effect of emission dipole orientation in conjunction with optical aberrations on the localization accuracy of position estimators based on a gaussian model PSF. Simulated image spots, calculated with all effects of high numerical aperture, interfaces between media, polarization, dipole orientation and aberrations taken into account, were fitted with a gaussian PSF based Maximum Likelihood Estimator. For freely rotating dipole emitters it is found that the gaussian works fine. The same, theoretically optimum, localization accuracy is found as if the true PSF were a gaussian, even for aberrations within the usual tolerance limit of high-end optical imaging systems such as microscopes (Marechal's diffraction limit). For emitters with a fixed dipole orientation this is not the case. Localization errors are found that reach up to 40 nm for typical system parameters and aberration levels at the diffraction limit. These are systematic errors that are independent of the total photon count in the image. The gaussian function is therefore inappropriate, and more sophisticated PSF models are a practical necessity.


Effects of changes in health insurance reimbursement level on outpatient service utilization of rural diabetics: evidence from Jiangsu Province, China.
Outpatient reimbursement levels of the New Rural Cooperative Medical Scheme have changed in recent years in China, and those changes may have a greater impact on patients with chronic diseases due to their higher outpatient expenses. This study represents the first attempt to identify the effects of reimbursement level on outpatient service utilization for chronic patients in rural China and it also gives strong estimation results by conducting a tracer illness study in order to control for possible biases associated with studying several diseases together. This study used difference-in-differences models to examine how changes in yearly maximum reimbursement amount and outpatient reimbursement rates affected rural residents with type 2 diabetes in three counties in Jiangsu Province, China. Other factors, such as sex, age and severity of illness, were also included in the model estimations. To make sure the treated group and control group are comparable, Propensity Score Match (PSM) was used to analysis the gender, age and severity of illness of the two groups. The results indicate that an increase in yearly maximum reimbursement amount for outpatient visits could cause an increase in yearly total outpatient expenses for patients with type 2 diabetes mellitus. However, changes in outpatient reimbursement rates between 2010 and 2011 did not significantly affect the utilization of different types of health institution. The reimbursement rates of village clinics should be substantially increased from the existing basis and the gap of reimbursement rates among different institutions should be further widened. It is also important for village clinics to improve their services. Moreover, measures to improve the quality of care and scope of services at lower-level healthcare institutions, and promote the health service utilization of rural women should be considered.


A nitrate-inducible ferredoxin in maize roots. Genomic organization and differential expression of two nonphotosynthetic ferredoxin isoproteins.
We have identified and characterized a nitrate-inducible ferredoxin (Fd) in maize (Zea mays L.) roots by structural analysis of the purified protein and by cloning of its cDNA and gene. In maize Fd isoproteins are encoded by a small multigene family, and the nitrate-inducible Fd was identified as a novel isoprotein, designated Fd VI, which differed from an Fd I to Fd V identified to date. In the roots of seedlings cultured without nitrate, Fd VI was undetectable. However, during the induction of the capacity for nitrate assimilation, the amount of Fd VI increased markedly within 24 h. Concurrently, the level of transcript for Fd VI increased, but more quickly, reaching a maximal level within 2 h with kinetics similar to those of nitrite reductase and Fd-NADP+ reductase. Fd III was constitutively expressed in roots, and no such changes at the protein and mRNA levels were observed during the nitrate induction. In the 5' flanking region of the gene for Fd VI only, we identified NIT-2 motifs, which are widely found in genes for enzymes related to nitrogen metabolism. These data indicate that Fd VI is co-induced with the previously characterized enzymes involved in nitrate assimilation, and they suggest that the novel Fd isoprotein, distinct from the constitutively expressed Fd, might play an important role as an electron carrier from NADPH to nitrite reductase and other Fd-dependent enzymes in root plastids.


Decursin attenuates hepatic fibrogenesis through interrupting TGF-beta-mediated NAD(P)H oxidase activation and Smad signaling in vivo and in vitro.
We studied that a potent antifibrotic effect of decursin on in vivo liver damage model and the mechanism in inhibiting which transforming growth factor (TGF)-β1-induced human hepatic stellate cells (HSCs) activation. Liver injury was induced in vivo by intraperitoneal injection of carbon tetrachloride (CCl4) with or without decursin for 4weeks in mice. Human hepatic stellate cell line, an immortalized human HSC line, was used in in vitro assay system. The effects of decursin on HSC activation were measured by analyzing the expression of α-smooth muscle actin (α-SMA) and collagen I in liver tissue and human HSCs. Decursin treatment significantly reduced the ratio of liver/body weight, α-SMA activation, and type I collagen overexpression in CCl4 treated mice liver. The elevated serum levels, including ALT, AST, and ALP, were also decreased by decursin treatment. Treatment of decursin markedly proved the generation of reactive oxygen species, NAD(P)H oxidase (NOX) protein (1, 2, and 4) upregulation, NOX activity, and superoxide anion production in HSCs by TGF-β1. It also significantly reduced TGF-β1-induced Smad 2/3 phosphorylation, nuclear translocation of Smad 4, and association of Smad 2/3-Smad 4 complex. Consistent with in vitro results, decursin treatment effectively blocked the levels of NOX protein, and Smad 2/3 phosphorylation in injured mice liver. Decursin blocked CCl4-induced liver fibrosis and inhibited TGF-β1-mediated HSC activation in vitro. These data demonstrated that decursin exhibited hepatoprotective effects on experimental fibrosis, potentially by inhibiting the TGF-β1 induced NOX activation and Smad signaling.


Nurturing Meaningful Intergenerational Social Engagements to Support Healthy Brain Aging for Anishinaabe Older Adults.
The emergence of Alzheimer's disease and related dementias (ADRD) in Indigenous populations across Canada is of rising concern, as prevalence rates continue to exceed those of non-Indigenous populations. The Intergenerativity Model, guided by Indigenous Ways of Knowing, nurtures a psychosocial approach to promoting healthy brain aging and quality of life. Community-based participatory action methods led by interviews, focus groups, and program observations aid in identifying the barriers to and facilitators of success for intergenerational social engagements in the Anishinaabe community of Wiikwemkoong in northwestern Ontario. A qualitative thematic analysis guides future recommendations for programming opportunities that foster traditional roles of older First Nation adults and support intergenerational relationships. The results of this project elicit culturally appropriate recommendations for community-driven supports that address healthy brain aging. These outcomes are relevant to other Indigenous communities as the framework for determining that culturally appropriate health supports can be adapted to the unique context of many communities.


Cortical Thickness Changes and Their Relationship to Dual-Task Performance following Mild Traumatic Brain Injury in Youth.
Mild traumatic brain injury (mTBI) is common in youth, especially in those who participate in sport. Recent investigations from our group have shown that asymptomatic children and adolescents with mTBI continue to exhibit alterations in neural activity and cognitive performance compared with those without a history of mTBI. This is an intriguing finding, given that current return-to-learn and return-to-play protocols rely predominately on subjective symptom reports, which may not be sensitive enough to detect subtle injury-related changes. As a result, youth may be at greater risk for re-injury and long-term consequences if they are cleared for activity while their brains continue to be compromised. It is currently unknown whether mTBI also affects brain microstructure in the developing brain, particularly cortical thickness, and whether such changes are also related to cognitive performance. The present study examined cortical thickness in 13 asymptomatic youth (10-14 years old) who had sustained an mTBI 3-8 months prior to testing compared with 14 age-matched typically developing controls. Cortical thickness was also examined in relation to working memory performance during single and dual task paradigms. The results show that youth who had sustained an mTBI had thinner cortices in the left dorsolateral prefrontal region and right anterior and posterior inferior parietal lobes. Additionally, cortical thinning was associated with slower reaction time during the dual-task condition in the injured youth only. The results also point to a possible relationship between functional and structural alterations as a result of mTBI in youth, and lend evidence for neural changes beyond symptom resolution.


Tissue distribution and retention of 5-thio-D-glucose in animal tumor models.
5-Thio-D-glucose (5-TDG) has demonstrated potential as a radiation sensitizer for hypoxic cells growing in monolayer culture and as multicell spheroids. We have studied the biodistribution and pharmacokinetics of radiolabeled (35S or 3H) 5-TDG prior to utilizing this bioisotere of D-glucose for radiation sensitization studies in vivo. At all times after administration, 5-TDG achieved higher tissue levels of radioactivity in hamster pancreatic tumors of ductal and acinar cell origin than in the normal pancreas. By 2 hours after intravenous injection, the duct tumor activity concentration exceeded that of any other tissue; a similar situation obtained after 4 hours in the acinar tumor model. However, the functioning insulinoma model exhibited similar radioactivity uptake and kinetics as did normal pancreas. In all models, metastases behaved similarly to the primary transplanted tumor in terms of uptake and retention of radioactivity. Addition of carrier 5-TDG appeared to abrogate much of the selective uptake of label into tumor relative to normal tissue, in terms of % dose/g tissue, but the total uptake into tumor was greater. The drug is rapidly excreted in the urine and the nature of this excretion was examined. Retention of activity in normal lung tissue was similar to that of pancreatic tumors and prompted a study of uptake into Lewis lung carcinoma. Again, tumor uptake in % dose/g approximated uptake into normal lung tissue, but total uptake into tumor was greater. These studies provided the basis for initiation of in vivo studies of potential cytotoxic and radiation-potentiating activity of 5-TDG.


Changes in brain volume and cognition in a randomized trial of exercise and social interaction in a community-based sample of non-demented Chinese elders.
Physical exercise has been shown to increase brain volume and improve cognition in randomized trials of non-demented elderly. Although greater social engagement was found to reduce dementia risk in observational studies, randomized trials of social interventions have not been reported. A representative sample of 120 elderly from Shanghai, China was randomized to four groups (Tai Chi, Walking, Social Interaction, No Intervention) for 40 weeks. Two MRIs were obtained, one before the intervention period, the other after. A neuropsychological battery was administered at baseline, 20 weeks, and 40 weeks. Comparison of changes in brain volumes in intervention groups with the No Intervention group were assessed by t-tests. Time-intervention group interactions for neuropsychological measures were evaluated with repeated-measures mixed models. Compared to the No Intervention group, significant increases in brain volume were seen in the Tai Chi and Social Intervention groups (p < 0.05). Improvements also were observed in several neuropsychological measures in the Tai Chi group, including the Mattis Dementia Rating Scale score (p = 0.004), the Trailmaking Test A (p = 0.002) and B (p = 0.0002), the Auditory Verbal Learning Test (p = 0.009), and verbal fluency for animals (p = 0.01). The Social Interaction group showed improvement on some, but fewer neuropsychological indices. No differences were observed between the Walking and No Intervention groups. The findings differ from previous clinical trials in showing increases in brain volume and improvements in cognition with a largely non-aerobic exercise (Tai Chi). In addition, intellectual stimulation through social interaction was associated with increases in brain volume as well as with some cognitive improvements.


Genotyping of Cryptosporidium spp. isolated from human stool samples in Switzerland.
In a study to estimate the frequency of Cryptosporidium infections in Switzerland, stool samples from patients found to be positive for Cryptosporidium spp. by modified Ziehl-Neelson staining and fluorescence microscopy were used for genotyping experiments. With 9 of 12 samples, DNA extraction and subsequent genotyping was successful. All Cryptosporidium-isolates belonged to the bovine genotype. In one stool sample, two strains of Cryptosporidium were demonstrated, suggesting a mixed infection. In comparison with reference strains from calves, one of the isolates showed a full sequence identity and the other a similarity of 97.5%. The fact that only bovine genotypes were detected suggests, that cryptosporidiosis must primarily be considered as a zoonotic disease in Switzerland. This is in contrast to other countries, where the human genotype of C. parvum was shown to dominate the epidemiological situation. The results of our study are supported by the previous finding, that two of the analysed strains originated from patients who used to consume raw milk or raw cream, a known risk factor for cryptosporidiosis.