Migration of cyclo-diBA from coatings into canned food: method of analysis, concentration determined in a survey and in silico hazard profiling.
Cyclo-diBA, the cyclic product formed from bisphenol A and bisphenol A diglycidyl ether during production of epoxy resins, was measured in canned food using reversed phase HPLC with fluorescence detection. Half (9 of 17) of the samples of canned fish in oil collected in April 2010 contained cyclo-diBA with an average concentration of 1025 μg/kg and a maximum of 1980 μg/kg. In September 2012, cyclo-diBA was detectable (>25 μg/kg) in merely 13 from 44 such products; the average concentration in these was 807 μg/kg and the maximum now reached 2640 μg/kg. Fish in brine contained far less cyclo-diBA. The majority of the canned meat products contained cyclo-diBA at a mean concentration of 477 μg/kg and a maximum of 1050 μg/kg. All prepared meals, such as ravioli or soups, contained cyclo-diBA, with a mean at 287 μg/kg. In canned tomatoes, peas and other vegetables in water or fruits in syrup, no cyclo-diBA was detected (<25 μg/kg). Since no experimental toxicity data are available except for its cytotoxicity, an in silico hazard profiling was performed. Cyclo-diBA seems to be stable and of low reactivity. There is indication for considerable oral bioavailability and for the potential to accumulate in the human body. Cyclo-diBA can be metabolized into cyclic and acyclic compounds. Based on SAR assessment for cyclo-diBA and read-across from BADGE to linear cyclo-diBA metabolites, genotoxic effects are improbable. Specific binding of cyclo-diBA to nuclear receptors, such as ERβ, can be predicted, indicating a potential endocrine-disrupting potency. The limit by the EFSA guidelines of 50 μg/person/d for compounds shown not to be genotoxic as well as the TTC-based Cramer structural class III value of 90 μg/person/d could be exceeded several fold by high consumers of canned fish in oil with high brand loyalty. As a consequence, risk reduction measures were taken.


Remote care: bidding our ICD patients a fond farewell?
Remote follow-up has proven its safety and efficacy in the literature, and indeed in daily use. It saves the patient time and travel, allows earlier detection of events, and reduces inappropriate shocks. At the same time, it is not infallible and definitely not a universal substitute for in-office visits, as the 5 cases below demonstrate. It saves time, but also takes some time, with problems and challenges we were hitherto not familiar with. Hopes for the future include uniform definitions regarding policy and responsibilities, reimbursement, even better connectivity, better alert management and remote programmability. To end on a more utopic note, it would be a huge step forward if devices and leads were so reliable that we did not need remote care to detect malfunctions.


Indocyanine green-based adjunctive antimicrobial photodynamic therapy for treating chronic periodontitis: A randomized clinical trial.
The objective was to evaluate the efficiency of indocyanine green (ICG)-based adjunctive antimicrobial photodynamic therapy (aPDT) in a prospective clinical study regarding non-surgical treatment of chronic periodontitis. Affected teeth of twenty patients were treated with scaling and root planing (control group). Using a split-mouth design, two quadrants received additional ICG-based (perio green®, 0.1 mg/ml) aPDT (test group) with a diode laser at 808 nm (100 mW at 2 kHz). Clinical assessment of bleeding on probing (BOP), sulcus fluid flow rate (SFFR) and microbiological analysis were performed at baseline, two weeks, three and six months after treatment. Relative attachment level (RAL), probing depths (PD) and gingival recession (GR), were also analyzed. At baseline, none of the assessed parameters showed significant differences between the test and control groups. Median values for BOP, RAL, PD, decreased significantly in both groups after three months of treatment (p ≤ 0.05) without significant difference between the groups. Two weeks after treatment, the SFFR showed significantly lower mean values in the test group (aPDT). Within the study limits, the only significant difference between the control group and the aPDT group was a transient smaller amount of SFFR in the latter during the first follow-up. With the applied parameters, this study does not conclusively support ICG-based aPDT, though it is promising because no adverse effects occurred. The precise modes of action of ICG must be elucidated, and further clinical trials are needed.


Rhodamine B induces oxidative stress and cervical epithelial cell proliferation in the uterus.
This study aimed to investigate the effect of Rhodamine B exposure on oxidative stress and cervical epithelial cells proliferation in the uterus. Twenty eight female Wistar albino rats were divided into four groups (n = 7 each): one control (untreated) group; and three Rhodamine B groups at several doses (4.5, 9, 18 mg/200 g body weight/day) for 36 days. Colometric analysis of malondialdehyde (MDA) level as a marker of lipid peroxidation and histological analysis of the cervical epithelial cells proliferation was performed. The MDA levels and proliferation of epithelial cells were significantly higher in all Rhodamine B groups compared to control group (P < 0.05). The MDA levels were increased in a dose-dependent manner in the Rhodamine B groups. Moreover, the proliferation of epithelial cells was also increased by Rhodamine B in a dose-dependent manner. In conclusion, subchronic Rhodamine B administration induces lipid peroxidation and cervical epithelial cells proliferation in a dose dependent manner.


Comparative studies on the role of calcium in triggering subcellular damage in cardiac muscle.
Isolated cardiac muscle strips from amphibians and mammals, together with isolated frog hearts, have been used as model systems for studying the action of elevated [Ca2+]i in promoting severe damage. A23187 and caffeine are believed to cause a rise in [Ca2+]i. Elevated [Ca2+]i causes characteristic damage which has been categorized and includes hypercontraction, Z-line damage and myofilament dissolution. The damage closely resembles that described in the isolated mammalian heart and in skeletal muscle preparations when [Ca2+]i is raised dramatically. Damage can therefore be triggered by releasing Ca2+ from intracellular sites, as distinct from increasing Ca2+ entry (as in the Ca2+-paradox). DNP and ruthenium red also cause identical damage and the results suggest that whilst the fall in pHi associated with ischaemia is probably the consequence of Ca2+/2H+ exchange at the mitochondria, coupled with ATP hydrolysis, lowered pHi by mitochondrial action is probably not the only cause of myofilament dissolution. Damage is not prevented by pretreatment with leupeptin, an inhibitor of Ca2+-activated neutral proteases, and it is concluded that the latter are probably not implicated in rapid and dramatic damage. The possible involvement of lysosomal enzymes in damage triggered by high [Ca2+]i is discussed.


Femoral nerve injury following inguinal hernia repair.
Insidious leg pain and weakness followed inguinal hernia repair. Despite the temporal relationship of the surgery and the symptoms, the diagnosis proved elusive, in part, because of the rarity of the complication.


Reliability and validity of the adapted Turkish version of the Spinal Appearance Questionnaire.
The Spinal Appearance Questionnaire (SAQ), scoliosis specific quality of life questionnaire, was developed to assess the spinal appearance in adolescent idiopathic scoliosis (AIS) patients. The aim of this study is to evaluate the adaptation, validity, and reliability of the Turkish version of the Spinal Appearance Questionnaire (Tr-SAQ). Tr-SAQ and already validated Turkish SRS-22 were applied to 75AIS patients (56 females) twice within a 2-week interval for test-retest reliability. Validity of the Tr-SAQ was assessed with factor analysis, convergent validity, and discriminant validity. Convergent validity was evaluated by calculating Spearman correlation coefficients between Tr-SAQ and SRS-22 self-image domain. Internal consistency and intraclass correlation coefficient (ICC) were evaluated for the determination of reliability. Factor analysis indicated that Tr-SAQ had two factors as appearance (items 1-10) and expectations (items 12-15). Convergent validity test showed a significant negative correlation between the Tr-SAQ appearance score and SRS-22 self-image score (Spearman's r = - 0.6).Test-retest was conducted within a mean of 16.7 (range 14-28) days. Both ICC and Cronbach's α were found to be high (0.98, 0. 91, respectively). The correlations with the major curve magnitude were stronger for the Tr-SAQ Appearance (r = 0.7) and Tr-SAQ Total (r = 0.6) scores than the correlations between the SRS-22 self-image (r = - 0.5) and SRS-22 Total (r = - 0.4) scores. The Turkish version of the SAQ was reliable and valid for clinical use for AIS patients who are native speakers of Turkish. Level I- diagnostic studies.


The metabolic transition speed between backward walking and running.
Although the metabolic transition speed for forward exercise has already been determined, the walk-run transition speed for backward exercise has not been investigated before. The aim of this study was to determine the speed at which it becomes metabolically more efficient to run backwards than to walk backwards. Eighteen healthy volunteers, who successfully completed three backward exercise practice sessions, participated in the study. All subjects randomly performed two exercise tests: backward walking and backward running. Both protocols started at a treadmill speed of 5 km(.)h(-1). Every minute the speed was increased by 0.5 km(.)h(-1) until 8 km(.)h(-1) was reached. Cardiorespiratory variables were continuously measured and blood lactate concentration [La] was determined every 2 min, using the Accusport lactate analyser. At each work load subjects rated their perceived exertion (RPE), using the Borg scale. There were no statistically significant differences in oxygen consumption, minute ventilation and heart rate between 6 and 7 km(.)h(-1), for backward walking and backward running ( P>0.05). There was no statistically significant difference in blood [La] between walking and running at 7.5 km(.)h(-1) ( P>0.05). According to the RPE values, subjects rated running at speeds less than 6 km(.)h(-1) more difficult than walking at similar speeds. We conclude that the metabolic transition speed between backward walking and running is between 6 and 7 km(.)h(-1), which is lower than the metabolic transition speed for forward locomotion (7.2-7.9 km(.)h(-1)).


Fear recognition impairment in early-stage Alzheimer's disease: when focusing on the eyes region improves performance.
A decline in the ability to identify fearful expression has been frequently reported in patients with Alzheimer's disease (AD). In patients with severe destruction of the bilateral amygdala, similar difficulties have been reduced by using an explicit visual exploration strategy focusing on gaze. The current study assessed the possibility of applying a similar strategy in AD patients to improve fear recognition. It also assessed the possibility of improving fear recognition when a visual exploration strategy induced AD patients to process the eyes region. Seventeen patients with mild AD and 34 healthy subjects (17 young adults and 17 older adults) performed a classical task of emotional identification of faces expressing happiness, anger, and fear in two conditions: The face appeared progressively from the eyes region to the periphery (eyes region condition) or it appeared as a whole (global condition). Specific impairment in identifying a fearful expression was shown in AD patients compared with older adult controls during the global condition. Fear expression recognition was significantly improved in AD patients during the eyes region condition, in which they performed similarly to older adult controls. Our results suggest that using a different strategy of face exploration, starting first with processing of the eyes region, may compensate for a fear recognition deficit in AD patients. Findings suggest that a part of this deficit could be related to visuo-perceptual impairments. Additionally, these findings suggest that the decline of fearful face recognition reported in both normal aging and in AD may result from impairment of non-amygdalar processing in both groups and impairment of amygdalar-dependent processing in AD.


Preoperative radiation response evaluated by 18-fluorodeoxyglucose positron emission tomography predicts survival in locally advanced rectal cancer.
This study focuses on the prognostic survival value of postirradiation metabolic activity in primary rectal cancer as measured with 18-fluorodeoxyglucose positron emission tomography. From July 1995 to March 2002, all 59 patients underwent two series of fluorodeoxyglucose positron emission tomography: one before preoperative radiation (standardized uptake values-1), and the other two to three weeks after radiation (standardized uptake values-2). Standardized uptake values-1 and standardized uptake values-2 correspond to before and after radiation, respectively. In univariate analysis, the following emerged as significant prognostic variables: with or without residual tumor, pathologic differentiation, with or without recurrence, standardized uptake values-2, and with or without lymph node metastases. In multivariate analysis, residual tumor and standardized uptake values-2 were significant prognostic factors for survival. The median survival and the five-year overall survival rate comparing standardized uptake values-2 values <5 vs. >5 were 95 vs. 42 months and 70 vs. 44 percent, respectively (P = 0.042). A significant survival benefit was observed in patients with low fluorodeoxyglucose uptake after preoperative radiotherapy in primary tumors of rectal cancer.


Prox1 is differentially localized during lens development.
Prox1, the vertebrate cognate of Drosophila Prospero, is a homeodomain protein essential for the development of the lens, liver and lymphatic system. While it is well established that the subcellular distribution of Prospero changes during development, this had not been demonstrated for Prox1. Here, high-resolution confocal microscopy demonstrated that Prox1 protein is predominately cytoplasmic in the lens placode as well as the lens epithelium and germinative zone throughout development. However during fiber cell differentiation, Prox1 protein redistributes to cell nuclei. Finally, as lens fiber cells condense their chromatin in response to lens denucleation, Prox1 remains in the nucleus but does not appear to interact with DNA. Thus, it appears that the function of Prox1, like that of its Drosophila cognate Prospero, is at least partially controlled by changes in its subcellular distribution during development.


Long-term spatial memory across large spatial scales in Heliconius butterflies.
Locating food in heterogeneous environments is a core survival challenge. The distribution of resources shapes foraging strategies, imposing demands on perception, learning and memory, and associated brain structures. Indeed, selection for foraging efficiency is linked to brain expansion in diverse taxa, from primates1 to Hymenopterans2. Among butterflies, Heliconius have a unique dietary adaptation, actively collecting and feeding on pollen, providing a source of essential amino acids as adults, negating reproductive senescence and facilitating an extended longevity3. Several lines of evidence suggest that Heliconius learn the spatial location of pollen resources within an individual's home range4, and spatial learning may be more pronounced at these large spatial scales. However, experimental evidence of spatial learning in Heliconius, or any other butterfly, is so far absent. We therefore tested the ability of Heliconius to learn the spatial location of food rewards at three ecologically-relevant spatial scales, representing multiple flowers on a single plant, multiple plants within a locality, and multiple localities. Heliconius were able to learn spatial information at all three scales, consistent with this ability being an important component of their natural foraging behaviour.


Purification of pertussis toxin substrates from bovine heart membranes.
Pertussis toxin substrates were purified from bovine heart membranes by means of a sequential column chromatographic techniques. The elution pattern from the final ion-exchange chromatography and relative mobilities of these peptides on SDS-polyacrylamide gel electrophoresis revealed that the bovine heart membranes contained two distinct pertussis toxin substrates: Gi2 and Go.


Influence of a metaphylactic treatment with Baycox(®) Bovis on the reproductive performances of Fresian heifers: a preliminary study.
A blind, randomised, controlled, multicentric field trial was conducted to assess the influence of a metaphylactic treatment with an oral solution of toltrazuril on some reproductive parameters of Italian Fresian heifers during the first 18-20 months of life. For this goal 40 calves were selected from two dairy farms and randomly divided into two homogeneous groups: MTol, treated with toltrazuril and NegC, left untreated. The calves were clinically and coprologically examined over the entire study period. The body condition scores, the body weights and the age at the first service were recorded and compared between the two groups, in addition to some other reproductive parameters including number of pregnancies, average service per pregnancy, conception rate, conception rate at first service and post first service conception rate. The analysis of the results showed that the metaphylactic treatment with toltrazuril influenced positively the average age of the first service (MTol 461.4 days versus NegC 485.45 days), the overall conception rate (MTol 95 % versus NegC 85 %), the success at first (MTol 60 % versus NegC 45 %) and second (MTol 75 % versus NegC 45 %) services and, consequently, the mean number of services to be carried out for each animal (MTol 1.4 ± 0.6 versus NegC 1.6 ± 0.79). Furthermore, the results confirmed that toltrazuril treatment, applied in accordance with the epidemiological aspects of each farm, is highly efficacious in persistent reduction of oocyst excretions with particular reference to Eimeria zuernii, Eimeria bovis, considered to be mainly responsible for clinical coccidiosis.


Electron microscopic investigation of intracellular events after ingestion of Rhodococcus equi by foal alveolar macrophages.
It has been suggested that R. equi causes pulmonary disease in foals by persisting within the lung as a facultative intracellular parasite of alveolar macrophages. This paper describes an ultrastructural study of the intracellular events after ingestion of R. equi by foal alveolar macrophages, in an attempt to determine the mechanism of intracellular survival of R. equi. Secondary lysosomes of alveolar macrophages recovered from foals by bronchoalveolar lavage were labelled with electron-dense ferritin, and the cells were challenged with either viable or formalin-killed R. equi. After 0-, 3-, 8- or 24-h incubation, the cells were fixed and processed for electron microscopy. There was no evidence of phagosome-lysosome fusion after ingestion of either viable or non-viable R. equi by foal alveolar macrophages. Rhodococcus equi persisted and multiplied within dilated phagosomes, which were often lined by elongate microvillous structures. After 24-h incubation, 75% of the ingested bacteria were still structurally intact. Macrophages with ingested viable R. equi were irreversibly damaged and released intracellular bacteria into the surrounding medium. These data confirm that R. equi is a facultative intracellular parasite of foal alveolar macrophages and is able to persist and multiply within the phagosome, apparently inhibiting phagosome-lysosome fusion by some as yet unknown mechanism.


Cocaine psychosis and AIDS: a contemporary diagnostic dilemma.
This article examines some of the differential diagnostic considerations associated with the interaction between advanced cocaine psychosis and the neuropsychiatric manifestations of AIDS-Spectrum Disorders. A clinical case is presented to illustrate the potential for institutional countertransference in the treatment of substance abusing patients. The role of a multi-dimensional hypotheses testing model in cocaine related cases is discussed.


Effect of endurance training on diurnal rhythms of superoxide dismutase activity, glutathione and lipid peroxidation in plasma of pinealectomized rats.
Melatonin deficit is characterized by disturbed circadian rhythms of many physiological and biochemical parameters including markers of oxidative stress. Moderate endurance training exerts protection against oxidative stress. In the present study, we aimed to explore the impact of endurance treadmill training on disturbed rhythmic fluctuations of some markers of oxidative stress in pinealectomized rats. Animals were divided into four groups: sham-operated sedentary rats (sham-sed), a sham group with exercise (sham-ex), pinealectomized sedentary rats (pin-sed) and pin rats with exercise (pin-ex). Animals were sacrificed by decapitation at 4-h intervals for biochemical analysis of plasma melatonin and markers of oxidative stress. The activity of superoxide dismutase (SOD) and the levels of glutathione (GSH) and lipid peroxidation demonstrated diurnal variations in the sham-sed group. The peak values of SOD were detected during the dark period that coincided with the peak plasma levels of melatonin in the sham-sed rats. The malondialdehyde (MDA) levels also showed a tendency to a progressive raise during the dark period. Pinealectomy was characterized by a remarkable melatonin deficit in plasma of sedentary rats, compromised fluctuations with decreased SOD activity and increased lipid peroxidation. While endurance training was unable to restore the melatonin deficit, it partly prevented the oxidative stress at selected time points in the pinealectomised rats. Our findings indicate the important role of endurance training against oxidative stress both in physiological conditions and melatonin deficit.


Hypothesis: A Novel Neuroprotective Role for Glucose-6-phosphatase (G6PC3) in Brain-To Maintain Energy-Dependent Functions Including Cognitive Processes.
The isoform of glucose-6-phosphatase in liver, G6PC1, has a major role in whole-body glucose homeostasis, whereas G6PC3 is widely distributed among organs but has poorly-understood functions. A recent, elegant analysis of neutrophil dysfunction in G6PC3-deficient patients revealed G6PC3 is a neutrophil metabolite repair enzyme that hydrolyzes 1,5-anhydroglucitol-6-phosphate, a toxic metabolite derived from a glucose analog present in food. These patients exhibit a spectrum of phenotypic characteristics and some have learning disabilities, revealing a potential linkage between cognitive processes and G6PC3 activity. Previously-debated and discounted functions for brain G6PC3 include causing an ATP-consuming futile cycle that interferes with metabolic brain imaging assays and a nutritional role involving astrocyte-neuron glucose-lactate trafficking. Detailed analysis of the anhydroglucitol literature reveals that it competes with glucose for transport into brain, is present in human cerebrospinal fluid, and is phosphorylated by hexokinase. Anhydroglucitol-6-phosphate is present in rodent brain and other organs where its accumulation can inhibit hexokinase by competition with ATP. Calculated hexokinase inhibition indicates that energetics of brain and erythrocytes would be more adversely affected by anhydroglucitol-6-phosphate accumulation than heart. These findings strongly support the paradigm-shifting hypothesis that brain G6PC3 removes a toxic metabolite, thereby maintaining brain glucose metabolism- and ATP-dependent functions, including cognitive processes.


The poor use of methadone in Italian hospices.
A hospice may provide an ideal setting to use methadone because of a more protective environment in comparison with a home care setting, where it is seldom used. The aim of this survey was to investigate the frequency of the use of methadone in Italian hospices and how it is used. The Italian registry of palliative care institutions, named Osservatorio Federazione cure palliative, was consulted at its official website. Phone interviews were planned with physicians who were delegated to provide the answers. Data regarding the hospice and experience of physicians were collected. Questions regarding the attitudes in using methadone were posed, including the frequency of use and how this drug was used. Of 165 centers contacted, information was available for 138 hospices (83.6 %). The majority of interviewed physicians considered methadone as a difficult drug to use, and only 38 responders (27.5 %) were using methadone as an analgesic in a minority of patients (less than 5 % in more than half of responders). When methadone is used as first opioid, all the patients receive less than 10 mg/day. Almost all physicians switched from morphine to methadone and used a conversion ratio inversely proportional to the morphine dose. Physicians working in hospices should improve the knowledge regarding the use of opioids, including methadone, as well as complex treatments which can be the principal reason for hospice admission.


HRS Clinical Document Development Methodology Manual and Policies: Executive summary.
The Heart Rhythm Society (HRS) has been developing clinical practice documents in collaboration and partnership with other professional medical societies since 1996. The HRS formed a Scientific and Clinical Documents Committee (SCDC) with the sole purpose of managing the development of these documents from conception through publication. The SCDC oversees the process for developing clinical practice documents, with input and approval from the HRS Executive Committee and the Board of Trustees. As of May 2017, the HRS has produced more than 80 publications with other professional organizations. This process manual is produced to publicly and transparently declare the standards by which the HRS develops clinical practice documents, which include clinical practice guidelines, expert consensus statements, scientific statements, clinical competency statements, task force policy statements, and proceedings statements. The foundation for this process is informed by the Institute of Medicine's standards for developing trustworthy clinical practice guidelines; the new criteria from the National Guidelines Clearinghouse, effective June 2014; SCDC member discussions; and a review of guideline policies and methodologies used by other professional organizations.


Affinity chromatography on immobilized hyaluronate and its application to the isolation of hyaluronate binding properties from cartilage.
Partially degraded hyaluronate was coupled to AH-Sepharose 4B using carbodiimide. Approximately 1 mg of hyaluronate was incorporated per ml of wet gel. The derivatized gel was used to purify components of the hyaluronate-proteoglycan complex of cartilage. Two link-proteins were isolated from a crude cartilage extract by affinity binding to the gel and eluted with 4 M guanidinium chloride. By the same procedure one link-protein and the globular portion of the proteoglycan monomer were isolated from a trypsin-treated cartilage extract and were separated from each other by subsequent gel chromatography on Sepharose 6B and Sephacryl S-200. The affinity technique was also used for the preparation of these proteins labelled with dansyl groups.


3-methoxyapigenin modulates β-catenin stability and inhibits Wnt/β-catenin signaling in Jurkat leukemic cells.
Aberrant activation of Wnt/β-catenin signaling has been implicated in carcinogenesis. Identification of inhibitors of this pathway may help in cancer therapy. The purpose of this study is to investigate the inhibitory effect of 3-methoxyapigenin (3-MA) with β-catenin/LEF reporter system. The anti-cancer mechanisms in Jurkat leukemic cells were also examined. HEK 293-TOP/FOP reporter cells were used to determine the inhibitory effect of 3-MA on Wnt/β-catenin pathway. We also used Jurkat-TOP reporter cells to confirm the inhibitory effect and the action mechanisms of 3-MA. Target genes and cell proliferation were analyzed by RT-PCR and (3)H-thymidine uptake assay. The effects of 3-MA on β-catenin phosphorylation was determined by Western blotting and by in vitro kinase assays. β-catenin translocation and its transactivation were verified by cellular fractionation and EMSA. 3-MA inhibited Wnt-3A-induced luciferase activity in the HEK 293-TOP/FOP reporter system. Western blotting analysis showed that phosphorylation sites in β-catenin by glycogen synthase kinase-3β (GSK-3β) and casein kinase 2 (CK2) were inhibited by 3-MA in Jurkat. In parallel, in vitro kinase assays verified this effect. As a result, total β-catenin turnover remained balanced by this dual inhibitory effect of 3-MA. Although the β-catenin protein level remained unchanged, 3-MA did inhibit β-catenin translocation. Finally, we found that the β-catenin/LEF transcriptional activity, expression of c-myc and cyclin-D3, and cell proliferation were inhibited by 3-MA. 3-MA modulates the turnover of β-catenin and suppresses the Wnt/β-catenin signaling pathway through inhibition of β-catenin translocation. We suggested that 3-MA has potential as an anti-cancer drug.


Decreased GABA receptor in the cerebral cortex of epileptic rats: effect of Bacopa monnieri and Bacoside-A.
Gamma amino butyric acid (GABA), the principal inhibitory neurotransmitter in the cerebral cortex, maintains the inhibitory tones that counter balances neuronal excitation. When this balance is perturbed, seizures may ensue. In the present study, alterations of the general GABA, GABAA and GABAB receptors in the cerebral cortex of the epileptic rat and the therapeutic application of Bacopa monnieri were investigated. Scatchard analysis of [3H]GABA, [3H]bicuculline and [3H]baclofen in the cerebral cortex of the epileptic rat showed significant decrease in Bmax (P < 0.001) compared to control. Real Time PCR amplification of GABA receptor subunits such as GABAAά1, GABAAγ, GABAAδ, GABAB and GAD where down regulated (P < 0.001) in epileptic rats. GABAAά5 subunit and Cyclic AMP responsible element binding protein were up regulated. Confocal imaging study confirmed the decreased GABA receptors in epileptic rats. Epileptic rats have deficit in radial arm and Y maze performance. Bacopa monnieri and Bacoside-A treatment reverses epilepsy associated changes to near control suggesting that decreased GABA receptors in the cerebral cortex have an important role in epileptic occurrence; Bacopa monnieri and Bacoside-A have therapeutic application in epilepsy management.


Expression of activation-induced cytidine deaminase enhances the clearance of pneumococcal pneumonia: evidence of a subpopulation of protective anti-pneumococcal B1a cells.
We describe a protective early acquired immune response to pneumococcal pneumonia that is mediated by a subset of B1a cells. Mice deficient in B1 cells (xid), or activation-induced cytidine deaminase (AID(-/-) ), or invariant natural killer T (iNKT) cells (Jα18(-/-) ), or interleukin-13 (IL-13(-/-) ) had impaired early clearance of pneumococci in the lung, compared with wild-type mice. In contrast, AID(-/-) mice adoptively transferred with AID(+/+) B1a cells, significantly cleared bacteria from the lungs as early as 3 days post infection. We show that this early bacterial clearance corresponds to an allergic contact sensitivity-like cutaneous response, probably due to a subpopulation of initiating B1a cells. In the pneumonia model, these B1a cells were found to secrete higher affinity antigen-specific IgM. In addition, as in contact sensitivity, iNKT cells were required for the anti-pneumococcal B1a cell initiating response, probably through early production of IL-13, given that IL-13(-/-) mice also failed to clear infection. Our study is the first to demonstrate the importance of AID in generating an appropriate B1a cell response to pathogenic bacteria. Given the antibody affinity and pneumonia resistance data, natural IgM produced by conventional B1a cells are not responsible for pneumonia clearance compared with the AID-dependent subset.


The activation and distribution of GABA and L-glutamate receptors on goldfish Mauthner neurones: an analysis of dendritic remote inhibition.
1. The sensitivity of the membrane of the cell-body and lateral dendrite of the Mauthner neurone to iontophoretically applied GABA and L-glutamic acid has been investigated in goldfish. The cells were afterwards examined histologically. The results, taken together with physiological observations, help to explain some functional properties of dendritic synapses.2. GABA inhibits by increasing the membrane conductance, and has little effect on resting potential. L-Glutamate depolarizes, and its effects could sum with subthreshold excitatory post-synaptic potentials (e.p.s.p.s) to cause impulse firing. Neither substance was effective when released intracellularly, but only when applied to the outside of the membrane.3. Using a roving double-barrelled micropipette to apply brief iontophoretic pulses very close to the cell, and recording from the cell-body, the sensitivity to GABA was found to be uniform over a region extending from the axon-hillock to about half-way along the lateral dendrite; it then appeared to fall progressively as the drug was pipetted further and further out on the dendrite. The sensitivity to L-glutamate however seemed relatively unaffected by varying the site of application.4. In contrast to this, when the recording electrode was inserted into the same region of the cell where the drugs were applied, then the sensitivity to GABA, like that to L-glutamate, was found to be similar everywhere, including the distal part of the lateral dendrite.5. The antidromic spike, which does not actively invade the lateral dendrite, was used as a convenient test signal for detecting electrotonic decrement as well as changes in membrane conductance. The conductance changes due to small doses of GABA applied to the dendrite could be detected only over relatively short distances (tens of microns). But over these same distances very little decrement of potential change took place.6. The concept of dendritic remote inhibition has been put on a firm experimental basis. It occurs in the Mauthner cell, and is shown to be capable of providing a high spatial selectivity of action. The e.p.s.p.s most affected by it are those generated out on the same dendrite, and it follows that many of the e.p.s.p.s produced far out on other dendrites could well be influenced by it, though to a lesser extent. Least affected are large e.p.s.p.s generated in the cell-body and axon-hillock regions. The synaptic grouping provided by the distal parts of dendrites and their associated nerve endings is discussed.7. The results of a theoretical treatment (Appendix) of the effects of a localized membrane resistance change on electrotonic potentials recorded nearer to the source have been applied to results obtained on the Mauthner cell. The experimentally observed properties of the ;remote inhibition' caused by localized GABA application to the lateral dendrite, accord well with those predicted both for an infinitely long uniform cable and for a cable having a closed end at the region of the conductance change. The possible implications of these results for other central neurones are considered.8. Evidence is adduced showing that the synapses responsible for collateral inhibition in the Mauthner cell include at least part of the group of small nerve endings found on the axon-hillock, cell-body, proximal and distal lateral dendrite. (The latter distally-located endings cause dendritic remote inhibition.) All these regions, some of which certainly have excitatory synapses, thus receive an inhibitory input as well, and are sensitive to both GABA and L-glutamate.9. E.p.s.p.s recorded in the cell-body were reduced by about the same amount as was the antidromic spike when GABA was applied there, but considerably more than the spike when the GABA application was to the lateral dendrite near the origin of the e.p.s.p. Even when the recording was made from the affected region of the dendrite, the e.p.s.p. seemed more sensitive to GABA than was the electrotonically conducted spike. This, and other observations, suggests that GABA may act presynaptically on the excitatory endings as well as post-synaptically.10. The detection of effects, and the recognition of site of action, of drugs applied iontophoretically in the central nervous system is discussed on the basis of the present findings.


New partially degradable and bioactive acrylic bone cements based on starch blends and ceramic fillers.
This work reports the development of new partially biodegradable acrylic bone cements based on corn starch/cellulose acetate blends (SCA), prepared by the free radical polymerization of methyl methacrylate and acrylic acid at low temperature. Amounts of biocompatible, osteoconductive and osteophilic mineral component such as hydroxylapatite (sintered and non-sintered), were incorporated in different percentages to confer a bone-bonding character to the bone cements in this type of applications. All cement formulations were characterized by 1H NMR spectroscopy. Curing parameters and mechanical properties were determined finding formulations which complete the ASTM legislation. Hydration degree, degradation studies, as well as bioactivity tests were performed in all prepared formulations. The developed systems show a range of properties that might allow for their application as self-curing bone cements, exhibiting several advantages with respect to other commercially available bone cements.


Preparation and evaluation of SnO2-based 68Ge/68Ga generator made from 68Ge produced through (nat)Zn(α,xn) reaction.
(68)Ge was produced by (nat)Zn(α,xn)(68)Ge reaction and its production yield was 31.82 kBq/μAh (0.86 μCi/μAh) at the end of irradiation (EOI). A simple chromatographic method using a SnO2 column was employed to separate (68)Ge from the target material and the co-produced non-isotopic radioisotope impurities. (68)Ge retained in the column served as the (68)Ge/(68)Ga generator. Elution efficiency of the column was about 60%. First 2 ml of the eluate contained more than 95% of the elutable activity. Post-elution purification cum concentration was done with a small cation exchange resin column. The presence of the inactive tin ions in the (68)Ga eluate was determined by the ICP-OES technique and was found to be about 0.03 ppm. Radiochemical purity of the final (68)Ga preparation was more than 99.99% and it was found to be suitable for making complex with ethylenediamine-N,N,N',N'-tetrakis(methylene phosphonic acid) (EDTMP).


Thermogenic hydrocarbon biodegradation by diverse depth-stratified microbial populations at a Scotian Basin cold seep.
At marine cold seeps, gaseous and liquid hydrocarbons migrate from deep subsurface origins to the sediment-water interface. Cold seep sediments are known to host taxonomically diverse microorganisms, but little is known about their metabolic potential and depth distribution in relation to hydrocarbon and electron acceptor availability. Here we combined geophysical, geochemical, metagenomic and metabolomic measurements to profile microbial activities at a newly discovered cold seep in the deep sea. Metagenomic profiling revealed compositional and functional differentiation between near-surface sediments and deeper subsurface layers. In both sulfate-rich and sulfate-depleted depths, various archaeal and bacterial community members are actively oxidizing thermogenic hydrocarbons anaerobically. Depth distributions of hydrocarbon-oxidizing archaea revealed that they are not necessarily associated with sulfate reduction, which is especially surprising for anaerobic ethane and butane oxidizers. Overall, these findings link subseafloor microbiomes to various biochemical mechanisms for the anaerobic degradation of deeply-sourced thermogenic hydrocarbons.


Eco-friendly detoxification of hazardous Congo red dye using novel fungal strain Trametes flavida WTFP2: Deduced enzymatic biomineralization process through combinatorial in-silico and in-vitro studies.
Growing textile industry is a major global concern, owing to the presence of recalcitrant hazardous pollutants, like synthetic dyes in discharged effluents. To explore new bioresources for mycoremediation, a high laccase-producing novel white-rot fungus (WRF), Trametes flavida WTFP2, was employed. T. flavida is an underexplored member of Polyporales. Using bioinformatic tools, 8 different cis-acting RNA elements were identified in the 5.8 S ITS gene sequence, where CRISPR (CRISPR-DR15), sRNA (RUF1), and snoRNA (ceN111) are uniquely present. Molecular docking was adopted to predict the catalytic interaction of chosen toxic diazo colorant, Congo red (CR), with four dye-degrading enzymes (laccase, lignin peroxidase, azoreductase, and aryl alcohol oxidase). With 376.41 × 103 U/L laccase production, novel WRF exhibited dye-decolorization potential. WTFP2 effectively removed 99.48 ± 0.04% CR (100 mg/L) and demonstrated remarkable recyclability and persistence in consecutive remediation trials. Mycelial dye adsorption was not only substantial driver of colorant elimination; decolorization using active T. flavida was regulated by enzymatic catalysis, as outlined by in-vitro growth, induction of extracellular enzymes, and FESEM. Fifteen metabolites were identified using HRLCMS-QTOF, and novel CR degradation pathway was proposed. Furthermore, microbial and phyto-toxicity tests of metabolites suggested complete detoxification of toxic dye, making the process clean, green, and economically sustainable.


Trichorhinophalangeal syndrome type 1 (Giedion syndrome): A case report with literature review.
Trichorhinophalangeal syndrome (TRPS) is a rare multisystem disorder characterized by abnormalities in the hair (tricho), nose (rhino), and digits (phalangeal). A variety of nonspecific intraoral findings have been reported in the literature, including hypodontia, delayed tooth eruption, malocclusion, a high-arched palate, mandibular retrognathia, midface hypoplasia, and multiple impacted teeth. In addition, supernumerary teeth have been detected in several persons with TRPS, especially type 1. This report describes the clinical manifestations and dental management of a TRPS 1 patient with multiple impacted supernumerary and permanent teeth. A 15-year-old female patient visited our clinic with a known medical history of TRPS 1 with laceration of the tongue caused by teeth eruption in the palate. Radiographic images showed a total of 45 teeth: two deciduous, 32 permanent, and 11 supernumerary teeth. Six permanent teeth and 11 supernumerary teeth in the posterior quadrants were impacted. Four impacted third molars, supernumerary teeth, retained deciduous teeth, and impacted maxillary premolars were removed under general anaesthesia. This case suggests that all patients with TRPS should undergo full clinical and radiographic oral examination and should be informed about the disease and the importance of dental counselling.


5-HT2A and 5-HT2C/2B receptor subtypes modulate dopamine release induced in vivo by amphetamine and morphine in both the rat nucleus accumbens and striatum.
In vivo microdialysis and single-cell extracellular recordings were used to assess the involvement of serotonin(2A) (5-HT(2A)) and serotonin(2C/2B) (5-HT(2C/2B)) receptors in the effects induced by amphetamine and morphine on dopaminergic (DA) activity within the mesoaccumbal and nigrostriatal pathways. The increase in DA release induced by amphetamine (2 mg/kg i.p.) in the nucleus accumbens and striatum was significantly reduced by the selective 5-HT(2A) antagonist SR 46349B (0.5 mg/kg s.c.), but not affected by the 5-HT(2C/2B) antagonist SB 206553 (5 mg/kg i.p.). In contrast, the enhancement of accumbal and striatal DA output induced by morphine (2.5 mg/kg s.c.), while insensitive to SR 46349B, was significantly increased by SB 206553. Furthermore, morphine (0.1-10 mg/kg i.v.)-induced increase in DA neuron firing rate in both the ventral tegmental area and the substantia nigra pars compacta was unaffected by SR 46349B (0.1 mg/kg i.v.) but significantly potentiated by SB 206553 (0.1 mg/kg i.v.). These results show that 5-HT(2A) and 5-HT(2C) receptors regulate specifically the activation of midbrain DA neurons induced by amphetamine and morphine, respectively. This differential contribution may be related to the specific mechanism of action of the drug considered and to the neuronal circuitry involved in their effect on DA neurons. Furthermore, these results suggest that 5-HT(2C) receptors selectively modulate the impulse flow-dependent release of DA.


Arginase II inhibitory activity of flavonoid compounds from Scutellaria indica.
Arginase II has recently reported as a novel therapeutic target for the treatment of cardiovascular diseases such as atherosclerosis. In the course of screening plants used in natural medicines as arginase II inhibitory activity, a methanol extract of Scutellaria indica showed significant inhibitory effect. Further fractionation and repeated column chromatography led to the isolation of a new flavan-type (1), and seven known compounds (2-8). The chemical structures of isolated compounds were elucidated based on extensive 1D and 2D NMR spectroscopic data. The isolates 1-8 were investigated in vitro for their arginase II inhibitory activity using enzyme solution prepared from kidney of anesthetized C57BL/6 mice. Compounds 3 and 5 significantly inhibited arginase II activity with IC₅₀ values of 25.1 and 11.6 μM, respectively, whereas the other compounds were apparently inactive.


Management of air quality in the vicinity of congested area in Kuwait.
An assessment of air quality in the vicinity of a selected school has been carried out by monitoring the concentrations of primary pollutants. The results has shown that during the school hours, the measured pollutants emitted from the cars next to the selected school, such as CO and NO(2), are always under the allowable limits for Kuwaiti air quality standards. On the other hand, the concentrations of non methane hydrocarbon pollutant (nm-HC), some of which are considered to be cancergenic, are found to be above the Kuwaiti standard most of the times. A traffic counter is used to record the number of cars in the main road next to the school in 15 min intervals for 10 days during the monitoring period for air quality. Statistical analysis is performed to develop a relation for prediction of the necessary reduction in traffic, based on CO concentrations, during morning and afternoon periods on working days. A computer dispersion model (CALINE4) is also used to assess the CO concentrations based on recorded flow of traffic and emission inventory with the prevailing meteorological conditions existed at the specified time. After the validation of model, different scenarios have been evaluated to provide an acceptable solution to resolve the traffic congestion problem near the schools in the early morning hours with substantial reduction in pollution levels. The optimal solution for CO concentration reduction by managing smooth traffic flow is to reduce the traffic intensity by half in early morning and afternoon rush hours. The results of the predicted CO concentration in the vicinity of the school for the model and the statistical analysis has shown reduction of 30% and 42% respectively, for approximately 50% decrease in the car use. On the other hand the predicted CO concentration for the model and the statistics reached 24% and 33% respectively when 50% of students opted for buses instead of using private cars.


Comparison of early events during infection of human and chimpanzee peripheral blood mononuclear cells with HIV-1.
Differences in early events during entry and integration of HIV-1 into peripheral blood mononuclear cells (PBMC) might contribute to the absence of AIDS-like disease in chimpanzees as compared to humans. To address this question, we first tested the in vitro susceptibility of human and chimpanzee PBMC for infection with the two HIV-1 isolates III B and RF. The results of these studies revealed that chimpanzee PBMC had a slightly lower capability to support the growth of HIV-1 as compared to human PBMC. This was accompanied by a delayed accumulation of proviral HIV-1 DNA in cultures of HIV-1-infected chimpanzee PBMC. However, no differences between cells of the two species were observed when very early events of HIV-1 infection were studied. Shortly (20 h) after infection chimpanzee and human cells harbored similar amounts of proviral HIV-1 DNA and PBMC of both species behaved comparably with respect to pre-integration latency (i.e. the ability to activate extrachromosomal HIV-1 intermediates in HIV-1 infected quiescent cells at various times after infection). These results strongly suggest that the absence of AIDS-like disease in chimpanzees cannot be correlated with defects in early events of the HIV-1 replicative cycle.


Impact of the COVID-19 pandemic on the glycemic control, eating habits, and body compositions of people with diabetes mellitus: A retrospective longitudinal observational study.
To evaluate the impact of the COVID-19 pandemic on the glycemic control, eating habits, and body composition of people with diabetes mellitus; to identify the determinants of worsening glycemic control in people with diabetes mellitus. This retrospective, longitudinal observational study was performed in outpatients with diabetes mellitus who visited our hospital between April 2019 and March 2020 (pre-COVID-19 period) and continued for follow up from April 2020 to March 2021 (COVID-19 period). We compared the glycemic control, nutritional intakes, and body composition of people with diabetes mellitus between the two periods. The changes in the HbA1c values (ΔHbA1c) and other study variables were compared between the two periods. Logistic regression analysis was performed to identify the factors associated with the increase of HbA1c levels. A significant increase of HbA1c was observed during the COVID-19 period. The percent fat mass (FM) also increased, while the percent skeletal muscle mass (SMM) decreased during the COVID-19 period. After adjustments for age and sex, the ΔBMI (OR:2.33), ΔFM (OR:1.45), and ΔSMM (OR:0.51) were identified as being associated with elevated levels of HbA1c. The COVID-19 pandemic had a negative impact on the glycemic control and body composition of people with diabetes mellitus. The increased body weight and FM and decreased SMM observed during the pandemic were associated with poor glycemic control in people with diabetes mellitus.


The role of putative glutamatergic neurons and their connections in the locomotor central pattern generator of the mollusk, Clione limacina.
In the pteropod mollusk Clione limacina, locomotor rhythm is produced by the central pattern generator (CPG), due mainly to the activity of interneurons of groups 7 (active in the phase of the dorsal flexion of the wings) and 8 (active in the phase of the ventral flexion). Each of these groups excites the neurons active in the same phase of the locomotor cycle, and inhibits the neurons of the opposite phase. In this work, the nature of connections formed by group 7 interneurons was studied. Riluzole (2-amino-6-trifluoro-methoxybenzothiazole), which is known to inhibit the presynaptic release of glutamate, suppressed the action of the type 7 interneurons onto the follower neurons of the same and of the antagonistic phase of the locomotor cycle. The main pattern of rhythmic activity of CPG with alternation of two phases could be maintained after suppression of inhibitory connections from group 7 interneurons to antagonistic neurons. This suggests redundancy of the mechanisms controlling swimming rhythm generation, which ensures the reliable operation of the system.


Efficacy of methylprednisolone administration in a case of continuous abnormal muscle activity syndrome.
We report a 20-year-old woman with continuous abnormal muscle activity syndrome in which pharmacological blockade of different sites of the motor unit demonstrates a peripheral nerve origin for the abnormal muscle activity. The case showed no markers of autoimmune disease. A rapid recovery after high dose i.v. methylprednisolone was observed supporting the hypothesis of an autoimmune pathogenesis.


Curcumin-loaded graphene oxide quantum dots enhance otoprotective effects via blocking cuproptosis.
Background: Cisplatin (CIS) is widely used to treat various cancers but can cause ototoxicity and sensory hair cell loss in the inner ear. Copper induces an excessive production of reactive oxygen species (ROS) in hair cells, leading to the development of various antioxidants. Methods and results: This study aimed to evaluate the potential antioxidant properties of curcumin (CUR) in the inner ear organ of corti-1 cells (OC1) and animal models (zebrafish and guinea pigs). Graphene oxide quantum dots (GOQDs) enabled CUR to penetrate the round window membrane (RWM) and maintain the concentration in the perilymph after inner ear administration. The results showed that CUR/GOQDs had favorable biocompatibility and strongly affected ROS generation induced by CIS in OC1 cells. DCFHDA Green staining demonstrated that CUR/GOQDs successfully reversed the decrease in mitochondrial membrane potential induced by CIS in vitro and rescued cells from early cuproptosis, which was confirmed by FDX1 staining. Additionally, the experiment found that CUR decreased the expression of cuproptosis proteins (FDX1, LIAS, and LIPT1) and increased the expression of the Bcl-2 protein. Conclusion: The results demonstrate that CUR/GOQDs is a promising therapeutic agent that can prevent CIS-induced ototoxicity by blocking the cuproptosis signal pathway.


Afterdischarge thresholds and kindling rates in dorsal and ventral hippocampus and dentate gyrus.
Electrodes were implanted to dorsal hippocampus (CA1), ventral CA1, DOrsal dentate gyrus or ventral dentate gyrus. Epileptiform afterdischarge (AD) thresholds were lower in dorsal areas than in ventral areas. Dorsal areas, however, required a greater number of stimulations to develop ("kindle") a fully generalized convulsion than did ventral areas. Thresholds and kindling rates in the dentate gyrus were intermediate between dorsal and ventral CA1, except for the ventral dentate which had higher AD thresholds than ventral CA1. Secondary sites within the hippocampus subsequently kindled within a few stimulations following completion of kindling in the primary site, regardless of which hippocampal area served as the primary site.


Retropleural Thoracic Approach.
The lateral retropleural thoracic approach offers minimally invasive access for the treatment of thoracic spine pathology, specifically thoracic herniated discs. Alternatives to the retropleural approach traditionally included posterolateral or anterior approaches, which carry increased morbidity. The retropleural approach affords lateral access to the thoracic spine that allows for addressing pathology such as herniated discs, corpectomy, tumor, or trauma. This article outlines preoperative workup and planning, intraoperative steps, tips, and postoperative care.


Long-Term Survival and Causes of Death in Children with Trisomy 21 After Congenital Heart Surgery.
To evaluate long-term transplant-free survival and causes of death in the trisomy 21 (T21) population after surgery for congenital heart disease (CHD) in comparison with patients who are euploidic. This is a retrospective cohort study from the Pediatric Cardiac Care Consortium, enriched with prospectively collected data from the National Death Index and the Organ Procurement and Transplantation Network for patients with sufficient direct identifiers. Kaplan-Meier survival plots were generated and multivariable Cox proportional hazards models were used to examine risk factors for mortality between patients with T21 and 1:1 matched patients with comparable CHD who are euploidic. A long-term survival analysis was completed for 3376 patients with T21 (75 155 person-years) who met inclusion criteria. The 30-year survival rate for patients with T21 ranged from 92.1% for ventricular septal defect to 65.3% for complex common atrioventricular canal. Of these, 2185 patients with T21 were successfully matched with a patient who was euploidic. After a median follow-up of 22.86 years (IQR, 19.45-27.14 years), 213 deaths occurred in the T21 group (9.7%) compared with 123 (5.6%) in the euploidic comparators. After adjustment for age, sex, era, CHD complexity, and initial palliation, the hazard ratio of CHD-related mortality was 1.34 times higher in patients with T21 (95% CI, 0.92-1.97; P = .127). CHD-related mortality for patients with T21 after cardiac surgical intervention is comparable with euploidic comparators. Children with T21 require lifelong surveillance for co-occurring conditions associated with their chromosomal abnormality.


Influence of supercoiling and sequence context on operator DNA binding with lac repressor.
The dissociation of the repressor-operator complex from a series of negatively supercoiled plasmid DNAs was examined as a function of the sequence context, orientation, and spacing. The plasmids were grouped into four classes, each with common sequence context. The highest dissociation rate constants were observed for the plasmids containing only a single operator (or pseudooperator) sequence, while approximately 10-fold lower rate constants were measured for plasmids with the I gene pseudooperator in conjunction with either the Z gene pseudooperator or the primary operator. Comparison of the behavior of these two classes of plasmids demonstrated the importance of two operator sequences and supported a model of DNA loop formation to stabilize the repressor-operator complex (Whitson, P. A., and Matthews, K. S. (1986) Biochemistry 25, 3845-3852; Whitson, P. A., Olson, J. S., and Matthews, K. S. (1986) Biochemistry 25, 3852-3858; Whitson, P. A., Hsieh, W. T., Wells, R. D., and Matthews, K. S. (1987) J. Biol. Chem. 262, 4943-4946; Krämer, H., Niemöller, M., Amouyal, M., Revet, B., von Wilcken-Bergmann, B., and Müller-Hill, B. (1987) EMBO J. 6, 1481-1491). The third class, with intermediate dissociation rate constants, was comprised of plasmids which contained the primary operator and the higher affinity pseudooperator normally located in the Z gene. Neither the additional presence of the I gene pseudooperator nor the orientation of the primary operator relative to the Z gene pseudooperator significantly affected the dissociation rate constants. The binding characteristics of this group of plasmids demonstrated the essential role of the Z gene pseudooperator in the formation of intramolecular ternary complex and suggested an in vivo function for this pseudooperator. Plasmids containing two primary operator sequences were the class with lowest dissociation rate constants from lac repressor, and minimal effects of salt or spacing on dissociation of this class were observed. These data are consistent with formation of an intramolecular complex with a looped DNA segment stabilized by the combination of increased local concentration of binding sites and torsional stresses on the DNA which favor binding in supercoiled DNA.


Reduction of donor mononuclear phagocytes with clodronate-liposome during ex vivo lung perfusion attenuates ischemia-reperfusion injury.
Clodronate-liposome is used for depleting mononuclear phagocytes associated with ischemia-reperfusion injury. We hypothesized that administration of clodronate-liposome into the perfusate during ex vivo lung perfusion could reduce mononuclear phagocytes and attenuate ischemia-reperfusion injury. First, the number of mononuclear phagocytes in flushed grafts (minimum cold ischemic time, 6-hour cold ischemic time, 15-hour cold ischemic time, and 18-hour cold ischemic time; n = 6 each) was determined using flow cytometry. Second, grafts (15-hour cold ischemic time) were allocated to control or clodronate (n = 5 each). In the clodronate group, clodronate-liposome is administered into the perfusate. After 4 hours of ex vivo lung perfusion, the number of mononuclear phagocytes in the perfusate and lung tissues was measured. Third, grafts (15-hour cold ischemic time) were allocated to control or clodronate (n = 6 each). After 4 hours of ex vivo lung perfusion, the left lungs were transplanted and reperfused for 2 hours. Lung function was evaluated, and samples were analyzed. First, mononuclear phagocytes remain in flushed grafts after prolonged cold ischemia. Second, the number of mononuclear phagocytes in lung tissues after ex vivo lung perfusion was significantly reduced in the clodronate group (P = .008). Third, lung compliance and vascular resistance during ex vivo lung perfusion were significantly improved in the clodronate group (P < .001 for both). Blood oxygenation and pulmonary edema were significantly improved in the clodronate group after 2 hours of reperfusion (P = .015 and P = .026, respectively). Histological findings showed reduced lung injury in the clodronate group (P = .013). Administration of clodronate-liposome into the perfusate during ex vivo lung perfusion resulted in a significant reduction of mononuclear phagocytes in donor lungs, leading to attenuation of ischemia-reperfusion injury.


Relative body fat and anthropometric prediction of body density of female athletes.
Ninety-one percent (n = 182) of the female members of South Australian representative squads in 14 sports volunteered to act as subjects. Twenty-seven percent of them had represented Australia. The underwater weighing method together with the measurement of residual volume (RV) by helium dilution were used to determine body density (BD); the percent body fat (% BF) was then computed according to Siri. A stepwise multiple regression analysis yielded a correlation coefficient (R) of 0.863 between the criterion (BD) and the best weighted sum of predictors (anthropometric variables): BD (g X cm-3) = 1.14075-0.04959 (log10 sigma triceps, subscapular, supraspinale and calf skinfolds in mm) + 0.00044 (age in decimal years)-0.000612 (waist girth in cm) + 0.000284 (height in cm)-0.000505 (gluteal girth in cm) + 0.000331 (breast girth in cm). Only those predictors which resulted in a statistically significant increase in R (p less than or equal to 0.05) were included. The standard error of estimate of 0.00597 g X cm-3 was equivalent to 2.7% BF at the mean. This equation was shown to be largely population specific. There was a range of 7.6-35.8% of BF and the overall mean 18.5% was significantly lower (p less than 0.001) than that of 23.4% obtained on a moderately active reference sample of similar age (n = 135). If group sizes of only one or two are regarded as too small for meaningful comparison, then the lowest mean of 13.5% was achieved by the long-distance runners (n = 14). The highest averages were registered by the heavyweight rowers (24.2%; n = 7) and soccer players (22.0%; n = 11). The overall average for games players (n = 107) was 19.4%.


Thymic Parenchymal Hyperplasia.
Thymic hyperplasia is a rare condition generally caused by lymphoid follicular hyperplasia associated with autoimmune disorders. True thymic parenchymal hyperplasia unassociated with lymphoid follicular hyperplasia is extremely rare and may give rise to difficulties in diagnosis. We have studied 44 patients with true thymic hyperplasia (38 females and 6 males) aged 7 months to 64 years (mean, 36 years). Eighteen patients presented with symptoms of chest discomfort or shortness of breath; in 20 patients, the lesions were discovered incidentally. Imaging studies demonstrated enlargement of the mediastinum by a mass lesion suspicious for malignancy. All patients were treated with complete surgical excision. The tumors measured from 3.5 to 24 cm (median, 10 cm; mean, 10.46 cm). Histologic examination showed lobules of thymic tissue displaying well-developed corticomedullary architecture, with scattered Hassall corpuscles separated by mature adipose tissue and bounded by a thin fibrous capsule. No cases showed evidence of lymphoid follicular hyperplasia, cytologic atypia, or confluence of the lobules. Immunohistochemical studies showed a normal pattern of distribution for keratin-positive thymic epithelial cells against a background rich in CD3/TdT/CD1a+ lymphocytes. Twenty-nine cases had an initial clinical or pathological diagnosis of thymoma or thymoma vs thymic hyperplasia. Clinical follow-up in 26 cases showed that all patients were alive and well between 5 and 15 years after diagnosis (mean, 9 years). Thymic parenchymal hyperplasia causing significant enlargement of the normal thymus that is sufficient to cause symptoms or worrisome imaging findings should be considered in the differential diagnosis of anterior mediastinal masses. The criteria for distinguishing such lesions from lymphocyte-rich thymoma are presented.


Are animals shrinking due to climate change? Temperature-mediated selection on body mass in mountain wagtails.
Climate change appears to affect body size of animals whose optimal size in part depends on temperature. However, attribution of observed body size changes to climate change requires an understanding of the selective pressures acting on body size under different temperatures. We examined the link between temperature and body mass in a population of mountain wagtails (Motacilla clara) in KwaZulu-Natal, South Africa, between 1976 and 1999, where temperature increased by 0.18 [Formula: see text]C. The wagtails became lighter by 0.035 g per year. Partitioning this trend, we found that only a small part of the effect (0.009 g/year) was due to individuals losing weight and a large part (0.027 g/year) was due to lighter individuals replacing heavier ones. Only the latter component was statistically significant. Apparently, the wagtails were reacting to selection for reduced weight. Examining survival, we found that selection was temperature-mediated, i.e., lighter individuals survived better under high temperatures, whereas heavier individuals survived better under low temperatures. Our results thus support the hypothesis that temperature drove the decline in body mass in this wagtail population and provides one of the first demonstrations of the selective forces underlying such trends.


Human embryos commonly form abnormal nuclei during development: a mechanism of DNA damage, embryonic aneuploidy, and developmental arrest.
What is the prevalence and developmental significance of morphologic nuclear abnormalities in human preimplantation embryos? Nuclear abnormalities are commonly found in human IVF embryos and are associated with DNA damage, aneuploidy, and decreased developmental potential. Early human embryonic development is complicated by genomic errors that occur after fertilization. The appearance of extra-nuclear DNA, which has been observed in IVF, may be a result of such errors. However, the mechanism by which abnormal nuclei form and the impact on DNA integrity and embryonic development is not understood. Cryopreserved human cleavage-stage embryos (n = 150) and cryopreserved blastocysts (n = 105) from clinical IVF cycles performed between 1997 and 2008 were donated for research. Fresh embryos (n = 60) of poor quality that were slated for discard were also used. Immunohistochemical, microscopic and cytogenetic analyses at different developmental stages and morphologic grades were performed. Embryos were fixed and stained for DNA, centromeres, mitotic activity and DNA damage and imaged using confocal microscopy. Rates of abnormal nuclear formation were compared between morphologically normal cleavage-stage embryos, morphologically normal blastocysts, and poor quality embryos. To control for clinical and IVF history of oocytes donors, and quality of frozen embryos within our sample, cleavage-stage embryos (n = 52) were thawed and fixed at different stages of development and then analyzed microscopically. Cleavage-stage embryos (n = 9) were thawed and all blastomeres (n = 62) were disaggregated, imaged and analyzed for karyotype. Correlations were made between microscopic and cytogenetic findings of individual blastomeres and whole embryos. The frequency of microscopic nuclear abnormalities was lower in blastocysts (5%; 177/3737 cells) than in cleavage-stage embryos (16%, 103/640 blastomeres, P < 0.05) and highest in arrested embryos (65%; 44/68 blastomeres, P < 0.05). DNA damage was significantly higher in cells with microscopic nuclear abnormalities (γH2AX (phosphorylated (Ser139) histone H2A.X): 87.1%, 74/85; replication protein A: 72.9%, 62/85) relative to cells with normal nuclear morphology (γH2AX: 9.3%, 60/642; RPA: 5.6%, 36/642) (P < 0.05). Blastomeres containing nuclear abnormalities were strongly associated with aneuploidy (Fisher exact test, two-tailed, P < 0.01). The embryos used were de-identified, and the clinical and IVF history was unknown. This study explores a mechanism of abnormal embryonic development post-fertilization. While most of the current data have explored abnormal meiotic chromosome segregation in oocytes as a primary mechanism of reproductive failure, abnormal nuclear formation during early mitotic cell division in IVF embryos also plays a significant role. The detection of abnormal nuclear formation may have clinical application in noninvasive embryo selection during IVF. The study was supported by Columbia University and the New York Stem Cell Foundation. Authors declare no competing interest.


Uptake, tissue distribution and accumulation of microcystin-RR in Corydoras paleatus, Jenynsia multidentata and Odontesthes bonariensis. A field and laboratory study.
The uptake and accumulation of microcystin-RR (MC-RR) in fish was investigated under laboratory conditions and in wild fish. Jenynsia multidentata and Corydoras paleatus were exposed for 24h to 50mug/L MC-RR dissolved in water. After exposure, liver, gill, brain, intestine, gall bladder, blood and muscle were analyzed for MC-RR by HPLC and analysis confirmed by LC-ESI-TOF-MS spectrometry. Furthermore, wild individuals of Odontesthes bonariensis were sampled from the eutrophic, cyanobacteria-containing San Roque reservoir, and analyzed for the presence of MC-RR in liver, gill, intestine, and muscle. MC-RR was found in liver, gills, and muscle of all exposed and wild fish, while in C. paleatus MC-RR was also present in the intestine. Moreover, we found presence of MC-RR in brain of J. multidentata. Results indicate that MC-RR uptake might occur at two different organs: intestine and gills, through either feeding (including drinking) or respiratory activities. This suggests that MC-RR is taken into the blood stream after absorption, and distributed to different tissues. The liver showed the major bioaccumulation of MC-RR in both experimentally exposed and wild individuals, with muscle of wild fish showing relative high amounts of this toxin in comparison with those exposed in the laboratory; though MC-RR was present in muscle of fish exposed for 24h. The amount of MC-RR in muscle of O. bonariensis exceeded the value suggested by WHO to be safe, thus causing a health risk to persons consuming fish as a result of chronic exposure to microcystin. Gills also showed bioaccumulation of MC-RR, raising questions on the mechanism involved in the possible uptake of MC-RR through gills as well as on its accumulation in this organ. Although MC-LR has been reported in brain of fish, this is the first report confirming the presence of MC-RR in this organ, which means that both toxins are able to cross the blood-brain barrier. These findings also raise questions on the probable neurotoxicity of microcystins.


Accidental, Non-masturbatory, Non-intercourse Related, Self-Inflicted Penile Fracture: Case Report and Review of Literature.
Penile fracture is a relatively rare condition arising from a rupture of the corpus cavernosum. In the western world, it is most commonly associated with sexual intercourse. We hereby, report a rare case of accidental, non-masturbatory, self-inflicted penile fracture. It was diagnosed promptly based on clinical assessment, confirmed radiologically and managed surgically, resulting in a good long-term clinical outcome. Our case highlights the importance of maintaining a high index of suspicion to diagnose this rare condition, even in the absence of a typical mechanism of injury, in order to ensure that such an injury is not missed.


Development of mass allelic exchange, a technique to enable sexual genetics in Escherichia coli.
Despite dramatic advances in genomics, connecting genotypes to phenotypes is still challenging. Sexual genetics combined with linkage analysis is a powerful solution to this problem but generally unavailable in bacteria. We build upon a strong negative selection system to invent mass allelic exchange (MAE), which enables hybridization of arbitrary (including pathogenic) strains of Escherichia coli. MAE reimplements the natural phenomenon of random cross-overs, enabling classical linkage analysis. We demonstrate the utility of MAE with virulence-related gain-of-function screens, discovering that transfer of a single operon from a uropathogenic strain is sufficient for enabling a commensal E. coli to form large intracellular bacterial collections within bladder epithelial cells. MAE thus enables assaying natural allelic variation in E. coli (and potentially other bacteria), complementing existing loss-of-function genomic techniques.


Neonatal pneumococcal colonisation caused by Influenza A infection alters lung function in adult mice.
There is emerging epidemiological data to suggest that upper respiratory tract bacterial colonisation in infancy may increase the risk of developing respiratory dysfunction later in life, and respiratory viruses are known to precipitate persistent colonisation. This study utilized a neonatal mouse model of Streptococcus pneumonia (SP) and influenza A virus (IAV) co-infection, where bronchoalveolar leukocyte infiltration had resolved by adulthood. Only co-infection resulted in persistent nasopharyngeal colonisation over 40 days and a significant increase in airway resistance in response to in vivo methacholine challenge. A significant increase in hysteresivity was also observed in IAV and co-infected mice, consistent with ventilatory heterogeneity and structural changes in the adult lung. Airway hyper-responsiveness was not associated with a detectable increase in goblet cell transdifferentiation, peribronchial smooth muscle bulk or collagen deposition in regions surrounding the airways. Increased reactivity was not observed in precision cut lung slices challenged with methacholine in vitro. Histologically, the airway epithelium appeared normal and expression of epithelial integrity markers (ZO-1, occludin-1 and E-cadherin) were not altered. In summary, neonatal co-infection led to persistent nasopharyngeal colonisation and increased airway responsiveness that was not associated with detectable smooth muscle or mucosal epithelial abnormalities, however increased hysteresivity may reflect ventilation heterogeneity.


Nopaline synthase gene is expressed in Escherichia coli and in cucumber cells under a hybrid promoter.
Fusion of the nopaline synthase gene (nos) to the Escherichia coli trp promoter gave rise to a hybrid promoter (tros). Under control of this hybrid element, synthesis of nopaline was observed in E. coli as well as in cucumber cells transformed with the described vector.


Glucocorticoids and Their Receptor Isoforms: Roles in Female Reproduction, Pregnancy, and Foetal Development.
Alterations in the hypothalamic-pituitary-adrenal (HPA) axis and associated changes in circulating levels of glucocorticoids are integral to an organism's response to stressful stimuli. Glucocorticoids acting via glucocorticoid receptors (GRs) play a role in fertility, reproduction, placental function, and foetal development. GRs are ubiquitously expressed throughout the female reproductive system and regulate normal reproductive function. Stress-induced glucocorticoids have been shown to inhibit reproduction and affect female gonadal function by suppressing the hypothalamic-pituitary-gonadal (HPG) axis at each level. Furthermore, during pregnancy, a mother's exposure to prenatal stress or external glucocorticoids can result in long-lasting alterations to the foetal HPA and neuroendocrine function. Several GR isoforms generated via alternative splicing or translation initiation from the GR gene have been identified in the mammalian ovary and uterus. The GR isoforms identified include the splice variants, GRα and GRβ, and GRγ and GR-P. Glucocorticoids can exert both stimulatory and inhibitory effects and both pro- and anti-inflammatory functions in the ovary, in vitro. In the placenta, thirteen GR isoforms have been identified in humans, guinea pigs, sheep, rats, and mice, indicating they are conserved across species and may be important in mediating a differential response to stress. Distinctive responses to glucocorticoids, differential birth outcomes in pregnancy complications, and sex-based variations in the response to stress could all potentially be dependent on a particular GR expression pattern. This comprehensive review provides an overview of the structure and function of the GR in relation to female fertility and reproduction and discusses the changes in the GR and glucocorticoid signalling during pregnancy. To generate this overview, an extensive non-systematic literature search was conducted across multiple databases, including PubMed, Web of Science, and Google Scholar, with a focus on original research articles, meta-analyses, and previous review papers addressing the subject. This review integrates the current understanding of GR variants and their roles in glucocorticoid signalling, reproduction, placental function, and foetal growth.


Effect of papaverine on solute transport in peritoneal dialysis.
Since vasodilators enhance the peritoneal solute transport, the influence of intraperitoneal papaverine was studied. To verify the action of papaverine on peritoneal transport we selected 33 patients on acute peritoneal dialysis and performed two 2-hour cycles with 2000 cc of a 1.5% solution, adding 40 mg of papaverine to the fourth cycle. At the end of the third and fourth cycles blood and dialysate were drawn for urea, creatinine, glucose and protein levels, using peritoneal clearances of urea and creatinine, glucose absorption and net protein loss to compare the two cycles. We found no significant change in solute transport (urea clearance p greater than 0.0.5; creatinine clearance, protein loss and glucose absorption p greater than 0.1).


Ecologic study of influenza vaccination uptake and COVID-19 death rate in New York City.
The aim of this ecological study was to assess the area-level relationship between cumulative death rate for COVID-19 and historic influenza vaccination uptake in the New York City population. Predictors of COVID-19 death included self-reported influenza vaccination in 2017, as well as four CDC-defined risk factors of severe COVID-19 infection available at the ecological level, which were diabetes, asthma, BMI 30-100 (2 kg/m2) and hypertension, in addition to race and age (65 + years). After adjusting for potential confounders, for every one-unit increase in influenza vaccination uptake for each zip code area, the rate of COVID-19 deaths decreased by 5.17 per 100,000 residents (p < 0.0001). Zip codes with a higher prevalence of influenza vaccination had lower rates of COVID-19 mortality, inciting the need to further explore the relationship between influenza vaccination uptake and COVID-19 mortality at the individual level.


Are the aged a minority group?
The aged resemble minority groups in three ways, i.e. they suffer from prejudice, discrimination, and deprivation. On the other hand, they are not born into their age group, and they have little sense of group identity or political unity. The most descriptive term would seem to be "quasi-minority group." However, the aged are becoming less and less like minority groups as society seems to be moving toward an "age-irrelevant" era.


Enzyme classification and the entanglement of values and epistemic standards.
This paper investigates the case of enzyme classification to evaluate different ideals for regulating values in science. I show that epistemic and non-epistemic considerations are inevitably and untraceably entangled in enzyme classification, and argue that this has significant implications for the two main kinds of views on values in science, namely, Epistemic Priority Views and Joint Satisfaction Views. More precisely, I argue that the case of enzyme classification poses a problem for the usability and descriptive accuracy of these two views. The paper ends by suggesting that these two views provide different but complementary perspectives, and that both are useful for evaluating values in science.


Migration and morbidity: implications for geographical studies of disease.
Many studies have examined associations between the geographical patterns of disease and possible causal factors. Underlying such studies is the assumption that residence in an area can be equated with exposure to conditions that pertain there. Migration upsets this assumption. Using data on self-reported morbidity from the 1981 Census for Great Britain it is shown that the health status of migrants differs considerably from that of non-migrants. Young migrants, particularly those moving longer distances are relatively healthy. Area of net out-migration of this age group are therefore likely to become characterised by a less healthy population whereas the reverse will be true of areas of net in-migration. Amongst older people migration tends to be selective of those in poor health moving shorter distances to avoid environmental health hazards or to be closer to medical care. This has the curious effect of increasing morbidity and mortality rates in areas with favourable environmental conditions and good medical services. Migration is therefore likely to be a significant source of error in geographically based studies of the association between disease and the environment. Sometimes the effect will be to obscure real environmental causes of disease. In other circumstances spurious association will be produced. These problems will be greatest for diseases with long latent periods which allow plenty of time for mobility of the population. Problems are also likely to increase with the growing mobility of the population.


Is glaucoma associated with an increased risk of cataract?
The risk of developing cataract in patients with untreated glaucoma or with ocular hypertension was evaluated by comparing the values of lenticular autofluorescence and light transmission in 16 patients with primary open angle glaucoma and 22 patients with ocular hypertension with those of 24 healthy controls. Increase of lenticular autofluorescence and decrease of transmission values in comparison with controls were considered to be precursors of cataract. The values of both variables were determined by fluorophotometry. Each value was normalised for age by dividing it by the value for a healthy control of the same age. The mean age normalised autofluorescence and transmission values of all patients did not differ significantly from those of the controls (difference < 5%; p = 0.6 and p = 0.2, respectively). Also the mean age normalised autofluorescence and transmission values between glaucoma and ocular hypertension patients did not differ significantly (p = 0.8 and p = 0.9, respectively). The study indicates that untreated primary open angle glaucoma or untreated ocular hypertension do not seem to increase significantly the risk of developing cataract.


Association between high risk for preterm birth and changes in gingiva parameters during pregnancy-a prospective cohort study.
The objective of this study was to investigate clinical and microbiological gingival changes during pregnancy in women without periodontal disease. Additionally, these parameters were to be compared in women with high risk for preterm birth and women with a normal course of pregnancy. Group I consisted of 40 subjects at high risk for preterm birth, while group II involved 49 subjects with a normal course of pregnancy. The control group (III) was made up of 50 non-pregnant women. Clinical parameters (plaque index, gingival index, probing pocket depths, gingival swelling, bleeding on probing) and microbiological changes were monitored during pregnancy and 2-4 weeks after parturition. In the high-risk preterm group (I), 19 women could be included in data analysis. This group was compared to 41 women in the normal pregnancy group (II) and 50 non-pregnant women (III). Gingival inflammation was significantly higher in women with high risk for preterm birth (I) compared to non-risk pregnant women (II, p < 0.05). In addition, in this group (I), the subgingival amounts of Fusobacterium nucleatum (> 105) were found to be significantly higher after childbirth compared to non-pregnant women (p < 0.05). Even without having periodontal disease, women with high risk for preterm birth showed worse clinical values compared to non-risk pregnant and non-pregnant women and an increased detection of Fusobacterium nucleatum after delivery. High risk for preterm birth might be associated with the occurrence of increased gingival inflammation.


19q distal trisomy due to a de novo (19;22)(q13.2;p11) translocation.
A 2 4/12-year-old girl whith a de novo 46,XX,-22 + der(22), t(19;22)(q13.2;p11) karyotype is described. From this and other eight similar cases previously published, a typical phenotype in distal 19q trisomy is concluded.


Accessibility of selenomethionine proteins by total chemical synthesis: structural studies of human herpesvirus-8 MIP-II.
The determination of high resolution three-dimensional structures by X-ray crystallography or nuclear magnetic resonance (NMR) is a time-consuming process. Here we describe an approach to circumvent the cloning and expression of a recombinant protein as well as screening for heavy atom derivatives. The selenomethionine-modified chemokine macrophage inflammatory protein-II (MIP-II) from human herpesvirus-8 has been produced by total chemical synthesis, crystallized, and characterized by NMR. The protein has a secondary structure typical of other chemokines and forms a monomer in solution. These results indicate that total chemical synthesis can be used to accelerate the determination of three-dimensional structures of new proteins identified in genome programs.


A multimodal approach to understanding motor impairment and disability after stroke.
Many different measures have been found to be related to behavioral outcome after stroke. Preclinical studies emphasize the importance of brain injury and neural function. However, the measures most important to human outcomes remain uncertain, in part because studies often examine one measure at a time or enroll only mildly impaired patients. The current study addressed this by performing multimodal evaluation in a heterogeneous population. Patients (n = 36) with stable arm paresis 3-6 months post-stroke were assessed across 6 categories of measures related to stroke outcome: demographics/medical history, cognitive/mood status, genetics, neurophysiology, brain injury, and cortical function. Multivariate modeling identified measures independently related to an impairment-based outcome (arm Fugl-Meyer motor score). Analyses were repeated (1) identifying measures related to disability (modified Rankin Scale score), describing independence in daily functions and (2) using only patients with mild deficits. Across patients, greater impairment was related to measures of injury (reduced corticospinal tract integrity) and neurophysiology (absence of motor evoked potential). In contrast, (1) greater disability was related to greater injury and poorer cognitive status (MMSE score) and (2) among patients with mild deficits, greater impairment was related to cortical function (greater contralesional motor/premotor cortex activation). Impairment after stroke is most related to injury and neurophysiology, consistent with preclinical studies. These relationships vary according to the patient subgroup or the behavioral endpoint studied. One potential implication of these results is that choice of biomarker or stratifying variable in a clinical stroke study might vary according to patient characteristics.


Oxidative stress and psoriasis: the effect of antitumour necrosis factor-α inhibitor treatment.
Psoriasis is a chronic, inflammatory skin condition associated with a high frequency of cardiovascular events. Modifications of plasma lipids, and an increase in the levels of biochemical markers of inflammation and lipid peroxidation have been reported in subjects with psoriasis, suggesting a relationship between psoriasis, inflammation and oxidative damage. To investigate whether modulation of inflammatory activity by tumour necrosis factor-α inhibitors in patients with psoriasis is associated with modification of lipid profiles, oxidative stress and paraoxonase (PON)1 activity. The levels of plasma lipids and lipoprotein(a), and the levels of the markers of inflammation and lipid peroxidation were evaluated in subjects with psoriasis (n=23) before and after 24 weeks of treatment with etanercept. In the same subjects plasma total antioxidant capacity and the activity of PON1, an antioxidant and anti-inflammatory enzyme associated with the high-density lipoproteins (HDLs), were investigated. The results showed that clinical improvement in patients with psoriasis treated with etanercept is associated with a reduction in the levels of inflammatory markers [C-reactive protein (CRP)] and lipid peroxidation, and also with increased antioxidant capacity in the serum of patients with psoriasis. These modifications are associated with a significant increase in the activity of PON1. A significant increase in the PON1/CRP ratio has also been observed in patients with psoriasis after treatment. The significant inverse correlation between CRP and PON1 activity suggests a relationship between PON1 activity and inflammation. Treatment with etanercept is associated with a reduction in lipid peroxidation and an improvement in HDL antioxidant and anti-inflammatory properties.


RIN4 interacts with Pseudomonas syringae type III effector molecules and is required for RPM1-mediated resistance in Arabidopsis.
In Arabidopsis, RPM1 confers resistance against Pseudomonas syringae expressing either of two sequence unrelated type III effectors, AvrRpm1 or AvrB. An RPM1-interacting protein (RIN4) coimmunoprecipitates from plant cell extracts with AvrB, AvrRpm1, or RPM1. Reduction of RIN4 protein levels inhibits both the hypersensitive response and the restriction of pathogen growth controlled by RPM1. RIN4 reduction causes diminution of RPM1. RIN4 reduction results in heightened resistance to virulent Peronospora parasitica and P. syringae, and ectopic defense gene expression. Thus, RIN4 positively regulates RPM1-mediated resistance yet is, formally, a negative regulator of basal defense responses. AvrRpm1 and AvrB induce RIN4 phosphorylation. This may enhance RIN4 activity as a negative regulator of plant defense, facilitating pathogen growth. RPM1 may "guard" against pathogens that use AvrRpm1 and AvrB to manipulate RIN4 activity.


Hepatocyte growth factor promotes cancer cell migration and angiogenic factors expression: a prognostic marker of human esophageal squamous cell carcinomas.
Hepatocyte growth factor/scatter factor (HGF/SF) and its receptor, c-Met, play important roles in tumor development and progression. In this study, we measured the serum HGF levels in patients with esophageal squamous cell carcinoma (ESCC) to evaluate its relationships with clinicopathologic features and the role of HGF in ESCC. One hundred and forty-nine patients with ESCC were studied. Pretherapy serum was collected and ELISA was used to detect the concentrations of HGF, vascular endothelial growth factor (VEGF), and interleukin 8 (IL-8). The function of HGF was shown by invasion chamber assay. Pretherapy serum HGF was found to be significantly higher in patients with ESCC than in control subjects. The levels of HGF correlated significantly with advanced tumor metastasis stage and survival. Multivariate analyses showed that serum HGF level in cell migration was an independent prognostic factor. Increased HGF serum levels correlated positively with serum levels of VEGF and IL-8. Our results also showed that HGF was overexpressed in ESCC tissues and cell lines. In vitro study showed that HGF could stimulate ESCC cell to express VEGF and IL-8 and markedly enhance invasion and migration of ESCC cells. Furthermore, HGF-induced IL-8 and VEGF expression was dependent on extracellular signal-regulated kinase signaling pathways. The inhibition of extracellular signal-regulated kinase activation reduced HGF-mediated IL-8 and VEGF expression. Our results suggest that serum HGF may be a useful biomarker of tumor progression and a valuable independent prognostic factor in patients with ESCC. HGF may be involved in the progression of ESCC as an autocrine/paracrine factor via enhancing angiogenesis and tumor cell invasion and migration.


Epidermal growth factor modulates claudins and tight junctional functions in ovarian cancer cell lines.
Ovarian adenocarcinomas, like human ovarian surface epithelial cells, form functional tight junctions. Tight junction molecules claudin-3 and claudin-4, which are the receptors of Clostridium perfringens enterotoxin (CPE), are abnormally upregulated in epithelial ovarian cancers of all subtypes including, mucinous cystadenocarcinoma and serous cystadenocarcinoma. Clostridium perfringens enterotoxin may be a novel tumor-targeted therapy for ovarian cancers. In epithelial ovarian cancers, overexpression of epidermal growth factor receptor has been observed and the exogenous ligand EGF induces epithelial-mesenchymal transition in ovarian surface epithelium. Epidermal growth factor (EGF) signaling modulates expression of claudins with changes of fence and barrier functions in various cell types. However, the regulation of tight junctions by EGF in ovarian cancers remains unclear. In the present study, to investigate the mechanisms of the regulation of tight junctions in ovarian cancers, ovarian cancer cell lines mucinous cystadenocarcinoma (MCAS) and serous cystadenocarcinoma (HUOA) were treated with EGF. Epidermal growth factor downregulated claudin-3 in MCAS and claudin-4 in HUOA by inducing degradation of the proteins with changes in structures and functions of tight junctions via the MEK/ERK or PI3K/Akt signaling pathway. In addition, in HUOA but not MCAS, EGF downregulated the cytotoxic effect of CPE via claudin-4. Thus, there were different mechanisms for regulation of claudins by EGF between subtypes of epithelial ovarian cancer cells in vitro. These results indicate that EGF may affect claudins and tight junctional functions in ovarian cancer cells during cancer progression.


Molecular analysis of infectious bronchitis viruses isolated in Slovenia between 1990 and 2005: a retrospective study.
Fifteen infectious bronchitis viruses (IBV) isolated from broiler and broiler breeder flocks in Slovenia between 1990 and 2005 were molecularly characterised. IBV strains were divided into four genotypes by the analysis of the S1 gene region. Four strains belonged to the Massachusetts genotype, one strain was placed into the QX genotype, one strain formed a cluster together with the B1648 strain and nine strains were classified into the 624/I genotype. Nine Slovenian strains of the 624/I genotype formed two subgroups independently of the time of isolation and the geographical origin. Phylogenetic analysis of the partial N gene sequences revealed lower sequence variability and different clustering of the Slovenian IBV. Fourteen strains were grouped together with the strains H120 and D1466. One strain formed a cluster with the strain 793/B.


A critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo.
Recombinant adenoviruses have been used widely for gene therapy due to their high transduction efficiency in vivo. However, the attendant innate immune response to adenoviral vectors has limited their applications for in vivo gene therapy. Recent studies have shown that adenoviruses activate the innate immunity through both Toll-like receptor-dependent (TLR-dependent) and TLR-independent pathways, leading to the production of type I interferons (IFNs) and other inflammatory cytokines. Furthermore, type I IFNs play a pivotal role in innate immune elimination of adenoviral vectors in vivo. It remains to be defined how type I IFNs regulate innate immune clearance of adenoviral vectors. In this study, we showed in vivo that natural killer (NK) cells were activated and accumulated in the liver upon intravenous administration of adenoviral vectors, leading to the loss of adenoviral genome and the reduction of transgene expression. We further demonstrated that type I IFNs were critical for the activation of NK cells. This was achieved by direct action of type I IFNs on NK cells. Overall, our observations reveal a critical role for type I IFN-dependent NK cell activation in innate immune elimination of adenoviral vectors in vivo and may help design effective strategies to improve the outcome of adenovirus-mediated gene therapy.