Nutritive value of full-fat sunflower seeds in broiler diets.
A study was conducted to evaluate the nutritive value and inclusion level of sunflower seed (SFS) in broiler diets. SFS contained 38.7% ether extract (EE), 16.9% CP, 14.9% crude fiber (CF), 3.5% ash, 0.57% lysine, and 0.46% methionine (89.2% DM basis). The AME (kcal/ kg) content of SFS in roosters was 5,225 and in broilers at 4, 18, and 35 d of age was 3,493, 5,132, and 5,162, respectively. The CP, EE, and CF digestibilities were 80.4, 71.2, and 11.4%, respectively. In an isocaloric and isonitrogenous diet containing SFS at 0, 5, 10, 15, and 20%, SFS up to 20% did not affect weight gain and feed consumption, but the feed conversion ratio was improved (P < 0.05) when broilers were fed 15 or 20% SFS in the starter and finisher diets. CF digestibility of starter diet was significantly lower when 15 or 20% SFS was included. CF digestibility of the finisher diet and digestibility of other nutrients in starter and finisher diets were comparable in all treatment groups. Liver and muscle lipid content, plasma total and high-density lipoprotein cholesterol content, muscle cholesterol content, dressing percentage, liver weight, and giblet weight (as % live weight) were comparable among all treatment groups. Abdominal fat was increased in birds fed 20% SFS. Percentage skin was decreased in broilers fed > or = 10% SFS.


Prevalence of vestibular symptoms in individuals with auditory neuropathy spectrum disorder - A retrospective study.
The objective of the study was to retrospectively determine the prevalence of vestibular symptoms in individuals with auditory neuropathy spectrum disorder (ANSD). It was also attempted to determine the prevalence of vestibular symptoms and factors (gender and age of reported hearing loss) that could affect the prevalence in individuals with ANSD. The vestibular symptoms reported in the case history were analyzed in individuals diagnosed with ANSD. The symptoms reported by a total of 316 individuals (185 females and 131 males) with ANSD were analyzed. The result of the study showed that one in five individuals with ANSD reported at least one of the vestibular symptom. The vestibular symptoms were in more females and in individuals with earlier onset of hearing loss. The result of the study supports that there is a vestibular damage in individuals with ANSD. However, it is essential to carry out prospective studies validating these vestibular symptoms with objective vestibular tests before generalizing the results.


The therapy of osteomyelitis in outpatient settings.
Because of rising hospital costs, the use of outpatient antibiotic therapy, both oral and parenteral, is increasing. A program for the intravenous administration of antibiotics to outpatients has been established as a freestanding outpatient clinic (Intracare). Some diseases lend themselves to treatment in such a setting better than others. Osteomyelitis accounted for one-third of the diagnoses in the first 1,000 patients treated at Intracare. While patients are usually begun on intravenous antibiotics following initial surgical debridement in the hospital, hospitalization may be avoided entirely in some patients who undergo surgery in outpatient surgical centers and are begun on intravenous antibiotics immediately afterwards at Intracare. For outpatient use, cephalosporins are most frequently used because of their efficacy, safety, and pharmacokinetics, which allows less-frequent administration. Both cefazolin and ceftriaxone were used most frequently in the Intracare program, although the once-daily administration of ceftriaxone was considered most convenient for outpatients and allowed them to return to normal activities even while receiving antibiotic therapy.


Vasoactive intestinal peptide stimulates alkali excretion in turtle urinary bladder.
The turtle urinary bladder possesses an active transport mechanism for the electrogenic secretion of alkali. This process is independent of exogenous Cl and Na, induced by cyclic AMP (cAMP), and potentiated in bladders from NaHCO3-loaded (alkalotic) turtles. In the present study, it is shown that the serosal addition of vasoactive intestinal peptide (VIP) induces rapidly developing parallel increases in alkali secretion and in the short-circuiting current carried by this secretion. The VIP-induced increment in alkali secretion is greater in the presence than in the absence of an exogenously added phosphodiesterase inhibitor. Additions of a cAMP analog subsequent to the VIP-induced alkali secretion fail to induce any further increase in alkalinization. These results provide evidence for the action of VIP as a hormonal up regulator of alkali excretion in the turtle urinary bladder.


Triggering of atypical hemolytic uremic syndrome by influenza A (H1N1).
We report a case of atypical hemolytic uremic syndrome (aHUS) triggered by influenza A (H1N1) in a 17-year-old boy with a mutation in the gene (CD46) encoding the transmembrane complement regulator membrane cofactor protein. The patient recovered completely following treatment with oseltamivir, plasma exchange, and hemodialysis. We describe the case and discuss this unusual association of diseases.


AMPKα2 is a skeletal muscle stem cell intrinsic regulator of myonuclear accretion.
Due to the post-mitotic nature of skeletal muscle fibers, adult muscle maintenance relies on dedicated muscle stem cells (MuSCs). In most physiological contexts, MuSCs support myofiber homeostasis by contributing to myonuclear accretion, which requires a coordination of cell-type specific events between the myofiber and MuSCs. Here, we addressed the role of the kinase AMPKα2 in the coordination of these events supporting myonuclear accretion. We demonstrate that AMPKα2 deletion impairs skeletal muscle regeneration. Through in vitro assessments of MuSC myogenic fate and EdU-based cell tracing, we reveal a MuSC-specific role of AMPKα2 in the regulation of myonuclear accretion, which is mediated by phosphorylation of the non-metabolic substrate BAIAP2. Similar cell tracing in vivo shows that AMPKα2 knockout mice have a lower rate of myonuclear accretion during regeneration, and that MuSC-specific AMPKα2 deletion decreases myonuclear accretion in response to myofiber contraction. Together, this demonstrates that AMPKα2 is a MuSC-intrinsic regulator of myonuclear accretion.


Multicancer Early Detection: Learning From the Past to Meet the Future.
Multicancer early detection (MCED) tests may soon be available to screen for many cancers using a single blood test, yet little is known about these tests beyond their diagnostic performance. Taking lessons from the history of cancer early detection, we highlight 3 factors that influence how performance of early detection tests translates into benefit and benefit-harm trade-offs: the ability to readily confirm a cancer signal, the population testing strategy, and the natural histories of the targeted cancers. We explain why critical gaps in our current knowledge about each factor prevent reliably projecting the expected clinical impact of MCED testing at this point in time. Our goal is to communicate how much uncertainty there is about the possible effects of MCED tests on population health so that patients, providers, regulatory agencies, and the public are well informed about what is reasonable to expect from this potentially important technological advance. We also urge the community to invest in a coordinated effort to collect data on MCED test dissemination and outcomes so that these can be tracked and studied while the tests are rigorously evaluated for benefit, harm, and cost.


Retroviral vectors pseudotyped with severe acute respiratory syndrome coronavirus S protein.
The worldwide outbreak of severe acute respiratory syndrome (SARS) was shown to be associated with a novel coronavirus (CoV) now called SARS CoV. We report here the generation of SARS CoV S protein-pseudotyped murine leukemia virus (MLV) vector particles. The wild-type S protein pseudotyped MLV vectors, although at a low efficiency. Partial deletion of the cytoplasmic tail of S dramatically increased infectivity of pseudotypes, with titers only two- to threefold lower than those of pseudotypes generated in parallel with the vesicular stomatitis virus G protein. S-pseudotyped MLV particles were used to analyze viral tropism. MLV(SARS) pseudotypes and wild-type SARS CoV displayed similar cell types and tissue and host restrictions, indicating that the expression of a functional receptor is the major restraint in permissiveness to SARS CoV infection. Efficient gene transfer could be detected in Vero and CaCo2 cells, whereas the level of gene marking of 293T, HeLa, and HepG2 cells was only slightly above background levels. A cat cell line and a dog cell line were not susceptible. Interestingly, PK-15, a porcine kidney cell line, and primary porcine kidney cells were also highly permissive for SARS S pseudotypes and wild-type SARS CoV. This finding suggests that swine may be susceptible to SARS infection and may be a source for infection of humans. Taken together, these results indicate that MLV(SARS) pseudotypes are highly valuable for functional studies of viral tropism and entry and, in addition, can be a powerful tool for the development of therapeutic entry inhibitors without posing a biohazard to human beings.


[Expediency of application of miniinvasive access in prosthesis of aortal valve].
The literature data, concerning expediency of miniinvasive access in aortal valve prosthesis for its failures of various etiologies, are analyzed. Advantages and failures, occurring while its application, are discussed. Own clinical observation of successful conduction of operative intervention for aortal valve failure, using miniinvasive access, is adduced.


[Integrin beta1 mediates hepatocellular carcinoma cells adhesion & chemotaxis to type IV collagen].
A micropipette technique was adopted to investigate the effect of blockade of integrin betal on adhesion of hepatocellular carcinoma (HCC) cells onto type IV collagen (Col IV) coated surfaces and pseudopod protrusion of HCC cells in response to Col IV stimulation. Adhesion strength was expressed as an adhesion force, which was defined as the product of the cross sectional area and critical negative pressure needed to detach single cell away from the substrate. Chemotactic pseudopod protrusion of an HCC cell was evaluated using a dual-pipette set-up, in which two pipettes filled with Col IV solution were positional in close contact with the same cell and pseudopod protrusion into each pipette was viewed dynamically and recorded with a tape recorder. The lengths of pseudopods were measured and plotted against time to obtain a pseudopod growth curve. The integrin beta1 subunit on the surfaces of HCC cells was analyzed by flow cytometry. The results showed that the adhesion forces for HCC cells adhering on 5 microg/ml Col IV coated surfaces were 932 +/- 134 (x 10(-10) N, n = 60). Upon treatment of HCC cells with Anti-CD29 in a protein concentration of 5 microg/ml and 10 microg/ml, the value decreased significantly to 449 +/- 119 (x 10(-10) N, n = 60) and 220 +/- 78 (x 10(-10) N, n = 55), respectively. In dual pipette chemotaxis experiment, when the two pipettes were filled with Col IV in an identical concentration of 600 microg/ml, pseudopods extended from the HCC cell into each of the pipettes nearly symmetrically, i.e., with nearly identical maximum pseudopod length and similar pseudopod growth curves. Upon addition of Anti-CD29 to one of the pipettes in a protein concentration of 20 microg/ml, pseudopod protrusion was blocked nearly completely while protrusion into the opposite pipette became more evidently, with larger maximum length. Expression of integrin beta1 was up to 95.78% to cells chosen in the experiment. These results suggested that integrin beta1 subunit was important constituent receptor subunit for mediating HCC cell adhesion and chemotactic pseudopod protrusion to Col IV.


Permanent endocardial pacing in pediatric patients.
Improved technology with smaller pulse generators and passive and active fixation endocardial lead systems has the potential for facilitating safe, reliable pacing in pediatric patients. Of 18 pediatric patients (mean age 11.1 years) undergoing permanent transvenous cardiac pacing during the period April, 1977, to January, 1981, two (11%) required reoperation during a mean follow-up of 18.8 months. Satisfactory stable pacing was maintained even in patients with dual-chamber pacing systems and in patients with transposition of the great vessels in whom a lead was placed in a morphologically left atrium. This reoperation rate of 11% contrasts with a reoperation rate of 32% in 22 pediatric patients (mean age 8.7 years) who underwent placement of ventricular-demand epicardial pacing systems during the same period. In our experience, transvenous endocardial pacing is the preferred route in pediatric patients because of the improved lead system survival and the potential for atrial or atrioventricular sequential pacing.


Human adipose tissue possesses a unique population of pluripotent stem cells with nontumorigenic and low telomerase activities: potential implications in regenerative medicine.
In this study, we demonstrate that a small population of pluripotent stem cells, termed adipose multilineage-differentiating stress-enduring (adipose-Muse) cells, exist in adult human adipose tissue and adipose-derived mesenchymal stem cells (adipose-MSCs). They can be identified as cells positive for both MSC markers (CD105 and CD90) and human pluripotent stem cell marker SSEA-3. They intrinsically retain lineage plasticity and the ability to self-renew. They spontaneously generate cells representative of all three germ layers from a single cell and successfully differentiate into targeted cells by cytokine induction. Cells other than adipose-Muse cells exist in adipose-MSCs, however, do not exhibit these properties and are unable to cross the boundaries from mesodermal to ectodermal or endodermal lineages even under cytokine inductions. Importantly, adipose-Muse cells demonstrate low telomerase activity and transplants do not promote teratogenesis in vivo. When compared with bone marrow (BM)- and dermal-Muse cells, adipose-Muse cells have the tendency to exhibit higher expression in mesodermal lineage markers, while BM- and dermal-Muse cells were generally higher in those of ectodermal and endodermal lineages. Adipose-Muse cells distinguish themselves as both easily obtainable and versatile in their capacity for differentiation, while low telomerase activity and lack of teratoma formation make these cells a practical cell source for potential stem cell therapies. Further, they will promote the effectiveness of currently performed adipose-MSC transplantation, particularly for ectodermal and endodermal tissues where transplanted cells need to differentiate across the lineage from mesodermal to ectodermal or endodermal in order to replenish lost cells for tissue repair.


Cancer incidence in the Swedish leather tanning industry: updated findings 1958-99.
To assess how a 10 year extension of the follow up period affected cancer incidence in the Swedish leather tanning cohort. A cohort of 2027 tannery workers (of which 482 were women) who had been employed for at least one year between 1900 and 1989 at one of three Swedish leather tanneries, was established. The start of observation varied between 1958 and 1966 for the three plants. Through linkage with the Swedish Cancer Registry, incident cancer cases were recorded up to 1999. Cause specific expected cancer incidence was calculated for 1958-99 based on calendar year, sex, and five year age group specific incidence rates for the counties where the plants had been located. Altogether 56,022 person-years at risk were generated. A total of 351 incident cancer cases were observed compared to 302 expected, which resulted in an increased standardised incidence ratio (SIR) of 1.16 (95% CI 1.04 to 1.29). An enhanced risk for prostate cancer was observed (SIR 1.44, 95% CI 1.10 to 1.86), mainly attributable to the later part of the observation period (1990-99). In this updated analysis the previously observed risk excess for soft tissue sarcomas was no longer significant (SIR 2.62, 95% CI 0.96 to 5.70). For multiple myelomas and sinonasal cancer the slight non-significant excesses remained, still based on very few cases. The increased risk for prostate cancer in the present study might be a chance finding, but is noteworthy, since it is in acccordance with the finding of increased SIR for prostate cancer among leather workers in another recent Swedish study. Moreover, excess risks for prostate cancer among farmers have been reported, indicating pesticides as possible causative agents. Leather tanners have also been exposed to pesticides.


An anesthetic method compatible with (18)F-FDG-PET studies in mice.
The purpose of this study was to establish an experimental setting and an anesthetic method compatible with future sequential studies using (18)F-FDG-PET single scans, i.e. autoradiographic measurements, for the estimation of metabolic rate of glucose (MRglc) in mice. In this study we had no access to a small animal PET scanner and therefore focus was on the anesthetic setting and optimization of the input function as a preparation for the future tumor metabolic studies. Initially, four combinations of intraperitoneal (ip) anesthesia were tested on tumor bearing mice. Fentanyl-fluanisone plus diazepam yielded low and stable blood glucose levels and kept the animals sedated for approximately 2 h. The anesthesia was also tested in a longitudinal (18)F-FDG study, where tumor bearing mice were anesthetized, injected with (18)F-FDG, and sampled for blood, before, one day after, and 8 days after treatment with cisplatin. The animals were in good condition during the entire study period. To validate the method, average MRglc of whole brain and cerebellum in mice were calculated and compared with the literature. The average MRglc in the whole brain and cerebellum were 46.2±4.4 and 39.0±3.1 µmol 100g(-1) min(-1). In the present study, we have shown that an ip anesthesia with a combination of fentanyl-fluanisone and diazepam is feasible and provides stable and low blood glucose levels after a fasting period of 4 h in experiments in nude mice with xenografted human tumors. We have also verified that (18)F-FDG, intraperitoneally administrated, results in an expected plasma activity uptake and clearance. The method doesn't alter the uptake in brain which is an indirect indication that the anesthesia doesn't alter the uptake in other organs. In combination with meticulous animal handling this set-up is reliable and future sequential tumor studies of early metabolic effects with calculation of MRglc following cytotoxic therapy are made possible.


Distribution of microtubules and microfilaments in developing vestibular sensory epithelium of mouse otocysts grown in vitro.
Otocysts explanted from 12th-gestation-day mice and maintained in organ culture under went a series of developmental changes which paralleled those that occurred in vivo and which resulted in the formation of a sensory epithelium of the vestibular type. At the time of explantation presumptive vestibular sensory epithelium consisted of cells that were undifferentiated, pseudostratified and rapidly proliferating. The only microtubules present were those of the mitotic apparatus. After 4 days of in vitro development cells comprising the presumptive vestibular sensory epithelium were less pseudostratified and more elongate; their nuclei had assumed a basal orientation and there was a clear maginal velum. Longitudinally oriented cytoplasmic microtubules were present at the apices of some cells; they were often grouped around a centriole which may have served as a nucleation centre for their assembly. After 7 days of in vitro development mitosis had ceased and supporting cells had innervated hair cells were present: both types of cells were always longer than they were broad and were often highly asymmetrical. Hair cells were flask- or columnar-shaped, with a nucleus situated in the basal third of the cell. Most mitochondria in hair cells were located in the apical third of the cell. The same distribution of mitochondria and nuclei was evident in supporting cells. Microtubules occurred throughout the length of the supporting cell and were always parallel to its longitudinal axis. In hair cells microtubules were more frequent than in supporting cells: the majority were parallel to the longitudinal axis of the cell but there were two exceptions. First, at the apex of hair cells some microtubules were oriented transversely and diagnonally: these were probably involved in the development and maintenance of the constricted apex of these cells. Secondly, microtubules appeared to be randomly arranged in the narrow region of the cytoplasm between the ventral surface of the nucleus and the base of the hair cells. Microfilaments were confined to the basal third of hair cells where their orientation paralleled that of microtubules. The possible functions of microtubules and microfilaments in the development of hair cells and supporting cells of the mouse vestibular epithelium are discussed.


Small blob identification in medical images using regional features from optimum scale.
Recent advances in medical imaging technology have greatly enhanced imaging-based diagnosis which requires computational effective and accurate algorithms to process the images (e.g., measure the objects) for quantitative assessment. In this research, we are interested in one type of imaging objects: small blobs. Examples of small blob objects are cells in histopathology images, glomeruli in MR images, etc. This problem is particularly challenging because the small blobs often have in homogeneous intensity distribution and an indistinct boundary against the background. Yet, in general, these blobs have similar sizes. Motivated by this finding, we propose a novel detector termed Hessian-based Laplacian of Gaussian (HLoG) using scale space theory as the foundation. Like most imaging detectors, an image is first smoothed via LoG. Hessian analysis is then launched to identify the single optimal scale on which a presegmentation is conducted. The advantage of the Hessian process is that it is capable of delineating the blobs. As a result, regional features can be retrieved. These features enable an unsupervised clustering algorithm for postpruning which should be more robust and sensitive than the traditional threshold-based postpruning commonly used in most imaging detectors. To test the performance of the proposed HLoG, two sets of 2-D grey medical images are studied. HLoG is compared against three state-of-the-art detectors: generalized LoG, Radial-Symmetry and LoG using precision, recall, and F-score metrics.We observe that HLoG statistically outperforms the compared detectors.


Illegal road crossing behavior of pedestrians at overpass locations: Factors affecting gap acceptance, crossing times and overpass use.
The aim of designing overpasses is to provide safe road crossings for pedestrians by helping them to avoid conflicts with motor vehicles. However, the number of pedestrians who do not use overpasses to cross the road is very high. An observational survey of illegal road crossings was conducted at four overpass locations in Izmir, Turkey to determine the crossing time, crossing speed of the pedestrians and their distance and time gap perception for safe road-crossing within 25 m of the overpasses in both directions. Crossing time is the time needed for a pedestrian to cross a particular road. Time gap is strongly related with safety margin. If a pedestrian chooses a larger time gap, then the arrival time of the oncoming vehicle to the crossing point of the pedestrian increases thus, the possibility of a collision decreases. Each overpass was observed on weekdays during peak afternoon (12.30-13.30) and evening hours (17.00-18.00). At all overpass locations 454 illegal crossings were observed. ANOVA results revealed that age had a significant effect both on safety margin and crossing time. During the observations a survey was conducted among pedestrians who completed their crossings either using the overpass or at street level within 25 m of the overpass (n=231). Factors affecting the crossing choice of pedestrians were specified in the surveys. The major part of the respondents (71.7%) indicated that time saving was the main reason for crossing at street level. Pedestrians' crossing speeds were extracted from the video recordings to observe the effect of speed limit on pedestrian behavior. As a result, at locations where the speed limit was 70 km/h, pedestrians' average crossing speed was found to be 1.60 m/s and 1.73 m/s while at locations where the speed limit was 50 km/h, pedestrians' average crossing speed was found to be 1.04 m/s and 0.97 m/s. This shows that pedestrians feel safer while crossing when the vehicle speed is low.


Biocompatibility analysis of the Sundt-Kees booster clip and the Drake aneurysm clip. Technical note.
A combination of the Sundt-Kees reinforcing aneurysm clip applied to a Drake aneurysm clip in a piggyback fashion was studied for possible defects due to corrosion and or tissue toxicity. These two clips, which are made of similar metal (301 stainless steel), showed little or no defects when immersed in 5% saline or when implanted in rats for 6 months. This study demonstrates that clips made of similar metals can be used piggyback in patients without ill effects provided that they are carefully handled to avoid any abrasion or misbends which could conceivably lead to corrosion.


A cause of excessive daytime sleepiness. The upper airway resistance syndrome.
Subjects with isolated complaints of chronic daytime sleepiness are usually classified as "idiopathic hypersomniacs" and treated symptomatically. A group of these subjects was investigated during nocturnal sleep and daytime naps. In a subgroup of them, sleep was fragmented by very short alpha EEG arousals throughout the sleeping period. These short arousals are usually ignored in sleep analyses, but their impact is significant (in the 15 subjects identified with the syndrome, the mean sleep latency in multiple sleep latency tests was 5.1 +/- 1 min). These arousals are directly related to an abnormal increase in respiratory efforts during sleep (the mean peak inspiratory esophageal pressure measured in our subjects in the respiratory cycle just preceding a transient arousal was -33 +/- 7 cm H2O). Typically, an arousal occurs within one to three breaths of flow limitation associated with abrupt but limited reduction in tidal volume (ie, abnormal increase in upper airway resistance during sleep). The arousal restores normal breathing. Snoring was noted in association with these transient arousals in 10 of the 15 subjects; however, snoring was neither sufficient nor necessary for the identification of the clinical syndrome. Both sexes were equally represented in the affected group. All studied subjects had upper airway anatomy that was mildly abnormal. Nasal continuous positive airway pressure, used as an experimental tool, eliminated the daytime sleepiness (multiple sleep latency mean score = 13.5 min), the transient arousals (mean alpha EEG arousal index decreased from 31.3 +/- 12.4 to 8 +/- 2 per hour of sleep), and the abnormal upper airway resistance. Chronic daytime sleepiness is a major cause of social, economic, and medical impairment. Recognition of this syndrome and its cause is important, as specific treatments can be developed to eliminate the problem.


Diversity in emotion regulation strategy use: Resilience against posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD) is associated with impaired emotion regulation (ER). ER diversity, the variety, prevalence, and relative abundance of ER strategy use, may provide resilience against PTSD. This study examined the prospective relation between ER diversity and PTSD, while accounting for negative and positive life events, in interpersonal violence (IPV) survivors. IPV-exposed women with PTSD onset (PTSD; n = 22), without PTSD onset (IPV; n = 37), and non-traumatized control participants (NTC; n = 41) rated their ER strategy use and experience of negative and positive life events. The ER diversity index differentiated the participant groups. Importantly, group differences in ER diversity depended on the experience of life events. When experiencing fewer positive life events and more negative life events, the IPV and NTC groups, but not the PTSD group, demonstrated higher ER diversity. Thus, greater ER diversity during periods with more negative life events and fewer positive life events may play a protective role against PTSD onset for IPV survivors.


Deprivation of corticosterone does not prevent onset of obesity in Zucker fa/fa pups.
Adrenalectomy has been shown to reduce the development of obesity in adult Zucker fatty rat. In this study, we examined whether adrenalectomy could prevent the emergence of obesity and correct any of the first abnormalities to develop in fa/fa pups. Four-day-old Zucker pups were adrenalectomized and fed by adrenalectomized wet nurses until 11 days of age. The frequency distribution curves of fat cell volume clustered in two groups as they do in control litters, providing evidence that two phenotypes were present. Oxygen consumption measured at 8 days of age was significantly lower in fa/fa than in Fa/fa. Adrenalectomy did not restore the decreased oxygen consumption of fa/fa. In control litters, the GDP binding to brown adipose tissue mitochondria was twofold lower, whereas fatty acid synthase activity of this tissue was significantly increased in fa/fa pups. In inguinal adipose tissue of fa/fa pups, fatty acid synthase, and lipoprotein lipase activities were twice as active as in the tissue of lean pups. In adrenalectomized fa/fa pups, none of these metabolic abnormalities was corrected. The results demonstrate that adrenalectomy early in life did not prevent the emergence of obesity in suckling fa/fa rats.


Role of matrix Gla protein in angiotensin II-induced exacerbation of vascular calcification.
Vascular calcification predicts an increased risk for cardiovascular events in atherosclerosis, diabetes, and end-stage kidney diseases. Matrix Gla protein (MGP), an inhibitor of calcification, limits calcium phosphate deposition in the vessel wall. There are many factors contributing to the progression of atherosclerosis, including hypertension, hyperlipidemia, the renin-angiotensin system, and inflammation. Angiotensin II (ANG II) plays a crucial role in the atherogenic process through not only its pressor responses but also its growth-promoting and inflammatory effects. In this study, we investigated the role of MGP in ANG II-induced exacerbation of vascular calcification in human vascular smooth muscle cells (VSMCs). The expression of MGP, calcification, and apoptosis in human VSMCs were examined by Western blot analysis, real-time PCR, in situ terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling, and enzyme-linked immunosorbent assay, respectively. Increase in VSMC calcification in human atherosclerotic plaques upregulates MGP expression and apoptosis in a negative feedback manner. ANG II inhibited MGP expression in VSMCs via and in vitro in a dose- and time-dependent manner through ANG II type 1 receptor and NF-κB signaling pathway. Meanwhile, MGP inhibited the calcification, caspase-3 activity, activation of runt-related transcription factor 2, and release of inflammatory cytokines by VSMCs induced by calcification medium (2.5 mM P(i)) and ANG II in vitro. These observations provide evidence that ANG II exacerbates vascular calcification through activation of the transcription factors, runt-related transcription factor 2 and NF-κB, and regulation of MGP, inflammatory cytokines expression in human VSMCs.


Dimethyl fumarate mitigates optic neuritis.
Dimethyl fumarate (DMF) has been approved by the U.S. Food and Drug Administration (FDA) for the treatment of relapsing-remitting multiple sclerosis (RRMS), a demyelinating autoimmune disease characterized by acute episodes of motor, sensory, and cognitive symptoms. Optic neuritis is an episodic sequela experienced by some patients with RRMS that typically presents as acute, monocular vision loss. Episodes of optic neuritis damage and kill retinal ganglion cells (RGCs), and can culminate in permanent vision loss. The purpose of these studies was to evaluate the capacity of DMF to mitigate optic neuritis. The work presented combines studies of a mouse model of MS and a retrospective chart analysis of files of patients with RRMS treated at the MS Center of Excellence within the Oklahoma Medical Research Foundation. Experimental autoimmune encephalomyelitis (EAE) is a well-established mouse model that recapitulates cardinal features of somatic and visual MS pathologies. EAE was induced in female C57BL/6J mice by inoculation with myelin oligodendrocyte glycoprotein peptide (residues 35-55; MOG35-55). DMF or vehicle was administered twice a day by oral gavage. Visual acuity was measured longitudinally with optokinetic tracking. Post-mortem analyses included quantification of RGCs in retinal flatmounts and quantitative PCR (qPCR) of Nrf2 target genes and regulators of myelin. Retrospective chart analyses were performed using data obtained from deidentified files of patients with RRMS. In the EAE mouse studies, DMF decreased optic neuritis severity, preserved vision and RGCs, and concomitantly reduced motor deficits when administered by two different treatment regimens (prevention or interventional). DMF was more efficacious when administered as an interventional therapy, and the beneficial effects occurred independently of the induction of Nrf2 target genes. A complementary retrospective chart analysis demonstrated that DMF increased the time to a recurrence of optic neuritis, and protected against subsequent bouts of optic neuritis. This work underscores the potential of DMF to mitigate the severity and recurrence of optic neuritis episodes in patients with RRMS.


The role of phosphoinositide 3-kinase C2alpha in insulin signaling.
The members of the class II phosphoinositide 3-kinase (PI3K) family can be activated by several stimuli, indicating that these enzymes can regulate many intracellular processes. Nevertheless, to date, there has been no definitive identification of their in vivo product, their mechanism(s) of activation, or their precise intracellular roles. By metabolic labeling, we here identify phosphatidylinositol 3-phosphate as the sole in vivo product of the insulin-dependent activation of PI3K-C2alpha, confirming the emerging role of such a phosphoinositide in signaling. We demonstrate that activation of PI3K-C2alpha involves its recruitment to the plasma membrane and that activation is mediated by the GTPase TC10. This is the first report showing a membrane targeting-mediated mechanism of activation for PI3K-C2alpha and that a small GTP-binding protein can activate a class II PI3K isoform. We also demonstrate that PI3K-C2alpha contributes to maximal insulin-induced translocation of the glucose transporter GLUT4 to the plasma membrane and subsequent glucose uptake, definitely assessing the role of this enzyme in insulin signaling.


Hyperglycemia induces epithelial-mesenchymal transition in the lungs of experimental diabetes mellitus.
Diabetes mellitus (DM) reduces lung function and increases the risk of asthma, chronic obstructive pulmonary disease, pneumonia, and pulmonary fibrosis. Epithelial-mesenchymal transition (EMT) plays a crucial role in the development of pulmonary fibrosis. The pathogenesis of pulmonary fibrosis in diabetes remains unknown. We investigated the effects of hyperglycemia on EMT in the lungs of gerbils with streptozotocin (STZ)-induced diabetes. Diabetic gerbils exhibited a significantly lower volume fraction of the alveolar airspace and significantly higher septal thickness, volume fraction of the alveolar wall, and lung injury scores than did nondiabetic gerbils. The percentage of 8-hydroxy-2'-deoxyguanosine-positive cells and transforming growth factor-β-positive cells was significantly higher, the expression of E-cadherin was significantly lower, and the expression of N-cadherin was significantly higher in diabetic gerbils than in nondiabetic gerbils. These EMT characteristics were associated with a significant increase in α-smooth muscle actin (SMA) expression and collagen deposition in the lungs of diabetic gerbils. The increased α-SMA expression was co-localized with surfactant protein-C in alveolar type II cells in hyperglycemic animals. In conclusion, our study demonstrates that hyperglycemia induces EMT and contributes to lung fibrosis in an experimental DM model.


Effects of orthotic intervention and nine holes of simulated golf on club-head velocity in experienced golfers.
This study was an initial investigation evaluating the effects of orthotic intervention on club-head velocity (CHV) among a group of experienced golfers before and after 9 holes of simulated golf. Northwestern College of Chiropractic, Bloomington, Minnesota. Twelve experienced golfers were included in the study. CHV was measured with a device used by many Professional Golf Association and Ladies Professional Golf Association teaching professionals before and after wearing orthotics and before and after completing 9 holes of simulated golf. Subjects wore custom-made, flexible orthotics daily for 6 weeks and then were retested with the same objective measurement parameters. CHV (swing speed in miles per hour) was measured in all subjects before and after wearing custom-fit, flexible orthotics for 6 weeks and before and after completing 9 holes of simulated golf. There was an approximate increase in CHV of between 3 and 5 mph, or a relative increase in CHV by up to 7%, after subjects had worn custom-made, weight-bearing, flexible orthotics daily for 6 weeks. A 5-mph increase in CHV is equivalent to an approximate increase in golf ball travel distance of 15 yards, a significant increase for the tour player for whom small increases in performance can reflect large position changes on the roster board. In addition, the use of these custom of those eliminated the effects of fatigue associated with playing 9 holes of golf (relative to CHV) and therefore may improve the likelihood for more consistent golf performance. The use of the custom-fit, flexible orthotics in this study had a positive influence on CHV in experienced golfers.


A classification for the interferon-tau.
An attempt has been made to provide a rational organization for the many interferon-tau (IFN-tau) sequences entered in GenBank based on phylogenetic analysis and common amino acid substitutions, which might form the basis for a universal nomenclature scheme. Over the 13 years since these genes were first discovered, large numbers of cDNA and gene sequences have been reported, and there is reason to suspect that representatives of all the major ovine and bovine forms have now been described. The data are consistent with the presence of many genes and also allelic variants in sheep and cattle analogous to what has been observed for the IFN-alpha in the human. Future variants should be easily accommodated into the scheme outlined here. A flexible system of nomenclature, based on that used for HuIFN, is needed to provide a common base for comparison between research done in different laboratories and to assign relative biologic potencies to these molecules.


Individual monitoring for internal exposure in Europe and the integration of dosimetric data.
The European Radiation Dosimetry Group, EURADOS, established a working group consisting of experts whose aim is to assist in the process of harmonisation of individual monitoring as part of the protection of occupationally exposed workers. A catalogue of facilities and internal dosimetric techniques related to individual monitoring in Europe has been completed as a result of this EURADOS study. A questionnaire was sent in 2002 to services requesting information on various topics including type of exposures, techniques used for direct and indirect measurements including calibration and sensitivity data and the methods employed for the assessment of internal doses. Information relating to Quality Control procedures for direct and indirect measurements, Quality Assurance Programmes in the facilities and legal requirements for "approved dosimetric services" were also considered. A total of 71 completed questionnaires were returned by internal dosimetry facilities in 26 countries. This results in an overview of the actual status of the processes used in internal exposure estimation in Europe. In many ways harmonisation is a reality in internal dose assessments, especially when taking into account the measurements of the activity retained or excreted from the body. However, a future study detailing the estimation of minimum detectable activity in the laboratories is highly recommended. Points to focus on in future harmonisation activities are as follows: the process of calculation of doses from measured activity, establishment of guidelines, similar dosimetric tools and application of the same ICRP recommendations. This would lead to a better and more harmonised approach to the estimation of internal exposures in all European facilities.


Competing for consciousness: prolonged mask exposure reduces object substitution masking.
In object substitution masking (OSM) a sparse, temporally trailing 4-dot mask impairs target identification, even though it has different contours from, and does not spatially overlap with the target. Here, we demonstrate a previously unknown characteristic of OSM: Observers show reduced masking at prolonged (e.g., 640 ms) relative to intermediate mask durations (e.g., 240 ms). We propose that with prolonged exposure, the mask's visual representation is consolidated, which allows processing of the lingering target icon to be reinitiated, thereby improving performance. Our findings suggest that when the visual system is confronted with 2 temporally contiguous stimuli, although one may initially gain access to consciousness above the other, the "losing" stimulus is not irreversibly lost to awareness.


Genetic analysis of the beta-lactamases of Mycobacterium tuberculosis and Mycobacterium smegmatis and susceptibility to beta-lactam antibiotics.
Mycobacteria produce beta-lactamases and are intrinsically resistant to beta-lactam antibiotics. In addition to the beta-lactamases, cell envelope permeability and variations in certain peptidoglycan biosynthetic enzymes are believed to contribute to beta-lactam resistance in these organisms. To allow the study of these additional mechanisms, mutants of the major beta-lactamases, BlaC and BlaS, were generated in the pathogenic Mycobacterium tuberculosis strain H37Rv and the model organism Mycobacterium smegmatis strain PM274. The mutants M. tuberculosis PM638 (DeltablaC1) and M. smegmatis PM759 (DeltablaS1) showed an increase in susceptibility to beta-lactam antibiotics, as determined by disc diffusion and minimal inhibitory concentration (MIC) assays. The susceptibility of the mutants, as assayed by disc diffusion tests, to penicillin-type beta-lactam antibiotics was affected most, compared to the cephalosporin-type beta-lactam antibiotics. The M. tuberculosis mutant had no detectable beta-lactamase activity, while the M. smegmatis mutant had a residual type 1 beta-lactamase activity. We identified a gene, blaE, encoding a putative cephalosporinase in M. smegmatis. A double beta-lactamase mutant of M. smegmatis, PM976 (DeltablaS1DeltablaE : : res), had no detectable beta-lactamase activity, but its susceptibility to beta-lactam antibiotics was not significantly different from that of the DeltablaS1 parental strain, PM759. The mutants generated in this study will help determine the contribution of other beta-lactam resistance mechanisms in addition to serving as tools to study the biology of peptidoglycan biosynthesis in these organisms.


Daily regeneration of rat adrenocortical cells: circadian and zonal variations in cytogenesis.
Daily regeneration of rat adrenocortical cells were investigated in terms of circadian and zonal variations by following the cells at the DNA-synthesizing stage. An S-phase was assessed by 5-bromo-2'-deoxyuridine (BrdU) incorporation into the cell-nuclei and/or by visualizing proliferating cell nuclear antigen. The BrdU-positive cells were observed throughout the day mainly in two regions of the adrenal cortex, i.e. the innermost portion of the zona glomerulosa and the outermost portion of the zona fasciculata. Cells only in a latter region showed a distinct circadian rhythm of cell proliferation with a peak at 3-4 a.m. A remarkable rise in the plasma adrenocorticotropin (ACTH) concentration preceded such an increase in the cell proliferation by about 4 hours. This phenomenon could be mimicked by raising the plasma ACTH concentration by the administration of Cortrosyn Z or metyrapone. Angiotensin II-stimuli induced by Na-deficiency increased the proliferation of zona glomerulosa cells in the former region at 6-7p.m without significant effects on that of the zona fasciculata cells in the latter region. Thus at least two sites, which respond differentially to the day/night cycle and circulating hormone levels, exist in rat adrenal cortex being responsible for the cytogenesis in this endocrine organ.


[Initial washout time of inhaled anesthetic agents].
Recovery of inhalation anesthesia is generally considered the inverse of induction. While other factors remain unchanged, the rate of elimination of an inhaled anesthetic agent with a lower solubility such as nitrous oxide (N2O) is faster than one with a high solubility. Therefore, at the end of surgery it is a common practice to discontinue the volatile agent prior to discontinuing N2O. However, the results of our routine monitoring of end-tidal concentrations of anesthetics and physiological gases by employing a mass spectrometer lead us to question the practice. We studied 24 non-premedicated ASA I/II patients requiring endotracheal intubation with an anesthesia time over 2 h. All patients were mechanically ventilated at an appropriate tidal volume and respiratory rate to maintain an ETCO2 of 35-38 mmHg with an anesthetic mixture of 40% O2 and 60% N2O and isoflurane (Forane). At the end of surgery, both N2O and isoflurane were discontinued simultaneously and total oxygen flow was increased to 6 or 10 l/min by random. The exhaled partial pressure of both inhaled anesthetic agents were assessed at the time of anesthetic termination, after 1, 2 min, and then every 2 min up to 20 min or at the time of extubation. The results obtained were plotted against real time using two different scales. Despite the marked difference in their plasma solubilities, these two inhaled anesthetic agents reached to low exhaled partial pressure that had no clinical significance about ten minutes after discontinuation.(ABSTRACT TRUNCATED AT 250 WORDS)


Magnetic properties of the RbMnPO4 zeolite-ABW-type material: a frustrated zigzag spin chain.
The crystal structure and magnetic properties of the RbMnPO4 zeolite-ABW-type material have been studied by temperature-dependent neutron powder diffraction, low-temperature magnetometry, and heat capacity measurements. RbMnPO4 represents a rare example of a weak ferromagnetic polar material, containing Mn(2+) ions with TN = 4.7 K. The neutron powder diffraction pattern recorded at T = 10 K shows that the compound crystallizes in the chiral and polar monoclinic space group P2(1) (No. 4) with the unit cell parameters: a = 8.94635(9), b = 5.43415(5), and c = 9.10250(8) Å and β = 90.4209(6)°. A close inspection of the crystal structure of RbMnPO4 shows that this material presents two different types of zigzag chains running along the b axis. This is a unique feature among the zeolite-ABW-type materials exhibiting the P2(1) symmetry. At low temperature, RbMnPO4 exhibits a canted antiferromagnetic structure characterized by the propagation vector k1 = 0, resulting in the magnetic symmetry P2(1)'. The magnetic moments lie mostly along the b axis with the ferromagnetic component being in the ac plane. Due to the geometrical frustration present in this system, an intermediate phase appears within the temperature range 4.7-5.1 K characterized by the propagation vector k2 = (kx, 0, kz) with kx/kz ≈ 2. This ratio is reminiscent of the multiferroic phase of the orthorhombic RMnO3 phases (R = rare earth), suggesting that RbMnPO4 could present some multiferroic properties at low temperature. Our density functional calculations confirm the presence of magnetic frustration, which explains this intermediate incommensurate phase. Taking into account the strongest magnetic interactions, we are able to reproduce the magnetic structure observed experimentally at low temperature.


A Loss-of-Function HCN4 Mutation Associated With Familial Benign Myoclonic Epilepsy in Infancy Causes Increased Neuronal Excitability.
HCN channels are highly expressed and functionally relevant in neurons and increasing evidence demonstrates their involvement in the etiology of human epilepsies. Among HCN isoforms, HCN4 is important in cardiac tissue, where it underlies pacemaker activity. Despite being expressed also in deep structures of the brain, mutations of this channel functionally shown to be associated with epilepsy have not been reported yet. Using Next Generation Sequencing for the screening of patients with idiopathic epilepsy, we identified the p.Arg550Cys (c.1648C>T) heterozygous mutation on HCN4 in two brothers affected by benign myoclonic epilepsy of infancy. Functional characterization in heterologous expression system and in neurons showed that the mutation determines a loss of function of HCN4 contribution to activity and an increase of neuronal discharge, potentially predisposing to epilepsy. Expressed in cardiomyocytes, mutant channels activate at slightly more negative voltages than wild-type (WT), in accordance with borderline bradycardia. While HCN4 variants have been frequently associated with cardiac arrhythmias, these data represent the first experimental evidence that functional alteration of HCN4 can also be involved in human epilepsy through a loss-of-function effect and associated increased neuronal excitability. Since HCN4 appears to be highly expressed in deep brain structures only early during development, our data provide a potential explanation for a link between dysfunctional HCN4 and infantile epilepsy. These findings suggest that it may be useful to include HCN4 screening to extend the knowledge of the genetic causes of infantile epilepsies, potentially paving the way for the identification of innovative therapeutic strategies.


Clostridium difficile infection (CDI) severity and outcome among patients infected with the NAP1/BI/027 strain in a non-epidemic setting.
Determine whether the NAP1 strain identified by polymerase chain reaction (PCR)-based stool assay is correlated with CDI severity and clinical outcomes. Medical records of adult patients with positive stool Xpert® Clostridium difficile PCR assay for an initial episode of CDI between January 2012 and January 2013 at a tertiary care hospital in Chicago were reviewed. Two patients diagnosed with CDI caused by a non-NAP1 strain (positive Xpert® C. difficile assay but negative Xpert® C. difficile Epi assay) were included for each patient diagnosed with CDI caused by a NAP1 strain (positive Epi assay). Patient charts were reviewed for markers of severity, risk factors, treatment regimens, and outcomes. Of 494 stool specimens, 90 (18%) that were positive for C. difficile by PCR were positive for NAP1 strain. In total, 37 patients with CDI due to NAP1 were matched with 74 patients with CDI due to non-NAP1 strains. Multivariable model revealed individuals ≥65 years old were 3 times more likely to have NAP1 strain than individuals <65 (P=.02). Residents of a nursing home prior to hospitalization were 10 times more likely to have NAP1 strain than patients residing in their homes (P=.001). More NAP1 cases had a change in treatment from metronidazole to oral vancomycin plus intravenous metronidazole (P=.01). The severity of CDI, incidence of mortality and recurrent CDI were similar between groups. In a nonepidemic setting, NAP1 strains were more common in older patients and individuals admitted from nursing homes. Identification of NAP1 by PCR of stool specimens was associated in a change of therapy but did not predict worse outcomes. Reporting strain results may not be clinically useful in routine settings.


Revisiting the polytopal rearrangements in penta-coordinate d7-metallocomplexes: modified Berry pseudorotation, octahedral switch, and butterfly isomerization.
This paper provides a first-principles theoretical investigation of the polytopal rearrangements and fluxional behavior of five-coordinate d7-transition metal complexes. Our work is primarily based on a potential energy surface analysis of the iron tetracarbonyl hydride radical HFe˙(CO)4. We demonstrate the existence of distorted coordination geometries in this prototypical system and, for the first time, introduce three general rearrangement mechanisms, which account for the non-ideal coordination. The first of these mechanisms constitutes a modified version of the Berry pseudorotation via a square-based pyramidal C4v transition state that connects two chemically identical edge-bridged tetrahedral stereoisomers of C2v symmetry. It differs from the classical Berry mechanism, which involves two regular D3h equilibrium structures and a C4v transition state. The second mechanism is related to the famous "tetrahedral jump" hypothesis, postulated by Muetterties for a number of d6 HML4 and H2ML4 complexes. Here, our study suggests two fluxional rearrangement pathways via distinct types of C2v transition states. Both pathways of this mechanism can be described as a single-ligand migration to a vacant position of an "octahedron", thus interchanging (switching) the apical and basal ligands of the initial quasi-square pyramidal isomer, which is considered as an idealized octahedron with a vacancy. Accordingly, we call this mechanism "octahedral switch". The third mechanism follows a butterfly-type isomerization featuring a key-angle deformation, and we thus call it "butterfly isomerization". It connects the quasi-square pyramidal and edge-bridged tetrahedral isomers of HFe˙(CO)4 through a distorted edge-bridged tetrahedral transition state of Cs symmetry. Our paper discusses the overall features of the isomers and rearrangement mechanisms as well as their implications. We rationalize the existence of each stationary point through an electronic structure analysis and argue their relevance for isolobal analogues of HFe˙(CO)4.


Monitoring sinew contraction during formation of tissue-engineered fibrin-based ligament constructs.
The ability to study the gross morphological changes occurring during tissue formation is vital to producing tissue-engineered structures of clinically relevant dimensions in vitro. Here, we have used nondestructive methods of digital imaging and optical coherence tomography to monitor the early-stage formation and subsequent maturation of fibrin-based tissue-engineered ligament constructs. In addition, the effect of supplementation with essential promoters of collagen synthesis, ascorbic acid (AA) and proline (P), has been assessed. Contraction of the cell-seeded fibrin gel occurs unevenly within the first 5 days of culture around two fixed anchor points before forming a longitudinal ligament-like construct. AA+P supplementation accelerates gel contraction in the maturation phase of development, producing ligament-like constructs with a higher collagen content and distinct morphology to that of unsupplemented constructs. These studies highlight the importance of being able to control the methods of tissue formation and maturation in vitro to enable the production of tissue-engineered constructs with suitable replacement tissue characteristics for repair of clinical soft-tissue injuries.


Early Predictors of Eating Problems in Preadolescence-A Prospective Birth Cohort Study.
The epidemiology of childhood eating problems is far from being fully described. The present study aims to explore early predictors of eating behavior problems in preadolescence. The study sample comprised 1,939 children from the birth cohort study, the Copenhagen Child Cohort (CCC2000). Logistic regression models were used to investigate associations among infancy health, developmental and relational factors, maternal mental health problems, socioeconomic factors, parental reported eating behavior patterns in preschool age and eating behavior problems in preadolescence. A number of factors expressing socioeconomic disadvantage across childhood were associated with an increased risk of eating behavior problems at age 11-12 years. In addition, overeating patterns at age 5-7 years predicted restrained eating in preadolescence (odds ratio [OR] = 2.77; 95% confidence interval [CI] = 1.13-6.77; p = .03), with overweight at age 11-12 years and low annual household income as strong explanatory factors (OR = 4.79; 95% CI = 2.81-8.17; p < .0001 and OR = 2.06; 95% CI = 1.19-3.58; p = .02, respectively). No significant associations between perinatal, early child- and relational factors, or maternal mental disorder and eating behavior problems in preadolescence were found. Our results suggest that overeating at age 5-7 years is prospectively associated with restrained eating in preadolescence, with contemporaneous socioeconomic disadvantages and overweight as strong explanatory factors. Our findings might reflect successful public health interventions toward childhood obesity or might reflect a developmental course of problematic eating fluctuating between over- and undereating. Future studies should focus on the possible pathways from overeating to restrained eating and more severe eating pathology, including possible negative side effects of otherwise successful interventions aimed at reducing childhood obesity.


Trial of retinol and isotretinoin in skin cancer prevention: a randomized, double-blind, controlled trial. Southwest Skin Cancer Prevention Study Group.
The objective of this study was to examine the effect of retinol and isotretinoin on the incidence of nonmelanoma skin cancer in high-risk subjects. A total of 525 participants with a history of at least four basal cell carcinomas (BCCs) and/or cutaneous squamous cell carcinomas (SCCs) were entered into a randomized, double-blind, placebo-controlled trial, performed in free-standing study clinics. Participants were randomly assigned to receive oral retinol (25,000 units), isotretinoin (5-10 mg), or placebo supplementation daily for 3 years. The time to first new occurrence of BCC or cutaneous SCC was used as the outcome measure. During the study period, 319 BCCs and 125 cutaneous SCCs were diagnosed clinically and pathologically. There were no differences between those who received retinol, isotretinoin, or the placebo, with regard to the time to first occurrence or to the total number of tumors noted. No beneficial effects were noted with regard to the prevention of nonmelanoma skin cancer with either retinol or isotretinoin.


High levels of CD2 expression identify HIV-1 latently infected resting memory CD4+ T cells in virally suppressed subjects.
Resting memory CD4(+) T cells are the largest reservoir of persistent infection in HIV-1-positive subjects. They harbor dormant, stably integrated virus despite suppressive antiretroviral therapy, posing an obstacle to a cure. Surface markers that identify latently infected cells remain unknown. Microarray analyses comparing resting latently infected and uninfected CD4(+) T cells generated in vitro showed profound differences in the expression of gene programs related to transcriptional and posttranscriptional regulation, cell proliferation, survival, cycle progression, and basic metabolism, suggesting that multiple biochemical and metabolic blocks contribute to preventing viral production in latently infected cells. We identified 33 transcripts encoding cell surface markers that are differentially expressed between latently infected and uninfected cells. Quantitative reverse transcriptase PCR (RT-QPCR) and flow cytometry analyses confirmed that the surface marker CD2 was expressed at higher levels on latently infected cells. To validate this result in vivo, we sorted resting memory CD4(+) T cells expressing high and low surface levels of CD2 from six HIV-1-infected subjects successfully treated with antiretroviral drugs for at least 3 years. Resting memory CD4(+) CD2(high) T cells from all subjects harbored higher HIV-1 DNA copy numbers than all other CD4(+) T cell subsets. Moreover, after ex vivo viral reactivation, robust viral RNA production was detected only from resting memory CD4(+) CD2(high) T cells but not from other cell subsets. Altogether, these results show that a high CD2 expression level is a hallmark of latently infected resting memory CD4(+) T cells in vivo.


Racial and ethnic variation in presentation and outcomes of high-grade soft tissue sarcoma at a Southeastern United States comprehensive cancer center.
High-grade soft tissue sarcoma is rare and associated with poor prognosis. This study examines racial and ethnic variation in presentation and outcomes at a Southeastern US cancer center. Among an institutional cohort of patients seen between January 2016-December 2021, racial and ethnic differences were evaluated using chi-squared tests, Kaplan Meier curves, and Cox proportional hazards models. There were 295 patients (71 % Non-Hispanic White, 24 % Black, 3 % Hispanic White, 2 % Other). Black representation was greater than national cohorts (24 % vs. 12 %). Histological subtype varied by race/ethnicity (p = 0.007). Adjusting for histology and stage, survival was worse for Black vs. White patients (HR 1.71, 95 % CI 1.07-2.76) and those with metastatic disease (5.47, 3.54-8.44). In non-metastatic patients, survival differences for Black vs. White patients were attenuated by receipt of multi-modal treatment (1.53, 0.82-2.88). Observed racial disparities in survival of high-grade sarcoma may be addressed by early, multidisciplinary management.


Protection against lethal challenge of BALB/c mice by passive transfer of monoclonal antibodies to five glycoproteins of herpes simplex virus type 2.
Monoclonal antibodies secreted by six hybridomas and recognizing antigenic sites on glycoproteins gC, gAB, gD, gE, and gF of herpes simplex virus type 2 were examined for their ability to protect BALB/c mice from lethal infection by the virus. Administration of monoclonal antibodies to individual glycoproteins intraperitoneally 3 h before footpad challenge with 10 times the 50% lethal dose of virus protected between 35 and 75% of the mice, except for one of two monoclonal antibodies recognizing antigens on gC. The antibodies did not neutralize virus in vitro and protected A/J mice deficient in the fifth component of complement as efficiently as complement-sufficient BALB/c mice. A good correlation was found between protection and titers of monoclonal antibodies assessed by antibody-dependent cell-mediated cytolysis. The results indicate that any of the glycoproteins can serve as antigens for a protective immune response. In addition, the data are compatible with protection being mediated by an antibody-dependent cell-mediated cytolysis mechanism.


Salivary duct carcinoma of the parotid gland.
Salivary duct carcinoma of the parotid gland is an uncommon tumor, highly aggressive. About 200 cases have been reported in the English literature. Pathomorphologically, these tumors showed great similarities to ductal carcinoma of the female breast, which is why they described this tumor as "salivary duct carcinoma." The authors describe a new case of salivary duct carcinoma of the parotid gland. We present the case of a 50-year-old patient with progressive facial paralysis. The MRI examination of the head showed two ill-defined formations. A malignant tumor was strongly suspected, so that a total left parotidectomy with excision of the adjacent facial nerve and left lymph node dissection was performed. Microscopic examination concluded to a salivary duct carcinoma of the left parotid gland negative with Her2/neu antibody with lymph node metastasis. There were no recurrences or metastases within 3 years of follow-up. Salivary duct carcinoma of the parotid gland is a rare tumor with an aggressive behavior. This is due to its propensity to infiltrate distant organs. The diagnosis is based on microscopic examination. Treatment modalities are non-consensual, but some authors advocate the necessity of aggressive approach, especially in tumors negative with Heur2/neu antibody. This is due to the fact that the overexpression of this antigen was reported to be associated with a poor prognosis.


Suitability of multiple Mediterranean oak species as a food resource for Reticulitermes grassei Clément (Isoptera: Rhinotermitidae).
The subterranean termite Reticulitermes grassei Clément causes lesions in the trunk of Quercus suber L. by constructing feeding galleries, but no information is available regarding other Quercus species from the Mediterranean region. This work aimed to study the suitability of the other main oak species of Mediterranean forests as a food resource for R. grassei. Two experiments, choice and non-choice feeding, were conducted lasting for 15, 30, and 45 days each. In the non-choice experiment, termites were offered one of the following food types: Quercus suber, Quercus ilex L., Quercus faginea Lam, cork or Pinus pinea L., which was considered the control. The choice feeding experiment used all the same food types listed above, supplied simultaneously in the same container. Food selection was examined by analysing the relationships over time between surviving termites and food consumption. The results indicated that R. grassei could be considered a generalist species, as it consumed the cork and wood of all oak species, as well as displaying a clear preference for soft wood (pine). Correlation analysis indicated that consumption was not dependent on wood density. Survival of R. grassei was influenced by the time of exposure to different oak species, but a high survival rate was maintained over time in the pine treatment (upper 70% in the three experiments). Given these results, it can be concluded that all the oak species are a suitable food source for R. grassei.


Co-combustion of coal and sewage sludge: chemical and ecotoxicological properties of ashes.
The co-combustion of sewage sludge (SS) and coal is widely used for the treatment and thermal valorization of SS produced in wastewater treatment plants. The chemical and ecotoxicological properties of the ashes produced in this thermal treatment have not been fully studied. Two combustion tests were performed in a fluidized bed combustor. Colombian coal was used as fuel in test A. A blend (1+1) of this coal and a stabilized SS (Biogran) was used in a second test B. Samples of the bottom and fly ashes trapped in two sequential cyclones were collected. The characterization of the ashes was focused on two main aspects: (1) the bulk content of a set of metals and (2) the characterization of eluates produced according to the European Standard leaching test EN 12457-2. The eluates were submitted to an ecotoxicological characterization for two bio-indicators. In what concerns the bulk content of ashes, both combustion tests have produced ashes with different compositions. The ashes formed during the co-combustion test have shown higher concentrations of metals, namely Cr, Cu, Ni, Pb, Zn and Fe for all ashes. The leaching test has shown low mobility of these elements from the by-products produced during the combustion and co-combustion tests. Cr and Cr(VI) were mainly detected in the eluates of the 1st cyclone ashes produced in both combustion tests and in the 2nd cyclone ashes produced in the co-combustion test. Considering the ecotoxicity assays, the eluates of bottom and fly ashes for both combustion and co-combustion tests have shown low ecotoxic levels. The micro-crustacean Daphnia magna was generally more sensitive than the bacterium Vibrio fischeri. CEMWE criterion has allowed to classify the bottom ashes for both combustion and co-combustion tests as non-toxic residues and the fly ashes collected in both cyclones as toxic.


Inhibition by flavonoids of amyloid-like fibril formation by Plasmodium falciparum merozoite surface protein 2.
Merozoite surface protein 2 (MSP2) is a glycosylphosphatidylinositol (GPI)-anchored protein expressed abundantly on the surface of Plasmodium falciparum merozoites. The results of a phase 2 trial in Papua New Guinean children showed MSP2 to be a promising malaria vaccine candidate. MSP2 is intrinsically unstructured and forms amyloid-like fibrils under physiological conditions. Oligomers containing beta-strand interactions similar to those in amyloid fibrils may be a component of the fibrillar surface coat on P. falciparum merozoites. As the propensity of MSP2 to form fibrils in solution also has the potential to impede its development as a vaccine candidate, finding an inhibitor that specifically inhibits fibrillogenesis may enhance vaccine development. In this study, we tested the ability of three flavonoids, EGCG, baicalein, and resveratrol, to inhibit MSP2 fibrillogenesis and found marked inhibition with EGCG but not with the other two flavonoids. The inhibitory effect and the interactions of the flavonoids with MSP2 were characterized using NMR spectroscopy, thioflavin T fluorescence assays, electron microscopy, and other biophysical methods. EGCG stabilizes soluble oligomers and blocks fibrillogenesis by preventing the conformational transition of MSP2 from a random coil to an amyloidogenic beta-sheet structure. Structural comparison of the three flavonoids indicates an association between their propensity for autoxidation and their fibril inhibitory activity; the activity of EGCG can be attributed to the vicinal hydroxyl groups present in this flavonoid and their ability to form quinones. The molecular mechanism of fibril inhibition by EGCG appears to be complex and involves noncovalent binding followed by covalent modification of the protein. Although the addition of EGCG appears to be an effective means of stabilizing MSP2 in solution, the covalent modification of MSP2 would most likely not be acceptable in a vaccine formulation. However, these small molecule inhibitors of MSP2 fibril formation will be useful as mechanistic probes in studying oligomerization and fibril assembly of MSP2.


Quantitation of eumelanin and pheomelanin markers in diverse biological samples by HPLC-UV-MS following solid-phase extraction.
Eumelanin and pheomelanin are well known and common pigments found in nature. However, their complex polymer structure and high thermostability complicate their direct chemical identification. A widely used analytical method is indirect determination using HPLC with UV detection of both types of melanin by their most abundant oxidation products: pyrrole-2,3-dicarboxylic acid (PDCA), pyrrole-2,3,5-tricarboxylic acid (PTCA), thiazole-4,5-dicarboxylic acid (TDCA), and thiazole-2,4,5-tricarboxylic acid (TTCA). An increasing interest in pigmentation in biological research led us to develop a highly sensitive and selective method to identify and quantify these melanin markers in diverse biological samples with complex matrices. By introducing solid-phase extraction (SPE, reversed-phase) following alkaline oxidation we could significantly decrease background signals while maintaining recoveries greater than 70%. Our HPLC-UV-MS method allows for confident peak identification via exact mass information in corresponding UV signals used for quantitation. In addition to synthetic melanin and Sepia officinalis ink as reference compounds eumelanin markers were detected in brown human hair and a brown bivalve shell (Mytilus edulis). Brown feathers from the common chicken (Gallus g. domesticus) yielded all four eumelanin and pheomelanin markers. The present method can be easily adapted for a wide range of future studies on biological samples with unknown melanin content.


Colloid mobilization and arsenite transport in soil columns: effect of ionic strength.
Colloid generation and transport in soils is of significance because of suspected colloid-facilitated transport of contaminants to the groundwater. In this study, colloid mobilization and its effect on the transport of arsenite [As(III)] were investigated in Olivier (fine-silty, mixed, active, thermic Aquic Fraglossudalfs) and Windsor (mixed, mesic typic Udipsamments) soil columns. Input solution of 10 mg L(-1) As(III) in 0.01 M NaCl was applied to water-saturated columns, and followed by leaching with deionized water (DIW). Flow interruptions were performed during the As(III) input and DIW leaching phases. Turbidity, electrical conductivity (EC), and pH of column effluents were monitored with time. Total and dissolved concentrations of As, Fe, and Al were analyzed. Effluent results demonstrated that colloid-facilitated transport contributed little to arsenic movement when the solution ionic strength was maintained constant. Mobilization of colloidal amorphous material and enhanced transport of As(III) were observed as a result of changes in ionic strength of the input solution. The peak of colloid generation coincided with peak concentrations of Fe, suggesting mobilization of Fe oxides and facilitated transport of As(III) adsorbed on oxide surfaces. Colloid mobilization was enhanced due to flow interruption in the Olivier column, which suggests slow dissociation of aggregated colloidal particles. Moreover, effluent results indicate significant effect of organic matter in stabilizing aggregates of colloidal particles.


Potential Founder Variants in COL4A4 Identified in Bukharian Jews Linked to Autosomal Dominant and Autosomal Recessive Alport Syndrome.
Alport syndrome is a hereditary disorder caused by pathogenic variants in the COL4A gene, which can be inherited in an autosomal recessive, dominant, or X-linked pattern. In the Bukharian Jewish population, no founder pathogenic variant has been reported in COL4A4. The cohort included 38 patients from 22 Bukharian Jewish families with suspected Alport syndrome who were referred the nephrogenetics clinic between 2012 and 2022. The study collected demographic, clinical, and genetic data from electronic medical records, which were used to evaluate the molecular basis of the disease using Sanger sequencing, and next-generation sequencing. Molecular diagnosis was confirmed in 20/38 patients, with each patient having at least one of the three disease-causing COL4A4 variants detected: c.338G<A (p.Gly113Asp), c.3022G>A (p.Gly1008Arg), and c.871-6T>C. In addition, two patients were obligate carriers. Overall, there were 17 heterozygotes, 2 compound heterozygotes, and 3 homozygotes. Each variant was detected in more than one unrelated family. All patients had hematuria with/without proteinuria at referral, and the youngest patient with proteinuria (age 5 years) was homozygous for the c.338G>A variant. End-stage renal disease was diagnosed in two patients at the age of 38 years, a compound heterozygote for c.338G>A and c.871-6T>C. Hearing deterioration was detected in three patients, the youngest aged 40 years, all of whom were heterozygous for c.338G>A. This study unveils three novel disease-causing variants, c.3022G>A, c.871-6T>C, and c.338G>A, in the COL4A4 gene that are recurrent among Jews of Bukharian ancestry, and cause Alport syndrome in both dominant and recessive autosomal inheritance patterns.


Role of AcrR and ramA in fluoroquinolone resistance in clinical Klebsiella pneumoniae isolates from Singapore.
The MICs of ciprofloxacin for 33 clinical isolates of K. pneumoniae resistant to extended-spectrum cephalosporins from three hospitals in Singapore ranged from 0.25 to >128 microg/ml. Nineteen of the isolates were fluoroquinolone resistant according to the NCCLS guidelines. Strains for which the ciprofloxacin MIC was >or=0.5 microg/ml harbored a mutation in DNA gyrase A (Ser83-->Tyr, Leu, or IIe), and some had a secondary Asp87-->Asn mutation. Isolates for which the MIC was 16 microg/ml possessed an additional alteration in ParC (Ser80-->IIe, Trp, or Arg). Tolerance of the organic solvent cyclohexane was observed in 10 of the 19 fluoroquinolone-resistant strains; 3 of these were also pentane tolerant. Five of the 10 organic solvent-tolerant isolates overexpressed AcrA and also showed deletions within the acrR gene. Complementation of the mutated acrR gene with the wild-type gene decreased AcrA levels and produced a two- to fourfold reduction in the fluoroquinolone MICs. None of the organic solvent-tolerant clinical isolates overexpressed another efflux-related gene, acrE. While marA and soxS were not overexpressed, another marA homologue, ramA, was overexpressed in 3 of 10 organic solvent-tolerant isolates. These findings indicate that multiple target and nontarget gene changes contribute to fluoroquinolone resistance in K. pneumoniae. Besides AcrR mutations, ramA overexpression (but not marA or soxS overexpression) was related to increased AcrAB efflux pump expression in this collection of isolates.


Repopulation with IgA-containing cells of bronchial and intestinal lamina propria after transfer of homologous Peyer's patch and bronchial lymphocytes.
Transfer of 50 million rabbit allogeneic lymphocytes from either bronchus-associated lymphoid tissue (BALT) or Peyer's patches into 1000 R x-irradiated recipients results, 6 days later, in predominant repopulation of gut and bronchial lamina propria, as well as spleen with IgA-containing cells. After repopulation with BALT or Peyer's patch cells, lymphoid follicles in both gut and lung showed peripheral cellular membrane type of fluorescence with fluorescein-conjugated anti-IgA antisera only. Six days after x-irradiation alone, little evidence of repopulation was seen and immunofluorescent qualitative observations of gut and lung, and quantitative data in the spleen, confirmed these findings. After transfer of 50 million lymph node cells, very few immunoglobulin-containing cells were seen in the gut or bronchial lamina propria. These results suggest that there may be a common mucosal immunologic system, and that repopulation of gut and lung lamina propria may be through the organized lymphoid tissue therein.


ORF6 from the clavulanic acid gene cluster of Streptomyces clavuligerus has ornithine acetyltransferase activity.
The clinically used beta-lactamase inhibitor clavulanic acid is produced by fermentation of Streptomyces clavuligerus. The orf6 gene of the clavulanic acid biosynthetic gene cluster in S. clavuligerus encodes a protein that shows sequence homology to ornithine acetyltransferase (OAT), the fifth enzyme of the arginine biosynthetic pathway. Orf6 was overexpressed in Escherichia coli (at approximately 15% of total soluble protein by SDS/PAGE analysis) indicating it was not toxic to the host cells. The recombinant protein was purified (to > 95% purity) by a one-step technique. Like other OATs it was synthesized as a precursor protein which underwent autocatalytic internal cleavage in E. coli to generate alpha and beta subunits. Cleavage was shown to occur between the alanine and threonine residues in a KGXGMXXPX--(M/L)AT (M/L)L motif conserved within all identified OAT sequences. Gel filtration and native electrophoresis analyses implied that the ORF6 protein was an alpha2beta2 heterotetramer and direct evidence for this came from mass spectrometric analyses. Although anomalous migration of the beta subunit was observed by standard SDS/PAGE analysis, which indicated the presence of two bands (as previously observed for other OATs), mass spectrometric analyses did not reveal any evidence for post-translational modification of the beta subunit. Extended denaturation with SDS before PAGE resulted in observation of a single major beta subunit band. Purified ORF6 was able to catalyse the reversible transfer of an acetyl group from N-acetylornithine to glutamate, but not the formation of N-acetylglutamate from glutamate and acetyl-coenzyme A, nor (detectably) the hydrolysis of N-acetylornithine. Mass spectrometry also revealed the reaction proceeds via acetylation of the beta subunit.


HuR's post-transcriptional regulation of Death Receptor 5 in pancreatic cancer cells.
Apoptosis is one of the core signaling pathways disrupted in pancreatic ductal adenocarcinoma (PDA). Death receptor 5 (DR5) is a member of the tumor necrosis factor (TNF)-receptor superfamily that is expressed in cancer cells. Binding of TNF-related apoptosis-inducing ligand (TRAIL) to DR5 is a potent trigger of the extrinsic apoptotic pathway, and numerous clinical trials are based on DR5-targeted therapies for cancer, including PDA. Human antigen R (HuR), an RNA-binding protein, regulates a select number of transcripts under stress conditions. Here we report that HuR translocates from the nucleus to the cytoplasm of PDA cells upon treatment with a DR5 agonist. High doses of DR5 agonist induce cleavage of both HuR and caspase 8. HuR binds to DR5 mRNA at the 5'-untranslated region (UTR) in PDA cells in response to different cancer-associated stressors and subsequently represses DR5 protein expression; silencing HuR augments DR5 protein production by enabling its translation and thus enhances apoptosis. In PDA specimens (n = 53), negative HuR cytoplasmic expression correlated with elevated DR5 expression (odds ratio 16.1, p < 0.0001). Together, these data demonstrate a feedback mechanism elicited by HuR-mediated repression of the key apoptotic membrane protein DR5.


Structure of a double-domain phosphagen kinase reveals an asymmetric arrangement of the tandem domains.
Tandem duplications and fusions of single genes have led to magnificent expansions in the divergence of protein structures and functions over evolutionary timescales. One of the possible results is polydomain enzymes with interdomain cooperativities, few examples of which have been structurally characterized at the full-length level to explore their innate synergistic mechanisms. This work reports the crystal structures of a double-domain phosphagen kinase in both apo and ligand-bound states, revealing a novel asymmetric L-shaped arrangement of the two domains. Unexpectedly, the interdomain connections are not based on a flexible hinge linker but on a rigid secondary-structure element: a long α-helix that tethers the tandem domains in relatively fixed positions. Besides the connective helix, the two domains also contact each other directly and form an interdomain interface in which hydrogen bonds and hydrophobic interactions further stabilize the L-shaped domain arrangement. Molecular-dynamics simulations show that the interface is generally stable, suggesting that the asymmetric domain arrangement crystallographically observed in the present study is not a conformational state simply restrained by crystal-packing forces. It is possible that the asymmetrically arranged tandem domains could provide a structural basis for further studies of the interdomain synergy.


Ferric citrate controls phosphorus and delivers iron in patients on dialysis.
Patients on dialysis require phosphorus binders to prevent hyperphosphatemia and are iron deficient. We studied ferric citrate as a phosphorus binder and iron source. In this sequential, randomized trial, 441 subjects on dialysis were randomized to ferric citrate or active control in a 52-week active control period followed by a 4-week placebo control period, in which subjects on ferric citrate who completed the active control period were rerandomized to ferric citrate or placebo. The primary analysis compared the mean change in phosphorus between ferric citrate and placebo during the placebo control period. A sequential gatekeeping strategy controlled study-wise type 1 error for serum ferritin, transferrin saturation, and intravenous iron and erythropoietin-stimulating agent usage as prespecified secondary outcomes in the active control period. Ferric citrate controlled phosphorus compared with placebo, with a mean treatment difference of -2.2±0.2 mg/dl (mean±SEM) (P<0.001). Active control period phosphorus was similar between ferric citrate and active control, with comparable safety profiles. Subjects on ferric citrate achieved higher mean iron parameters (ferritin=899±488 ng/ml [mean±SD]; transferrin saturation=39%±17%) versus subjects on active control (ferritin=628±367 ng/ml [mean±SD]; transferrin saturation=30%±12%; P<0.001 for both). Subjects on ferric citrate received less intravenous elemental iron (median=12.95 mg/wk ferric citrate; 26.88 mg/wk active control; P<0.001) and less erythropoietin-stimulating agent (median epoetin-equivalent units per week: 5306 units/wk ferric citrate; 6951 units/wk active control; P=0.04). Hemoglobin levels were statistically higher on ferric citrate. Thus, ferric citrate is an efficacious and safe phosphate binder that increases iron stores and reduces intravenous iron and erythropoietin-stimulating agent use while maintaining hemoglobin.


Shock treatment of corn stover.
Corn stover digestibility was enhanced via shock treatment. A slurry of lime-treated corn stover was placed in a partially filled closed vessel. From the ullage space, either a shotgun shell was fired into the slurry, or a gas mixture was detonated. Various conditions were tested (i.e., pressures, depth, solids concentrations, gas mixtures). A high pressurization rate (108,000 MPa/s shotgun shells; 4,160,000 MPa/s hydrogen/oxygen detonation) was the only parameter that improved enzymatic digestibility. Stoichiometric propane/air deflagration had a low pressurization rate (37.2 MPa/s) and did not enhance enzymatic digestibility. Without shock, enzymatic conversion of lime-treated corn stover was 0.80 g glucan digested/g glucan fed with an enzyme loading of 46.7 mg protein/g glucan. With shock, the enzyme loading was reduced by ∼2× while maintaining the same conversion. Detonations are extraordinarily fast; rapidly cycling three small vessels (0.575 m3 each) every 7.5 s enables commercially relevant shock treatment (2,000 tone/day). © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:815-823, 2017.


Flexor reflex control of the external sphincter of the urethra in paraplegia.
In paraplegics and quadriplegics a profound paralysis of skeletal muscles occurs below the level of the spinal lesion. Unexplained in this state is the development of an overactive external urethral sphincter, which interferes with emptying of the bladder and may lead to infection of the urinary tract. Studies of cats show that the discharge of motoneurons causing this contraction has all the characteristics of a flexor reflex.


Role of TLR4 in the gut-brain axis in Parkinson's disease: a translational study from men to mice.
Recent evidence suggesting an important role of gut-derived inflammation in brain disorders has opened up new directions to explore the possible role of the gut-brain axis in neurodegenerative diseases. Given the prominence of dysbiosis and colonic dysfunction in patients with Parkinson's disease (PD), we propose that toll-like receptor 4 (TLR4)-mediated intestinal dysfunction could contribute to intestinal and central inflammation in PD-related neurodegeneration. To test this hypothesis we performed studies in both human tissue and a murine model of PD. Inflammation, immune activation and microbiota composition were measured in colonic samples from subjects with PD and healthy controls subjects and rotenone or vehicle-treated mice. To further assess the role of the TLR4 signalling in PD-induced neuroinflammation, we used TLR4-knockout (KO) mice in conjunction with oral rotenone administration to model PD. Patients with PD have intestinal barrier disruption, enhanced markers of microbial translocation and higher pro-inflammatory gene profiles in the colonic biopsy samples compared with controls. In this regard, we found increased expression of the bacterial endotoxin-specific ligand TLR4, CD3+ T cells, cytokine expression in colonic biopsies, dysbiosis characterised by a decrease abundance of SCFA-producing colonic bacteria in subjects with PD. Rotenone treatment in TLR4-KO mice revealed less intestinal inflammation, intestinal and motor dysfunction, neuroinflammation and neurodegeneration, relative to rotenone-treated wild-type animals despite the presence of dysbiotic microbiota in TLR4-KO mice. Taken together, these studies suggest that TLR4-mediated inflammation plays an important role in intestinal and/or brain inflammation, which may be one of the key factors leading to neurodegeneration in PD.


The proliferation potential protein-related (P2P-R) gene with domains encoding heterogeneous nuclear ribonucleoprotein association and Rb1 binding shows repressed expression during terminal differentiation.
Terminal differentiation is associated with repression in the expression of the proliferation potential proteins (P2P) subset of heterogeneous nuclear ribonucleoprotein (hnRNP) proteins. We report here the cloning and characterization of a 5173-bp P2P-related (P2P-R) cDNA that contains a 4214-bp open reading frame. Probes to this cDNA detect a single 8-kb mRNA in multiple murine tissues and in proliferating 3T3T cells, but not in terminally differentiated 3T3T adipocytes. Evidence that this cDNA can encode peptides with domains for hnRNP association was established by showing that such peptides are recognized by two monoclonal antibodies known to detect core hnRNP proteins, and by showing that the C130 monoclonal antibody, produced against a cDNA-derived fusion protein, also selectively detects native P2P hnRNP proteins. In addition, P2P-R cDNA-derived fusion proteins bind single-stranded nucleic acids, and a P2P-R cDNA-derived antisense oligonucleotide selectively represses P2P expression. Because terminal differentiation is associated with modulation in Rb1 function, we assayed if products of this cDNA might interact with Rb1. Evidence that the P2P-R cDNA encodes a protein domain that binds Rb1 was established using a glutathione S-transferase fusion protein to selectively precipitate Rb1 from cellular extracts. Data also show that this binding is reduced by competition with the adenovirus E1a protein, indicating that binding occurs through the "pocket" domain of Rb1. These results establish that the P2P-R cDNA encodes protein domains involved in both hnRNP association and Rb1 binding and complement recent reports that localize Rb1 to sites of RNA processing in the nucleus.


Ion transport in backbone-embedded polymerized ionic liquids.
We use atomistic computer simulations to examine ion-transport phenomena for backbone polymerized cationic liquids with bistrifluoromethylesulfonylimide (TFSI-) counterions. We consider a system in which the polymerized cation moiety is the imidazolium ring and study the structural characteristics and ion mobilities for cases in which the cations are separated by four, six, and eight methylene units on the backbone. A pendant polymerized ionic liquid, 1-butyl-3-vinylimidazolium, is compared to the backbone series across ion coordination and hopping features. The anion diffusivity in backbone polymerized cationic liquids is found to decrease with increasing spacer length, which is shown to result from a decrease in intramolecular and intermolecular hopping frequencies due to an increasing distance separating imidazolium moieties. In comparison with pendant polymerized ionic liquids, we observe that the participation rates of intermolecular hopping events in the backbone polymers far exceed that of the pendant, and the intrapolymeric ionic coordination profile shows the TFSI- of the pendant polymer with a high propensity for coordination by multiple imidazolium, compared with one monomer from a given polymer for the backbone series. Despite these differences, backbone polymerized ionic liquids are seen to possess correlated diffusivity and ion-association relaxation times, in a manner similar to the results observed in past studies for pendant variants.


Controlled release drug delivery in farmed animals: commercial challenges and academic opportunities.
Currently there is a lack of new active pharmaceutical ingredients (APIs) appearing on the veterinary market. In the short term this problem can be offset by developing controlled release drug delivery technologies to extend the commercial life of existing drugs. However, such a commercial opportunity does not come without its challenges. These generally revolve around financial factors, which include limited budgets assigned to conduct veterinary R&D, the cost-competitive amount that can be charged for the finished product and the expensive time-consuming registration process. In addition, the gap between the perceived and actual market needs makes the return on investment hard to defend. It is not surprising therefore that few controlled release products appear on the market for farmed animals, despite their potential advantages to that sector. The landscape for an academic veterinary pharmaceutical scientist is quite different from that of the industrial one. When you remove the commercial requirement associated with product development, there are numerous fundamental and applied research opportunities, with the outcome of demonstrating the potential worth, or otherwise, of an approach being sufficient to achieve the major goal of academics, publication in peer-reviewed journals. A further opportunity arises when controlled release dosage forms are used as research tools to forward knowledge in the area of animal science. The aim of this review is to provide a perspective of the current animal health industry through examination of the commercial challenges per se, along with the potential for academic collaboration that lie within this demanding area of pharmaceutical science.


High latitude changes in ice dynamics and their impact on polar marine ecosystems.
Polar regions have experienced significant warming in recent decades. Warming has been most pronounced across the Arctic Ocean Basin and along the Antarctic Peninsula, with significant decreases in the extent and seasonal duration of sea ice. Rapid retreat of glaciers and disintegration of ice sheets have also been documented. The rate of warming is increasing and is predicted to continue well into the current century, with continued impacts on ice dynamics. Climate-mediated changes in ice dynamics are a concern as ice serves as primary habitat for marine organisms central to the food webs of these regions. Changes in the timing and extent of sea ice impose temporal asynchronies and spatial separations between energy requirements and food availability for many higher trophic levels. These mismatches lead to decreased reproductive success, lower abundances, and changes in distribution. In addition to these direct impacts of ice loss, climate-induced changes also facilitate indirect effects through changes in hydrography, which include introduction of species from lower latitudes and altered assemblages of primary producers. Here, we review recent changes and trends in ice dynamics and the responses of marine ecosystems. Specifically, we provide examples of ice-dependent organisms and associated species from the Arctic and Antarctic to illustrate the impacts of the temporal and spatial changes in ice dynamics.


Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing.
CRISPR/Cas9 genome editing has gained rapidly increasing attentions in recent years, however, the translation of this biotechnology into therapy has been hindered by efficient delivery of CRISPR/Cas9 materials into target cells. Direct delivery of CRISPR/Cas9 system as a ribonucleoprotein (RNP) complex consisting of Cas9 protein and single guide RNA (sgRNA) has emerged as a powerful and widespread method for genome editing due to its advantages of transient genome editing and reduced off-target effects. In this review, we summarized the current Cas9 RNP delivery systems including physical approaches and synthetic carriers. The mechanisms and beneficial roles of these strategies in intracellular Cas9 RNP delivery were reviewed. Examples in the development of stimuli-responsive and targeted carriers for RNP delivery are highlighted. Finally, the challenges of current Cas9 RNP delivery systems and perspectives in rational design of next generation materials for this promising field will be discussed.


The Political Realignment of Health: How Partisan Power Shaped Infant Health in the United States, 1915-2017.
The US two-party system was transformed in the 1960s when the Democratic Party abandoned its Jim Crow protectionism to incorporate the policy agenda fostered by the civil rights movement, and the Republican Party redirected its platform toward socioeconomic and racial conservatism. The authors argue that the policy agendas promoted by the two parties through presidents and state legislatures codify a racially patterned access to resources and power detrimental to the health of all. To test the hypothesis that fluctuations in overall and race-specific infant mortality rates (IMRs) shift between the parties in power before and after the political realignment (PR), the authors apply panel data analysis methods to state-level data from the National Center for Health Statistics for the period 1915 through 2017. Net of trend, overall, and race-specific IMRs were not statistically different between presidential parties before the PR. This pattern, however, changed after the PR, with Republican administrations consistently underperforming Democratic ones. Net of trend, non-Southern state legislatures controlled by Republicans underperform Democratic ones in overall and racial IMRs in both periods.