Oscillations without cortex: Working memory modulates brainwaves in the endbrain of crows.
Complex cognition requires coordinated neuronal activity at the network level. In mammals, this coordination results in distinct dynamics of local field potentials (LFP) central to many models of higher cognition. These models often implicitly assume a cortical organization. Higher associative regions of the brains of birds do not have cortical layering, yet single-cell correlates of higher cognition are very similar to those found in mammals. We recorded LFP in the avian equivalent of prefrontal cortex while crows performed a highly controlled and cognitively demanding working memory task. We found signatures in local field potentials, modulated by working memory. Frequencies of a narrow gamma and the beta band contained information about the location of target items and were modulated by working memory load. This indicates a critical involvement of these bands in ongoing cognitive processing. We also observed bursts in the beta and gamma frequencies, similar to those that play a vital part in 'activity silent' models of working memory. Thus, despite the lack of a cortical organization the avian associative pallium can create LFP signatures reminiscent of those observed in primates. This points towards a critical cognitive function of oscillatory dynamics evolved through convergence in species capable of complex cognition.


Somatostatin decreases somatostatin messenger ribonucleic acid levels in the rat periventricular nucleus.
Growth hormone (GH) secretion from the pituitary is known to be under the dual control of GH-releasing factor (GRF) and somatostatin (SRIF). Hypothalamic SRIF, the major inhibitor of pituitary growth hormone secretion, inhibits its own release by a negative ultrashort-loop feedback mechanism. However, it is not known whether this negative regulation is mediated by inhibition of SRIF mRNA production. GRF may also inhibit its own release, thereby modifying pituitary GH secretion, possibly through an ultrashort-loop feedback mechanism. Thus, SRIF production and GRF release are both regulated by SRIF. Periventricular nucleus (PeN) and mediobasal hypothalamus (MBH) from adult male rats were incubated for 6 h in Waymouth's medium with either SRIF or the SRIF agonist analog RC 160 (10(-9) to 10(-6) M). Levels of SRIF mRNA were determined by an S1 nuclease protection assay using a 32[P]-labeled rat SRIF riboprobe. SRIF (10(-7) M) and RC 160 (10(-8), 10(-7) M) significantly (p< or =0.01) decreased SRIF mRNA levels in the PeN. The levels of SRIF mRNA in the MBH were not modified by either SRIF or RC 160. SRIF (10(-7) and 10(-6) M) significantly (p < or = 0.01 and p < or = 0.001, respectively) inhibited the release of GRF at 30 min in the MBH. Likewise, the release of GRF was slightly decreased by 10(-7) M RC 160, and significantly inhibited by 10(-6) M (p < or = 0.001) at 30 min. At 6 h, the levels of GRF were significantly reduced by 10(-7) M SRIF (p < or = 0.05) and by RC 160 (10(-7), 10(-6) M; p < or = 0.001 and p < or = 0.05, respectively). In contrast with these results, the SRIF analog was unable to alter SRIF release at 30 min. At 6 h incubation, RC 160 (10(-7) M) significantly (p < or = 0.001) reduced SRIF release from MBH fragments. These results demonstrate that SRIF and a SRIF analog decrease SRIF mRNA levels in the PeN and inhibit the release of SRIF from the nerve terminals of the MBH. Thus, SRIF appears to regulate its own gene expression by negative ultrashort-loop feedback. Therefore, when SRIF is secreted from these neurons in response to GRF, it down-regulates the preceding stimulatory input as well as its own secretion.


Interneurons in the tritocerebrum of the crayfish.
In isolated head preparations of the freshwater crayfish Orconectes limosus 268 local and projecting interneurons with branches in the tritocerebrum have been penetrated with glass microelectrodes and characterized for their sensory inputs. Using 3 criteria (sensory modality, site of receptors, response type of interneurons), the interneurons found were divided into 16 classes. The interneurons were either unimodal mechanoreceptive (89%) or bimodal (9% responding to mechanical and chemical stimuli, 2% responding to mechanical and visual stimuli). No trimodal interneurons were found. Within each modality the neurons received mostly bilateral input (70% of all interneurons responding to antennal stimulation, 84% of all chemosensitive interneurons). If the input was lateralized it was more often ipsilateral. The types of interneuronal responses evoked by sensory stimulation were: neurons that were exclusively excited (84%), those that were exclusively inhibited (10%), those that were excited or inhibited depending on the modality or laterality of the stimulus (6%), those showing long lasting excitatory aftereffects (3%), and those showing excitation or inhibition upon identical stimulation depending on the state of the neurons while being stimulated (1%). Interneurons that responded to mechanical antennal stimulation responded best either to low (10 Hz) or to high (100 Hz) stimulus frequencies. Six neurons responded best to a certain phase relationship between the movements of both antennal flagellae.


Effects of highly selective kappa-opioid agonists on EEG power spectra and behavioural correlates in conscious rats.
The present study compares the electroencephalographic (EEG) and behavioural effects of highly selective kappa-opioid agonists, spiradoline (U62066), enadoline (CI-977), BRL 52656, and BRL 53001, after SC administration to conscious rats. All compounds caused a distinctive dose- and time-related EEG effect and behavioural profile. The EEG power spectra (PS) of treated animals were characterised by power reduction indicative of CNS activation and an unusual spectral peak at 4-7 Hz, compared to controls. Behaviourally, however, the animals appeared to be sedated, with a reduction of locomotor activity, loss of postural tone, ataxia, and unusual hyperreactivity. The compounds differed in their liability to produce these CNS effects, with kappa-opioid analgesics such as enadoline and BRL 53001 showing a reduced propensity at equivalent antinociceptive doses. Where tested, these effects were inhibited by the nonselective opioid antagonist naloxone. The non-brain penetrating kappa-opioid agonist, BRL 52974, failed to produce alterations in EEG PS or behaviour, up to a dose of 10 mg/kg, indicating that the effects depend on activation of centrally located kappa-receptors.


Effects of 2-[1-(ethoxyimino)propyl]-3-hydroxy-5-(2,4,6-trimethylphenyl) cyclohex-2-enone on hepatic haem biosynthesis: species differences in hepatic porphyria.
2-[1-(Ethoxyimino)propyl]-3-hydroxy-5-(2,4,6-trimethylphenyl) cyclohex-2-enone (ETC) is a novel alkyl ketone herbicide. Continuous administration of ETC to mice for 28 days resulted in marked liver enlargement and severe intrahepatic cholestasis. These effects have been shown to result directly from a rapid and marked accumulation of porphyrin in the liver. The porphyrin which accumulates in the liver has been identified as protoporphyrin IX and dose response and time course studies confirm prior inhibition of mitochondrial ferrochelatase as the causal lesion. ETC was a very potent porphyrinogenic compound in mice, with a no-effect level for a single oral dose of 1 mg/kg. Rats and hamsters were insensitive to this type of hepatotoxicity following single oral doses of up to 750 mg/kg or following repeated, and indeed prolonged administration. The sensitivity of different species to ETC-induced porphyria correlated with the effect of ETC on hepatic ferrochelatase activity. The inhibition of ferrochelatase activity and the hepatic porphyria in mice were both found to be readily reversible upon withdrawal of ETC.


Platelet aggregability in humans: contrasting in vivo and in vitro findings during sympatho-adrenal activation and relationship to serum lipids.
Platelet aggregability was studied in 18 healthy volunteers during mental stress (a colour word test; CWT) and low- and high-dose adrenaline infusions using an ex vivo technique (filtragometry) and conventional in vitro aggregometry. CWT and high-dose adrenaline (3.4 nmol l-1 in plasma) shortened filtragometry readings, suggesting increased platelet aggregability in vivo. Low-dose adrenaline had no effect despite higher adrenaline levels in plasma (0.9 nmol l-1) than during CWT (0.4 nmol l-1). Platelet sensitivity to ADP in vitro was reduced following CWT and further reduced following adrenaline infusions. In vitro, adrenaline (50 nmol l-1) had little effect on platelet aggregation per se, but enhanced aggregability evoked by ADP (at ED50). Adrenaline potentiation of ADP-induced aggregation was enhanced after CWT, but was not related to filtragometry responsiveness to stress in vivo. Serum LDL-cholesterol levels were inversely correlated to filtragometry readings at rest, suggesting an adverse influence on platelet aggregability in vivo. HDL-cholesterol levels were inversely correlated to platelet sensitivity to ADP in vitro, suggesting a positive influence. Thus, sympatho-adrenal activation enhances platelet aggregability in vivo (as assessed by ex vivo filtragometry), but adrenaline alone cannot explain the pro-aggregatory effect of mental stress. Serum lipoprotein alterations associated with increased risk for atherosclerosis seem to enhance platelet aggregability. The conventional in vitro technique may poorly reflect platelet aggregability in vivo.


LSD, afterglow and hangover: Increased episodic memory and verbal fluency, decreased cognitive flexibility.
Psychedelics acutely impair cognitive functions, but these impairments decline with growing experiences with psychedelics and microdoses may even exert opposing effects. Given the recent evidence that psychedelics induce neuroplasticity, this explorative study aimed at investigating the potential of psychedelics to sub-acutely change cognition. For this, we applied a randomized, double-blind, placebo-controlled, crossover study with 24 healthy volunteers receiving 50 μg lysergic acid diethylamide (LSD) or an inactive placebo. Sub-acute changes in cognition were measured 24 h after dosing, including memory (Rey-Osterrieth Complex Figure, ROCF; 2D Object-Location Memory Task, OLMT; Rey Auditory-Verbal Learning Test, RAVLT), verbal fluency (phonological; semantic; switch), design fluency (basic; filter; switch), cognitive flexibility (Wisconsin Card Sorting Test, WCST), sustained and switching attention (Trail Making Test, TMT), inhibitory control (Stroop Task) and perceptual reasoning (Block Design Test, BDT). The results show that when compared to placebo and corrected for Body Mass Index (BMI) and abstinence period from psychedelics, LSD sub-acutely improved visuospatial memory (ROCF immediate recall points and percentage, OLMT consolidation percentage) and phonological verbal fluency and impaired cognitive flexibility (WCST: fewer categories achieved; more perseveration, errors and conceptual level responses). In conclusion, the low dose of LSD moderately induced both "afterglow" and "hangover". The improvements in visuospatial memory and phonological fluency suggest that LSD-assisted therapy should be explored as a novel treatment perspective in conditions involving memory and language declines such as brain injury, stroke or dementia.


Anterior cruciate ligament transection of rabbits alters composition, structure and biomechanics of articular cartilage and chondrocyte deformation 2 weeks post-surgery in a site-specific manner.
Anterior cruciate ligament (ACL) injury often leads to post-traumatic osteoarthritis (OA) and articular cartilage degradation, changing biomechanics of the tissue and chondrocytes, and altering the fixed charged density (FCD) and collagen network. However, changes in these properties are not known at a very early time point after ACL rupture, but recognizing early changes might be crucial for successful intervention. We investigated the effects of ACL transection (ACLT) in rabbits on the site-specific biomechanical properties of articular cartilage and chondrocytes, FCD content and collagen network organization, two weeks post-surgery. Unilateral ACLT was performed in eight rabbits, and femoral condyles, tibial plateaus, femoral grooves and patellae were harvested from experimental and contralateral knee joints. An intact control group was used as a reference. We analyzed chondrocyte morphology under pre- and static loading, cartilage biomechanical properties, FCD content and collagen fibril orientation. ACLT caused FCD loss in the lateral and medial femoral condyle, lateral tibial plateau, femoral groove and patellar cartilage (p < 0.05). Minor changes in the collagen orientation occurred in the femoral groove and lateral and medial femoral condyle cartilage (p < 0.05). Cartilage stiffness was reduced in the lateral and medial femoral condyles, and chondrocyte biomechanics was altered in the lateral femoral condyle and patellar cartilage (p < 0.05). We observed loss of FCD from articular cartilage two weeks after ACLT at several joint locations. These changes may have led to decreased cartilage stiffness and altered cell deformation behavior, especially in the femoral condyles.


Weight-height ratios and parameters of body posture in 7-9-year-olds with particular posture types.
Small and relatively balanced anteroposterior curvatures and symmetric positioning of the shoulders, scapulae, waist triangles, anterior superior iliac spines, knees and feet are the elements of a good posture. The aim of our study was to determine which somatic features and parameters of spinal curvatures in the sagittal plane show statistically significant differences among children with particular types of body posture. The study was carried out in selected primary schools of the Subcarpathian and Silesian regions. We examined a total of 563 randomly selected first- to third-graders whose parents had submitted written informed consent. The study group included 278 boys (49.38%) and 285 girls (50.62%). Twenty-four parameters describing the body posture were determined in every participant using the photogrammetric method. Gender-independent parameters helpful in differentiating postural types comprise the compensation indicator, inclination angle of the superior thoracic spine, C7-PL length (length of kyphosis), and inclination angle of the thoracolumbar spine. The inclination angle of the lumbosacral spine, total curvature length, position of the apex of lordosis relative to S1, and S1-PL length (length of lordosis), appeared to be the least helpful in differentiating among postural types.


[SARS-CoV-2: the experience of the Legouest armed forces teaching hospital].
The Legouest military training hospital is one of the eight hospitals of the armed forces health service. Situated in the Grand-Est region, one of the regions most affected by the COVID-19 epidemic in spring, it had to reorganise itself within a few days with its regional and national partners. While continuing to support forces sent abroad, to overseas territories or located in the East of France, the armed forces hospital had three major missions: the support of other military hospital facilities, the continued care of non-COVID patients and the care of patients affected by COVID-19 requiring non-intensive hospital care.


Red nucleus: past and future.
The red nucleus has greatly interested scientists for almost a century. This can be explained by the fact that problems of general interest are encountered when studying this nucleus. Some of them are outlined in this paper, such as the phylogenetic evolution of the rubrospinal tract, the respective roles of the rubrospinal and pyramidal tracts in the execution of various types of movements, and the respective roles of these two tracts in movement automatization.


Single-stage surgical repair in a complex case of aberrant right subclavian artery aneurysm and common carotid trunk.
Aberrant right subclavian artery with coexisting common carotid trunk is an extremely rare congenital anomaly affecting <0.1% of the population. We report the case of a 77-year-old Caucasian man presenting with dysphagia and dyspnea secondary to an aberrant right subclavian artery aneurysm and describe our technique for open surgical repair.


New pi-extended water-soluble squaraines as singlet oxygen generators.
[graph: see text] Condensation of squaric acid with a number of differently substituted 2-pyrrolyl derivatives afforded three new classes of squaraines. Their sharp and intense absorption bands in the biological window (700-900 nm), inherent singlet oxygen generation capabilities, together with proper functionalization allowing good water solubility make them suitable candidates as new non-porphyrinic singlet oxygen photosensitizers for photodynamic therapy (PDT).


Body surface profile in ambient and hot temperatures during a rectangular test in race walker champions of the World Cup in Oman 2022.
There is current interest in infrared thermography as a method to assess changes in body surface temperature to determine thermoregulatory mechanisms, especially in endurance sports. The aim of this study was to evaluate the effect of two environmental temperatures (17 and 28°C) on body surface temperature in different anterior and posterior aspects of the body during a rectangular test in international walkers of the Spanish National Team. Three international walkers performed a rectangular test, where body temperature was measured at rest, and after the 5th, 10th and 15th run using an infrared thermographic camera in room temperatures at 17 and 28°C. In addition, oxygen consumption was measured simultaneously. ANOVA detected a group × time interaction in the chest and abdomen (right and left), left back and right calf (p = < 0.05), with a trend in the right hamstring (p = 0.053) when comparing 17°C and 28°C. ANOVA detected no significant group × time interaction (p = 0.853) but there was a significant group effect (p = 0.022). The eleven degrees increase in ambient temperature (17 to 28°C) produces changes in almost all anatomical zones, but not homogeneously in international walkers during a rectangular test. This indicates that metabolic and blood flow changes are different depending on the anatomical zone measured.


Genistein contributes to cell cycle progression and regulates oxidative stress in primary culture of osteoblasts along with osteoclasts attenuation.
The present study was designed to examine the role of isoflavone genistein (GS) on bone formation, regulating oxidative stress and cell cycle in primary osteoblasts, as well as attenuation of osteoclast formation. Primary calvaria osteoblasts were isolated from 2 to 3 days old neonatal rat pups (n = 6-8) of Sprague Dawley rats. Osteoblasts were incubated with varying concentrations of GS and different assays viz. cell proliferation, differentiation, calcium deposition, cell cycle progression, antioxidant ability, and osteogenic gene expression were performed. Tartrate-resistant acid phosphatase (TRAP) staining and immunolocalization of cathepsin K protein were assessed in bone marrow-derived osteoclasts. Results revealed that GS markedly induced cell growth and osteoblast differentiation depending upon dose. The fluorescent dye DCFH-DA staining data proved the antioxidant ability of GS, which reduced the H2O2- induced intracellular oxidative stress in osteoblasts. Quantitative real-time PCR analysis revealed that GS treatment upregulated the expression of osteoblastic genes of Runt-related transcription factor 2 (Runx2), bone morphogenetic proteins 2 (BMP2), and osteocalcin. Immunolocalization of BMP2 also indicated the osteogenic efficacy of GS. Furthermore, TRAP staining and cathepsin K expression depicted that GS inhibited multinucleated osteoclasts formation. In conclusion, GS isoflavone might impart protective effects against oxidative stress-induced bone loss and thus, could maintain skeletal growth.


Prediction of porcine semen fertility by homologous in vitro penetration (hIVP) assay.
A field trial was conducted to compare the fertility predicting capacity of different sperm assays applying classical semen analysis, sperm function and the homologous in vitro penetration test (hIVP) to 60 ejaculates from four boars collected over a period of 15 weeks. No differences were found between the groups of fertility (Low Fertility: < 20%; Intermediate: 40-60% and High: > 80%) for sperm-rich fraction volume collection, sperm concentration, total sperm number, cationic contents in seminal plasma and ATP concentration. Partial differences were found in the parameters of motility, normal morphology, normal apical ridge (NAR), viability with eosin-nigrosin stain, hypo-osmotic swelling test (HOS), osmotic resistance test (ORT) and functional membrane integrity (with carboxyfluorescein diacetate, DCF). These parameters would be useful for detecting sperm with poor fertility, but they are not precise enough to discriminate an ejaculate with higher fertility than the herd median. Only the penetration percentage (10.24 +/- 1.45 vs. 55.13 +/- 3.35 vs. 84.72 +/- 1.73) and sperm number per oocyte (1.29 +/- 0.07 vs. 11.29 +/- 1.79 vs. 25.86 +/- 1.43) in a hIVP system were parameters with a predictive capacity to discriminate between the three fertility groups. Consequently, hIVP was found to be the best seminal assay and it may improve the in vitro assessment of sperm fertilizing ability.


Liver resection with the Sonocut ultrasonic knife.
Previous reports and our findings suggest that resection by ultrasound is of appreciable merit in liver surgery. Blood can be saved and bile leakage diminished as the larger vessels and bile ducts can be skeletonized unharmed, whereas the dissector selectively carries away the parenchyma. Otherwise virtually inaccessible tumours can be approached. The ultrasonic dissector/aspirator does not harm the tissue in depth beyond the resection surface and thus not much devitalized tissue is left in the wound.


IgM rheumatoid factor elaboration by blood, bone marrow, and synovial mononuclear cells in patients with rheumatoid arthritis.
IgM rheumatoid factor (RF) elaboration by rheumatoid arthritis (RA) synovial, bone marrow, and blood mononuclear cells (MNC) is reported. IgM RF was prepared from RF-positive sera by sequential euglobulin precipitation, Sephacryl S300 gel filtration, and IgG-Sepharose affinity chromatography. Purified material, which contained no detectable IgG or IgA, was used in an enzyme-linked immunosorbent assay (ELISA) to quantitate cellular elaboration of IgM RF. Excellent standard curves (r2 = 0.98) were obtained without nonspecific binding of samples or antisera to IgG-coated microtiter plates and without cross-reactivity of standards with antisera other than anti-IgM. We found RA blood MNC (11 patients) spontaneously averaged 15 ng/ml IgM RF (6% of total IgM produced), but elaborated 254 ng/ml IgM RF following pokeweed mitogen (PWM) stimulation (22 patients), exceeding that of 13 normal controls. Bone marrow MNC spontaneously (4 patients) produced 71 ng/ml IgM RF and secreted 78 ng/ml IgM RF with PWM stimulation (9 patients). In contrast synovial fluid MNC (5 patients) spontaneously elaborated 6652 ng/ml IgM RF, significantly (P less than 0.05) more than blood or bone marrow MNC; PWM-stimulated synovial fluid MNC (5 patients) produced 5472 ng/ml IgM RF. These observations confirm selective localization of activated, IgM RF-producing cells to the rheumatoid synovial space.


Plasminogen Kanagawa-I, a novel missense mutation, is caused by the amino acid substitution G732R.
A new dysplasminogen, plasminogen Kanagawa-I, was identified in a healthy male with no previous thrombotic episodes. His plasma plasminogen (PLG) activity was 51.4% of that of normal pooled plasma (reference interval 70-130%) and the antigen level was 94.2% of that of normal pooled plasma (reference interval 80-150%). Nucleotide sequencing revealed a heterozygous G to A transition in exon 18, which resulted in an amino acid substitution of G732R. Both the proband's father and paternal grandfather were heterozygous for this mutation. Interestingly, the grandfather was found to be a compound heterozygote for plasminogen Kanagawa-I and Tochigi (A601T), so that his plasminogen activity and antigen level was 7.7% and 87.2% of that of normal pooled plasma, respectively. However, he has never been affected by significant thrombosis.


[Transarterial embolization of intracranial dural arteriovenous malformations with ethylene vinyl alcohol copolymer (Onyx18): report of three cases].
This is a report of the endovascular treatment of three intracranial dural arteriovenous malformations (DAVM) using Onyx. We analyzed the clinical and angiographic results in three patients with intracranial DAVM, revealed by a hemorrhagic event and treated by endovascular way by the use Onyx. Angiographic investigation showed the DAVM to be located in the lateral sinus in two cases and in the tentorium region in the third. In all three cases, embolization was performed by selective catheterization of the meningeal arterial feeder of the DAVM. The injection of Onyx resulted in complete anatomical exclusion of the DAVM, as demonstrated by posttreatment angiography. Clinically, the patients recovered partially or completely their neurological deficit. Onyx is a liquid embolization agent recently introduced for the treatment of DAVM. When used under optimal conditions, it offers a feasible alternative option in the treatment of such vascular malformations.


Physical learning environments' supportiveness to innovative pedagogies: students' and teachers' experiences.
Research into the relationship between innovative physical learning environments (PLEs) and innovative psychosocial learning environments (PSLEs) indicates that it must be understood as a network of relationships between multiple psychosocial and physical aspects. Actors shape this network by attaching meanings to these aspects and their relationships in a continuous process of gaining and exchanging experiences. This study used a psychosocial-physical, relational approach for exploring teachers' and students' experiences with six innovative PLEs in a higher educational institute, with the application of a psychosocial-physical relationship (PPR) framework. This framework, which brings together the multitude of PLE and PSLE aspects, was used to map and analyse teachers' and students' experiences that were gathered in focus group interviews. The PPR framework proved useful in analysing the results and comparing them with previous research. Previously-identified relationships were confirmed, clarified, and nuanced. The results underline the importance of the attunement of system aspects to pedagogical and spatial changes, and of a psychosocial-physical relational approach in designing and implementing new learning environments, including the involvement of actors in the discourse within and between the different system levels. Interventions can be less invasive, resistance to processes could be reduced, and innovative PLEs could be used more effectively.


Genetic susceptibility, homocysteine levels, and risk of all-cause and cause-specific mortality: A prospective cohort study.
The associations of homocysteine (Hcy) and gene-Hcy interactions with the risk of all-cause and cause-specific mortality remain unclear. A total of 19,826 middle-aged and elderly Chinese adults were included from the Dongfeng-Tongji cohort in 2013-2014 and were followed-up to 31 December 2018. Cox regression was used to examine the association between Hcy and mortality. We selected 18 well-established Hcy-associated genetic variants to constructed the weighted genetic risk score (GRS) among 15,434 participants with genetic data, and interactions between genetic susceptibility and Hcy on mortality were assessed. After multivariate adjustment, elevated serum Hcy levels were associated with higher risk of mortality from all-cause, CVD, coronary heart disease (CHD), stroke, and cancer. We also observed a significant interaction between GRS and Hcy on CHD mortality. Moreover, the rs7130284 and rs957140 on NOX4 modified the association between Hcy and mortality from CVD and CHD, and rs154657 on DPEP1 modified the association between Hcy and CHD mortality. Elevated Hcy levels were associated with increased risk of all-cause and cause-specific mortality among middle-aged and elderly Chinese. Hcy-related genetic variants on NOX4 and DPEP1 might modify the associations of Hcy with CVD mortality or CHD mortality.


Evaluation of recombinant factor VIIa treatment for massive hemorrhage in patients with multiple traumas.
Recent studies and case reports have shown that recombinant factor VIIa (rFVIIa) treatment is effective for reversing coagulopathy and reducing blood transfusion requirements in trauma patients with life-threatening hemorrhage. The purpose of this study is to evaluate the effect of rFVIIa treatment on clinical outcomes and cost effectiveness in trauma patients. Between January 2007 and December 2010, we reviewed the medical records of patients who were treated with rFVIIa (N=18) or without rFVIIa (N=36) for life-threatening hemorrhage due to multiple traumas at the Emergency Department of Pusan National University Hospital in Busan, Korea. We reviewed patient demographics, baseline characteristics, initial vital signs, laboratory test results, and number of units transfused, and then analyzed clinical outcomes and 24-hr and 30-day mortality rates. Thromboembolic events were monitored in all patients. Transfusion costs and hospital stay costs were also calculated. In the rFVIIa-treated group, laboratory test results and clinical outcomes improved, and the 24-hr mortality rate decreased compared to that in the untreated group; however, 30-day mortality rate did not differ between the groups. Thromboembolic events did not occur in both groups. Transfusion and hospital stay costs in the rFVIIa-treated group were cost effective; however, total treatment costs, including the cost of rFVIIa, were not cost effective. In our study, rFVIIa treatment was shown to be helpful as a supplementary drug to improve clinical outcomes and reduce the 24-hr mortality rate, transfusion and hospital stay costs, and transfusion requirements in trauma patients with life-threatening hemorrhage.


Highly sensitive biotin-avidin sandwich ELISA for the rapid detection of pneumococcal capsular polysaccharide antigens.
The immunological detection of soluble pneumococcal polysaccharide antigens in pathological products is of importance in the direct diagnosis of meningitis or pulmonary infections. We have developed a double antibody sandwich ELISA method using a biotin-avidin system using antibodies constituted with a mixture of IgGs from pooled and/or monospecific antipneumococcal sera provided by the Danish Statens Seruminstitut. The sensitivity of this rapid ELISA method was optimized with purified capsular polysaccharides of the 24 main pneumococcal serotypes. With incubation steps of 30 min at 37 degrees C for the antigens and the conjugates, the detection limit was close to 1 ng/ml for 75% of the purified polysaccharides. A retrospective study of 46 CSF samples established the validity of the assay. This type of modified ELISA system represents a specific, sensitive and rapid procedure for the potential detection of capsular soluble antigens of all pneumococcal serotypes.


Diagnostic and predictive factors of significant liver fibrosis and minimal lesions in patients with persistent unexplained elevated transaminases. A prospective multicenter study.
In patients with unexplained elevated transaminases, prognosis of the liver disease and factors associated with increased risk of liver fibrosis and normal/subnormal liver are unknown. The aim of this prospective study was to identify diagnosis and clinical and biological factors associated with significant (bridging) fibrosis and minimal lesions of the liver in patients with persistent unexplained elevated ALT levels. From July 2002 through October 2004, all consecutive asymptomatic patients with unexplained chronically elevated ALT levels were included. All patients had clinical, biological, ultrasonographic examination and a liver biopsy. 272 patients (60.3% males, mean age 46.4 years, BMI 26.7) were included. Pathological findings were: minimal lesions (18.7%), steatosis (26.8%), NASH (32.7%), and miscellaneous (21.7%). Significant fibrosis was found in 27.4% of cases, including 9 cases of cirrhosis. By multivariate analysis, independent predictors of significant fibrosis were tobacco use (OR 2.5, 95% CI 1.34-4.74 p=0.04), BMI>25 (2.49, 1.31-4.73 p=0.005) and diabetes (4.41, 1.73-11.29 p=0.002). Independent factors associated with minimal lesions were female gender (OR 3.4 95% CI 1.73-6.75 p<0.0001) and BMI<25 (3.55, 1.8-6.98, p<0.0001). In patients with unexplained chronically elevated transaminases, significant fibrosis is statistically associated with tobacco use, BMI>25 and diabetes, and minimal lesions are significantly associated with female gender and BMI<25.


Carbamazepine metabolism in humans: effect of concurrent anticonvulsant therapy.
Free and total carbamazepine (CBZ) and carbamazepine-epoxide (CBZ-EP) plasma levels were obtained on 113 patients with epilepsy (18-61 years old) controlled on either monotherapy or coadministration with either phenobarbital (PB), phenytoin (PHT), valproic acid (VPA), or all three. A subset of patients were administered tetradeuterium labeled CBZ to evaluate the effects of autoinduction and coadministration of VPA on the kinetics of CBZ and its metabolite CBZ-EP. Polytherapy had variable effect on free and total CBZ plasma levels compared to monotherapy. Coadministered PHT (co-PHT), or all three anticonvulsants together (PHT, PB, and VPA: co-AEDs) decreased free and total CBZ plasma levels. No change was noted for coadministered VPA (co-VPA). Compared to monotherapy the free and total CBZ-EP levels increased with co-VPA, less with coadministered PB (co-PB), and no change with co-PHT or co-AEDs. Protein binding of CBZ and CBZ-EP was not affected by any antiepileptic drugs studied. The free and total CBZ-EP/CBZ ratio was tripled with co-VPA or co-AED's, and doubled with co-PHT or co-PB. Isotope labeling did not demonstrate any differences in half-life (t1/2), plasma clearance (Cl), or volume of distribution (Vd). Compared to naive controls, monotherapy and co-VPA decreased CBZ t1/2 by 50%, and more than doubled the CBZ Cl without a significant change in the Vd. Autoinduction is one explanation for these changes with chronic CBZ therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


A pictorial key to the species of the Aedes (Zavortinkius) in the Afrotropical Region (Diptera: Culicidae).
Six species of the subgenus Zavortinkius of Aedes Meigen in the Afrotropical Region are treated in a pictorial key based on diagnostic morphological features. Images of the diagnostic morphological structures of the adult thorax and leg are included.


Retrograde ejaculation owing to ectopic erectile tissue.
We report a case of retrograde ejaculation caused by ectopic erectile tissue in the posterior urethra. Transurethral resection of the tissue resulted in antegrade ejaculation.


Assessment of Breast Cancer Mortality Trends Associated With Mammographic Screening and Adjuvant Therapy From 1986 to 2013 in the State of Victoria, Australia.
Diagnosis of early breast cancer (EBC) in women by mammographic screening and postsurgical adjuvant endocrine therapy and chemotherapy (termed adjuvant therapy) began simultaneously in many countries in the 1990s. Subsequent breast cancer mortality declines were variously attributed to mammographic screening and/or adjuvant therapy. To determine the relative mortality reductions associated with these 2 interventions in women with EBC who had been exposed to both. This secondary analysis of cross-sectional studies assessed groups of women with invasive breast cancer in the State of Victoria, Australia, from January 1, 1982, to December 31, 2013, who were included in the Victorian Cancer Registry (VCR). The population consisted of participants in population-based studies of female breast cancer from 1986 to 2013 using data from 4 VCR population-based surveys of breast cancer treatment from 1986 to 1999; VCR data on breast cancer incidence, mortality, and TNM stage at diagnosis from 1986 to 2013; and Victorian mammographic screening program (BreastScreen Victoria) data from 1992 to 2007. Breast cancer incidence and mortality data were analyzed for all 76 630 women registered with invasive breast cancer with the VCR from January 1, 1982, to December 31, 2013, and breast cancer treatment and screening data were analyzed additionally for the groups of surveyed women as described above. Participation in BreastScreen Victoria and receipt of adjuvant therapy after surgery for EBC. Data were analyzed for associations between crude breast cancer mortality trends and uptake of adjuvant therapy and downstaging by mammographic screening. Of all 76 630 women registered with breast cancer with the VCR from January 1, 1982, to December 31, 2013. Joinpoint analyses of the time trend in crude mortality showed an increase from 31.6 per 100 000 women in 1982 to 34.3 per 100 000 women in 1994, with a single joinpoint at 1994, followed by a significant declining trend to 23.9 per 100 000 women in 2013 (annual percentage change, -1.3%; 95% CI, -1.6% to -0.9%). By 1999, 74% of all Victorian women with EBC (737 of 1001) had commenced adjuvant endocrine therapy, and 72% (187 of 260) of premenopausal and 29% (215 of 741) of postmenopausal women with EBC had commenced adjuvant chemotherapy. Crude incidence of advanced-stage breast cancer almost doubled from 12.2 per 100 000 women in 1986 to 23.9 per 100 000 women in 2013. This study found that mammographic screening did not downstage breast cancer in Victoria from advanced to early, so population mortality benefit is lacking. Adjuvant therapy uptake was associated with all of the decline in Victorian breast cancer mortality since 1994. Given these findings, monitoring the relative contributions of mammographic screening and adjuvant therapy for EBC to breast cancer mortality reductions in populations of women exposed to both should be mandatory.


Influence of Biometric and Seasonal Parameters on the Metal Content of Scomber colias in Northwestern African Waters.
Studies of the content of heavy metals and trace elements in fish and other marine organisms are of great importance to know the state of the marine ecosystem. This study has been carried out in northwestern African waters, Scomber colias being the target species, sampling a total of 345 specimens over 2 years. For the determination of metals and trace elements (Al, B, Cd, Cr, Cu, Fe, Li, Ni, Pb, V, and Zn) in the muscle, the ICP-OES (inductively coupled plasma optical emission spectrometry) was used. In the statistical study, the factors (oceanographic and maturity in the seasons, size of the specimens) were used. The samples sampled in the cold season that had high concentrations of metals may be due to the influence that African upwelling has on the Canary Islands. The metallic contents of our samples are lower than those found in the studies in Mauritania and Morocco due to the greater influence that upwelling has in these areas.


Proteasome-mediated protein processing by bidirectional degradation initiated from an internal site.
The proteasome is a barrel-shaped protease that conceals its active sites within its central cavity. Proteasomes usually completely degrade substrates into small peptides, but in some cases, degradation yields biologically active protein fragments. Some transcription factors are generated from precursors by this activity, but the mechanism of proteasomal protein processing remains unclear. Here we show that proteasomal processing of the yeast NFkappaB-related transcription factors Spt23 and Mga2 is initiated by an internal cleavage, followed by bidirectional proteolysis of the polypeptides. Studies with protein fusions indicate that stable proteolytic fragments are generated if the protein contains tightly folded structures that prevent the complete degradation of the protein. Furthermore, we provide evidence that the ability of the proteasome to initiate proteolysis from an internal site and to proceed via bidirectional polypeptide degradation may be relevant for the complete degradation of proteins as well.


Chromatographic separation of R/S-enantiomers of amphetamine and methamphetamine: Pathways of methamphetamine synthesis and detection in blood samples by qualitative enantioselective LC-MS/MS analysis.
Methamphetamine can be synthesized either enantiopure or in its racemic form. We separated (R)- and (S)-enantiomers of methamphetamine and amphetamine by a fast LC-MS/MS-method using a Lux® 3μm AMP 150×3.0mm analytical column after simple protein precipitation with methanol. Sufficient resolution could be achieved. Method validation for qualitative detection showed limits of quantification <5ng/mL while only little (maximum 14.5%) ion suppression could be shown. Stability in the processed sample could be achieved using isotopically labelled internal standards. Plasma samples of police cases from the german regions of Franconia and Northrhine revealed that in the majority of 106 tested samples (>99%) only (S)-methamphetamine was detected which leads to the conclusion that, in Germany, predominantly enantiopure (S)-methamphetamine is consumed which is synthesized via (1R,2S)-ephedrine or (1S,2S)-pseudoephedrine. However, racemic methamphetamine seems also to be on the market.


Why do bacteria divide?
The problem of not only how but also why cells divide can be tackled using recent ideas. One idea from the origins of life - Life as independent of its constituents - is that a living entity like a cell is a particular pattern of connectivity between its constituents. This means that if the growing cell were just to get bigger the average connectivity between its constituents per unit mass - its cellular connectivity - would decrease and the cell would lose its identity. The solution is division which restores connectivity. The corollary is that the cell senses decreasing cellular connectivity and uses this information to trigger division. A second idea from phenotypic diversity - Life on the Scales of Equilibria - is that a bacterium must find strategies that allow it to both survive and grow. This means that it has learnt to reconcile the opposing constraints that these strategies impose. The solution is that the cell cycle generates daughter cells with different phenotypes based on sufficiently complex equilibrium (E) and non-equilibrium (NE) cellular compounds and structures appropriate for survival and growth, respectively, alias 'hyperstructures.' The corollary is that the cell senses both the quantity of E material and the intensity of use of NE material and then uses this information to trigger the cell cycle. A third idea from artificial intelligence - Competitive Coherence - is that a cell selects the active subset of elements that actively determine its phenotype from a much larger set of available elements. This means that the selection of an active subset of a specific size and composition must be done so as to generate both a coherent cell state, in which the cell's contents work together harmoniously, and a coherent sequence of cell states, each coherent with respect to itself and to an unpredictable environment. The solution is the use of a range of mechanisms ranging from hyperstructure dynamics to the cell cycle itself.


Operating room fires: how to prevent and minimize spread.
To prevent fires, a simple rule to remember is the "fire triangle" of oxygen, heat, and fuel. Minimizing oxygen enrichment will reduce the fire hazard. Proper protocols need to be established for the handling of heat-bearing units, such as electrosurgical pencils, electrocautery units, and lasers.


[Abnormal respiration with dullness over the thorax in cattle. Due to an extensive lesion of the liver (author's transl)].
The case of a four-year-old cow is reported, in which a very extensive necropurulent lesion was localized in the left half of the liver. When the animal was studied clinically and by X-ray, a space-occupying lesion in the thorax was strongly suspected. The importance of including diseases of the liver in the differential diagnosis of thoracic disorders is stressed.


The antitumor effect of TIG3 in liver cancer cells is involved in ERK1/2 inhibition.
Tazarotene-induced gene 3 (TIG3) was first characterized in tazarotene-treated human keratinocytes and identified as a retinoic acid responder gene, an important mediator of antitumor effects by retinoids. In this study, we aim to investigate the inhibitory effect of TIG3 on the growth of liver cancer and explore its underlying mechanism. Human hepatocellular carcinoma (HCC) Hep3B cells were transfected with plasmid GV141 carrying full-length TIG3 complementary DNA (cDNA). The effects of TIG3 on cell proliferation, apoptosis, and migration were determined in vitro. The suppressor effect of TIG3 on tumor growth was evaluated in vivo in a nude mouse HCC model. We observed that TIG3 expression is decreased in the Hep3B cell line as well as primary HCC tumors, and TIG3 expression inversely correlates with Ki-67 expression. Overexpression of TIG3 suppresses tumor growth in HCC both in vitro and in vivo via ERK1/2 inhibition by promoting apoptosis and inhibiting proliferation and migration. These findings identify TIG3 as an attractive therapeutic target for HCC.


Significance of angiogenic factors in liver metastatic tumors originating from colorectal cancers.
We examined the expression of vascular endothelial growth factor (VEGF) and microvessel counts expressed by CD34 staining in 39 patients with primary and 44 patients with metastatic liver tumors of metastatic colorectal carcinoma, and 29 patients with nonmetastatic colorectal carcinoma as control in order to determine their value in the evaluation of prognosis and recurrence after hepatectomy. Microvessel counts in primary colorectal carcinomas of the metastatic group were significantly higher than those in control (P<0.05). Neither factor correlated with any clinicopathological feature of primary or metastatic liver carcinomas. Higher microvessel counts in metastatic liver tumors tended to be associated with a shorter disease-free interval to second recurrence in the remaining liver (P = 0.069) and were significantly associated with poor prognosis after hepatectomy (P<0.05). We conclude that microvessel count is an important marker of liver metastasis and prognosis in patients with colorectal carcinoma treated with hepatectomy.


Evaluation of nasal airway alterations associated with septorhinoplasty by both objective and subjective methods.
The aim of functional septorhinoplasty is to create an esthetically elegant nose and harmony in the face by preserving nasal function as well as maintaining or restoring adequate airway. Since nasal complaints are usually subjective, it may be difficult to evaluate the functions objectively. In the present study, we aimed to investigate the alterations in nasal function associated with septorhinoplasty by using both objective and subjective methods. The study population consisted of 40 patients who underwent septorhinoplasty and 40 healthy controls. Before and after the operation, visual analog scale, acoustic rhinometry, rhinomanometry, and Odiosoft-Rhino test were applied to all patients and controls. There were significant differences in all parameters both before and after the operation. While a significant difference was obtained between the patient and control groups in terms of preoperative values, no significant difference was found between postoperative values of these groups. Both objective and subjective methods are important in evaluations.


Mosaic disease induced by turnip yellow mosaic Tymovirus.
Mosaic disease is one of the most common consequences of infection of plants by viruses. Most such mosaics include islands of dark green tissue, which are normal in appearance, contain little or no virus and are resistant to superinfection at least for a period of weeks. The mosaic pattern that develops in successive expanded leaves of larger plants is variable and unpredictable. Therefore it has not been possible to investigate the early events in leaf primordia that lead to the formation of dark green tissue. Here we describe an experimental system which should allow the application of molecular techniques to the study of the nature of dark green tissue.


Exploring the role of empathy in prolonged grief reactions to bereavement.
Grief reactions to the bereavement of a close individual could involve empathy for pain, which is fundamental to social interaction. To explore whether grief symptoms interact with social relatedness to a person to whom one directs empathy to modulate the expression of empathy, we administered an empathy task to 28 bereaved adults during functional magnetic resonance imaging, in which participants were subliminally primed with facial stimuli (e.g., faces of their deceased or living relative, or a stranger), each immediately followed by a visual pain stimulus. Individuals' grief severity promoted empathy for the pain stimulus primed with the deceased's face, while it diminished the neural response to the pain stimulus primed with the face of either their living relative or a stranger in the medial frontal cortex (e.g., the right dorsal anterior cingulate cortex). Moreover, preliminary analyses showed that while the behavioral empathic response was promoted by the component of "longing" in the deceased priming condition, the neural empathic response was diminished by the component of "avoidance" in the stranger priming condition. Our results suggest an association between grief reactions to bereavement and empathy, in which grief symptoms interact with interpersonal factors to promote or diminish empathic responses to others' pain.


Identification of receptors in the liver that mediate endocytosis of circulating tissue kallikreins.
The liver plays an important role in the clearance, by receptor-mediated endocytosis, of circulating glycoproteins. It has been demonstrated that tissue kallikreins, which are acid glycoproteins, circulate in plasma, where they are poorly inhibited by plasma proteins. We have shown that the liver is the main organ that clears tissue kallikreins from the circulation. We now report the identification of receptors involved in this clearance. Using a perfused rat-liver system, and as models, pig pancreatic (PPK) and horse urinary (HoUK) kallikreins, we have found that: (a) the binding of PPK to the perfused liver was inhibited by 50 mM methyl alpha-D-mannoside and 20 microM mannan, was partially inhibited by 50 mM mannose and was unaffected by 1.5 microM asialofetuin; (b) binding of HoUK to the perfused liver was inhibited by 1.5 microM asialofetuin, 50 mM galactose and 50 mM lactose and was unaffected by 50 mM mannose; (c) the clearance rate of both kallikreins followed the equation y = a.xb; (d) their binding was Ca2+-dependent and their clearance was inhibited by 3 mM chloroquine and 10 mM methylamine. Using isolated liver cells and tritiated HoUK, we calculated that 500,000 receptors/cell were present and the Scatchard plot showed that there were two apparent affinity constants: 0.24.10(9) l/M) (high-affinity) and 0.3.10(8) l/M (low-affinity). These results show that PPK is recognized by a liver mannose receptor and HoUK by the galactose receptor. The liver uptake of native and circulating tissue kallikreins thus emerges as a mechanism by which their levels in plasma are regulated.


Calculating nominal group statistics in collaboration studies.
In many areas of psychology researchers compare the output of pairs of people with people working individually. This is done by calculating estimates for nominal groups, the output of two individuals if they had worked together. The way this is often done is by creating a single set of pairs either randomly or based on their location in a data file. This paper shows that this approach introduces unnecessary error. Two alternatives are developed and described. The first calculates statistics for all permissible sets of pairs. Unfortunately the number of sets is too large for modern computers for moderate sample sizes. The second alternative calculates statistics on all possible pairs. Several simulations are reported which show that both methods provide good estimates for the mean and trimmed mean. However, the all pairs procedure provides a biased estimate of the variance. Based on simulations, an adjustment is recommended for estimating the variance. Functions in S-Plus/R are provided in an appendix and are available from the author's Web page along with updates and alternatives (www.sussex.ac.uk/users/danw/s-plus/ngstats.htm).


Characterization of radioiodinated recombinant human TGF-beta 1 binding to bone matrix within rabbit skull defects.
Bone healing is regulated in part by the local production of TGF-beta 1 and other growth factors produced by cells at the site of injury. The single application of recombinant human TGF-beta 1 (rhTGF-beta 1) to calvarial defects in rabbits induces an accelerated recruitment and proliferation of osteoblasts within 3 days. This ultimately results in the formation of new bone and the complete closure of the defect within 28 days. The persistence and localization of [125I]rhTGF-beta 1 within an osseous defect was investigated after applying a single dose of [125I]rhTGF-beta 1 formulated in a 3% methylcellulose vehicle. Normal bone encompassing the defect site, the periosteum, and the gel film covering the dura were harvested at 0, 4, 8, and 24 h and 3, 7, and 16 days after [125I]rhTGF-beta 1 application. The defect site-associated radioactivity was quantitated, visualized by autoradiography, and characterized by TCA precipitation and SDS-PAGE. Radioactivity was observed in autoradiographs of gross specimens, histologic sections of the bone matrix, and periosteal tissue surrounding the defect. There was a time-dependent decrease in TCA-precipitable radioactivity; however, radioactivity was still associated with the bone matrix 16 days after application of [125I]rhTGF-beta 1. SDS-PAGE and autoradiography of the radioactivity in homogenized bone and periosteal samples revealed a 25 kD band, suggesting that the radioactivity remaining at the defect site represented intact [125I]rhTGF-beta 1.(ABSTRACT TRUNCATED AT 250 WORDS)


Simultaneous appraisals of pathway and probable health risk associated with trace metals contamination in groundwater from Barapukuria coal basin, Bangladesh.
In this study, we analyzed 33 groundwater samples from the Barapukuria coal basin (BCB), Bangladesh for 10 trace metals (TMs) using Atomic Absorption Spectroscopy. Pathways and associated probable health risk were appraised by employing multivariate statistical approaches, health risk model and Monte-Carlo simulation. Except for the Cu, Cr and Zn concentrations, the mean concentrations of all TMs in the basin were above the permissible water quality limits set by Bangladesh and international standards. Correlation coefficient and principal component analysis, supported by cluster analysis indicated that anthropogenic inputs were more contributed to the elevated concentrations of TMs compared to geogenic sources as the major reasons of groundwater pollution in the basin. The results of non-carcinogenic risk appraisal depicted that hazard index (HI) values for both adults and children were exceeded the safe limits (>1.0) except for few locations, indicating serious health risks on the human via oral and dermal absorption pathways. However, the carcinogenic risk values of Cd and Cr exceeded the US EPA range of 1 × 10-6 to 1 × 10-4, with higher risk for children than adults, with oral intake as the key exposure pathway. A sensitivity study identified the concentration of Cr, exposure frequency and ingestion rate for carcinogenic effect as the most sensitive parameters influencing the probable health risk. Overall, the results suggest that Cr in drinking water could cause detrimental effects to exposed local residents; thus, strict health regulation and groundwater management should concentrate on Cr contamination in groundwater from the coal basin.


Gut sensing of potassium intake and its role in potassium homeostasis.
Extracellular K(+) homeostasis has been explained by feedback mechanisms in which changes in extracellular K(+) concentration drive renal K(+) excretion directly or indirectly via stimulating aldosterone secretion. However, this cannot explain meal-induced kaliuresis, which often occurs without increases in plasma K(+) or aldosterone concentrations. Recent studies have produced evidence supporting a feedforward control in which gut sensing of dietary K(+) increases renal K(+) excretion (and extrarenal K(+) uptake) independent of plasma K(+) concentrations, namely, a gut factor. This review focuses on these new findings and discusses the role of gut factor in acute and chronic regulation of extracellular K(+) as well as in the beneficial effects of high K(+) intake on the cardiovascular system.


[Ultrastructural analysis of the cortical substances of the adrenal glands of rats under conditions of bilateral subdiaphragmatic vagotomy].
Electron-microscopic study of zona fasciculata and zona reticularis of the adrenal cortex showed that 7 days after bilateral subphrenic vagotomy there occurred a diatation of the perinuclear space of adrenocorticocytes and endothelial cells; there was also a dilatation of the tubules in the smooth cytoplasmic reticulum, edema of the mitochondria, reduction of their cristae, and increase in the number and size of lipid drops. In 45 days there are encountered mitochondria with the beginning myelinization, lipid drops form accumulations, and electron-transparent vacuoles are revealed in them. The functional activity of adrenocorticocytes in zona fasciculata and zona reticularis of the adrenal cortex diminished in rats after vagotomy.


Interplay between rumen digestive disorders and diet-induced inflammation in dairy cattle.
In this review, an overview is provided on the current achievements regarding the interplay between rumen digestive disorders and diet-induced inflammation in dairy cattle. It starts with a review of factors favoring the disturbances in the rumen metabolism, which culminate with development of sub-acute rumen acidosis (SARA). The latter digestive disorder is often linked to greater metabolic stress of gastrointestinal (GI) microbiota and lowered fiber digestion, as well as with disruption of the barrier functions of the GI epithelia, which open the route of deleterious molecules to translocate from the GI lumen into the portal system. A model is suggested to illustrate the mechanisms of the involvement of digestive disorders in the disruption of the host's inner homeostasis leading to activation of acute phase response (APR). The latter is part of multifaceted innate immune and metabolic responses of the host. According to this model, endotoxin, its toxicity, and other metabolic compounds of microbial origin are regarded as important immunogenic components of GI tract, which when favored by disruption of host barriers triggers a systemic APR. Although the activation of an APR is viewed as a protective reaction aiming to reestablish the disturbed homeostasis, the presence of inflammatory state over long periods might be associated with negative consequences for the host. The review concludes that prolonged systemic inflammation can: (1) cause significant changes in the energy and lipid metabolism in different body tissues, (2) lead to the development of refractory states associated with immune suppression and increased susceptibility to various diseases, and (3) artificially increase host's requirements in energy and nutrients, lowering the efficiency of energy and feed use by the animal. The paper emphasizes the critical role that formulation of healthy diets plays for curbing down inflammation and enhancing metabolic health of dairy cows.


A fast, cheap, and easy protocol for celiac disease HLA haplotype typing using buccal swabs.
Celiac disease (CeD) is a complex autoimmune disorder characterized by intestinal immune-derived injury that develops in response to dietary gluten consumption. Human Leucocyte Antigen (HLA) complex haplotype typing is one of the main tests for CeD diagnosis, together with anti-endomysium and anti-transglutaminase autoantibody detection in blood and inflammation observation in the intestine, being the former mainly used for the initial discarding of the pathogenesis. Among the many types of HLA proteins, HLA-DQ2.5 and HLA-DQ8 are considered essential for CeD development. These receptors are only expressed when specific alleles are present, which can be accurately predicted by the presence of the tagging SNPs rs2187668 and rs7454108, respectively. Taking advantage of this premise, we present here an easy workflow to assess HLA genotyping in saliva by a quick and cheap isopropanol-ethanol precipitation-based DNA extraction method followed by the genotyping of two tagging SNPs for the most frequent CeD risk-associated HLA haplotypes. All the actual diagnostic methods for CeD are performed after acquisition of intestine biopsies or blood samples by invasive techniques. Therefore, the development of non-invasive methods would be of a great improvement and advantage for patients, especially children, as an alternative method for initial CeD screening.


Retracted: RAGE Regulating Vascular Remodeling in Diabetes by Regulating Mitochondrial Dynamics with JAK2/STAT3 Pathway.
[This retracts the article DOI: 10.1155/2022/2685648.].


Evaluation of approach to predict the contribution of multiple cytochrome P450s in drug metabolism using relative activity factor: effects of the differences in expression levels of NADPH-cytochrome P450 reductase and cytochrome b(5) in the expression system and the differences in the marker activities.
The concept of relative activity factor (RAF) to extrapolate data obtained with recombinant cytochrome P450(CYP)s to human liver microsomes has been proposed. To evaluate the approach to predict the contribution of multiple CYPs using RAF, we investigated the effects of the differences in the expression levels of NADPH-cytochrome P450 reductase (OR) and cytochrome b(5) (b(5)) in recombinant CYPs from baculovirus-infected insect cells and the differences in the marker activities. Because we previously clarified that azelastine, an antiallergy and antiasthmatic drug, is N-demethylated by CYP1A2, CYP2D6, and CYP3A4 in humans, the reaction was used as a model. For calculation of RAF, three lots of recombinant CYP1A2, CYP2D6, and CYP3A4 from baculovirus-infected insect cells with different expression levels of OR and b(5) were used. The OR/CYP ratios for recombinant CYP1A2, CYP2D6, and CYP3A4 were 3.9-4.8, 5.1-8.7, and 8.0-11.3, respectively. The b(5)/CYP ratio for recombinant CYP3A4 was 2.1-18.7. As marker activities, ethoxyresorufin O-deethylation and phenacetin O-deethylation for CYP1A2, bufuralol 1'-hydroxylation and debrisoquin 4-hydroxylation for CYP2D6, testosterone 6beta-hydroxylation and midazolam 1'-hydroxylation for CYP3A4 were compared. Our results indicated that the differences in the expression levels of OR and b(5) coexpressed in baculovirus-infected insect cells would not be a critical factor for the quantitative prediction using RAF. In addition, we confirmed that differences in the marker activities did not significantly affect the calculation of RAF values, when the marker activities are specific for a certain CYP isoform. It was suggested that the RAF approach using recombinant CYPs from baculovirus-infected insect cells coexpressing OR (and b(5) if required) could be valuable for the prediction of the contribution of each CYP in drug metabolism.


The evolution of metapopulation dynamics and the number of stem cells in intestinal crypts and other tissue structures in multicellular bodies.
Carcinogenesis is a process of somatic evolution. Previous models of stem and transient amplifying cells in epithelial proliferating units like colonic crypts showed that intermediate numbers of stem cells in a crypt should optimally prevent progression to cancer. If a stem cell population is too small, it is easy for a mutator mutation to drift to fixation. If it is too large, it is easy for selection to drive cell fitness enhancing carcinogenic mutations to fixation. Here, we show that a multiscale microsimulation, that captures both within-crypt and between-crypt evolutionary dynamics, leads to a different conclusion. Epithelial tissues are metapopulations of crypts. We measured time to initiation of a neoplasm, implemented as inactivation of both alleles of a tumor suppressor gene. In our model, time to initiation is dependent on the spread of mutator clones in the crypts. The proportion of selectively beneficial and deleterious mutations in somatic cells is unknown and so was explored with a parameter. When the majority of non-neutral mutations are deleterious, the fitness of mutator clones tends to decline. When crypts are maintained by few stem cells, intercrypt competition tends to remove crypts with fixed mutators. When there are many stem cells within a crypt, there is virtually no crypt turnover, but mutator clones are suppressed by within-crypt competition. If the majority of non-neutral mutations are beneficial to the clone, then these results are reversed and intermediate-sized crypts provide the most protection against initiation. These results highlight the need to understand the dynamics of turnover and the mechanisms that control homeostasis, both at the level of stem cells within proliferative units and at the tissue level of competing proliferative units. Determining the distribution of fitness effects of somatic mutations will also be crucial to understanding the dynamics of tumor initiation and progression.


Structural insight into the mechanism of synergistic autoinhibition of SAD kinases.
The SAD/BRSK kinases participate in various important life processes, including neural development, cell cycle and energy metabolism. Like other members of the AMPK family, SAD contains an N-terminal kinase domain followed by the characteristic UBA and KA1 domains. Here we identify a unique autoinhibitory sequence (AIS) in SAD kinases, which exerts autoregulation in cooperation with UBA. Structural studies of mouse SAD-A revealed that UBA binds to the kinase domain in a distinct mode and, more importantly, AIS nestles specifically into the KD-UBA junction. The cooperative action of AIS and UBA results in an 'αC-out' inactive kinase, which is conserved across species and essential for presynaptic vesicle clustering in C. elegans. In addition, the AIS, along with the KA1 domain, is indispensable for phospholipid binding. Taken together, these data suggest a model for synergistic autoinhibition and membrane activation of SAD kinases.


Screening for hypertension: results of the Munich Blood Pressure Program.
The Munich Blood Pressure Program (MBP) is a community hypertension control program for the metropolitan area of Munich. It tries to improve "incidental screening" in doctors' offices in the city through continuing education programs for physicians and auxiliary personnel and, in addition, emphasizes worksite screening to bring high-risk hard-to-reach groups under treatment and control. Blood pressure (BP) is measured at the worksite under standardized conditions. Those with values greater than or equal to 140 mm Hg systolic BP and/or greater than or equal to 90 mm Hg diastolic BP at the first visit have BPs measured again within 1-5 days. If their BP values are still elevated, they are referred to their family physicians for further assessment. At the same time, they are given a postage-paid card on which their doctors are asked to note the BP value measured in their offices. This postcard is returned to the MBP for evaluation. A computer-based reminder system informs all MBP participants at certain time intervals to have their BP checked. The text of the letters and the time interval between them depend on the most recent BP and treatment status data available to the MBP. In 1983, screenings were performed in seven firms. Participation rates of up to 57% were achieved for this entirely voluntary examination (3,948 participants). Age- and sex-specific prevalence rates (for the pooled data of the seven firms) were similar to those of a random sample of the Munich population (Munich Blood Pressure Study I). Seventy-four percent of those with elevated BP at first visit participated in the remeasurement at a second visit. Only 58% of these showed elevated BP values the second time. More than half (n = 101) of those second-visit hypertensives (greater than or equal to 160/95 mm Hg; n = 183) returned at least one completed postcard. The examining physician confirmed the hypertensive BP values in 60 of those cases. Of the 101, 57 were under treatment, 19 controlled and 38 uncontrolled. The follow-up period considered here lasted until May 1984 and varied from 5 to 15 months according to the date of screening in the respective firm. Only the first returned card has been evaluated thus far.


Can follow-up radiography for acute scaphoid fracture still be considered a valid investigation?
To determine whether follow-up radiography is a valid diagnostic investigation in patients with suspected acute scaphoid fractures and normal initial radiographs. Fifty sets of radiographs (initial and follow-up) were independently assessed by four expert observers for the presence or absence of a scaphoid fracture. Magnetic resonance imaging (MRI), performed in all cases, was used as the gold standard to determine the sensitivity, specificity, positive and negative predictive values of the observers' assessment of the follow-up radiograph. In addition, the reliability among observers of the follow-up radiograph was determined by reliability variance analysis. Of the 50 sets of radiographs, 35 patients had a scaphoid fracture and 15 were normal, as established from MRI report. For individual observer's assessment of the follow-up radiograph, sensitivities of 11, 9, 43 and 49%; specificities of 93, 93, 87 and 80%; positive predictive values of 80, 75, 88 and 85%; and negative predictive values of 31, 30, 39 and 40%. A reliability coefficient of more than 60% is needed for a diagnostic test to be considered reliable. Overall, the inter-observer reliability coefficient was 33%, with pair-wise inter-observer coefficients ranging from 18-53%. With poor sensitivity, poor negative predictive value and poor reliability, follow-up radiography cannot be considered a valid diagnostic examination for the detection of scaphoid fracture in patients with normal initial radiographs.


Functional analysis of synthetic DELLA domain peptides and bioactive gibberellin assay using surface plasmon resonance technology.
DELLA proteins and phytohormone gibberellin act together to control convergence point of plant development. A gibberellin-bound nuclear receptor that interacts with the N-terminal domain of DELLA proteins is required for gibberellin induced degradation of DELLA proteins. N-terminal DELLA domain includes two conserved motifs: DELLA and VHYNP. However, their respective functions remain unclear. Meanwhile, the identification and detection of several bioactive gibberellins from the more than 100 gibberellin metabolites are overwhelmingly difficult for their similar structures. Using in vitro biochemical approach, our work demonstrates for the first time that the synthetic GAI N-terminal DELLA domain peptides have similar bioactive function as the expressed protein to interact with AtGID1a receptor. Furthermore, our results reveal that DELLA motif is vitally important region and DELLA segment is essentially required region to recognize AtGID1a receptor. Finally, based on bioactive GA-dependent of the interaction between AtGID1a and DELLA protein, we generated a new method that could identify and detect bioactive GAs accurately and rapidly with surface plasmon resonance assays.


Dopamine transporter is enriched in filopodia and induces filopodia formation.
Dopamine transporter (DAT, SLC6A3) controls dopamine (DA) neurotransmission by mediating re-uptake of extracellular DA into DA neurons. DA uptake depends on the amount of DAT at the cell surface, and is therefore regulated by DAT subcellular distribution. Hence we used spinning disk confocal microscopy to demonstrate DAT localization in membrane protrusions that contained filamentous actin and myosin X (MyoX), a molecular motor located in filopodia tips, thus confirming that these protrusions are filopodia. DAT was enriched in filopodia. In contrast, R60A and W63A DAT mutants with disrupted outward-facing conformation were not accumulated in filopodia, suggesting that this conformation is necessary for DAT filopodia targeting. Three independent approaches of filopodia counting showed that DAT expression leads to an increase in the number of filopodia per cell, indicating that DAT can induce filopodia formation. Depletion of MyoX by RNA interference resulted in a significant loss of filopodia but did not completely eliminate filopodia, implying that DAT-enriched filopodia can be formed without MyoX. In cultured postnatal DA neurons MyoX was mainly localized to growth cones that displayed highly dynamic DAT-containing filopodia. We hypothesize that the concave shape of the DAT molecule functions as the targeting determinant for DAT accumulation in outward-curved membrane domains, and may also allow high local concentrations of DAT to induce an outward membrane bending. Such targeting and membrane remodeling capacities may be part of the mechanism responsible for DAT enrichment in the filopodia and its targeting to the axonal processes of DA neurons.


Increased CSF amino acids and ventricular enlargement in schizophrenia: a preliminary study.
We found significantly higher levels of cerebrospinal fluid alanine, glycine, leucine, and phenylalanine in schizophrenic patients compared to healthy controls. Ventricular enlargement was present in 4 of 11 schizophrenics, and elevated CSF alanine was highly correlated with ventricular enlargement. The implications of these findings are discussed.


Cost-utility of sentinel lymph node biopsy in cT1-T2N0 oral cancer.
To calculate the cost-utility of different strategies for the detection of occult lymph node metastases in cT1-T2N0 oral cancer. A decision tree followed by a Markov model was designed to compare the cost-utility of the following strategies: (a) USgFNAC (ultrasound guided fine needle aspiration cytology), (b) SLNB (sentinel lymph node biopsy), (c) USgFNAC and, if negative, SLNB (d) END (elective neck dissection). Data was collected from 62 patients in four Dutch head and neck centres. Utilities were measured with the EQ5D questionnaire and resource use was recorded from patient charts. Costs were calculated from a hospital perspective. Uncertainty was explored with scenario analyses and probabilistic sensitivity analyses. With a 5- or 10-year time horizon, SLNB results in the highest number of additional quality-adjusted life years (QALYs, 0.12 and 0.26, respectively) for the smallest additional costs (€56 and €74, respectively) compared to USgFNAC. With a lifetime horizon END results in the highest number of additional QALYs (0.55) for an additional €1.626 per QALY gained compared to USgFNAC. When we make different assumptions regarding the duration of disutilities (⩾5 years) or the improvement (⩾3%) of sensitivity of SLNB, SLNB is the most favourable strategy from all time horizons. SLNB is a good diagnostic strategy to evaluate cT1-T2N0 oral cancer. SLNB is the preferred strategy in a 5- or 10-year time horizon. From a lifetime horizon, END may be preferred. SLNB may become the optimal strategy from all time horizons if its sensitivity can be slightly improved.


The extent of mercury (Hg) exposure among Saudi mothers and their respective infants.
A total of 1016 healthy Saudi mothers and their respective infants (aged 3-12 months) were recruited from 57 Primary Health Care Centers (PHCCs) in Riyadh, Saudi Arabia, to evaluate the extent of mercury (Hg) exposure and predict its sources in the healthy Saudi population. Total Hg levels were measured in maternal urine, breast milk, blood, and hair and in the infants' urine and hair. Only 1.9% of the mothers had urinary Hg (UHg)>10 μg/l, the limit for asymptomatic adults recommended by the World Health Organization, but the median (0.99 μg/l) was higher than in other countries. Also, 49.3% of the mothers had UHg>1 μg/l, the German reference value for adults. Median infant UHg was 0.729 μg/l, and 77 and 93 % of the infants had levels higher than 0.4 and 0.1 μg/l, the reference values of the Centers for Disease Control and Prevention and for Germany, respectively. The median Hg level in breast milk was 0.884 μg/l. Even though 43.2% of the milk samples were above the background level for Hg in human milk (1 μg/l), our results were lower than those reported from other countries. Median maternal total Hg in blood was 0.637 μg/l, and only 0.4 and 6.9% of samples were higher than the Hg reference levels of 5.8 μg/l of the Environmental Protection Agency (EPA) and of 2 μg/l for Germany, respectively. Total Hg levels in hair (HHg) varied widely among mothers and infants, but only 3.9% of the mothers and 2.8% of the infants had HHg>1 μg/g (the EPA reference level). Median HHg values were 0.117 μg/g dry weight in mothers and 0.1 μg/g dry weight in infants; both were lower than in other countries. The Hg levels in mothers and their respective infants were relatively low, but our results were consistent with other studies indicating that dental amalgam fillings and fish consumption were the main predictors of maternal Hg exposure. Among the several biomarkers of Hg exposure, Hg levels in maternal hair and urine were the strongest predictors of infant exposure. The lack of an association between Hg in breast milk and Hg in infant urine and hair suggested that the infants were exposed to Hg predominately during pregnancy rather than during breastfeeding. We expect that our data can serve as a baseline for further biomonitoring and follow-up studies, particularly of the long-term impact of Hg on childhood neurodevelopment.


The Influence of Oligosaccharides when Measuring Lactose and Total Carbohydrates in Human Milk and Comparison of Methods.
Understanding how human milk impacts growth requires valid analytical methods for quantifying the composition. Lactose, the most abundant constituent in human milk and a predominant source of energy, is often assessed using methods borrowed from the bovine dairy industry. However, the carbohydrate matrices of bovine and human milk are quite different, especially as they relate to human milk oligosaccharides (HMOs), each with a terminal lactose unit that may influence analytical methods. Our goals were to determine the extent to which HMOs influence common analytical methods for measuring carbohydrates in human milk and to compare common methods for measuring lactose. Two sets of experiments were performed. In the first set, native and HMO-spiked human milk samples (n = 16 each) were assessed and compared using 4 methods: AOAC 2006.06 (based on the Megazyme enzymatic assay), BioVision enzymatic assay, ultraperformance LC with MS, and infrared analysis. In the second set, human milk samples (n = 20) were assessed using 2 methods approved for measuring lactose in bovine milk: AOAC 984.22 that uses high-performance LC and refractive index detection and AOAC 2006.06 prepared using both volume and weighted dilutions. Native and HMO-spiked samples were not significantly different in lactose using AOAC 2006.06 and ultraperformance LC with MS but were significantly different using BioVision (mean difference = 0.2 g/dL; 95% CI: 0.1, 0.4; P = 0.005). Total carbohydrate measurements assessed using infrared were also higher after HMO spiking (mean difference = 0.4 g/dL; 95% CI: 0.3, 0.6; P < 0.001). Only AOAC methods 984.22 and 2006.06 for measuring lactose were very highly correlated (r > 0.90, P < 0.001). AOAC methods 984.22 and 2006.06 are comparable for measuring lactose in human milk and are not influenced by HMOs. HMOs influence other enzymatic methods as well as infrared analysis, which leads to an overestimate of energy values. J Nutr 2023;x:xx.


The slippage of the Ca2+ pump and its control by anions and curcumin in skeletal and cardiac sarcoplasmic reticulum.
Ca(2+) transport by sarcoplasmic reticulum (SR) ATPase occurs with an optimal coupling ratio of 2 Ca(2+) per ATP in pre-steady state. However, slippage of the pump and lower coupling ratios are observed in steady state. Slippage depends on the presence of high Ca(2+) in the lumen of SR vesicles and high nucleotide in the medium. Thereby, Ca(2+) and/or nucleotide-bound phosphoenzyme intermediates accumulate and undergo uncoupled cleavage, before vectorial translocation of bound Ca(2+) in the forward direction of the cycle or before productive reversal to ATP synthesis. Transport efficiency and coupling ratios are improved by reduction of nucleotide concentration in the presence of ATP regenerating systems and/or complexation of luminal Ca(2+) with phosphate or oxalate. Curcumin (1-5 microm) lowers the concentration of phosphate or oxalate required to reduce slippage of the Ca(2+) pump. Thereby, under appropriate conditions, curcumin favors kinetic flow, completion of productive cycles, and improvement of coupling ratios. The findings obtained with isolated SR vesicles suggest that slippage is an important phenomenon under prevailing conditions of muscle fibers in situ. Ca(2+) transport and its slippage can be improved by curcumin in cardiac as well as in skeletal SR, raising the possibility of pharmacological interventions to correct defective Ca(2+) homeostasis. Higher curcumin concentrations (5-30 microm), however, inhibit overall ATPase activity and Ca(2+) transport by interfering with phosphoenzyme formation with ATP or P(i).


A bent-tipped endolaser probe.
I have developed a bent-tipped probe for endolaser photocoagulation that facilitates treatment near the entry site of the instrument.


Spin-resolved charge displacement analysis as an intuitive tool for the evaluation of cPCET and HAT scenarios.
We introduce here the spin-resolved version of the charge displacement function, which is applied to two competing pathways of proton-coupled electron transfer in oxidation catalysis (hydrogen-atom transfer, concerted proton-coupled electron transfer). The difference in charge displacement between the two mechanisms is directly observable and can be translated to electron flow using this new analysis tool.


Chronic lymphocytic leukemia: study of cell subsets in cerebrospinal fluid and peripheral blood.
Lymphocytic cells in a case of leptomeningeal involvement were studied by immunological methods during chronic lymphocytic leukemia. Conspicuous differences were found between blood and cerebrospinal fluid.


The effect of a health extension program on improving water, sanitation, and hygiene practices in rural Ethiopia.
To make basic primary health care services accessible, especially to the rural community, the government of Ethiopia launched the Health Extension Program (HEP) in 2004. Most of components of HEP are dedicated to hygiene and sanitation. Few studies have assessed the role of the Health Extension Program in improving water, hygiene, and sanitation (WASH) practices in Ethiopia. This study explored the role of health extension workers (HEWs) in influencing household water treatment practices, latrine ownership, latrine use and ownership, and the use of hand-washing facilities on the incidence of diarrheal diseases among the children under five years of age in rural Ethiopia. Using a cross sectional design, we conducted a national assessment that covered all nine regions of Ethiopia. We conducted face-to-face interviews among a sample of 6430 rural households using a structured questionnaire and an observation checklist to collect data from March 2018 to May 2019. Multilevel logistic regressions models were used to determine the relationships between the exposure of households to HEWs and WASH practice outcomes such as the use of water from an improved water source, household water treatment practices, availability of hand-washing and hand-washing with soap and water, availability of latrines, and use of latrines as well as the incidence of diarrheal diseases among children age 5 and younger. Our models were adjusted for covariates and confounders and P-values less than 5% were set to determine statistical significance. We found that 72.7% of rural households had some type of latrine and 27.3% reported practicing open defecation. A total of 71.5% of rural households had access to drinking water from improved water sources, but only 9.4% reported practicing household water treatment. Exposure to HEWs was positively associated with household water treatment practices (AOR: 1.46; 95% CI = 1.01-2.10) and latrine availability (AOR: 1.44; 95% CI = 1.15-1.80). Among the households who were either visited by HEWs at their home or the that visited health posts to meet with the HEWs, being exposed to WASH health education by HEWs was significantly associated with the availability of a hand-washing facility (AOR: 5.14; 95% CI = 4.11-6.42) and latrine availability (AOR: 1.48; 95% CI = 1.10-2.01). However, we did not find a relationship between the incidence of diarrhea among children age 5 and under and exposure to HEWs (AOR: 2.09; 95% CI = 0.73- 6.62). Our results show a significant association between exposure to the Health Extension Program/ HEWs and improved household water treatment practices, latrine construction, and the availability of hand-washing facilities in rural Ethiopia, suggesting the need to strengthen efforts to change WASH behavior through the Heath Extension Program. On the other hand, further investigation is needed regarding the spillover effect of latrine use practices and the reduction of the incidence of diarrheal diseases.


Partitioning of amino acids and nucleotides between water and micellar hexadecyltrimethylammonium halides. The prebiotic significance of cationic surfaces.
Using quantitative gel filtration techniques partition coefficients, Kp-values, have been determined between aqueous cationic micellar hexadecyltrimethylammonium bromide, CTAB, and several biomonomer. Kp-values for 5'-adenylic acid, 5'-cytidylic acid, 5'-guanylic acid, 5'-uridylic acid and 5'-thymidylic acid are 1,400 +/- 150. Nucleotides bind to CTAB micelles effectively, but nonselectively. Conversely, the binding of tRNAs to micellar CTAB is selective. Kp-values for glutamic acid II, tyrosine and phenylalanine tRNAs (in 1.0MNaCl) are 520, 3,100 and 5,600, respectively. Kp-values for the binding of alanine, arginine, aspartic acid, glutamic acid, glycine, histidine, phenylalanine, serine, threonine and tryptophan to micellar CTAB are less than 8. Conversion of unitless Kp-values for the binding of amino acids, nucleotides and nucleosides to both anionic and cationic micelles, to K (in 1/g) values allows the comparison of clays and micelles as prebiotic concentrating media. Using correlations between surface densities of the biomonomers and their binding constants, it is shown that aqueous micelles (at pH = 8) are a better concentrating media than are clays.


The effects of oral and transdermal hormone replacement therapy on C-reactive protein levels and other inflammatory markers in women with high risk of thrombosis.
In the estrogen in venous thromboembolism (EVTET) study of 140 women with a history of venous thromboembolism (VTE), oral hormone replacement therapy (HRT) was associated with strong activation of coagulation markers and increased risk of recurrent VTE. No such associations were observed in the estrogen women atherosclerosis (EWA) study of 118 women with established coronary artery disease who were given transdermal HRT. The aim of the present study was to evaluate the effects of oral and transdermal HRT on levels of C-reactive protein (CRP), which was assayed with a highly sensitive method. We also evaluated the effects on other inflammatory markers and the influence of possible confounding factors. Oral HRT was associated with a significant increase in CRP after 3 months as compared with placebo (median 79% [95% confidence interval 36-119%] versus -4% [-13 to 10%], p = 0.001). These changes sustained after 12 months. Among those allocated HRT, the median increase in CRP was higher in women who subsequently developed recurrent thrombosis (median 328%, n = 5, versus 54%, n = 60). TNF-alpha levels decreased significantly by mean -10% [-15 to -5%] versus 3% [-4 to 10%], p=0.004. Soluble VCAM-1 decreased in the HRT group compared to the placebo group (mean -13% [-18 to -8%] versus 1%, [-3 to 5%], p < 0.001). There were no significant changes in levels of IL-6, TGF-beta or P-selectin. On transdermal HRT no significant changes in CRP were observed after 3 and 12 months of treatment. Our findings substantiate that oral HRT containing estradiol is associated with a marked and rapid increase in CRP, whereas transdermal treatment is not. However, this increase on oral treatment was associated with no increases of other inflammatory markers.


Direct visualization of ligand-protein interactions using atomic force microscopy.
1. Streptavidin is a 60-kDa tetramer which binds four molecules of biotin with extremely high affinity (K(A) approximately 10(14) M(-1)). We have used atomic force microscopy (AFM) to visualize this ligand-protein interaction directly. 2. Biotin was tagged with a short (152-basepair; 50-nm) DNA rod and incubated with streptavidin. The resulting complexes were then imaged by AFM. The molecular volume of streptavidin calculated from the dimensions of the protein particles (105+/-3 nm(3)) was in close agreement with the value calculated from its molecular mass (114 nm(3)). Biotinylation increased the apparent size of streptavidin (to 133+/-2 nm(3)), concomitant with an increase in the thermal stability of the tetramer. 3. Images of streptavidin with one to four molecules of DNA-biotin bound were obtained. When two ligands were bound, the angle between the DNA rods was either acute or obtuse, as expected from the relative orientations of the biotin binding sites. The ratio of acute : obtuse angles (1 : 3) was lower than the expected value (1 : 2), indicating a degree of steric hindrance in the binding of the DNA-biotin. The slight under-representation of higher occupancy states supported this idea. 4. Streptavidin with a single molecule of DNA-biotin bound was used to tag biotinylated beta-galactosidase, a model multimeric enzyme. 5. The ability to image directly the binding of a ligand to its protein target by AFM provides useful information about the nature of the interaction, and about the effect of complex formation on the structure of the protein. Furthermore, the use of DNA-biotin/streptavidin tags could potentially shed light on the architecture of multi-subunit proteins.


The effect of carbodiimide on push-out bond strength of fiber posts and endogenous enzymatic activity.
To investigate the effect of 0.3 M 1-ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC) aqueous solution pretreatment on push-out bond strength (PBS) and matrix-metalloproteinases (MMPs) activity within radicular dentin when different post cementation strategies were employed. One hundred and twenty monoradicular human teeth were endodontically treated and randomly divided into six groups, depending on the cementation strategy and root dentin pretreatment (n = 20): EAR: cementation with an etch-and-rinse adhesive (LuxaBond Total Etch, DMG) and resin cement (LuxaCore Z Dual, DMG); EAR/EDC: 1 min EDC pretreatment after etching + EAR; SE: cementation with a self-etch primer (Multilink Primer, Ivoclar Vivadent) and corresponding cement (Multilink Automix, Ivoclar Vivadent); SE/EDC: self-etch primer + EDC pretreatment + SE; SA: cementation with a universal self-adhesive cement (RelyX Universal, 3 M); SA/EDC: EDC pretreatment + SA. Slices were submitted to PBS test and interfacial nanoleakage evaluation 24 h after cementation or after thermocycling (40.000 cycles, 5-55 °C). To investigate the effect of EDC on MMPs activity, 4 additional first maxillary premolars per group were processed for in situ zymography analysis. Multivariate ANOVA and post hoc Tukey tests were used to analyze PBS values. The data from in situ zymography were analyzed with Kruskal-Wallis test and Dunn's pairwise multiple comparison procedures (α = 0.05). The variables "EDC pretreatment", "root region" and "thermocycling" significantly influenced PBS (p < 0.05), while the variable "cementation strategy" had no influence (p > 0.05). Thermocycling significantly reduced PBS in SE and SA groups (p < 0.05). EDC was effective in preserving PBS after artificial aging. EDC pretreatment significantly reduced enzymatic activity at baseline in EAR and SE groups, and in SA group after thermocycling (p < 0.05). The use of EDC prevents the reduction of bond-strength values after artificial aging and silences endogenous enzymatic activity within radicular dentin when different cementation strategies were employed.


Jejunal lymphangioma causing intussusception in an adult: An unusual case with review of the literature.
Adult intussusception is rare, and 90% are due to a lead point secondary to a pathologic condition. Lymphangioma is an uncommon tumor of the lymphatic system and is rarely found within the small bowel. Small bowel lymphangioma causing intussuception in an adult is a rare occurrence, with three very distinct rare pathologies occurring simultaneously. A 70-year-old male patient with multiple pre-existing pathologies such as advanced ADPKD, multiple persistent tubulovillous colon polyps and colon cancer in situ, was hospitalized due to rapid weight loss of 20 lbs, hematemesis, and abdominal pain. He was subsequently found to have jejunal intussusception caused by two lymphangiomas of the small bowel. The portion of intussuscepted jejunum was resected and final diagnosis on pathology was two jejunal lymphangiomas. Lymphangiomas of the small bowel are rare, but increasing in incidence due to the accessibility of endoscopic evaluations. A hypothetical connection between lymphangioma and ADPKD is unknown, but both diseases are built on a foundation of cystogenesis. There is little known about the effect ADPKD on cystogenesis and tumor formation extra-renally, and there is a possible correlation between genetic mutations in polycystin and cystic tumors such as lymphangiomas. Lymphangioma, although rare in the small bowel, is a possible cause of intussusception and should be considered on the differential of abdominal pain in adults. The pathogenesis of polycystic kidney disease has implications that could predispose to cystic development beyond the kidney, and more research into the genetic mechanism behind the disease is necessary to support or deny this claim.


A new system for substaging pT1 papillary bladder cancer: a prognostic evaluation.
Superficially invasive (pT1) papillary urothelial cell carcinomas (UCCs) may run a variable course. Several attempts have been made for the substaging of UCC to identify the clinically aggressive tumors. We present a new substaging system, based on the extent of invasion. From a series of 53 primary pT1 UCC, 24 cases showed invasion of the subepithelial stroma by an invasive front extending more than a maximum length of 0.5 mm (pT1mic), and 29 showed extensively (>0.5 mm) infiltrating UCC (pT1ext). We tested diagnostic reproducibility between 2 pathologists and found 81% agreement. Furthermore, all cases were analyzed for mutations in the fibroblast growth factor receptor 3 (FGFR3) gene, which represents the favorable pathway of urothelial cell carcinogenesis. Mutant FGFR3 was commonly observed in pT1mic UCC (15/24, 63%), but rarely (2/29, 7%) in pT1ext UCC (chi2 test, P < .001). The presence of pT1ext at initial diagnosis proved to be the strongest predictor for progression, also when adjusted for FGFR3 mutation status in a Cox regression model. If confirmed on a larger series of pT1 UCC, this relatively simple and new substaging system for pT1 UCC may prove to be of prognostic value and supportive to clinical decision-making.


A mutation in the GABAA receptor alpha 1 subunit linked to human epilepsy affects channel gating properties.
A genetic component is associated with the development of many forms of epilepsy. Recently, mutations in the GABAA receptor have been linked to several inherited epilepsies. One of these mutations is a non-conservative change of alanine to aspartate in the third transmembrane domain of the alpha1 subunit. To determine the functional consequences of this alteration, mutated alpha subunits were transiently transfected along with wild-type beta3 and gamma2L subunits into HEK-293T cells. The mutated alpha1(A294D) subunit reduced GABA sensitivity of the receptor, increased the deactivation rate and slowed desensitization. The mutation caused a reduction in channel open time but no change in single channel conductance. Studies with additional mutants, altering the charge and/or size of the side-chain, indicated that both size and hydrophobicity of the residue at this location influence channel gating. The effects on GABA sensitivity, deactivation rate and channel open time are consistent with a reduced efficacy of channel gating, and would be expected to decrease GABAergic neurotransmission. The alpha1 subtype is the most widely expressed of the alpha subunits, with expression increasing throughout development. Therefore, production of the mutated subunit could cause global hyperexcitability throughout the brain, leading to generalized seizures with juvenile onset.