A bifunctional enzyme complex in coenzyme A biosynthesis: purification of pantetheine phosphate adenylyltransferase and dephospho-CoA kinase.
Pantetheine phosphate adenylyltransferase (EC 188.8.131.52) and dephospho-CoA kinase (EC 188.8.131.52) were purified to near homogeneity from pig liver. The purification steps included the use of Sepharose-linked triazine dyes and affinity elution by CoA. Both activities co-purified at every stage of the 18 000-fold purification. An Mr of 115 000 was obtained by gel filtration on Sephadex G-150, and the final preparation yielded one major band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, with a subunit Mr of 57 000. It is concluded that pantetheine phosphate adenylyltransferase and dephospho-CoA kinase exist as a bifunctional dimeric protein, which could be designated CoA synthetase.


Glomerular and renal vascular structural changes in alpha8 integrin-deficient mice.
Integrins are matrix receptors that regulate cell-matrix interactions during development and in adult tissue. In the adult kidney, the alpha8 chain is specifically expressed in glomerular mesangial cells and vascular smooth muscle cells. alpha8-deficient (alpha8-/-) mice demonstrate reductions in renal mass, which can range from complete renal agenesis to the development of kidneys that are only slightly smaller than wild-type kidneys. No histologic abnormalities of these kidneys have been described. However, considering the prominent expression of alpha8 in glomeruli and renal vessels, it seemed unlikely that the kidneys of alpha8-/- mice would be completely normal. Therefore, the renal phenotype of adult alpha8-/- mice was investigated, for assessment of more subtle morphologic alterations in kidney tissue. alpha8-/- mice displayed a significant reduction in nephron number and an increase in glomerular volume, compared with wild-type control animals. Albuminuria was not different in wild-type and alpha8-/- mice. Quantitative morphologic analyses revealed that the glomeruli of alpha8-/- mice were hypercellular, with an increased number of mesangial cells, compared with wild-type mice. Mesangial matrix deposition (as demonstrated for collagen IV and the alpha8 ligand fibronectin) was expanded in alpha8-/- mice, compared with wild-type mice. Collagens I and III, which are not normally present in glomeruli, were detected in the glomeruli of alpha8-/- mice. Staining for other glomerular integrins demonstrated an increased abundance of the collagen receptor alpha2 integrin in alpha8-/- mice. The glomerular capillary length density was significantly greater in alpha8-/- mice than in wild-type mice. Cortical arterial vessel walls were not altered in alpha8-/- mice, but the capillaries of the peritubular network were widened. Despite the strong mesangial and vascular expression of alpha8, glomerular and renal vascular alterations in alpha8-/- mice were relatively mild. Only aged alpha8-/- mice demonstrated increased glomerular capillary widening, compared with control animals. The results suggest that the lack of alpha8 can be largely compensated for, at least in younger alpha8-/- mice. It is not yet clear whether the occurrence of collagens that are not normally present in glomeruli and the increased abundance of the collagen receptor alpha2 contribute to maintaining the glomerular structure in alpha8-/- mice. The compensatory mechanisms involved will be the subject of future research.


[Thymoma with Hemorrhage and Necrosis Presenting with Fever and Chest Pain;Report of a Case].
A 53-year-old man was admitted to our hospital for treatment of fever and chest pain. Chest computed tomography showed an anterior mediastinal cystic tumor 39×57 mm in size surrounded by fat with edema and left pleural effusion. After one course of antibiotic administration, the edema of fat surrounding the tumor disappeared and the patient underwent scheduled tumor resection with thymectomy through a median sternotomy. Postoperative pathological examination revealed a thymoma of type AB according to the World Health Organization (WHO) classification associated with hemorrhage and necrosis. Cases of thymoma with hemorrhage or necrosis may lead to atypical presentations such as fever, acute chest pain, pleural effusion, and spontaneous regression. Clinicians should be aware of these unusual presentations of chest pain and fever due to thymoma, and consider the possibility of a differential diagnosis of an anterior mediastinal tumor.


Experimental cancer cachexia: the role of host-derived cytokines interleukin (IL)-6, IL-12, interferon-gamma, and tumor necrosis factor alpha evaluated in gene knockout, tumor-bearing mice on C57 Bl background and eicosanoid-dependent cachexia.
MCG 101 tumors were implanted sc. on wild-type C57 Bl and gene knockout mice to evaluate the role of host-produced cytokines [interleukin (IL)-6, IL-12, IFNgamma, tumor necrosis factor (TNF) receptor 1, and TNF receptor 2] to explain local tumor growth, anorexia, and carcass weight loss in a well-defined model with experimental cachexia. Indomethacin was provided in the drinking water to explore interactions between host and tumor-derived prostaglandins and proinflammatory cytokines for tumor growth. Wild-type tumor-bearing mice developed cachexia because of rapid tumor growth, which were both attenuated in IL-6 gene knockouts. Similar findings were observed after provision of anti-IL-6 to wild-type tumor-bearing mice. Alterations in food intake were not directly related to systemic IL-6 but rather secondarily to IL-6-dependent tumor growth. The absence of host-derived IL-12, IFN-gamma, or the TNF receptor 1 or receptor 2 gene did not attenuate tumor growth or improve subsequent cachexia. Thus, carcass weight loss was not improved by the omission of host cytokine (TNF-alpha, IL-12, or IFN-gamma) except for IL-6. Systemic indomethacin provision decreased plasma prostaglandin E2 in five of six groups of gene knockout tumor-bearing mice, which was associated with improved carcass weight in these groups. Indomethacin seemed to improve food intake to a similar extent in both wild-type and gene knockouts, which agree with the speculation that eicosanoids are more important to explain anorexia than host cytokines. Our results demonstrate that host- and tumor-derived cytokines and prostaglandins interact with tumor growth and promote cachexia in a more complex fashion than usually presented based on previous information in studies on either anti-cytokine experiments in vivo or on gene knockouts with respect to a "single cytokine model." Overall, host cytokines were quantitatively less important than tumor-derived cytokines to explain net tumor growth, which indirectly explains subsequent cachexia and anorexia.


A wasp manipulates neuronal activity in the sub-esophageal ganglion to decrease the drive for walking in its cockroach prey.
The parasitoid Jewel Wasp hunts cockroaches to serve as a live food supply for its offspring. The wasp stings the cockroach in the head and delivers a cocktail of neurotoxins directly inside the prey's cerebral ganglia. Although not paralyzed, the stung cockroach becomes a living yet docile 'zombie', incapable of self-initiating spontaneous or evoked walking. We show here that such neuro-chemical manipulation can be attributed to decreased neuronal activity in a small region of the cockroach cerebral nervous system, the sub-esophageal ganglion (SEG). A decrease in descending permissive inputs from this ganglion to thoracic central pattern generators decreases the propensity for walking-related behaviors. We have used behavioral, neuro-pharmacological and electrophysiological methods to show that: (1) Surgically removing the cockroach SEG prior to wasp stinging prolongs the duration of the sting 5-fold, suggesting that the wasp actively targets the SEG during the stinging sequence; (2) injecting a sodium channel blocker, procaine, into the SEG of non-stung cockroaches reversibly decreases spontaneous and evoked walking, suggesting that the SEG plays an important role in the up-regulation of locomotion; (3) artificial focal injection of crude milked venom into the SEG of non-stung cockroaches decreases spontaneous and evoked walking, as seen with naturally-stung cockroaches; and (4) spontaneous and evoked neuronal spiking activity in the SEG, recorded with an extracellular bipolar microelectrode, is markedly decreased in stung cockroaches versus non-stung controls. We have identified the neuronal substrate responsible for the venom-induced manipulation of the cockroach's drive for walking. Our data strongly support previous findings suggesting a critical and permissive role for the SEG in the regulation of locomotion in insects. By injecting a venom cocktail directly into the SEG, the parasitoid Jewel Wasp selectively manipulates the cockroach's motivation to initiate walking without interfering with other non-related behaviors.


Small players with big roles: microRNAs as targets to inhibit breast cancer progression.
As modulators of gene expression, microRNAs (miRNAs) are essential for normal development. Not surprisingly, aberrant expression of miRNAs is associated with many diseases, including cancer. Studies of various breast cancer subtypes have demonstrated that, like gene expression profiles and pathological differences, miRNA profiles can distinguish various tumor subtypes. Over the last few years, roles for miRNAs during many stages of breast cancer progression have been established. This includes potential breast cancer associated polymorphisms in miRNA target sites or miRNAs themselves, miRNAs that can act as tumor suppressors or oncogenes, and miRNAs that can modulate metastatic spread. Recent studies have also suggested key roles for miRNAs in regulating cancer stem cells. Thus, miRNAs have now become important therapeutic targets. This can be achieved by replacing miRNA expression where it has been lost or decreased, or conversely by inhibiting miRNA expression where it has been amplified or overexpressed in cancers. Ultimately, miRNAs should provide both important prognostic biomarkers as well as new targetable molecules for the treatment of breast cancer.


MicroRNA-138 is a potential regulator of memory performance in humans.
Genetic factors underlie a substantial proportion of individual differences in cognitive functions in humans, including processes related to episodic and working memory. While genetic association studies have proposed several candidate "memory genes," these currently explain only a minor fraction of the phenotypic variance. Here, we performed genome-wide screening on 13 episodic and working memory phenotypes in 1318 participants of the Berlin Aging Study II aged 60 years or older. The analyses highlight a number of novel single nucleotide polymorphisms (SNPs) associated with memory performance, including one located in a putative regulatory region of microRNA (miRNA) hsa-mir-138-5p (rs9882688, P-value = 7.8 × 10(-9)). Expression quantitative trait locus analyses on next-generation RNA-sequencing data revealed that rs9882688 genotypes show a significant correlation with the expression levels of this miRNA in 309 human lymphoblastoid cell lines (P-value = 5 × 10(-4)). In silico modeling of other top-ranking GWAS signals identified an additional memory-associated SNP in the 3' untranslated region (3' UTR) of DCP1B, a gene encoding a core component of the mRNA decapping complex in humans, predicted to interfere with hsa-mir-138-5p binding. This prediction was confirmed in vitro by luciferase assays showing differential binding of hsa-mir-138-5p to 3' UTR reporter constructs in two human cell lines (HEK293: P-value = 0.0470; SH-SY5Y: P-value = 0.0866). Finally, expression profiling of hsa-mir-138-5p and DCP1B mRNA in human post-mortem brain tissue revealed that both molecules are expressed simultaneously in frontal cortex and hippocampus, suggesting that the proposed interaction between hsa-mir-138-5p and DCP1B may also take place in vivo. In summary, by combining unbiased genome-wide screening with extensive in silico modeling, in vitro functional assays, and gene expression profiling, our study identified miRNA-138 as a potential molecular regulator of human memory function.


Head, neck, and facial injuries in ice hockey: the effect of protective equipment.
To determine the factors and behaviors associated with facial, head, and/or neck injuries to those engaged in ice hockey; the use of protective equipment was also examined. Prospective case series. Emergency Department of Sudbury General Hospital. All patients presenting to the emergency department with a head, neck, or facial injury while playing hockey between the months of October and March 1993-94 and 1994-95 were included in the study. Physicians completed data forms on each patient. Information was validated by review of the emergency-room records; further information was obtained by telephone. A total of 226 patients were identified with ice-hockey-related head, neck, or facial injuries. Most injuries involved males (99%), and the mean age of patients was 23.9 (range, 4-63). Injuries occurred most frequently to the face [192 (85%)]. Many of the injuries were minor, with a mean injury severity score of 1.5 (range, 1-25). However, three patients (1%) required hospital admission, and one teenager suffered a serious spinal fracture. Protective facial hockey equipment use was low in our sample, except among younger injured players. Most of the facial injuries occurred in mature athletes playing recreational hockey. Full facial protection reduced the chance of upper facial injury (p = 0.0001), but the risk of such injury while wearing a half-visor was the same as while wearing no facial protection at all (p > 0.05). From the current study, we estimate that these hockey injuries result in approximately 2.7-3.0 million dollars of direct acute-care medical expenditure per year in emergency departments throughout Ontario. Head, neck, and facial injuries suffered during ice hockey participation are common problems presented to emergency departments. Moreover, serious injuries can occur while playing this sport. Most injuries appear to be preventable, and facial protection appears to be less frequently used, especially by older men, than is currently recommended. Prevention strategies are discussed.


Genetic divergence of a sympatric lake-resident-anadromous three-spined stickleback Gasterosteus aculeatus species pair.
The genetic relationship between sympatric, morphologically divergent populations of anadromous and lake-resident three-spined stickleback Gasterosteus aculeatus in the Jim Creek drainage of Cook Inlet, Alaska, was examined using microsatellite loci and mitochondrial d-loop sequence data. Resident samples differed substantially from sympatric anadromous samples in the Jim Creek drainage with the magnitude of the genetic divergence being similar to that between allopatric resident and anadromous populations in other areas. Resident samples were genetically similar within the Jim Creek drainage, as were the anadromous samples surveyed. Neighbour-joining and Structure cluster analysis grouped the samples into four genetic clusters by ecomorph (anadromous v. all resident) and geographic location of the resident samples (Jim Creek, Mat-Su and Kenai). There was no evidence of hybridization between resident and anadromous G. aculeatus in the Jim Creek drainage, which thus appear to be reproductively isolated.


Fear of cancer progression in patients with stage IA malignant melanoma.
We aimed to determine the prevalence and importance of fear of cancer progression (FoP) in melanoma patients with stage IA tumours to assess psychosocial and demographic factors associated with severity of FoP and to determine the relationship of FoP and quality of life (QoL). One hundred and thirty-six patients with stage IA melanoma completed the short version of the Fear of Progression Questionnaire (FoP-Q-SF), the Hospital Anxiety and Depression Scale (HADS) and the EORTC-QLQ-C30. We found a mean FoP-Q-SF sum score of 30.2 points (±8.4 points SD). In this study, 33% of patients reported high FoP at or above the cutoff-value of 34 points. Higher FoP was found in women (p < 0.01), young (p = 0.03) and employed (p = 0.02) patients. Being confronted with a cancer diagnosis in closely related persons predicted higher FoP (p < 0.01). FoP correlated positively with the HADS anxiety (r = 0.50, p < 0.01) and depression scales (r = 0.26, p < 0.01) and negatively with the EORTC-QLQ-C30 global health state (r = -0.32, p < 0.01). FoP is considerably prevalent in low-risk melanoma patients and associated with reduced QoL, cancer in related persons, women sex and participation in working life. Considerably high levels of FoP, even in patients with low-risk malignancies, underline the need for psychosocial support and psychotherapeutic interventions for melanoma patients.


Outcomes of Titanium Mesh Cranioplasty in Pediatric Patients.
Cranial defects in children have been repaired with various materials ranging from autologous bone to synthetic materials. There is little published literature on the outcomes of titanium mesh cranioplasty (TMC) in calvarial reconstruction in the pediatric population. This study evaluates a pediatric cohort who underwent calvarial defect reconstruction with titanium mesh and assesses the efficacy and outcomes of TMC. An Institutional Review Board approved retrospective review of patients ≤18 years of age who underwent cranioplasty from 1999 to 2014 at 2 centers was performed. The cohort undergoing TMC was studied. A total of 159 cranioplasties were performed. Autologous reconstruction included 84 bone flap replacements and 36 split calvarial bone graft reconstructions. Six patients underwent PEEK implant reconstruction. Titanium mesh cranioplasty was performed on 33 patients. Two patients underwent 2 separate cranioplasties. The median age of patients was 6 years (19 months to 18 years). The most common underlying etiologies were congenital syndromes/craniosynostosis (13 patients), and trauma (11). The majority of patients had prior cranial surgeries (85%). Various types of titanium mesh were used with sizes ranging from 2×3 cm to 19×20 cm, with some patients requiring distinct areas of defect reconstruction. Perioperative complications were noted in 2 patients that subsequently improved. Two patients had late soft tissue problems with complications of wound infections requiring resection of a portion of the mesh. Patients were followed an average of 4 years (range 13 days to 6.8 years), with 2 patients lost to follow-up. Overall, all patients with follow-up achieved a cranial contour with good symmetry to the unaffected side, as well as effective protection to the brain. Titanium mesh cranioplasty is an effective option for correcting pediatric cranial defects when autologous bone availability is limited and soft tissue coverage allows placement of an implant. The interim outcome for these patients is favorable with few complications and no evidence of growth restriction in the authors' series. Follow-up will be ongoing for these patients.


Effect of screw thread length on stiffness of proximal humerus locking plate constructs: A finite element study.
Plate-based treatment of proximal humerus fractures is associated with a high risk of complications such as screw perforation into glenohumeral joint. Smooth and threaded pegs were developed with the hope of minimising these risks. No consensus exists onto which threading profile achieves stiffest bone-plate construct. This study investigated the biomechanical effect of five percentages of threading on individual humeral head screws on a bone-plate construct. A finite element model simulating a two-part proximal humerus fracture treated with a Spatial Subchondral Support plate was developed and validated against in vitro biomechanical tests. The proportion of the humeral head screw length that was threaded was varied between 0%-100% in 25% increments. A 5-mm cantilever varus displacement was applied and the required load (F5) was calculated. Full (100%) threading achieved the stiffest construct for all six screws. Fully threading all smooth pegs at once increased F5 by 18%. Threading did not increase F5 equally in all screws. Inferior three plate screws exhibited a larger increase in stiffness than superior three. Most of the mechanical benefits of threading in inferior three screws can be achieved by using threaded pegs (50% threading) while the superior three screws need to be fully threaded. In practice, the smooth surface profile may also offer additional mechanical benefits if implanted with longer lengths and larger diameters. Threading is an effective way of increasing the varus bending stiffness of proximal humerus plates constructs.


Further observations on the B-type neurosecretory cells in the thoracic ganglion of the crab, Potamon magnum magnum (Pretzman).
Only four types of the secretory neurones were observed in the thoracic ganglion of the fresh water crab, Potamon magnum magnum (Pretzman). These cells are classified as A, A', B and C types. Our results also confirmed that the B and B' types of cells (BAID et al. 1968) are one and the same type, the B cells. The B cells are of two forms, with and without the axons. They clearly showed sequential histological patterns pointing out to their phases. The secretory material is discharged either through the periphery or through the axon and periphery both. The vacuoles were observed in both the forms of the B cells. The remarkable sign of the activity of the nuclear membrane especially at the beginning of the secretory cycle and the appearance of the first secretory granules around the nucleus strongly support the vital role of the nucleus in the initiation of the secretory cycle.


[Amyotrophic lateral sclerosis and frontotemporal dementia: overlapping characteristics].
There is an overlap between amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Some 5-10% of ALS patients show changes in their behaviour and personality that are characteristic of FTD and about 10% of FTD patients develop ALS. Mild cognitive impairment occurs in 30% of ALS patients. The progressive decline of muscle strength in ALS patients and social skills in FTD patients places severe demands on the patient and his or her contacts. In some ALS and FTD patients, ubiquitin-positive inclusions have been found in the hippocampus and anterior horn cells. In patients with familial FTD who have ubiquitin-positive inclusions, mutations have been found in the progranulin (PGRN) gene. TAR-DNA-binding protein-43, encoded by the TARDBP gene, has recently been identified as a constituent of the ubiquitin inclusions. TARDBP and PGRN mutations are found in patients with ALS. The overlapping characteristics provide clues for further research into the pathogenesis of ALS and FTD.


[Properties of stationary dissipative structures in mathematical models of morphogenesis].
Critical selection of the model of morphogenesis is analysed in relation to the properties of corresponding stationary dissipative structures (DS). In particular differences of DS satisfying Turing, Girer and Meinhard models in uni-, two- and three-dimensional cases were revealed. It has been concluded on the basis of DS stationary state that the shape of the "organism" satisfying different models of morphogenesis is changed with external influences.


Customers' satisfaction about prehospital emergency medical services in Lorestan, Iran.
Patient's satisfaction with health care in ambulance services is an important quality indicator and a helpful tool for managers of prehospital emergency services. This study aimed to measure patient satisfaction with health provided by prehospital emergency medical services (EMS) in Lorestan, Iran. This cross-sectional study was conducted on patients (n=450) transferred by EMS to hospitals of Lorestan University of Medical Sciences in a two-year period (2013-2014). Data collection was performed by patient questionnaire, which is a standard LKFR tool. Validity and reliability of the instrument was confirmed by scientific method. Collected data were analyzed by SPSS Version 19. Descriptive and inferential statistics such as Chi-square, paired-samples t-test, independent-samples t-test, ANOVA, Pearson's product-moment correlation coefficient, and Fisher's exact test were used. One hundred ninety-two (42.8%) and 257 (57.2%) patients were female and male, respectively (mean: 41 years, r: 37-83). Patient satisfaction of the dispatcher was good, and satisfaction level in regards to the technicians' performance, physical situation, and facilities inside the ambulance was moderate. The Wilcoxon test did not show any significant difference between pain severity before and after arriving EMS in the cardiac and respiratory patients (p=0.691), but severity of pain in orthopedic patients after arriving EMS was decreased (p=0.001). Cardiac and respiratory patients had low satisfaction of EMS, and the Chi-square test was significant (p=0.001). Orthopedic patients had the most satisfaction of EMS. Generally, patients' satisfaction of EMS was low. Satisfaction with pain relief in orthopedic patients was better than pain relief in cardiac and respiratory patients. It is recommended to take necessary actions to improve the level of patient satisfaction of EMS.


The immunotherapeutic effects of Astragalus polysaccharide in type 1 diabetic mice.
The present study investigated whether Astragalus polysaccharide (APS) possessed immunotherapeutic effects on type 1 diabetes mellitus. Diabetic mice induced by multiple low dose streptozotocin (MLD-STZ) were administered either APS (100, 200, 400 mg/kg body weight) or saline intraperitoneally daily, and sacrificed after 15 or 30 d of treatment. Meanwhile normal mice not treated with STZ nor with APS were offered into non-diabetic group. Blood glucose and serum insulin levels were measured, histologic and morphometric analyses of the pancreas were performed to determine the effect of APS on pancreatic islets. Further investigations on immune changes in spleens were tested by ELISA, semi-quantitative RT-PCR and Western blot. Downregulated blood glucose level, upregulated serum insulin concentration, increased beta cell mass, decreased apoptotic beta cell percentage, downregulation of Th1/Th2 cytokine ratio and upregulation of peroxisome proliferator-activated receptor gamma (PPARgamma) gene expression in spleens were significantly time- and dose-dependent on APS treatment, when compared to saline controls. These results show that APS seems to be helpful to attenuate insulitis and preserve beta cells from apoptosis, but it can't entirely rescue type 1 diabetes mellitus. APS ameliorates both the clinical and histological parameters of the MLD-STZ induced diabetic mice in a long-lasting fashion, most likely through immunoregulatory actions on Th1/Th2 cytokine ratio, strongly associated with PPARgamma gene expression in spleens.


Cellular and humoral immune responses following SARS-CoV-2 mRNA vaccination in patients with multiple sclerosis on anti-CD20 therapy.
SARS-CoV-2 messenger RNA vaccination in healthy individuals generates immune protection against COVID-19. However, little is known about SARS-CoV-2 mRNA vaccine-induced responses in immunosuppressed patients. We investigated induction of antigen-specific antibody, B cell and T cell responses longitudinally in patients with multiple sclerosis (MS) on anti-CD20 antibody monotherapy (n = 20) compared with healthy controls (n = 10) after BNT162b2 or mRNA-1273 mRNA vaccination. Treatment with anti-CD20 monoclonal antibody (aCD20) significantly reduced spike-specific and receptor-binding domain (RBD)-specific antibody and memory B cell responses in most patients, an effect ameliorated with longer duration from last aCD20 treatment and extent of B cell reconstitution. By contrast, all patients with MS treated with aCD20 generated antigen-specific CD4 and CD8 T cell responses after vaccination. Treatment with aCD20 skewed responses, compromising circulating follicular helper T (TFH) cell responses and augmenting CD8 T cell induction, while preserving type 1 helper T (TH1) cell priming. Patients with MS treated with aCD20 lacking anti-RBD IgG had the most severe defect in circulating TFH responses and more robust CD8 T cell responses. These data define the nature of the SARS-CoV-2 vaccine-induced immune landscape in aCD20-treated patients and provide insights into coordinated mRNA vaccine-induced immune responses in humans. Our findings have implications for clinical decision-making and public health policy for immunosuppressed patients including those treated with aCD20.


Transient anterior corneal deposits in a human immunodeficiency virus-positive patient.
To report findings of pigmented anterior corneal deposits in a human immunodeficiency virus-positive patient. Case report. A 49-year-old human immunodeficiency virus-positive patient was examined after the appearance of pigmented corneal deposits. Slit-lamp biomicroscopy, fundus photography, and laser-scanning in vivo confocal microscopy were performed to visually document the ocular condition. The patient had a history of Mycobacterium avium infection and was suspected to have recovery uveitis from a cytomegalovirus infection. Small, rounded, light brown-colored deposits were distributed across the anterior cornea from limbus to limbus, bilaterally. In vivo confocal microscopy revealed the deposits to be confined to the basal epithelium and Bowman layer, whereas the posterior stroma, Descemet membrane, and the endothelium appeared normal. Systemic steroid treatment was administered, and 2 weeks later, the deposits had vanished on slit-lamp examination, whereas remnants were observed at the microscopic level. The deposits were unusual for their anterior corneal location and pancorneal distribution. The response to systemic steroid treatment remains unexplained and illustrates the complexity of the underlying conditions, their treatment, and the associated pathways of ocular manifestation.


Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems.
We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The μ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.


[Tuberculin diagnosis in children with a family history of allergy].
Two hundred and eighteen Mycobacterium tuberculosis-infected children and adolescents aged 4 to 18 years were examined. Whether the patients and their relatives had a history of allergic reactions was ascertained. A complex of diagnostic studies is of great value in verifying the presence of concomitant allergic diseases. The rate of allergic reactions in the child's history does not itself determine the likelihood of development of allergic states. The Mantoux (tuberculin) test in allergic children does not lead to an exacerbation of the underlying disease at remission. The remission manifestations of paraallergy are noted in 15.8% of the patients with allergic diseases and in 42.9% of the children having allergic reactions in the history. This supports the quality of a follow-up and treatment by an allergist, as well as an adequate response to a Mantoux test with 2 TE of PPD-L in the bulk of allergic children. The use of antihistamines as preparation for the test allows an inference about true infection and the sensitivity to tuberculin in both the most of allergic patients and persons with a history of allergic manifestations.


Differential Spatial Gradients of Wheat Streak Mosaic Virus into Winter Wheat from a Central Mite-Virus Source.
The wheat curl mite (WCM), Aceria tosichella Keifer, transmits three potentially devastating viruses to winter wheat. An increased understanding of mite movement and subsequent virus spread through the landscape is necessary to estimate the risk of epidemics by the virus in winter wheat. Owing to the small size of WCMs, their dispersal via wind is hard to monitor; however, the viruses they transmit produce symptoms that can be detected with remote sensing. The objective of this study was to characterize the spatial dispersal of the virus from a central mite-virus source. Virus infection gradients were measured spatially by using aerial remote sensing, ground measurements, geostatistics, and a geographic information system between 2006 and 2009. The red edge position vegetation index as measured via aerial imagery was significantly correlated with in-field biophysical measurements. The occurrence of virus symptoms extended differentially in all directions from mite-virus source plots, and predictions from cokriging revealed an oval pattern surrounding the source but displaced to the southeast. The variable dispersal in different directions appeared to be influenced by the mite source density and wind direction and speed, but temperature also seemed likely to have affected mite spread. The spatial spread revealed in this study may be used to estimate the potential sphere of influence of mite-infested volunteer wheat in production fields. These risk parameter estimates require further validation, but they may potentially aid growers in making better virus management decisions regarding differential virus spread potential away from a central source.


Relationships among body size, blood meal size, egg volume, and egg production of Tabanus fuscicostatus (Diptera: Tabanidae).
Number of ovarioles, egg production, and weight of unfed and bloodfed Tabanus fuscicostatus Hine females were related linearly to wing length. Wing length did not change whether flies were freshly thawed, preserved in formalin solution, or oven dried. Left and right wings were similar in length. The length from the costa to the anterior cross vein or to the intersection of R4 and R5 veins was associated linearly to wing length. The number of eggs produced by flies bloodfed on one or two bovine hosts was similar when adjusted by wing length. The average number of ovarioles per female was 277 +/- 50 (+/- SD), and eggs developed in only 63% of the ovarioles after a bloodmeal. An estimated 3.1 eggs were produced for each milligram of blood ingested by a fly, and the average bloodmeal size was 110% of the unfed weight of the flies (49.7 mg). Egg volume was unrelated to body size, but was associated inversely with the number of eggs produced per female.


The effects of training and detraining on memory, neurotrophins and oxidative stress markers in rat brain.
In the current investigation we tested how swimming training (T) (8 week, 5 times/week, 2 h/day), and detraining (DT) affects brain functions and oxidative stress markers in rat brain. The free radical concentration, measured by electron paramagnetic resonance, decreased in brain of T and DT rats compared to controls (C). The level of brain-derived neurotrophic factor (BDNF) increased as a result of training, but decreased below the control level after 6 weeks of detraining. In addition, the concentration of nerve growth factor (NGF) also declined with DT. The passive avoidance test was used to assess the memory of rats, and training-induced improvement was observed but the enhancement disappeared with detraining. When the content of mitochondrial electron transport complexes, as a potent free radical generator, was evaluated by the blue native gel method, no significant alterations were observed. The repair of nuclear and mitochondrial 8-oxodeoxyguanosine, as measured by the activity of OGG1, showed no significant difference. Therefore, the results suggest that regular exercise training improves memory, decreases the level of reactive oxygen species, and increase the production of BDNF and NGF. On the other hand, it appears that the beneficial effects of training are reversible in the brain, since detraining down-regulates the neurotrophin level, and memory. It is suggested that exercise training is more likely to beneficially effect the production of reactive oxygen species and the related oxidative damage.


Inhibition of ICAM-1 gene expression, monocyte adhesion and cancer cell invasion by targeting IKK complex: molecular and functional study of novel alpha-methylene-gamma-butyrolactone derivatives.
The transcription factor nuclear factor-kappaB (NF-kappaB) is a regulator related to cellular inflammation, immune responses and carcinogenesis. Therefore, components of the NF-kappaB-activating singnaling pathways are frequent targets for the anti-inflammatory and anticancer agents. In this study, CYL-19 s and CYL-26z, two synthetic alpha-methylene-gamma-butyrolactone derivatives, were shown to inhibit the tumor necrosis factor-alpha (TNF-alpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression in human A549 alveolar epithelial cells and the adhesion of U937 cells to these cells. RT-PCR analysis also demonstrated their inhibitory effects on TNF-alpha-induced ICAM-1 mRNA expression. TNF-alpha-induced ICAM-1 and NF-kappaB-dependent promoter activities were attenuated by CYL-19 s and CYL-26z. ICAM-1 promoter activities induced by the over-expression of wild-type NF-kappaB-inducing kinase and IkappaB kinase beta (IKKbeta) were also inhibited by both compounds. Furthermore, CYL-19 s and CYL-26z inhibited the TNF-alpha-induced phosphorylation and degradation of IkappaBalpha and NF-kappaB-specific DNA-protein binding activity via targeting IKK complex directly, without any effect on the activations of other kinases such as ERK1/2 and p38. In addition to ICAM-1 expression, CYL-19 s and CYL-26z also suppressed other NF-kappaB-mediated gene expressions such as matrix metalloproteinase-9 (MMP-9) mRNA and cyclooxygnease-2 (COX-2) protein. In Matrigel assays, ICAM-1 and COX-2 expressions induced by TNF-alpha elicited A549 and NCI-H292 cell invasion, respectively, and these effects were inhibited by both compounds. In summary, our data demonstrated that CYL-19 s and CYL-26z down-regulate the TNF-alpha-induced inflammatory genes expression through suppression of IKK activity and NF-kappaB activation. These agents may be effective in the anti-inflammatory and anticancer therapy.


[Flexible vaginal cast covered with oxidated cellulose. Surgical option in neovagina for patients with Rokitansky-Kuster-Hauser syndrome].
The alterations in differentiation of the mullerian derivatives and urogenital sinus are a frequent cause of demanding attention in the gynecologist office. Vaginal agenesis and hypoplasia are the representative events of alterations of human sexual differentiation. Handling such conditions is in direct relationship to the magnitude of the anatomical damage and the biopsychosocial consequences of such pathologies. From historical perspective, several surgical techniques of neovagina to correct such alterations have been described. We are presenting our experience in surgical treatment of vaginal agenesis, as well as the aesthetic-functional outcomes obtained from a highly specialized hospital of the Health Sector in Mexico City. Our patients attended the gynecologist office mainly because of primary amenorrhea, anatomical alterations and sexual dysfunction. Tbe age range varied between 19 and 28 years. Basic laboratory tests were conducted. Urography, genetic studies, and a laparoscopic procedure were included. All the women received psychological support. The diagnosis in all the cases was Rokitansky Kuster-Hauser Syndrome with agenesis of the 2 proximal thirds of vagina. A neovagina assisted by laparoscopy using a technique previously modified was conducted. In such technique we use a flexible vaginal mould and oxide cellulose to prevent fibrosis and surgical retraction. Cosmetic outcome was satisfactory and dysfunction sexual was all improved. We conclude that our procedure that includes the neovagina assisted by laparoscopy and the use of a vaginal mould and oxide cellulose is a surgical option in those patients presenting with vaginal agenesis and hypoplasia. This procedure improves the aesthetic and sexual performances that are profoundly altered in such patients.


Targeting the heat shock factor 1 by RNA interference: a potent tool to enhance hyperthermochemotherapy efficacy in cervical cancer.
Carcinoma of the uterine cervix is one of the highest causes of mortality in female cancer patients worldwide, and improved treatment options for this type of malignancy are highly needed. Local hyperthermia has been successfully used in combination with systemic administration of cisplatin-based chemotherapy in phase I/II clinical studies. Heat-induced expression of cytoprotective and antiapoptotic heat shock proteins (HSP) is a known complication of hyperthermia, resulting in thermotolerance and chemoresistance and hindering the efficacy of the combination therapy. Heat shock transcription factor 1 (HSF1) is the master regulator of heat-induced HSP expression. In the present report, we used small interfering RNA (siRNA) to silence HSF1 and to examine the effect of HSF1 loss of function on the response to hyperthermia and cisplatin-based chemotherapy in HeLa cervical carcinoma. We have identified the 322-nucleotide to 340-nucleotide HSF1 sequence as an ideal target for siRNA-mediated HSF1 silencing, have created a pSUPER-HSF1 vector able to potently suppress the HSF1 gene, and have generated for the first time human cancer cell lines with stable loss of HSF1 function. We report that, although it surprisingly does not affect cancer cell sensitivity to cisplatin or elevated temperatures up to 43 degrees C when administered separately, loss of HSF1 function causes a dramatic increase in sensitivity to hyperthermochemotherapy, leading to massive (>95%) apoptosis of cancer cells. These findings indicate that disruption of HSF1-induced cytoprotection during hyperthermochemotherapy may represent a powerful strategy to selectively amplify the damage in cancer cells and identify HSF1 as a promising therapeutic target in cervical carcinoma.


Calf circumference as a screening tool for low skeletal muscle mass: Cut-off values in independent Thai older adults.
Calf circumference is recommended as a marker for low muscle mass and as a case finding in the diagnosis of sarcopenia. However, the cut-off value differed by ethic and region. Currently there is no study among Thai population. Therefore, we aimed to identify the optimal cutoff value of calf circumference as a screening tool for low skeletal muscle mass in independent Thai older adults. Subgroup analysis was performed for obesity and adults over 75 years. This cross-sectional cohort studied in an outpatient geriatric check-up clinic. Participants, aged 60 and above, needed to be independent in basic activities of daily living to meet the inclusion criteria. Exclusion criteria comprised active malignancy, cardiac, pulmonary, or neurovascular diseases necessitating hospitalization in the preceding three months, chronic renal diseases requiring renal replacement therapy, and unstable psychiatric disorders. We measured the maximum calf circumference and appendicular skeletal muscle mass (ASMI) using bioelectrical impedance analysis (BIA). Low muscle mass is defined according to the Asian Working Group of Sarcopenia (AWGS) 2019 consensus. We enrolled 6,404 elderly adults (mean age 67.3 ± 5.1 years), with a 47% prevalence of low muscle mass in women and 25% in men. Lower muscle mass significantly correlated with reduced BMI and waist circumference in both genders (p < 0.001). Optimal cut-off values for low muscle mass screening were < 33 cm (sensitivity 80.1%, specificity 60.5%) for women and < 34 cm (sensitivity 85.4%, specificity 70.2%) for men. Subgroup analysis for those with BMI ≥ 25 kg/m² suggested raising the cut-off for women to < 34 cm (sensitivity 80.6%, specificity 54.0%) and for men to < 35 cm (sensitivity 88.7%, specificity 55.2%) to enhance specificity without substantial sensitivity loss. In the older-old adult subgroup (≥ 75 years), optimal cut-off values were < 33 cm (sensitivity 84.6%, specificity 79.9%) for women and < 34 cm (sensitivity 75.6%, specificity 87.0%) for men. There is a strong correlation between calf circumference and ASMI in independent Thai older adults. Calf circumference can serve as a screening tool for identifying low muscle mass. The recommended cut-off values for men and women are 34 cm and 33 cm, respectively in alignment with AWGS 2019 recommendation. Incorporating a 1-cm higher cut-off value for obese older adults improves the accuracy of muscle mass screening. Thai clinical trial registry: TCTR20200511003.


Growth performance and reproductive function impairment of glyphosate-based herbicide in male guinea pig (Cavia porcellus).
Glyphosate formulations, widely applied non-selective systemic herbicides, are progressively becoming the most controversial pesticides on the market due the adverse effects they pose to humans and environment. The information on these potential hazardous effects to the handlers of the pesticide remains obscure. This study investigated effects of glyphosate-based herbicide on growth performance, seminal parameters and hemato-biochemical profiles in male guinea pig. Forty sexually mature male guinea pigs weighing between 393.3 and 418.4 g were divided into four groups of 10 animals each and orally administered 0, 186, 280 and 560 mg/kg body weight of WILLOSATE daily for 60 days. Daily feed intake and body weight gain were recorded. At the end of experimental period all animals were humanely sacrificed, and blood samples and vital organs were collected for appropriate analysis. Results showed a significant decrease (p < 0.05) in body weight gain (-102.2%), final body weight (-9.8%) and feed intake (-13.1%) of animals following sub-chronic exposure of WILLOSATE. The weights of the liver and kidney increased significantly (p < 0.05) by 25.4% and 28.8%, respectively, while testicular weights decreased (p < 0.05) by 24% with increasing doses of WILLOSATE. A decrease in sperm motility (-67.9%), viability (-52.7%) and concentration per vas deferens (-40.7%), and an increase in sperm major (28.1%) and minor (45.3%) morphological aberrations were recorded in WILLOSATE - exposed guinea pigs when compared to controls. There was a dose-dependent increase (p < 0.05) in MCV and WBC and a decrease in Hb content and RBC, as well as serum content in total protein (-11.8%). The serum content of cholesterol (37.8%), urea (87.1%), creatinine (22.4%), ALAT (74.2%) and ASAT (88.7%) were significantly higher in treated groups compared to controls. These results point toward the toxic effects of WILLOSATE on vital organs and reproductive function of the body at high doses and long-term exposure.


Two-Excitation Routing via Linear Quantum Channels.
Routing quantum information among different nodes in a network is a fundamental prerequisite for a quantum internet. While single-qubit routing has been largely addressed, many-qubit routing protocols have not been intensively investigated so far. Building on a recently proposed many-excitation transfer protocol, we apply the perturbative transfer scheme to a two-excitation routing protocol on a network where multiple two-receivers block are coupled to a linear chain. We address both the case of switchable and permanent couplings between the receivers and the chain. We find that the protocol allows for efficient two-excitation routing on a fermionic network, although for a spin-12 network only a limited region of the network is suitable for high-quality routing.


[On the question of interval application of topical corticoids/Practicability and efficacy (author's transl)].
Therapeutic efficacy and practicability of an interval treatment were investigated in a multi-center study in a total of 1573 patients with psoriasis or eczema, respectively. Fluocortolone monohydrate as Syracort and Syracerin as steroid-free "partner drug" were applied at 4-13-day intervals. The findings and established results substantiate equivalent therapeutic success in comparison with consistent treatment. Starting from the premise that the risk of steroid-induced skin lesions can be reduced, if not avoided, by interval application of topical corticoids one may consider steroid application at intervals as superior to consistent therapy.


Reducing Hospitalization in Mild Acute Pancreatitis: Results of Long-term Follow-up.
Acute pancreatitis (AP) is a common emergency department (ED) diagnosis, amounting to enormous costs. Our previous pilot study demonstrated the feasibility of reducing hospitalization using an ED-based observation pathway. In this follow-up study, we hypothesize that the pathway is durable in clinical practice, outside of research supervision, and patients can safely be managed without hospitalization. We reviewed patients prospectively enrolled in the observation pathway after the end of the pilot study. We compared outcomes to patients enrolled in our pilot study and with a historic cohort of patients admitted with mild AP. Our primary outcome was hospitalization rate during the enrollment period and secondary outcomes included length of stay, 30-day readmissions, mortality, and health care utilization. Over a 2-year period 165 patients met criteria for AP with 118 (71.5%) having mild AP. Fifty-four of 118 patients (45.8%) were enrolled in the observation pathway and of these, 45 patients were discharged from the ED, reducing hospitalization by 31.2%, compared with pilot study (22.2%) and historic cohort (0%) (P<0.05). Median length of stay was shorter [19.9 (observation) vs. 72.0 h (historic cohort), P<0.01]. There were fewer radiographic examinations in the observation cohorts (pilot and current study) than in the historic cohort (P<0.05), with similar 30-day readmissions, and no reported deaths. This follow-up study demonstrates the durability of an observation-based pathway to manage mild AP outside of a research protocol and maintain its ability to reduce hospitalizations without affecting readmission rates or mortality.


Clinical Observations, Plasma Retinol Concentrations, and In Vitro Lymphocyte Functions in Children With Sickle Cell Disease.
Children with sickle cell disease (SCD) often have infections, growth deficits, and impaired immunity, problems that also are observed in individuals with a vitamin A deficiency (plasma retinol concentration <20 μg/dL). The goal of this study was to investigate the association between vitamin A, health status, and the in vitro immune function of children with SCD. Fifty-nine children (40 SS, 11 SC, and 8 Sβthalassemia [Sβthal] hemoglobin genotypes) 9 months to 18 years old were investigated for plasma levels of retinol, retinol binding protein, C-reactive protein, alpha-1-acid glycoprotein, lymphocyte proliferation, and interleukin (IL)-2 activity in supernatant of phytohemagglutinin-treated lymphocytes. The plasma retinol concentrations of children with SCD (mean 57.6 μg/dL, range 4.6-116 μg/dL) were not different from those of 21 normal individuals (mean 62 μg/dL, range 28.7-162 μg/dL). Plasma retinol concentrations did not vary by hemoglobin genotype but were lower in boys than in girls (P < 0.05) and were also lower in children with inflammation (P = 0.1). Seven children (11.9%) (6 HbSS, 1 HbSβ0thal) were vitamin A-deficient, and 9 children (15.3%) had suboptimal vitamin A status (plasma retinol concentration of 20-29 μg/dL). Children with vitamin A deficiency had slightly lower height (P = 0.09) and weight mean percentiles, lymphocyte proliferative responses, and IL-2 activity (P > 0.1), but higher means of C-reactive protein (P = 0.05), pain crisis episodes and inflammation (P = 0.1), and health scores (P > 0.1) than children who were not vitamin A-deficient. Lymphocyte proliferative responses negatively correlated with health score, pain crisis episodes, and blood units received, but positively correlated with retinol binding protein (P < 0.05 to P = 0.1). Identification and correction of suboptimal vitamin A status in children with SCD may improve immunity and attenuate certain health complications associated with this disease.


Conservative surgical treatment in rectal cancer after complete pathological response to neoadjuvant treatment. Prospective observational study.
In 2012 we started a prospective observational study at San Giovanni Addolorata Hospital in Rome for patients with rectal cancer with complete response to neoadjuvant therapy (nCRT). In our, IRB approved protocol, patients are evaluated at time 0 by physical, endoscopic, pathological and radiological examinations. 6 weeks after completion of nCRT they are re-evaluated. In case of persistence or progression of disease patients undergo surgery with Total Mesorectal Excision. In case of complete or major clinical response they are re-evaluated at 12 weeks and subjected to transanal surgical excision to confirm complete pathological response (pCR). If tumor is found in the transanal excision specimen the patient is operated upon whereas patients with pCR are followed up at 3 months interval.


Electronic acute kidney injury alert at the Brandenburg Medical School - implementation and follow-up.
Introduction Acute kidney injury (AKI) substantially worsens the prognosis of hospitalized patients world-wide. In order to optimize early AKI recognition and therapeutic intervention, AKI alert systems have been implemented and evaluated in the past. Herein, we aimed to analyze outcome var-iables of AKI patients under the conditions of a de-novo-established AKI alert system at the Brandenburg Hospital of the Brandenburg Medical School. Methods Automated e-mail messages were generated and sent to the nephrologist in responsibility based on an electronic algorithm. The message was exclusively generated if one of the two first KDIGO criteria was fulfilled. During period 1, all alerts were ignored. During the second period, every alert was followed up, coupled with therapeutic management of respective indi-viduals according to an AKI care bundle. Endpoints were in-hospital death, need for dialysis, and renal recovery. Results In periods 1 and 2, 200 and 112 patients were included. In period 1, 150 out of 200 AKI alerts were identified as correct (75%); in the second period, 93 out of 112 AKI alerts were accepted as correct (83%) (p=0.16). Kidney replacement therapy (KRT) was initiated in 21 (14%) of all period 1 patients and in 32 (34.4%) of the period 2 patients (p=0.017). In-hospital mortality of affected patients was 24 (16%) in period 1 and 21 (22,5%) (p=0.4). Restoration of kidney function was 69 (46%) in period 1 and 45 (48.3%) (p=0.71) in period 2. Conclusions We finally conclude that an AKI alert system, as implemented and followed-up in our study, did not significantly improve clinically relevant endpoints in AKI patients. Potential weaknesses were: the lack of documentation of the time between receiving the alert and patient contact, and physicians in responsibility were not particularly informed about the alert system.


Inhibitory Effects of Eurotium cristatum on Growth and Aflatoxin B1 Biosynthesis in Aspergillus flavus.
Probiotic strain Eurotium cristatum was isolated from Chinese Fuzhuan brick-tea and tested for its in vitro activity against aflatoxigenic Aspergillus flavus. Results indicated that E. cristatum can inhibit the radial growth of A. flavus. Furthermore, this inhibition might be caused by E. cristatum secondary metabolites. The ability of culture filtrate of strain E. cristatum against growth and aflatoxin B1 production by toxigenic A. flavus was evaluated in vitro. Meanwhile, the influence of filtrate on spore morphology of A. flavus was analyzed by scanning electron microscopy (SEM). Results demonstrated that both radial growth of A. flavus and aflatoxin B1 production were significantly weakened following increases in the E. cristatum culture filtrate concentration. In addition, SEM showed that the culture filtrate seriously damaged hyphae morphology. Gas chromatography mass spectrometry (GC/MS) analysis of the E. cristatum culture supernatant revealed the presence of multiple antifungal compounds. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis showed that the expression of aflatoxin biosynthesis-related genes (aflD, aflQ, and aflS) were down-regulated. Importantly, this latter occurrence resulted in a reduction of the AflS/AflR ratio. Interestingly, cell-free supernatants of E. cristatum facilitated the effective degradation of aflatoxin B1. In addition, two degradation products of aflatoxin B1 lacking the toxic and carcinogenic lactone ring were identified. A toxicity study on the HepG2 cells showed that the degradation compounds were less toxic when compared with AFB1.


Insulin-like growth factor II signaling in neoplastic proliferation is blocked by transgenic expression of the metalloproteinase inhibitor TIMP-1.
Insulin-like growth factor (IGF) II is overexpressed in many human cancers and is reactivated by, and crucial for viral oncogene (SV40 T antigen, [TAg])-induced tumorigenesis in several tumor models. Using a double transgenic murine hepatic tumor model, we demonstrate that tissue inhibitor of metalloproteinase 1 (TIMP-1) blocks liver hyperplasia during tumor development, despite TAg-mediated reactivation of IGF-II. Because the activity of IGFs is controlled by IGF-binding proteins (IGFBPs), we investigated whether TIMP-1 overexpression altered the IGFBP status in the transgenic liver. Ligand blotting showed that IGFBP-3 protein levels were increased in TIMP-1-overexpressing double transgenic littermates, whereas IGFBP-3 mRNA levels were not different, suggesting that TIMP-1 affects IGFBP-3 at a posttranscriptional level. IGFBP-3 proteolysis assays demonstrated that IGFBP-3 degradation was lower in TIMP-1-overexpressing livers, and zymography showed that matrix metalloproteinases (MMPs) were present in the liver homogenates and were capable of degrading IGFBP-3. As a consequence of reduced IGFBP-3 proteolysis and elevated IGFBP-3 protein levels, dissociable IGF-II levels were significantly lower in TIMP-1-overexpressing animals. This decrease in bioavailable IGF-II ultimately resulted in diminished IGF-I receptor signaling in vivo as evidenced by diminished receptor kinase activity and decreased tyrosine phosphorylation of the IGF-I receptor downstream effectors, insulin receptor substrate 1 (IRS-1), extracellular signal regulatory kinase (Erk)-1, and Erk-2. Together, these results provide evidence that TIMP-1 inhibits liver hyperplasia, an early event in TAg-mediated tumorigenesis, by reducing the activity of the tumor-inducing mitogen, IGF-II. These data implicate the control of MMP-mediated degradation of IGFBPs as a novel therapy for controlling IGF bioavailability in cancer.


Orientated Guidance of Peripheral Nerve Regeneration Using Conduits with a Microtube Array Sheet (MTAS).
Material surface topography has been shown to affect the biological behavior of cells in vitro; however, the in vivo effect on peripheral nerve regeneration has not been explored. Here, we studied the potential of a microtube array sheet (MTAS) with a unique longitudinal surface topography to promote peripheral nerve regeneration efficiency, both in vivo and in vitro. Schwann cells, spinal cord motor neurons, and dorsal root ganglion neurons were seeded on the MTAS to study the effect of the construct on the biological properties and behaviors of neural cells. The MTAS guided the oriented migration of Schwann cells without affecting other critical biological properties, such as proliferation and neurotrophin expression. In addition, the MTAS guided the directed extension of neurites from both types of neurons. Next, we tested the capability of the MTAS to facilitate peripheral nerve regeneration by bridging a 10 mm sciatic nerve defect in rats with a nerve conduit equipped with an MTAS lining. The MTAS significantly promoted peripheral nerve regeneration, as suggested by the greater fiber caliber in the midconduit and the greater abundance of fibers in nerve segment distal to the conduit. Moreover, scanning electron microscopy (SEM) analysis suggested the orientated guidance of nerve regeneration by the MTAS, as indicated by the smaller eccentricity of the nerve fibers and the concordant arrangement of the collagen fiber in both the fibers and the matrix in the MTAS group. Our results collectively suggest that the conduits with the MTAS developed in this study have significant potential for facilitating peripheral nerve regeneration by modifying critical biological behaviors and guiding orientated nerve growth.


Induction of the BIM Short Splice Variant Sensitizes Proliferating NK Cells to IL-15 Withdrawal.
Adoptive transfer of allogeneic NK cells holds great promise for cancer immunotherapy. There is a variety of protocols to expand NK cells in vitro, most of which are based on stimulation with cytokines alone or in combination with feeder cells. Although IL-15 is essential for NK cell homeostasis in vivo, it is commonly used at supraphysiological levels to induce NK cell proliferation in vitro. As a result, adoptive transfer of such IL-15-addicted NK cells is associated with cellular stress because of sudden cytokine withdrawal. In this article, we describe a dose-dependent addiction to IL-15 during in vitro expansion of human NK cells, leading to caspase-3 activation and profound cell death upon IL-15 withdrawal. NK cell addiction to IL-15 was tightly linked to the BCL-2/BIM ratio, which rapidly dropped during IL-15 withdrawal. Furthermore, we observed a proliferation-dependent induction of BIM short, a highly proapoptotic splice variant of BIM in IL-15-activated NK cells. These findings shed new light on the molecular mechanisms involved in NK cell apoptosis following cytokine withdrawal and may guide future NK cell priming strategies in a cell therapy setting.


The prevalence and significance of a patent foramen ovale in pulmonary hypertension.
In order to determine the prevalence and significance of a patent foramen ovale (PFO) in pulmonary hypertension, 58 patients were studied with transesophageal echocardiography, right-sided heart catheterization, and exercise testing. In order to examine if a PFO might be associated with a better outcome, survival was estimated, based on a formula derived from the National Institutes of Health Primary Pulmonary Hypertension Registry, for the patients with primary pulmonary hypertension (PPH). A PFO was found in 26 percent (15 of 58) of all patients studied, 25 percent (10 of 40) of those with PPH, and 28 percent (5 of 18) of those with secondary pulmonary hypertension. We found no significant difference in any hemodynamic variable or exercise tolerance between the patients with and without a PFO, or for subsets of patients with primary and secondary pulmonary hypertension. We also found no significant difference in the 1-, 2-, 3-, 4-, or 5-year estimated survival, for the patients with PPH between those with and without a PFO. The prevalence of a PFO in pulmonary hypertension appears similar to the normal population. A PFO provides no detectable influence on resting hemodynamics or exercise tolerance in patients with pulmonary hypertension and is not clearly associated with patients expected to survive longer.


A pharmacy's journey toward the patient-centered medical home.
To describe the integration of a clinic pharmacy with a patient-centered medical home (PCMH). Primary care clinic in Monroe, WA, from 1981 to January 2011. Pharmacists and physicians with a previous working relationship in a family practice residency program established colocated practices in 1981. In addition to traditional pharmacy services, collaborative practice agreements were developed and clinical pharmacy services expanded over time. Reimbursement challenges for clinical pharmacy services existed in the fee-for-service environment. The acquisition of the clinic and pharmacy by Providence Health and Services created a new financial alignment with additional opportunities for collaboration. An internally funded grant established a PCMH pilot that included pharmacist participation. PCMH pharmacists and the care provider team identified areas to improve physician and clinic efficiencies and to enhance patient care. Clinical pharmacy services expanded under the PCMH model. Pharmacist activities included value-added refill authorization services, coordinated patient visits with the PCMH pharmacist and physicians, medication therapy management, diabetes and anticoagulation services, hospital discharge medication reconciliation, and participation in the shared medical appointment. Clinical pharmacy services are woven into the PCMH fabric of this clinic. New pharmacists will be challenged and rewarded in this evolving health care model.


Pattern of long-distance projections from fetal hippocampal field CA3 and CA1 cell grafts in lesioned CA3 of adult hippocampus follows intrinsic character of respective donor cells.
Fetal hippocampal grafts transplanted to the lesioned CA3 of adult hippocampus can extend axonal projections to many regions of the host brain. However, the identity of grafted cells that project to specific host regions is unknown. We hypothesize that the pattern of long-distance axonal projections from distinct fetal hippocampal cells grafted to lesioned CA3 is specified by the intrinsic nature of respective donor cells rather than characteristics of the host graft region. We grafted fetal hippocampal CA3 or CA1 cells into kainic acid lesioned CA3 of adult hippocampus at four days post-lesion. Neurons projecting to either the contralateral hippocampus or the ipsilateral septum were then measured in these grafts at four months post-grafting using Fluoro-Gold and DiI tract tracing. CA3 grafts located close to the degenerated CA3 cell layer showed a high propensity for establishing projections into the contralateral hippocampus (commissural projections) compared to similarly located CA1 grafts, which exhibited negligible commissural projections. Similar distinction was observed between the two graft types even when they were located only partially in the lesioned CA3. Among CA3 grafts, those placed near the degenerated CA3 cell layer established significantly greater commissural projections than those placed only partially in the CA3 region. Septal projections, in contrast, were robust from both CA3 and CA1 grafts. This differential projection pattern between CA3 and CA1 grafts resembles projections of CA3 and CA1 cells in intact hippocampus.These results demonstrate that the intrinsic character of grafted fetal cells determines the type of efferent projections from fetal grafts into different targets in the lesioned adult host brain. However, the extent of efferent projections from specific grafts is also influenced by the location of grafted cells within the host region. Thus, graft-mediated appropriate reconstruction of damaged circuitry in the lesioned brain may require grafting of homotopic donor cells. Further, the robust and specific projections observed from CA3 grafts is likely beneficial for functional recovery of hippocampus following CA3 injury and hence of significance towards developing a graft-mediated therapy for human temporal lobe epilepsy.


TOPLESS mediates brassinosteroid-induced transcriptional repression through interaction with BZR1.
Brassinosteroid (BR) regulates plant development by activating the transcription factor brassinazole resistant 1 (BZR1), which activates and represses different target genes to switch cellular programmes. The mechanisms that determine BZR1's transcriptional activities remain largely unknown. Here we show that BZR1 represses target genes by recruiting the Groucho/TUP1-like transcriptional corepressor TOPLESS (TPL). Specific deletion or mutation of an evolutionarily conserved ERF-associated amphiphilic repression (EAR) motif at the carboxy terminus abolishes BZR1's abilities to regulate gene expression and cell elongation, but these defects are rescued by TPL fusion to the EAR motif-mutated BZR1. The EAR motif in BZR1 mediates recruitment of TPL to BZR1-repressed promoters. A triple tpl mutant (tpl;tpr1;tpr4) shows reduced BR sensitivity and suppresses the gain-of-function bzr1-1D mutant phenotype. BR repression of gene expression also requires histone deacetylases that interact with TPL. Our study demonstrates key roles of the EAR motif and TPL in BR regulation of gene expression and plant growth.


Graft failure within 2 years of isolated anterior cruciate ligament reconstruction is associated with increased risk of secondary meniscus tears.
A debilitating complication following anterior cruciate ligament reconstruction is a secondary meniscus tear. Currently, the literature is mixed regarding the risk factors associated with the incidence of secondary meniscus tears. The aim of this study was to investigate risk factors associated with meniscus tears following an isolated primary anterior cruciate ligament reconstruction. ACL graft failure was hypothesized to be the strongest risk factor for secondary meniscal injury occurrence. A retrospective cohort analysis was performed using the PearlDiver Database. Patients with a primary anterior cruciate ligament reconstruction were identified in the database. Patients with concomitant knee ligament injury or meniscus injury present at the time the index procedure were excluded. Patients were grouped to those who had a secondary meniscus tear within 2 years following anterior cruciate ligament reconstruction and those who did not. Univariate analysis and multivariable regression analysis was conducted to identify significant risk factors for a secondary meniscus tear. There were 25,622 patients meeting criteria for inclusion in this study. Within 2 years from the primary anterior cruciate ligament reconstruction, there were 1,781 patients (7.0%) that experienced a meniscus tear. Graft failure had the highest odds of having a postoperative meniscus tear within 2 years (OR: 4.1; CI 3.5-4.8; p < 0.002). Additional significant risk factors included tobacco use (OR: 2.0; CI 1.0-3.1; p < 0.001), increased Charlson Comorbidity Index (OR: 1.2; CI 1.1-1.4), male gender (OR: 1.1; CI 1.1-1.2; p < 0.001), obesity (OR: 1.1; CI 1.1-1.2; p < 0.001), delayed surgery (OR:1.1; CI 1.1-1.2; p < 0.002), and patients age 30 and older (OR: 1.0; CI 1.0-1.0; p < 0.001). This study found that anterior cruciate ligament graft failure is the strongest predictor of post-operative meniscus tears. Other risk factors, including tobacco use, increased CCI, male gender, obesity, delayed surgery, and age 30 and older, were established, with several being modifiable. Therefore, targeted preoperative optimization of modifiable risk factors and postoperative protocols may reduce the risk of secondary meniscus tears. Level III, prognostic trial.


Mesenchymal stem cell-derived extracellular vesicles for metabolic syndrome therapy: Assessing efficacy with nuclear magnetic resonance spectroscopy.
Metabolic syndrome (MetS) represents a cluster of metabolic abnormalities. The prevalence of MetS has surged, transforming it into a pressing public health concern that could potentially affect around 20%-25% of the global population. As MetS continues its ascent, diverse interventions, pharmacological, nonpharmacological and combined have been deployed. Yet, a comprehensive remedy that fully eradicates MetS symptoms remains elusive, compounded by the risks of polypharmacy's emergence. Acknowledging the imperative to grasp MetS's intricate pathologies, deeper insights for future research and therapy optimisation become paramount. Conventional treatments often target specific syndrome elements. However, a novel approach emerges in mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) therapy, promising a holistic shift. MSC-EVs, tiny membranous vesicles secreted by mesenchymal stem cells, have garnered immense attention for their multifaceted bioactivity and regenerative potential. Their ability to modulate inflammation, enhance tissue repair and regulate metabolic pathways has prompted researchers to explore their therapeutic application in MetS. This review primarily aims to provide an overview of how MSC-EVs therapy can improve metabolic parameters in subjects with MetS disease and also introduce the usefulness of NMR spectroscopy in assessing the efficacy of MSC-EVs therapy for treating MetS.


Influence of Ti and Cr Adhesion Layers on Ultrathin Au Films.
Efficient adhesion of gold thin films on dielectric or semiconductor substrates is essential in applications and research within plasmonics, metamaterials, 2D materials, and nanoelectronics. As a consequence of the relentless downscaling in nanoscience and technology, the thicknesses of adhesion layer and overlayer have reached tens of nanometers, and it is unclear if our current understanding is sufficient. In this report, we investigated how Cr and Ti adhesion layers influence the nanostructure of 2-20 nm thin Au films by means of high-resolution electron microscopy, complemented with atomic force microscopy and X-ray photoelectron spectroscopy. Pure Au films were compared to Ti/Au and Cr/Au bilayer systems. Both Ti and Cr had a striking impact on grain size and crystal orientation of the Au overlayer, which we interpret as the adhesion layer-enhanced wetting of Au and the formation of chemical bonds between the layers. Ti formed a uniform layer under the Au overlayer. Cr interdiffused with the Au layer forming a Cr-Au alloy. The crystal orientation of the Au layers was mainly [111] for all thin-film systems. The results showed that both adhesion layers were partially oxidized, and oxidation sources were scrutinized and found. A difference in bilayer electrical resistivity between Ti/Au and Cr/Au systems was measured and compared. On the basis of these results, a revised and more detailed adhesion layer model for both Ti/Au and Cr/Au systems was proposed. Finally, the implications of the results were analyzed, and recommendations for the selection of adhesion layers for nano-optics and nanoelectronics applications are presented.


Remote ischemic postconditioning attenuates damage in rats with chronic cerebral ischemia by upregulating the autophagolysosome pathway via the activation of TFEB.
The transcription factor EB (TFEB) is known for its role in lysosomal biogenesis, and it coordinates this process by driving autophagy and lysosomal gene expression during ischemia. In the present study, we aimed to explore the role of the TFEB-regulated autophagolysosome pathway (ALP) in rats with chronic cerebral ischemia (CCI) that were treated with remote ischemic postconditioning (RIPC). A modified 2-vessel occlusion (2-VO) method was utilized to establish the CCI rat model, and the CCI rats were identified by the Morris water maze test and histological staining. After the CCI rats were treated with RIPC, the damage to the rat cortex and hippocampal tissues and the status of the ALP were determined. Western blot analysis and immunofluorescence assays were performed to observe the nuclear translocation of TFEB. The rats were injected with TFEB siRNA via the lateral ventricle to investigate the effect of TFEB siRNA on the RIPC-treated CCI rats. The results suggested that RIPC of the CCI rats alleviated nerve injury, induced TFEB translocation into the nucleus, upregulated autophagy-related protein expression, and activated ALP machinery. Furthermore, TFEB siRNA decreased the levels of TFEB and impaired the neuroprotective effects of RIPC on the CCI rats. Collectively, we highlighted that RIPC attenuates damage in CCI rats via the activation of the TFEB-mediated ALP.


Initial behavior outcomes for the peacebuilders universal school-based violence prevention program.
PeaceBuilders is a universal, elementary-school-based violence prevention program that attempts to alter the climate of a school by teaching students and staff simple rules and activities aimed at improving child social competence and reducing aggressive behavior. Eight matched schools (N > 4,000 students in Grades K-5) were randomly assigned to either immediate postbaseline intervention (PBI) or to a delayed intervention 1 year later (PBD). Hierarchical linear modeling was used to analyze results from assessments in the fall and spring of 2 consecutive school years. In Year 1, significant gains in teacher-rated social competence for students in Grades K-2, in childself-reported peace-building behavior in Grades K-5, and reductions in aggressive behavior in Grades 3-5 were found for PBI but not PBD schools. Differential effects in Year 1 were also observed for aggression and prosocial behavior. Most effects were maintained in Year 2 for PBI schools, including increases in child prosocial behavior in Grades K-2. Implications for early universal school-based prevention and challenges related toevaluating large-scale prevention trials are discussed.


Conservative management of asymptomatic spinal lipomas of the conus.
The natural history of spinal lipomas of the conus (SLCs) has not been well studied. Because of disappointing long-term results with early surgical treatment of asymptomatic children with SLCs, we have followed a protocol of conservative management for these patients. The results are presented in this report. Since 1994, all asymptomatic children with SLCs who were examined at Necker-Enfants Malades Hospital were subject to a protocol of conservative management. The records for those patients were reviewed, to determine the incidence and timing of neurological deterioration. The findings were compared with those for a previously reported historical cohort of asymptomatic patients who underwent early surgery at our institution. Fifty-three asymptomatic children (35 girls and 18 boys) with SLCs were monitored, with conservative management. During a mean follow-up period of 4.4 years (range, 12 mo to 9 yr), 13 patients (25%) exhibited neurological deterioration. At 9 years, the actuarial risks of deterioration, as determined with the Kaplan-Meier method, were 33% for the conservatively treated patients and 46% for the surgically treated patients. With a Cox proportional-hazards model, there was no significant difference in the risks of neurological deterioration for patients who were treated conservatively and those who underwent early surgery. The incidences and patterns of neurological deterioration seemed to be very similar, regardless of whether early surgery was performed. These results suggest that conservative treatment of asymptomatic patients is a reasonable option. A more definitive randomized study will be required to clarify the relative efficacy of early surgery for SLCs among asymptomatic patients.


Associations between sinus tachycardia and adverse cardiovascular outcomes and mortality in cancer patients.
Sinus tachycardia in cancer reflects a significant multi-system organ stressor and disease, with sparse literature describing its clinical significance. We assessed cardiovascular (CV) and mortality prognostic implications of sinus tachycardia in cancer patients. We conducted a case-control study of 622 cancer patients at a U.S. urban medical center from 2008 to 2016. Cases had ECG-confirmed sinus tachycardia [heart rate (HR) ≥100 bpm] in ≥3 different clinic visits within 1 year of cancer diagnosis excluding a history of pulmonary embolism, thyroid dysfunction, left ventricular ejection fraction <50%, atrial fibrillation/flutter, HR >180 bpm. Adverse CV outcomes (ACVO) were heart failure with preserved ejection fraction (HFpEF), HF with reduced EF (HFrEF), hospital admissions for HF exacerbation (AHFE), acute coronary syndrome (ACS). Regression analyses were conducted to examine the effect of sinus tachycardia on overall ACVO and survival. There were 51 cases, age and sex-matched with 571 controls (mean age 70±10, 60.5% women, 76.4% Caucasian). In multivariate analysis over a 10-year follow-up period, sinus tachycardia (HR ≥100 vs. <100 bpm) was an independent predictor of overall ACVO (OR 2.8, 95% CI: 1.4-5.5; P=0.006). There was increased incidence of HFrEF (OR 3.3, 95% CI: 1.6-6.5; P=0.004) and AHFE (OR 6.3, 95% CI: 1.6-28; P=0.023), but not HFpEF or ACS (P>0.05) compared with controls. Sinus tachycardia was a significant predictor of overall mortality after adjusting for significant covariates (HR 2.9, 95% CI 1.8-5; P<0.001). Independent of typical factors that affect cardiovascular disease, sinus tachycardia around the time of cancer treatment is associated with increased ACVO and mortality in cancer patients at 10 years of follow-up.


New perspectives on the effects of action on perceptual and cognitive development.
This special section was motivated by a resurgence in the view that it is impossible to investigate perceptual and cognitive development without considering how it is affected by, and intertwined with, infants' and children's action in the world. This view has long been foundational to the field, yet contemporary investigations of the effects of acting on cognition and perception have been limited. The research showcased in this section indicates that this trend is changing as researchers consider anew the ways in which cognition derives structure from action. The work presented here illustrates the breadth of these potential effects across ages and domains of development, and it highlights the breadth of methods that can be recruited to investigate them. This new research focus provides insight for the mechanisms by which action affects perception and cognition and at the same time reveals that much remains to be learned.


[Transfer of active oncogenes and promoters into the mouse cell genome].
Different cell DNA's (normal NIH 3T3 DNA; human osteosarcoma cell DNA; human malignant glioma cell DNA with amplified c-Ha-ras) were cotransfected onto NIH 3T3 cells with cloned long terminal repeat (LTR) sequences of Rous sarcoma virus. LTR RSV and normal NIH 3T3 DNA c-fos oncogen expression was detected in tumors induced in nude mice. In the same system human tumour cell DNA with amplified c-Ha-ras gene was used, that to the integration and amplification of LTP sequences with simultaneous maintenance of c-Ha-ras amplification. Nude mouse tumour DNA with integrated LTR sequences was active in successive rounds of transfection.


Microscopic Imaging of the Stress Tensor in Diamond Using in Situ Quantum Sensors.
The precise measurement of mechanical stress at the nanoscale is of fundamental and technological importance. In principle, all six independent variables of the stress tensor, which describe the direction and magnitude of compression/tension and shear stress in a solid, can be exploited to tune or enhance the properties of materials and devices. However, existing techniques to probe the local stress are generally incapable of measuring the entire stress tensor. Here, we make use of an ensemble of atomic-sized in situ strain sensors in diamond (nitrogen-vacancy defects) to achieve spatial mapping of the full stress tensor, with a submicrometer spatial resolution and a sensitivity of the order of 1 MPa (10 MPa) for the shear (axial) stress components. To illustrate the effectiveness and versatility of the technique, we apply it to a broad range of experimental situations, including mapping the stress induced by localized implantation damage, nanoindents, and scratches. In addition, we observe surprisingly large stress contributions from functional electronic devices fabricated on the diamond and also demonstrate sensitivity to deformations of materials in contact with the diamond. Our technique could enable in situ measurements of the mechanical response of diamond nanostructures under various stimuli, with potential applications in strain engineering for diamond-based quantum technologies and in nanomechanical sensing for on-chip mass spectroscopy.


The quantized DCT and its application to DCT-based video coding.
The two-dimensional (2-D) discrete cosine transform (DCT) and the subsequent quantization of the transform coefficients are two computationally demanding steps of any DCT-based video encoder. In this paper, we propose an efficient joint implementation of these two steps, where the precision in computing the DCT can be exchanged for a reduction in the computational complexity. First, the quantization is embedded in the DCT, thus eliminating the need to explicitly quantize the transform coefficients. A multiplierless integer implementation of the quantized DCT (QDCT) is then proposed that performs shift and add operations instead of full multiplications. A sequence of multiplierless QDCT algorithms is obtained with increasing precision and number of computations. Finally, further savings in computations are obtained by terminating the DCT computations whenever intermediate results indicate that the transform and quantization steps will likely result in a block of zero values. The proposed algorithms are applied to, and results are presented for, high-quality MPEG-2 and low bit rate H.263 video encoding.


Improving image segmentation performance and quantitative analysis via a computer-aided grading methodology for optical coherence tomography retinal image analysis.
We demonstrate quantitative analysis and error correction of optical coherence tomography (OCT) retinal images by using a custom-built, computer-aided grading methodology. A total of 60 Stratus OCT (Carl Zeiss Meditec, Dublin, California) B-scans collected from ten normal healthy eyes are analyzed by two independent graders. The average retinal thickness per macular region is compared with the automated Stratus OCT results. Intergrader and intragrader reproducibility is calculated by Bland-Altman plots of the mean difference between both gradings and by Pearson correlation coefficients. In addition, the correlation between Stratus OCT and our methodology-derived thickness is also presented. The mean thickness difference between Stratus OCT and our methodology is 6.53 microm and 26.71 microm when using the inner segment/outer segment (IS/OS) junction and outer segment/retinal pigment epithelium (OS/RPE) junction as the outer retinal border, respectively. Overall, the median of the thickness differences as a percentage of the mean thickness is less than 1% and 2% for the intragrader and intergrader reproducibility test, respectively. The measurement accuracy range of the OCT retinal image analysis (OCTRIMA) algorithm is between 0.27 and 1.47 microm and 0.6 and 1.76 microm for the intragrader and intergrader reproducibility tests, respectively. Pearson correlation coefficients demonstrate R(2)>0.98 for all Early Treatment Diabetic Retinopathy Study (ETDRS) regions. Our methodology facilitates a more robust and localized quantification of the retinal structure in normal healthy controls and patients with clinically significant intraretinal features.


PTHrP expression in chondrocytes, regulation by TGF-beta, and interactions between epiphyseal and growth plate chondrocytes.
Although PTHrP has been identified as a key regulator of chondrocyte differentiation in the growth plate, the factors directly regulating PTHrP expression have not been identified. Furthermore, while cells from the epiphysis are considered the physiologic source of PTHrP, the relative expression of PTHrP in epiphyseal and growth plate chondrocytes has not been defined. PTHrP expression was examined in chondrocytes isolated from 3- to 5-week-old chick long bones. The expression of PTHrP mRNA was 10-fold higher in epiphyseal chondrocytes compared to cells from the growth plate. Growth plate chondrocytes were isolated into populations with distinct maturational characteristics by countercurrent centrifugal elutriation and analyzed for PTHrP expression. The expression was highest in the least mature cells and progressively declined with the onset of maturation. The regulation of PTHrP expression was further examined in epiphyseal chondrocytes. Both TGF-beta1 and cis-retinoic acid stimulation markedly increased PTHrP mRNA levels, while BMP-2 and PTHrP stimulation decreased the expression of this transcript. The effects of TGF-beta1 (8.9-fold stimulation) and TGF-beta3 (9.2-fold) were slightly greater than the effects of TGF-beta2 (4.9-fold). The effect of TGF-beta was dose-dependent and increases could be detected after 68 h of treatment. To analyze the paracrine effect of epiphyseal and growth plate chondrocytes on each other, these cells were placed in coculture and the mRNA from each of the populations was harvested separately after 24 h. Following coculture the PTHrP mRNA levels increased in the epiphyseal cells while the expression of type X collagen and Indian hedgehog transcripts decreased in growth plate chondrocytes. The results demonstrate potentially important paracrine interactions between these cell populations, possibly mediated by TGF-beta and PTHrP.


The nature and origin of spontaneous noise in G protein-gated ion channels.
Arrival of agonist is generally thought to initiate the signal transduction process in G protein-receptor coupled systems. However, the muscarinic atrial K+ (K+[ACh]) channel opens spontaneously in the absence of applied agonist, giving a noisy appearance to the current records. We investigated the nature and origin of the noise by measuring single channel currents in cell-attached or excised, inside-out membrane patches. Guanosine triphosphate (GTP) produced identical single channel currents in a concentration- and Mg(2+)-dependent manner in the presence or absence of carbachol, but the requirements for GTP were greater in the absence of agonist. Hence the agonist-independent currents appeared to be produced by an endogenous G protein, Gk. This prediction was confirmed when an affinity-purified, sequence-specific Gi-3 alpha antibody or pertussis toxin (PTX) blocked the agonist-independent currents. Candidate endogenous agonists were ruled out by the lack of effect of their corresponding antagonists. Thus agonist-independent currents had the same nature as agonist-dependent K+[ACh] currents and seemed to originate in the same way. We have developed a hypothesis in which agonist-free, empty receptors prime Gk with GTP and Gk activates atrial K+ [ACh] channels producing basal currents or noise. Agonist-independent activation by G proteins of effectors including ion channels appears to be a common occurrence.


Technetium complexes with triazacyclononane.
[NBu4][TcOCl4] reacts with ethylene glycol and 1,4,7-triazacyclononane (tacn) in MeOH under the formation of the deep-blue oxotechnetium(V) cation [TcO(OC2H4O)(tacn)]+, which can readily be oxidized by air to give the stable technetium(VII) compound [TcO3(tacn)]+. The reaction with aqueous HCl results in reduction and the formation of the cationic technetium(III) complex [TcCl2(OH2)(tacn)]Cl. The products were isolated in crystalline form and studied spectroscopically and by X-ray diffraction.


Giant cavernous liver hemangiomas: effect of operative approach on outcome.
The choice of operative technique for resection of giant cavernous hepatic hemangiomas has an effect on outcome. Case series. Tertiary hepatobiliary surgery-liver transplantation service. Fifty-two adult patients who underwent resection of symptomatic hepatic hemangiomas (38 [73%] in the right lobe; average diameter, 10.9 cm) by means of lobectomy or enucleation, with or without Pringle inflow occlusion. Blood transfusion, morbidity, mortality, and length of hospitalization. Patient age and sex and the size of hemangiomas were similar for patients who underwent lobectomy and enucleation. Right-lobe lesions were more often treated by enucleation, and inflow occlusion was used more frequently; transfusion requirements and length of hospitalization were similar for both techniques. Complications were more frequent after lobectomy. There were no deaths. In the latter years of the series, enucleation was used in 22 (63%) and inflow occlusion in 24 (69%) of 35 patients. Outcome is related to the operative approach used for resection of giant cavernous liver hemangiomas. Although lobectomy and enucleation are curative, enucleation offers greater preservation of normal hepatic parenchyma and fewer complications and is the preferred technique for suitable lesions.


[Autoimmune neuropathies: diagnosis, treatment, and recent topics].
Here, we have reviewed the clinical patterns, diagnostic paradigms, etiopathogenesis, and therapeutic strategies of autoimmune neuropathies such as Guillain-Barré syndrome (GBS), chronic inflammatory demyelinating polyneuropathy (CIDP), multifocal motor neuropathy (MMN), and IgM paraproteinemic neuropathy. Antiganglioside antibodies are frequently present in the serum samples obtained during the acutephase of GBS and Miller Fisher syndrome (MFS), a subtype of GBS. Recently, we found that some patients with GBS and MFS have serum antibodies against antigenic epitopes formed by 2 different gangliosides (ganglioside complex). The antibodies against GD1a/GD1b and/or GD1b/GT1b complexes are associated with severe disability and a requirement for mechanical ventilation. Anti-GM1/GalNAc-GD1a antibodies are found to be associated with pure motor GBS with frequent conduction blocks. In GBS, corticosteroids given alone do not significantly hasten the recovery or affect the long-term treatment outcome. Intravenous immunoglobulin therapy (IVIg) or plasma exchange (PE) is equally effective. Combined treatment with corticosteroids and IVIg may be a promising therapy for GBS. On the basis of the EFNS/PNS guidelines, we describe the treatment of chronic autoimmune neuropathies such as CIDP, MMN, and IgM paraproteinemic neuropathy. In treating CIDP, corticosteroids, IVIg, and plasma exchange are equally effective. In MMN, IVIg is the first-choice therapy; corticosteroids and PE are ineffective or even detrimental. IgM paraproteinemic neuropathies are known to be intractable, and these patients often have anti-myelin-associated glycoprotein antibodies and may respond to immunosuppressive and immunomodulatory therapies. However, the potential therapeutic benefits should be balanced against their possible side effects and usual slow disease progression.


Condyloma Acuminata Presenting as Isolated Papillary Lesions in the Prostatic Urethra.
BACKGROUND A condyloma acuminatum is a sexually transmitted, human papillomavirus (HPV) associated, neoplasm. In men, it is predominantly found on external genitalia and rarely progresses more proximally than the distal penile urethra. Condyloma acuminata of the prostatic urethra are rare and are usually seen as an extension of, or in association with external lesions. Therefore, it is not typically considered in the differential diagnosis of isolated papillary lesions limited to the prostatic urethra. CASE REPORT A 62-year-old male with rheumatoid arthritis treated with abatacept presented to urology due to a history of intermittent bladder self-catheterization for urinary obstruction. He underwent a transurethral resection of the prostate and had incidental findings of papillary lesions restricted to the prostatic urethra that were presumed to be urothelial carcinoma. Microscopic examination established the diagnosis of condyloma acuminata, and low-risk HPV 6 and 11 were detected by in-situ hybridization. Subsequent cystoscopy showed marked growth and extension of condyloma acuminata to near the external meatus. After multiple treatments with intraurethral 5-fluorouracil, several small lesions remained in the bulbous urethra. With follow up for 2 years since diagnosis, the patient has not developed external condylomata. CONCLUSIONS A condyloma acuminatum might present as an isolated papillary growth in the prostatic urethra without clinical or historical evidence of a visible lesion on external genitalia. Immunosuppression and/or urethral instrumentation might be a risk factor for such a presentation. Urologists and pathologists should be aware of this rare possibility in order to avoid misdiagnosis, and ensure that the patient receives appropriate therapy.