Single-cell RNA-seq reveals abnormal differentiation of keratinocytes and increased inflammatory differentiated keratinocytes in atopic dermatitis.
Atopic dermatitis (AD) is a chronic and recurrent inflammatory skin disease characterized by severe pruritus and eczematous lesions. Heterogeneity of AD has been reported among different racial groups according to clinical, molecular and genetic differences. This study aimed to conduct an in-depth transcriptome analysis of AD in Chinese population. We performed single-cell RNA sequencing (scRNA-seq) analysis of skin biopsies from five Chinese adult patients with chronic AD and from four healthy controls, combined with multiplexed immunohistochemical analysis in whole-tissue skin biopsies. We explored the functions of IL19 in vitro. ScRNA-seq analysis was able to profile a total of 87,853 cells, with keratinocytes (KCs) in AD manifesting highly expressed keratinocyte activation and pro-inflammatory genes. KCs demonstrated a novel IL19+ IGFL1+ subpopulation that increased in AD lesions. Inflammatory cytokines IFNG, IL13, IL26 and IL22 were highly expressed in AD lesions. In vitro, IL19 directly downregulated KRT10 and LOR in HaCaT cells and activated HaCaT cells to produce TSLP. Abnormal proliferation and differentiation of keratinocytes contribute immensely to the pathogenesis of AD, whereas AD chronic lesions have witnessed significant presence of IL19+ IGFL1+ KCs, which may be involved in the disruption of the skin barrier, the connection and magnification of Th2 and Th17 inflammatory responses, and mediation of skin pruritus. Furthermore, progressive activation of multiple immune axes dominated by Type 2 inflammatory reaction occur in AD chronic lesions.


Interferon-gamma production specific for hepatitis B virus antigen by intrahepatic T lymphocytes in patients with acute and chronic hepatitis B.
Studies were undertaken to determine whether intrahepatic lymphocytes have a greater cellular immune response specific for hepatitis B virus (HBV) antigen than peripheral blood lymphocytes in HBV-infected man. HB nucleocapsid antigen-stimulated interferon-gamma (IFN-gamma) production of lymphocytes was measured in acute self-limited hepatitis (AH) (eight cases) and chronic hepatitis (CH) (14 cases). In both patient groups, basal IFN-gamma levels without any stimulation were higher in the hepatic T cells than in the peripheral T cells. The values in cultures from blood and liver were larger in AH than in CH. Alternatively, IFN-gamma response to HBcAg and HBeAg was less in hepatic T cells than in the corresponding blood cultures; T cell response was HLA class II restricted. Cell flow cytometry study showed that in all patient groups, percentages of CD3+, CD8+, and HLA-DR+ cells were significantly greater in liver than in peripheral blood, whereas the proportion of hepatic CD4+ cells was decreased, compared with that in peripheral CD4+ cells. These findings suggest that liver infiltrates are already activated in vivo to produce IFN-gamma, particularly in patients with AH. The changes in the proportion of lymphocyte subsets may be responsible for the altered IFN-gamma response of hepatic T cells.


Genotypic and Phenotypic Analysis of 34 Cases of Inherited Junctional Epidermolysis Bullosa caused by COL17A1 Mutations.
Inherited epidermolysis bullosa defines a heterogeneous group of genodermatoses characterized by skin and/or mucosa fragility resulting in blistering. The junctional variant (JEB) is associated with mutations affecting the genes expressing the components of the dermo-epidermal junction (DEJ) [1-2]. We report 34 JEB patients with COL17A1 genetic mutations diagnosed in our Center between 1993 and 2019. Medical and biological records were collected with a standardized questionnaire.


Mines in the End Zone: Are there Downsides to Team Performance?
Research on teams in organizations tends to focus on understanding the causes of team performance with a focus on how to enjoy the benefits of team success and avoid the negative consequences of team failure. This paper instead asks the question, 'what are some of the negative consequences of team success?' A review of the literature on teams is augmented with research from cognitive science, sociology, occupational psychology, and psychology to explore the potential negative long-term consequences of teamwork success. The general topics of groupthink, overconfidence bias, regression to the mean, role overload, and strategy calcification are reviewed while discussing the implications for future research streams and practical team management.


Effect of anti-tumor necrosis factor α treatment on radiographic progression in patient with ankylosing spondylitis: A systematic review and meta-analysis.
Efficacy of anti-tumor necrosis factor (anti-TNF)α treatment in patient with active ankylosing spondylitis (AS) had been proved by many clinical studies. Inflammation and new bone formation in spine were two pivotal aspects in AS. TNF α inhibitor could eliminate inflammation including clinical and laboratory inflammatory manifestation. Paradoxical results whether TNF α antagonist could delay radiographic progression in AS were often been reported simultaneously. To review the literature about the effect of TNF α inhibitor on radiographic progression and disease activity in patient with AS. We conducted a comprehensive search including Medline, EMBASE and the Cochrane Library from 1 January 2000 to 15 August 2017. Two reviewers independently supplemented with hand searching for the reference lists of inclusion. All trials focusing on radiographic progression or disease activity in patients with AS treated with anti-TNF α agents. Primary outcomes were modified Stokes AS Spinal Score (mSASSS), as well as Bath AS disease activity index (BASDAI) and Bath AS functional index (BASFI). Two reviewers independently selected studies and analyzed data. Methodological quality was assessed using the Newcastle-Ottawa scale (NOS). We pooled effects recorded on different scales as Standardized mean differences (SMDs) with 95% confidence intervals (CIs) using random-effects models. We included 14 studies of low to moderate risk of bias with 3,186 patients, compared with control group, there was no effect of mSASSS changes (SMD = -0.12, 95% CI: -1.17-0.93, p value = .82, I2 = 95%) and follow-up (SMD = 0.03, 95% CI: 0.21-0.26, p value = .82, I2 = 36%) estimation in anti-TNF α group. However anti-TNF α agent treatment led to remarkable improvements on both Bath AS disease activity index (BASDAI) (SMD = 1.06, 95% CI: 0.22-1.89, p value = .01, I2 = 96%) and Bath AS functional index (BASFI) (SMD = 0.93, 95% CI: 0.24-1.92, p value = .01, I2 = 97%) scores at 12 weeks. Our meta-analysis found no significant effect on delaying radiographic progression in AS treated with TNF α inhibitor, although TNF α inhibitor could do improve significantly disease activity and physical function in AS.


Distribution of dopamine D3 receptor expressing neurons in the human forebrain: comparison with D2 receptor expressing neurons.
The dopamine D2 and D3 receptors are members of the D2 subfamily that includes the D2, D3 and D4 receptor. In the rat, the D3 receptor exhibits a distribution restricted to mesolimbic regions with little overlap with the D2 receptor. Receptor binding and nonisotopic in situ hybridization were used to study the distribution of the D3 receptors and neurons positive for D3 mRNA in comparison to the D2 receptor/mRNA in subcortical regions of the human brain. D2 binding sites were detected in all brain areas studied, with the highest concentration found in the striatum followed by the nucleus accumbens, external segment of the globus pallidus, substantia nigra and ventral tegmental area, medial preoptic area and tuberomammillary nucleus of the hypothalamus. In most areas the presence of D2 receptor sites coincided with the presence of neurons positive for its mRNA. D3 binding sites and D3 mRNA positive neurons were most abundant in the limbic striatum and efferent structures, such as the nucleus accumbens, ventral striatum, substantia nigra, internal segment of the globus pallidus, anteroventral nucleus of the thalamus, and rostral pars reticulata of the substantia nigra. One important difference from the rat is that D3 receptors were virtually absent in the ventral tegmental area. D3 receptor and D3 mRNA positive neurons were observed in sensory, hormonal, and association regions such as the nucleus basalis, anteroventral, mediodorsal, and geniculate nuclei of the thalamus, mammillary nuclei, the basolateral, basomedial, and cortical nuclei of the amygdala. As revealed by simultaneous labeling for D3 and D2 mRNA, D3 mRNA was often expressed in D2 mRNA positive neurons. Neurons that solely expressed D2 mRNA were numerous and regionally widespread, whereas only occasional D3-positive-D2-negative cells were observed. The regions of relatively higher expression of the D3 receptor and its mRNA appeared linked through functional circuits, but co-expression of D2 and D3 mRNA suggests a functional convergence in many regions of the signals mediated by the two receptor subtypes.


Integrated transcriptomics and metabolomics analyses to investigate the anticancer mechanisms of cinobufagin against liver cancer through interfering with lipid, amino acid, carbohydrate, and nucleotide metabolism.
Liver cancer has characteristics of high morbidity, high mortality, and poor prognosis. Metabolic reprogramming is a prominent characteristic of tumors and plays a key role in promoting tumorigenesis. The metabolic process of liver cancer cells has undergone many significant changes including abnormal active glycolysis, enhanced de novo synthesis of fatty acids, and hyperactive metabolism of amino acids and nucleotides. Targeting metabolic reprogramming through regulation of anomalously expressed key metabolic enzymes and signaling molecules is considered to be an important strategy for liver cancer treatment. Multi-omics association analyses currently facilitate precise diagnosis, personalized clinical therapy, and revelation of mechanisms of drug action. Cinobufagin, as the major anti-tumor active ingredient of Chansu, the famous chinese medicine used in clinic for cancer treatment, has been reported to exert anticancer effects through many different kinds of mechanisms, but the effects of cinobufagin on metabolic reprogramming of cancer cells still remain unclear. In our study, we identify that cinobufagin exhibits anti-hepatoma effects through interfering with metabolic reprogramming (lipid, amino acid, carbohydrate, and nucleotide metabolism) based on integrated transcriptomics and metabolomics analyses. Furthermore, the results of integrated multi-omics analyses enrich various core regulatory mechanisms of anti-tumor effects of cinobufagin which are associated with metabolic pathway. In addition, some verifications of the enriched mechanisms related to intervention of lipid and carbohydrate metabolism in response to cinobufagin are also performed. This work will promote the innovation of the research model of TCM, and lay a solid theoretical foundation for the clinical application of cinobufagin and Chansu.


Neuronal beta-amyloid precursor protein gene expression: regulation by aurintricarboxylic acid.
beta-Amyloid precursor protein (beta-APP) and its derivative, amyloid beta-protein (beta-A4), may cause death of differentiated neurons and aurintricarboxylic acid (ATA), a metabolic inhibitor, improves neuronal survival. Therefore, we studied the effect of ATA on neuronal beta-APP gene expression. ATA decreased beta-APP mRNA levels by increasing its degradation, without changing the rate of transcription. ATA decreased both steady state and interleukin-1 (IL1)-induced increase in beta-APP mRNA levels. These effects of ATA were associated with rounding of cells suggestive of decreased cell adhesion or neurite retraction that was completely reversible when ATA was removed. However, beta-APP mRNA levels continued to remain suppressed in neurons that were actively regrowing neurites following discontinuation of ATA. In studies carried out upto 24 h, ATA did not damage cells as determined by Trypan blue exclusion, lactate dehydrogenase (LDH)-release and transmission electron microscopy. The findings suggest that constitutive or steady state levels of beta-APP mRNA may not be essential for the survival and growth of neurons and that ATA suppresses beta-APP expression without causing cell damage. These observations may be a basis for studying whether ATA or a related compound could beneficially regulate beta-APP levels in vivo.


Major sperm protein genes from Onchocerca volvulus.
Nematode spermatozoa, unlike their mammalian counterparts, are nonflagellated crawling cells. The pseudopod of these cells contains the major sperm protein (MSP) which comprises more than 15% of the protein in the sperm. MSP is presumed to function as a cytoskeletal element involved in motility. An Ascaris MSP cDNA sequence was used as a probe to identify and isolate Onchocerca volvulus MSP clones from a lambda gt11 genomic library. Two clones, OVGS-1 (765 bp) and OVGS-2 (1765 bp), were characterized by restriction endonuclease mapping and sequence analysis. Both genomic clones contain MSP protein coding regions of 99 and 282 bp separated by an intervening sequence of 153 bp. The genes OVGS-1 and OVGS-2 are 95% similar in nucleotide sequence in the protein coding regions, but only 79% similar in their intron sequences. A number of potential regulatory sequences in the flanking regions and at the exon/intron junctions of the O. volvulus MSP genes are in good agreement with consensus sequences in other eukaryotic cells. The nucleotide sequence of the O. volvulus MSP genes were over 80% similar to the Ascaris MSP cDNA sequence and 79% similar to the Caenorhabditis MSP-3 cDNA. The predicted amino acid sequence of the O. volvulus MSPs were 96% similar to each other, 90-91% similar to Ascaris MSP and 81-82% similar to Caenorhabditis MSP-3. These results offer evidence that the MSP sequences have been highly conserved throughout nematode evolution but are variable in their genomic organization and the presence of introns.


The place of conventional MRI and newly emerging MRI techniques in monitoring different aspects of treatment outcome.
Magnetic resonance imaging (MRI) is the most important paraclinical measure for assessing and monitoring the pathologic changes implicated in the onset and progression of multiple sclerosis (MS). Conventional MRI sequences, such as T1-weighted gadolinium (Gd) enhanced and spin-echo T2-weighted imaging, only provide an incomplete picture of the degree of inflammation and underlying neurodegenerative changes in this disease. Two- and three-dimensional fluid-attenuated inversion recovery and double inversion recovery sequences allow better identification of cortical, periventricular and infratentorial lesions. Ultra-high field strength MRI has the potential to detect subpial cortical and deep gray matter lesions. Unenhanced T1-weighted imaging can reveal hypointense black holes, a measure of chronic neurodegeneration. Magnetization transfer imaging (MTI) is increasingly used to characterize the evolution of MS lesions and normal-appearing brain tissue. Evidence suggests that the dynamics of magnetization transfer changes correlate with the extent of demyelination and remyelination. Magnetic resonance spectroscopy, which provides details on tissue biochemistry, metabolism, and function, also has the capacity to reveal neuroprotective mechanisms. By measuring the motion of water, diffusion imaging can provide information about the orientation, size, and geometry of tissue damage in white and gray matter. These advanced non-conventional MRI techniques relate better to clinical impairment, disease progression, accumulation of disability, and have the potential to detect neuroprotective effects of treatment. Although detecting the status of neuronal integrity using MRI techniques continues to improve, a "gold standard" model remains to be established.


Surgical resection versus open-approach radiofrequency ablation for small hepatocellular carcinomas within Milan criteria after successful transcatheter arterial chemoembolization.
The aim of this study was to compare the effectiveness and safety of hepatic resection versus open-approach RFA (ORFA) for small hepatocellular carcinomas (HCC) within Milan criteria after successful downstaging therapy by transcatheter arterial chemoembolization. Between February 2005 and February 2008, a total of 110 patients with advanced HCC met the Milan criteria after successful downstaging therapy; 58 patients then underwent hepatic resection and 52 received ORFA. Outcomes, including short- and long-term morbidity, 1-, 3-, and 5-year mortality and HCC-free survival, were analyzed and compared between the two groups. Patients in the hepatic resection and ORFA groups showed similar baseline characteristics and downstaging protocols. The ORFA group showed less blood loss, lower hospital costs, shorter surgical time, and fewer hospital stay days (P < 0.05). The 1-, 3-, and 5-year overall survival rates were 94.8, 86.2, and 79.3%, respectively, with liver resection and 96.2, 82.7, and 76.9% with ORFA (P=0.772). The 1-, 3-, and 5-year recurrence-free survival rates were 93.1, 81.0, and 77.6% with resection and 94.2, 76.9, and 73.1% with ORFA (P=0.705). The ORFA patients suffered fewer postoperative complications (P=0.09), particularly among the cases of central HCC (P=0.015). Resection and ORFA achieved similar survival benefits in the management of HCC within Milan criteria after successful downstaging. The decreased blood loss, hospital costs, surgical time, and hospital stay days, and lower complication rates in central cases render ORFA a preferred treatment option.


Effects of high and low flows on abundances of fish species in Central European headwater streams: The role of ecological species traits.
Hydrological variability is considered a major structuring factor for biotic and abiotic processes in freshwater ecosystems and is of particular importance to fish communities. We used hydrological indices to investigate the short intermediate-and long-term effects of high- and low-flow patterns on the population abundances of 17 fish species in headwater streams in Germany. Generalized linear models, on average, explained 54 % of the variability in fish abundance, whereas long-term hydrological indices performed better than indices based on shorter periods. Three clusters of species were differentiated in their response patterns to low-flow conditions. Cold stenotherm and demersal species were susceptible to high frequency and long duration but tolerant to the magnitude of low-flow events. In contrast, species with a more benthopelagic habitat preference and tolerance to warmer water were susceptible to magnitude but tolerated larger frequencies of low-flow events. The euryoecious chub (Squalius cephalus), tolerating both long durations and large magnitudes of low-flow events, formed its own cluster. Species responses to high flows were more complex and five clusters of species were differentiated. Species with an equilibrium life history strategy were positively affected by longer durations of high-flow conditions, which may allow them to take advantage of the extended floodplain, whereas opportunistic and periodic species significantly differed by thriving in events characterized by high magnitude and frequency. These response patterns of fish species to high and low flows help understand species-specific risks when hydrological conditions are altered by climate change or direct human intervention.


Aortic thrombosis as a dramatic vascular complication in COVID-19 disease.
To report clinical outcomes of COVID-19 related acute aortic thrombosis (AAT). Consecutive COVID-19 patients presenting with AAT between April 2020 and August 2021 were included retrospectively. Clinical and radiological data were prospectively collected. Ten patients (men, 90%; mean age, 64 ± 2 years) were included. At the time of AAT diagnosis, four patients were in intensive care unit. Median time between diagnosis of COVID-19 and AAT was 5 days [IQR 0-8.5]. Clinical presentation was acute lower limb ischaemia (n=9) and mesenteric ischaemia (n=2). Thrombus localization was the abdominal aorta (n=5), the thoracic aorta (n=2) or both (n=3), with the following embolic sites: lower limbs (n=9), renal arteries (n=3), superior mesenteric artery (n=2), splenic artery (n=1), cerebral arteries (n=1). Revascularization was performed in 9 patients, using open (n=6), endovascular (n=2) or hybrid techniques (n=1). Three patients required reinterventions. The 30-day mortality was 30%. Three major amputations were performed in two patients, resulting in a free-amputation survival rate of 50% after a median follow-up of 3,5 months [IQR 2-4.1]. AAT is a rare and devastating complication of COVID-19 disease, responsible for high mortality and amputation rates.


Recovery of cell volume and electrolytes of A6 cells after re-establishing isotonicity following hypotonic stress.
Cellular element concentrations and dry weight contents in A6 cells were determined using electron microprobe analysis to establish whether these cells exhibit a regulatory volume increase (post-RVD-RVI) when re-establishing isotonicity following a hypotonically induced regulatory volume decrease (RVD). Hypotonic stress was induced by reducing basolateral [NaCl], and hence, osmolarity fell from 260 to 140 mosmol/l. The alterations in cell volume after re-establishing isotonicity, calculated from the cellular dry weight changes, indicate within the first 2 min cell shrinkage from 120 to 76% of control, compatible with almost ideal osmometric behaviour of A6 cells, and thereafter a post-RVD-RVI to 94%. The cellular uptake of osmolytes necessary to explain the post-RVD-RVI could be accounted for solely by a gain in cellular K and Cl. The involvement of a Na-K-2Cl cotransporter in most of the KCl uptake seems plausible since basolateral bumetanide blocked KCl uptake and post-RVD-RVI. The net uptake of cations (K uptake of 185.2, Na loss of 8.2 mmol/kg dry wt) during the isotonic period exceeded the Cl uptake by 38.2 mmol/kg dry wt, suggesting the uptake of another anion and/or the alteration of cellular buffer capacity. The relatively low Na concentration maintained during the isotonic period (13.3 vs. 20.4 mmol/kg wet wt under control conditions) might favour electrolyte uptake via the Na-K-2Cl cotransporter.


The response of bovine beta-lactoglobulin-specific T-cell clones to single amino acid substitution of T-cell core epitope.
Cow's milk is one of the most common food allergens in the first year of life, with approximately 2.5% of infants experiencing an allergic reaction to it. Beta-lactoglobulin (BLG) is one of the major allergens in cow's milk. Previously, we reported that four of six T-cell clones (TCC) which were established from cow's milk allergy patients recognized BLGp97-117 as the core sequence and also recognized BLG in association with the human leucocyte antigen (HLA)-DRB1*0405 allele. Using two of these four TCCs, we evaluated the T-cell response to BLG peptides with single amino acid substitution or deletion and identified BLGp102-112 as the minimum essential region in BLGp97-117. In the alanine-scan assay, the proliferative responses of TCCs to pE108A disappeared, and the proliferative responses of TCCs to pC106A decreased. In the analog peptide proliferation assay, pY102S had retained some T-cell response to the two TCCs. Collecting these results, we propose a motif for the interaction between the HLA-DRB1*0405 allele and antigen peptide, and suggest that BLGp105-108 are important residues to retain the TCR/BLG-peptide/HLA complex. pY102A and pY102S are partial agonists for the T-cell receptor. These peptides might be considered as candidate peptides for the modification of the T-cell response to BLG in cow's milk allergy.


Variability in paediatric cardiac postoperative chest tube management.
Limited evidence exists to guide chest tube management following cardiac surgery in children. We assessed chest tube practice variation by surveying paediatric heart centres to prepare for a multi-site quality improvement project. We summarised management strategies highlighting variability in criteria for chest tube removal between and within centres. This lack of standardisation provides an opportunity for quality improvement.


Psychiatric aspects of posttraumatic epilepsy: A still unexplored area.
Traumatic brain injury (TBI) represents one of the most common causes of death and disability in young people, and posttraumatic epilepsy (PTE) accounts for 10% to 20% of all symptomatic epilepsies. However, PTE is still a relatively underappreciated condition. This paper aimed at reviewing current knowledge about psychiatric comorbidities of PTE, looking in particular at the nature of the relationship between TBI, psychiatric problems, and epilepsy, at the phenomenology of psychiatric disorders in PTE, and how to manage them. Data on psychiatric comorbidities of PTE are almost nonexistent, and this is a paradox considering that TBI itself is burdened by a number of cognitive and psychiatric sequelae, which can profoundly affect the everyday life of these patients. Preliminary data seem to suggest that the bidirectional relationship between epilepsy and psychiatric disorders is maintained in TBI and people with a psychiatric condition at the time of the TBI, or as a consequence of it, are at increased risk of developing PTE and vice versa. However, a number of questions are still unanswered concerning the genetic and environmental contributors, the phenomenology of psychiatric disorders in PTE, and how to prevent and address them properly. Further research in this area is urgently needed in order to provide the best possible care to people with PTE. Special Issue: Epilepsy & Behavior's 20th Anniversary.


Developmental Biology: Decapentaplegic Controls Growth at a Distance.
Decapentaplegic has long been thought to be a morphogen that controls patterning and growth in Drosophila wings, but hard evidence for the requisite long-range action has only now come from two new studies.


Postoperative lymphocysts after lymphadenectomy for gynaecological malignancies: preventive techniques and prospects.
Postoperative lymphocyst formation is an insufficiently recognised complication of lymphadenectomy for gynaecological malignancies. Lymphocysts are collections of lymph organised into cysts that develop in contact with lymphadenectomy compartments. There has been considerable debate about the relevance of lymphocyst prevention using surgical or pharmacotherapeutic methods. Here, we review the available studies about the impact of these methods on the incidence of lymphocysts. This review suggests that several techniques may decrease the incidence of lymphocysts when used in combination. On a literature basis, the peritoneum should be left open over the lymphadenectomy sites at the end of the procedure and drains should not be placed at the end of the procedure. Omentoplasty should be encouraged and further studies are needed to assess the potential benefits of new energies. Postoperative octreotide therapy seems beneficial but the role of this drug in pelvic oncological surgery remains to be determined.


Nasopharyngeal angiofibroma: a twenty year study.
A control rate of approximately 80% has been obtained in 45 cases of juvenile nasopharyngeal angiofibroma treated by a single moderate dose of radiotherapy at the Princess Margaret Hospital, Toronto. The remaining patients ultimately achieved control generally after further irradiation. Immediate side-effects of the treatment have been minimal, and no late complications have been observed. In 2-20 years' follow-up, no irradiation-induced tumors have been found. Superiority of irradiation in these cases over surgery is emphasized, as well as the importance of using sophisticated techniques to limit the volume of tissues irradiated and protect vulnerable radiosenstitive structures. Because tumor regression can be expected to continue for many months after treatment has been completed, a policy of observation following radiotherapy is recommended.


Effects of dimethyl sulfoxide on humoral immune responses to acetylcholine receptors in the rat.
The effects of dimethyl sulfoxide (DMSO) on the humoral immune responses to acetylcholine receptors (AChR) were studied in rats with experimental autoimmune myasthenia gravis. DMSO was administered during primary, ongoing, and secondary phases of the immune response to AChR. Anti-AChR antibody titers were measured to determine the effect of the treatment. DMSO had pronounced effects on the humoral immune response, which differed depending on the stage of the response and the strength of antigenic stimulation. When given during an ongoing immune response, DMSO suppressed anti-AChR antibody levels by an average of 53-76%. This effect was similar whether DMSO was given by oral, rectal, or intraperitoneal routes. DMSO treatment also suppressed the anti-AChR antibody response to a weak primary antigenic stimulus to a similar extent. In contrast, when administered during a secondary or a strong primary immunization, DMSO enhanced the anti-AChR antibody response 1.7- to 2.8-fold. These results show that DMSO may either enhance or suppress humoral immune responses. Further studies will be required to analyze the cellular mechanisms underlying the actions of DMSO.


[Bile dynamics in patients with postcholecystectomy syndrome].
Twenty-three cholecystectomized patients, asymptomatic or with recurrent biliary pain, were studied. Six patients were asymptomatic without organic complications of the biliary tree and negative cholestasis laboratory tests; six were symptomatic with gallstones or stenosis of the main biliary tree; eleven patients were symptomatic without organic diseases. All the groups of patients were matched for age and sex. The filling and emptying kinetics of the biliary tree were studied by sequential cholescintigraphy with trimetil-Br-IDA 99mTc and computized gamma-camera. The exam was performed for sixty minutes during fast and for sixty minutes after a standard cholecystokinetic meal. Hepatic clearance time of the compound, filling and emptying of the biliary tree were significantly longer in patients with organic obstruction than in the other groups. The differences between controls and patients with anorganic diseases were otherwise not significative. 1) in cholecystectomized patients the recurrent biliary pain is likely present independently of alterations of the biliary wash-out; 2) sequential cholescintigraphy is a very good method, as well as endoscopic procedures, to identify organic obstruction of the main biliary tree. However, it is not sufficiently sensitive to demonstrate functional alterations of the biliary tract.


Comparative anti-mitotic effects of lithium gamma-linolenate, gamma-linolenic acid and arachidonic acid, on transformed and embryonic cells.
The effects of gamma-linolenic acid (GLA), the lithium salt of gamma-linolenic acid (LiGLA) and arachidonic acid (AA) were compared at doses of 50 microg/ml for periods of 6 and 24 h on cell cycle progression and apoptosis induction in transformed and in normal cells. In WHCO3 (oesophageal cancer) cells and on primary embryonic equine lung cells, we found LiGLA to be the most effective in apoptosis induction. After 24 h, 94% of the WHCO3 cancer cells and 44% of the primary embryonic equine lung cells exposed to LiGLA were apoptotic. The WHCO3 cancer cells were also very susceptible to the apoptosis-inducing effects of AA (56%) and GLA (44%), whereas the embryonic equine lung cells were much less affected by these two fatty acids. After 6 h exposure to all three compounds, most of the cycling WHCO3 cancer cells were blocked in S-phase. After 24 h treatment, some of the S-phase cells exposed to AA and GLA were apparently able to move into the G2/M phase, the LiGLA exposed cells were mostly apoptotic and no cycling cells were present. The primary embryonic equine lung cells were fairly resistant to the cytotoxic effects of GLA and AA. From our studies we conclude that, although LiGLA was the most toxic to the cancer cells, it is apparently less selective, compared to AA and GLA, in the killing of cancer and normal cells. It would also appear that the lithium might have added to the cytotoxic effects of LiGLA. The mechanism needs to be clarified.


Effects of keratin on bone resorption in experimental otitis media.
Keratin debris is a constant feature in middle-ear cholesteatoma. Keratin prepared from rat skin induced a foreign-body granuloma in the subcutaneous space in the rat. In vitro this granuloma produced high levels of bone-resorbing factors: prostaglandin E2, osteoclast-activating factor, and leucine aminopeptidase. In the in vivo study, keratin-induced granuloma in the rat middle ear caused partial resorption of the cochlear wall. Macrophages, fibroblasts, and osteoclastlike cells were found at bone-resorption areas. These cells appeared to be responsible for bone resorption through production of prostaglandin E2, osteoclast-activating factor, and proteases.


Non-invasive method of measuring the resonant frequency of a human tibia in vivo. Part 2.
Part 1 describes the special instrumentation used in the non-invasive measurement of tibia resonance in vivo. Here we discuss some of the problems which arise when real fractures are measured and comment on accuracy repeatability and the effects of muscle tension.


Detection of cervical high-grade squamous intraepithelial lesions and assessing diagnostic performance of colposcopy among women with oncogenic HPV.
HPV screening tests may improve cervical cancer risk stratification and better guide decisions about follow-up with colposcopy/biopsy. This study aimed to estimate the risk of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) among women with oncogenic HPV types and evaluate the performance of colposcopy in the diagnosis of histologic CIN2 + at Putuo Hospital, Shanghai, China. This cross-sectional survey was conducted from February 2020 to December 2022 among women who were referred to colposcopy. Women with high-risk (HR) HPV-positive, cytology testing and colposcopy-directed biopsy were included. Univariate and multivariate analysis indicated that high-grade colposcopic impression ((OR, 17.61%, 95%CI: 11.54-26.85%) was associated with the highest risk for detecting CIN2+, followed by HSIL + cytology (OR, 6.90%, 95%CI: 3.56-13.37%) and HPV16/18 positive (OR, 2.91%, 95%CI: 2.12-3.99%). Overall, CIN2 + was detected in 14.6% of 2007 women. HPV16/18 had higher CIN2 + risks than other HR-HPV genotypes (30.1% vs. 10.2%, P<0.001). Among women with low-grade cytology, 24.1% had CIN2+, and the risks for HPV16/18 (58.2%) were higher than for other HR-HPV(16.8%). For those with high-grade cytology, there was no significant difference between HPV groups ( 75.0% vs. 72.9%, P > 0.05). The diagnostic performance of colposcopy in diagnosis of CIN2 + by senior and junior colposcopists was comparable. The results indicated that referral to colposcopy is recommended in managing women with HR-HPV positive, and colposcopic impressions provide key clues for identification CIN2+.


Measurement of GPCR-G protein activity in living cells.
G protein-coupled receptors (GPCRs) are the largest family of cell surface receptors in eukaryotic genomes. They control a variety of cellular and physiological processes such as hormone secretion and heart rate, and therefore are associated with a majority of pathological conditions including cancer and heart diseases. Currently established assays to measure ligand-induced activation of GPCRs and G proteins possess limitations such as being time consuming, indirect, and expensive. Thus, an efficient method to measure GPCR-G protein activation is required to identify novel pharmacological modulators to control them and gain insights about molecular underpinnings of the associated pathways. Activation of GPCRs induces dissociation of G protein heterotrimers to form GαGTP and free Gβγ. Free Gβγ subunits have been shown to translocate reversibly from the plasma membrane to internal membranes. Gβγ translocation therefore represents the GPCR-G protein activation, and thus, imaging of this process can be used to quantify the kinetics and magnitude of the pathway activation-deactivation in real time in living cells. The objective of this chapter is to elaborate the protocols of (i) generation and optimization of the required sensor constructs; (ii) development of cell culture, transient transfection, imaging, and optogenetic procedures; (iii) imaging and data analysis methods; and (iv) stable cell line generation, pertaining to this assay to measure GPCR-G protein activation.


Potential adult sibling stem cell donors' perceptions and opinions regarding an information and care model.
The first international recommendations and guidelines for the care of sibling stem cell donors were established in 2010, and have not yet been evaluated. However, a model for information and care of adult potential sibling stem cell donors (the IC model) developed and introduced at the Skåne University Hospital, Lund, Sweden, in 2005 conforms closely to them. The IC model aims to protect the privacy of potential donors, support and respect their free choice, and identify quickly those unwilling or unable to donate and thus minimize delay in seeking alternative donors. To evaluate the IC model a questionnaire survey in 2010 gathered the perceptions and views on information provision; influences over decision making; and care provision under the IC model of 148 adult siblings informed about SCT donation, and asked to undergo HLA-typing since September 2005 at the hospital. The results suggest the IC model works well but highlights areas for improvement, such as in delivery of HLA typing results to non-matched siblings, and a need to further prevent complicating influence from health professionals and relatives on the decision to undergo HLA typing. Thus improved, the IC model could provide the groundwork for other SCT units seeking to implement the recommendations and guidelines.


Allostatic load may not be associated with chronic stress in pregnant women, NHANES 1999-2006.
Pregnant women are generally excluded from studies that measure allostatic load (AL) because there is concern that the changing levels of AL-related biomarkers during pregnancy do not reflect a woman's true AL. The goal of this study was to determine whether AL can be measured in a meaningful way during pregnancy. The National Health and Nutrition Examination Survey (NHANES) is a nationally representative, cross-sectional survey of the U.S. civilian population. AL was based on the distributions of 10 biomarkers in pregnant (n = 1138) and nonpregnant (n = 4993) women aged 15 to 44 from NHANES (1999-2006). The distribution of each AL-related biomarker differed significantly between pregnant and nonpregnant women (P < .01). Among nonpregnant women, high AL findings were consistent with previous studies (e.g., higher AL in women who are black, are older, and who have lower incomes). However, these associations were not seen in pregnant women. Our results suggest that the various biomarkers that comprise AL may reflect proximal factors in pregnancy more strongly than they represent exposure to chronic stress over a woman's lifetime. Therefore, our approach to measuring AL may not provide meaningful information about chronic stress in pregnant women without further consideration of pregnancy-related factors.


A Comparison of Hyperextension versus Neutral Positions for Vertebroplasty to Treat Neurologically Intact Kümmell Disease.
To compare the efficacy between hyperextension position (HPVP) and neutral position for vertebroplasty (NPVP) in treating Kümmell disease. A Comparative descriptive study. Department of Orthopaedics, Yantaishan Hospital, China, from December 2017 to July 2018. This study retrospectively analysed demographic features, operative information, radiologic data, and complications of 58 consecutive patients with single-level Kummel disease (KD) who underwent NPVP (n=27) or HPVP (n=31). All patients were observed preoperatively and at 2 days (POD 2) and one year postoperatively for cement leakage, Cobb's angle, antenior body height ratio, pain and disability. The cement leakage rate was significantly lower in the HPVP group (p<0.01). The visual analogue scale (VAS) scores for pain, and Oswestry disability indices (ODIs) were lower in the HPVP group at one year postoperatively (both p<0.05). For the HPVP group, Cobb's angle was significantly smaller, and the anterior body height ratio (AR) was larger at POD 2 (p<0.05) and one year (p<0.05), postoperatively. This study demonstrated that HPVP could achieve a lower cement leakage rate with similar operative time, lower VAS and ODI scores, as well as better kyphosis restoration and AR recovery at the 1-year follow-up than NPVP. HPVP is thus superior to NPVP in treating KD. Key Words: Kyphosis, Cobb's angle, Cement leakage, Intervertebral stability, Surgical outcome, Kümmell disease, Vertebroplasty, Hyperextension position.


Preclinical evaluation of a novel EGFR&c-Met bispecific near infrared probe for visualization of esophageal cancer and metastatic lymph nodes.
This study aimed to establish a near infrared fluorescent (NIRF) probe based on an EGFR&c-Met bispecific antibody for visualization of esophageal cancer (EC) and metastatic lymph nodes (mLNs). EGFR and c-Met expression were assessed by immunohistochemistry. EGFR&c-Met bispecific antibody EMB01 was labeled with IRDye800cw. The binding of EMB01-IR800 was assessed by enzyme linked immunosorbent assay, flow cytometry, and immunofluorescence. Subcutaneous tumors, orthotopic tumors, and patient-derived xenograft (PDX) were established for in vivo fluorescent imaging. PDX models using lymph nodes with or without metastasis were constructed to assess the performance of EMB01-IR800 in differential diagnosis of lymph nodes. The prevalence of overexpressing EGFR or c-Met was significantly higher than single marker either in EC or corresponding mLNs. The bispecific probe EMB01-IR800 was successfully synthesized, with strong binding affinity. EMB01-IR800 showed strong cellular binding to both Kyse30 (EGFR overexpressing) and OE33 (c-Met overexpressing) cells. In vivo fluorescent imaging showed prominent EMB01-IR800 uptake in either Kyse30 or OE33 subcutaneous tumors. Likewise, EMB01-IR800 exhibited superior tumor enrichment in both thoracic orthotopic esophageal squamous cell carcinoma and abdominal orthotopic esophageal adenocarcinoma models. Moreover, EMB01-IR800 produced significantly higher fluorescence in patient-derived mLNs than in benign lymph nodes. This study demonstrated the complementary overexpression of EGFR and c-Met in EC. Compared to single-target probes, the EGFR&c-Met bispecific NIRF probe can efficiently depict heterogeneous esophageal tumors and mLNs, which greatly increased the sensitivity of tumor and mLN identification.


Influence of Pleurotus ostreatus inoculation on wood degradation and fungal colonization.
The influence of Pleurotus ostreatus inoculation on wood degradation and on fungal community structure was studied. The experiments were performed on an organically poor fly ash deposit covered with a 10 cm layer of beech wood chips inoculated with P. ostreatus isolate ZIM76. Compared to non-inoculated wood chips, inoculation increased the temperatures and relative humidities and, in the first 6 months, accelerated Klason lignin degradation by 9% and also, after 17 months, increased iron translocation into wood chips by 30%. After 6 months, PCR-DGGE showed 22-28 and 13-21 fungal taxa in non-inoculated and P. ostreatus-inoculated beech chips, respectively. The differences in number of taxa and in the fungal community structure (based on Dice coefficient) between non-inoculated and inoculated wood chips diminished with time. The results indicate that the naturally occurring processes of wood degradation are as efficient as those occurring in sites inoculated with P. ostreatus.


Imaging the development of an ischemic core following photochemically induced cortical infarction in rats using Laser Speckle Contrast Analysis (LASCA).
Laser Speckle Contrast Analysis (LASCA) has been used to image the development of an ischemic core following photochemically induced infarction in rats up to 1 h post-lesion. Using LASCA, we have been able to image a central ischemic core which had little or no perfusion surrounded by a penumbral region with reduced perfusion. In addition, we have shown the existence of a surrounding region of hyperemic tissue. A potential feature of this imaging approach is its capability to track cerebral blood flow (CBF) changes in the region within and outside the ischemic core besides demonstrating the real-time progression of the ischemic core into the penumbral region. We have demonstrated the continuous disruption of CBF to the ischemic core that eventually affected the blood supply to the surrounding regions. The penumbral flow is shown to exhibit a sudden increase post-ischemic induction followed by a slow decline to the final baseline level. Interestingly, we observed an interaction (P < 0.03) between penumbral flow peak and the time effects of increase in pixel area from the infarct region to the surrounding penumbral region. Using a paired-sample t test, we observed that the mean pixel area was larger for the infarct region than for the penumbral region (P < 0.004) during the time interval between the induction of ischemia and the time point of peak flow in the penumbral region.


CGRP inhibits human Langerhans cells infection with HSV by differentially modulating specific HSV-1 and HSV-2 entry mechanisms.
Herpes simplex virus (HSV) is widespread globally, with both HSV-1 and HSV-2 responsible for genital herpes. During sexual transmission, HSV targets epithelial cells, sensory peripheral pain neurons secreting the mucosal neuropeptide calcitonin gene-related peptide (CGRP), and mucosal immune cells including Langerhans cells (LCs). We previously described a neuro-immune crosstalk, whereby CGRP inhibits LCs-mediated human immunodeficiency virus type 1 (HIV-1) transmission. Herein, to further explore CGRP-mediated anti-viral function, we investigated whether CGRP affects LCs infection with HSV. We found that both HSV-1 and HSV-2 primary isolates productively infect monocyte-derived LCs (MDLCs) and inner foreskin LCs. Moreover, CGRP significantly inhibits infection with both HSV subtypes of MDLCs and langerinhigh, but not langerinlow, inner foreskin LCs. For HSV-1, infection is mediated via the HSV-1-specific entry receptor 3-O sulfated heparan sulfate (3-OS HS) in a pH-depended manner, and CGRP down-regulates 3-OS HS surface expression, as well as abrogates pH dependency. For HSV-2, infection involves langerin-mediated endocytosis in a pH-independent manner, and CGRP up-regulates surface expression of atypical langerin double-trimer oligomers. Our results show that CGRP inhibits mucosal HSV infection by differentially modulating subtype-specific entry receptors and mechanisms in human LCs. CGRP could turn out useful for prevention of LCs-mediated HSV infection and HSV/HIV-1 co-infection.


Increasing the accuracy of volume and ADC delineation for heterogeneous tumor on diffusion-weighted MRI: correlation with PET/CT.
To improve the accuracy of volume and apparent diffusion coefficient (ADC) measurements in diffusion-weighted magnetic resonance imaging (MRI), we proposed a method based on thresholding both the b0 images and the ADC maps. In 21 heterogeneous lesions from patients with metastatic gastrointestinal stromal tumors (GIST), gross lesion were manually contoured, and corresponding volumes and ADCs were denoted as gross tumor volume (GTV) and gross ADC (ADC(g)), respectively. Using a k-means clustering algorithm, the probable high-cellularity tumor tissues were selected based on b0 images and ADC maps. ADC and volume of the tissues selected using the proposed method were denoted as thresholded ADC (ADC(thr)) and high-cellularity tumor volume (HCTV), respectively. The metabolic tumor volume (MTV) in positron emission tomography (PET)/computed tomography (CT) was measured using 40% maximum standard uptake value (SUV(max)) as the lower threshold, and corresponding mean SUV (SUV(mean)) was also measured. HCTV had excellent concordance with MTV according to Pearson's correlation (r=0.984, P<.001) and linear regression (slope = 1.085, intercept = -4.731). In contrast, GTV overestimated the volume and differed significantly from MTV (P=.005). ADC(thr) correlated significantly and strongly with SUV(mean) (r=-0.807, P<.001) and SUV(max) (r=-0.843, P<.001); both were stronger than those of ADC(g). The proposed lesion-adaptive semiautomatic method can help segment high-cellularity tissues that match hypermetabolic tissues in PET/CT and enables more accurate volume and ADC delineation on diffusion-weighted MR images of GIST.


Examining Childhood Obesity From Infancy: The Relationship Between Tummy Time, Infant BMI-z, Weight Gain, and Motor Development-An Exploratory Study.
This exploratory study investigated the infant time spent in tummy time (TT) in relation to body mass index z score (BMI-z), weight gain, and motor development in infants from birth to 4 months. Mothers and their infants were telephone surveyed at 2 and 4 months. Mother demographics; TT; feeding practices; and infant length, and height, and development were collected each time. Results from Cochran-Mantel-Haenszel and single logistic regression showed a significant association between development, level of BMI-z, and time spent in TT at 2 months of age (p < .0001). The threshold model showed there was a decline in BMI-z at 4 months as daily time in TT increases past the threshold value of approximately 12 minutes per day. Mother education and TT at 2 months were significant predictors of BMI-z at 4 months. Study outcomes suggest that infant positioning and time in TT promote infant motor development and may moderate rapid infant weight gain.


Synaptic Plasticity: Close Encounters of the Tonic and Phasic Kind.
Neuronal circuits have the capacity to maintain relatively constant activity in the face of perturbations that alter excitability or synaptic properties. A new study demonstrates that different classes of neurons co-innervating the same postsynaptic target express homeostatic plasticity with unique presynaptic features.


Interobserver agreement in the reporting of colorectal polyp pathology among bowel cancer screening pathologists in Wales.
To assess the interobserver agreement in the reporting of colorectal polyps among histopathologists participating in the Welsh Bowel Cancer Screening (BCS) programme. Twelve benign polyps representative of BCS cases were identified from pathology files and reported by 28 BCS histopathologists using proforma sheets. The level of agreement between the participants and a gold standard was determined using kappa (κ) statistics. A moderate level of agreement was achieved in the reporting of polyp type [κ = 0.45; 95% confidence interval (CI) 0.34-0.59] and adenomatous lesions were distinguished from non-adenomatous lesions in 96% of cases. Substantial agreement was obtained in distinguishing low- and high-grade dysplasias (κ = 0.67; 95% CI 0.50-0.86), but there was only fair agreement in reporting excision margin status (κ = 0.24; 95% CI 0.07-0.43) with frequent use of the 'uncertain' category. Significant issues included categorizing serrated lesions, recognizing focal high-grade dysplasia and epithelial misplacement, and apparent overdiagnosis of villous change in adenomas. Interobserver variability in some aspects of reporting colorectal polyps by BCS pathologists is suboptimal, with a potential impact upon patient management and the efficient running of the screening service. Approaches to addressing this are discussed.


Ultrasound-mediated drug delivery using liposomes modified with a thermosensitive polymer.
Ultrasound-mediated drug delivery was established using liposomes that were modified with the thermosensitive polymer (TSP) poly(NIPMAM-co-NIPAM), which sensitized the liposomes to high temperatures. TSP-modified liposomes (TSP liposomes) released encapsulated calcein under 1 MHz ultrasound irradiation at 0.5 W/cm(2) for 120 s as well as the case under incubation at 42 °C for 15 min. In addition, uptake of the drug released from TSP liposomes by cancer cells was enhanced by ultrasound irradiation. In a cell injury assay using doxorubicin (DOX)-loaded TSP liposomes and ultrasound irradiation, cell viability of HepG2 cells at 6 h after ultrasound irradiation (1 MHz, 0.5 W/cm(2) for 30 s) with DOX-loaded TSP liposomes (TSP/lipid ratio=1) was 60%, which was significantly lower than that of the control conditions such as DOX-loaded TSP liposomes alone and DOX-loaded intact liposomes under ultrasound irradiation.


Treatment of multifocal pain with spinal cord stimulation.
We report a retrospective case study of combined treatment of cancer-related pain and chronic low back and lower extremity pain related to postlaminectomy syndrome (PLS) with one spinal cord stimulation (SCS) system. The patient underwent an uneventful SCS trial with percutaneous placement of two temporary eight-electrode leads (Medtronic Inc., Minneapolis, MN) placed at the level of T8-T9-T10 and T5-T6-T7. After successful trial, he was implanted with permanent leads and generator, reporting sustained pain relief at 12-month follow-up visit. SCS is a trialable, reversible, and interactive therapy permitting patients to control the level of stimulation they feel based on their degree of pain. SCS provides an effective, alternative treatment for select patients with cancer-related chest wall pain and pain related to PLS who have failed conservative treatment.


Brugia malayi galectin 2 is a tandem-repeat type galectin capable of binding mammalian polysaccharides.
Galectins are among the most abundant excretory/secretory (ES) products of filarial worms, but their role in filarial biology is poorly understood. Galectin-2 (Lec-2), a major component of Brugia malayi extracellular vesicles, is released by filarial worms, and was recently identified in the serum of persons with loiasis. We therefore sought to clone and characterize Lec-2, and to develop reagents to examine its potential as a biomarker and its role in parasite biology. We cloned and expressed recombinant B. malayi Lec-2 (rBmLec-2), generated a Lec-2-specific monoclonal antibody (4B4), and used it to confirm the presence of Lec-2 in B. malayi ES products and whole worm lysate. We show that Lec-2 is absent in B. malayi oocytes, and increases in concentration as embryos mature. Recombinant BmLec-2 hemagglutinates rabbit red blood cells at concentrations less than 1 μg/mL, and this is abrogated by single amino acid substitutions in the predicted carbohydrate recognition domains. rBmLec-2 binds multiple LacNAc oligosaccharides on a mammalian carbohydrate array. Sera from 17/23 (78 %) persons with microfilaremic loiasis and 4/10 (40 %) persons with bancroftian filariasis had detectable antibody to Lec-2 by western blot. Our studies confirm the functionality of BmLec-2 and indicate anti-Lec-2 antibody responses are common in persons with filariasis. These studies set the stage for further examination of the role of Lec-2 in filarial biology and in filarial-host interactions.


Diabetes mellitus in Swedish middle-aged men. The study of men born in 1913 and 1923.
As part of a study of the epidemiology of diabetes mellitus in middle-aged Swedish men, the present paper reports the prevalence and incidence of diabetes and the prevalence of impaired glucose tolerance. Two cohorts of 50-year-old men, representative of the corresponding male population of Gothenburg, Sweden, were examined in 1963 and 1973, respectively, and then followed until 1980. In the cohort of men born in 1913 (n = 855) the diabetes prevalence (WHO criteria), based on a questionnaire and fasting blood glucose, increased from 1.5% at age 50 to 7.6% at age 67. In the cohort of men born in 1923 (n = 226) the prevalence was 3.7% at age 50 and 4.0% at age 57. The overall prevalence of diabetes and impaired glucose tolerance was 25% among men born in 1913 (age 67) and 18% among men born in 1923 (age 57). The cumulative risk of developing diabetes from age 50 to 67 was 7.8%. Variables associated with impaired glucose tolerance and newly found diabetes, when degree of obesity was considered, were systolic blood pressure and triglycerides, well known risk factors for both coronary heart disease and diabetes. Uric acid, fasting insulin and glutamic puruvic transaminase, recently discussed as possible risk factors, were also associated with impaired glucose tolerance and newly found diabetes. Thus, both impaired glucose tolerance and newly found diabetes were associated with a clustering of risk factors, not only for diabetes but also for coronary heart disease.


Gut microbiota and atopic dermatitis in children: a scoping review.
Gut microbiota plays an important role in the development of atopic dermatitis (AD). We aimed to elucidate research trends in gut microbiota and AD in children, to provide evidence and insights to the clinical prevention and treatment of AD in children. A scoping literature review on the studies of gut microbiota and AD were conducted. Two authors independently searched Pubmed et al. databases for studies focused on gut microbiota and AD in children up to January 15, 2022. The literatures were screened and analyzed by two reviewers. A total of 44 reports were finally included and analyzed. Current researches have indicated that abnormal human microecology is closely associated with AD, and the disturbance of intestinal microbiota plays an important role in the occurrence and development of AD. Probiotics can correct the microbiota disorder, have the functions of regulating immunity, antioxidant, and help to restore the microecological homeostasis. However, there is still a lack of high-quality research reports on the efficacy and safety of probiotics in the prevention and treatment of AD in children. The changes of gut microbiota are essential to the development of AD in children, which may be an effective target for the prevention and treatment of AD. Future studies with larger sample size and rigorous design are needed to elucidate the effects and safety of probiotics in AD.


Clinical application of single-pulse transcranial magnetic stimulation for the treatment of depression.
Transcranial magnetic stimulation (TMS) has been recently suggested for the treatment of patients with major depression. Based on the results of the authors' pilot study showing a possible antidepressive effect of single-pulse TMS, a clinical trial was conducted involving patients with major depression. For the present study single-photon emission computed tomography (SPECT) was recorded for six of the target patients to study the effects of TMS on the local blood flow volume. Twenty-three inpatients meeting the Diagnostic and Statistical Manual of Mental Disorders (4th edn; DSM-IV) criteria for major depression were invited to participate in the study. Depressive symptoms were rated using the Hamilton Rating Scale for Depression (HAM-D). Patients were given 10 stimuli over the frontal area of both sides for a total of 20 stimuli in a session. The subjects had daily TMS session for 5 days as an add-on therapy. In addition, six patients had their quantitative (99m)Tc-ethyl cysteinate dimer SPECT images measured before and after TMS treatment. Compared with the value 2 days prior to the start of TMS therapy (24.2 +/- 4.9), the average HAM-D scale dropped significantly to 15.3 +/- 6.6 on the day after completion of such therapy. The results of SPECT showed that the regional cerebral blood flow (rCBF) of the bilateral frontal region had increased in four out of six patients when comparing before and after treatment. The present study shows that single-pulse TMS, which is widely used as a neurological test method, possesses a wide range of antidepressive effects without inducing adverse reactions. The results suggest that although repetitive TMS is steadily becoming the mainstay technique today, single-pulse TMS also possesses sufficient antidepressive effects.


Strategies for developing a recombinant butyrylcholinesterase medical countermeasure for Organophosphorus poisoning.
Organophosphorus nerve agents represent a serious chemical threat due to their ease of production and scale of impact. The recent use of the nerve agent Novichok has re-emphasised the need for broad-spectrum medical countermeasures (MCMs) to these agents. However, current MCMs are limited. Plasma derived human butyrylcholinesterase (huBChE) is a promising novel bioscavenger MCM strategy, but is prohibitively expensive to isolate from human plasma at scale. Efforts to produce recombinant huBChE (rBChE) in various protein expression platforms have failed to achieve key critical attributes of huBChE such as circulatory half-life. These proteins often lack critical features such as tetrameric structure and requisite post-translational modifications. This review evaluates previous attempts to generate rBChE and assesses recent advances in mammalian cell expression and protein engineering strategies that could be deployed to achieve the required half-life and yield for a viable rBChE MCM. This includes the addition of a proline-rich attachment domain, fusion proteins, post translational modifications, expression system selection and optimised downstream processes. Whilst challenges remain, a combinatorial application of these strategies demonstrates potential as a technically feasible approach to achieving a bioactive and cost effective bioscavenger MCM.


Electrophysiology and the magnetic sense: a guide to best practice.
Magnetoreception, sensing the Earth's magnetic field, is used by many species in orientation and navigation. While this is established on the behavioural level, there is a severe lack in knowledge on the underlying neuronal mechanisms of this sense. A powerful technique to study the neuronal processing of magnetic cues is electrophysiology but, thus far, few studies have adopted this technique. Why is this the case? A fundamental problem is the introduction of electromagnetic noise (induction) caused by the magnetic stimuli, within electrophysiological recordings which, if too large, prevents feasible separation of neuronal signals from the induction artefacts. Here, we address the concerns surrounding the use of electromagnetic coils within electrophysiology experiments and assess whether these would prevent viable electrophysiological recordings within a generated magnetic field. We present calculations of the induced voltages in typical experimental situations and compare them against the neuronal signals measured with different electrophysiological techniques. Finally, we provide guidelines that should help limit and account for possible induction artefacts. In conclusion, if great care is taken, viable electrophysiological recordings from magnetoreceptive cells are achievable and promise to provide new insights on the neuronal basis of the magnetic sense.


Adolescent problem drinking. Psychosocial correlates in a national sample study.
A profile, developed using multivariate analysis of psychosocial data from a national survey of adolescent drinking, indicated that problem drinking was part of a general adaptation to self, others and circumstances rather than an isolated activity.


The use of short-term tests to measure the preventive action of reducing agents on formation and activation of carcinogenic nitroso compounds.
The effect of reducing agents on the nitrosation of methylguanidine (MG) and on the in vitro activation of dimethylnitrosamine (DMN) was examined by measuring DNA-repair synthesis (unscheduled incorporation of [3h]TdR), shifts in alkaline sucrose gradients, frequency of chromosome aberrations, and clone-forming capacity of cultured human fibroblasts. The reducing agents examined were sodium ascorbate, cysteine, cysteamine, and propyl gallate. Since the short-term bioassays used can be quantitated, it has become relatively easy to detect the inhibitory action of reducing compounds on the nitrosation reaction of MG and metabolic activation (with S-9 preparation) of the precarcinogen DMN, to measure their effective dose range, and to establish the most effective ratios between inhibitory agent and reactant. The results indicate that DNA-repair synthesis is a suitable short-term test for studying the numerous combinations and premutations between several carcinogenic or non-carcinogenic agents, and for estimating the capacity of inhibitory agents to affect formation and activation of chemical carcinogens.


Deep brain stimulation in China: present and future.
We reviewed the clinical applications, academic communications, and specialized training required for deep brain stimulation (DBS) in China. Current problems and possible solutions also were discussed. We reviewed all literature on DBS by Chinese authors. The WANFANG and PUBMED (2000-2009) data bases were searched to collect publications on DBS by Chinese authors. To compare the Chinese academic publications on DBS with those by American authors, data from publications by American authors also were collected. Information on the DBS population as categorized by year, by province, and by indication was collected from the Medtronic Neuromodulation data base. From 2000 to 2009, there were 187 publications on DBS in Chinese journals and 23 in English journals by Chinese authors. American peers contributed 569 articles during the same period. In total, 2082 patients received DBS treatment in China by the end of 2009. DBS has evolved significantly during the past decade in China. There are still several problems. The advent of Chinese homemade DBS hardware, multidisciplinary cooperation, and the establishment of guidelines and regulations for DBS will improve the application of this surgical treatment in China.


Caring for Sexual and Gender Minority Adults with Cardiovascular Disease.
This article summarizes existing evidence on cardiovascular disease (CVD) risk and CVD diagnoses among sexual and gender minority adults and provides recommendations for providing nursing care to sexual and gender minority adults with CVD. More research is needed to develop evidence-based strategies to care for sexual and gender minority adults with CVD.


Susceptibility of human liver cell cultures to hepatitis C virus infection.
To develop a cell culture system susceptible to infection by hepatitis C virus (HCV), human fetal hepatocytes, grown in serum-free medium, were inoculated with serum samples from two HCV-infected patients. Viral RNA sequences were detected by polymerase chain reaction, using primers specific for the 5' noncoding region of HCV, in extracts prepared from the hepatocyte cultures as early as 5 days after inoculation. Virus was also released from the infected cells into the medium. The HCV strains could be serially passaged three times into fresh liver cell cultures using intracellular virus as inoculum. Evidence that HCV replication really took place in primary human fetal hepatocytes was also obtained by detection of minus-strand viral RNA (replication intermediate) in cell extracts and of viral antigens in the infected cells.


Spatio-temporal partitioning facilitates mesocarnivore sympatry in the Stara Planina Mountains, Bulgaria.
The top trophic level in many terrestrial food webs is typically occupied by mammalian carnivores (Order Carnivora) that broadly affect and shape ecosystems through trophic cascades. Their inter-specific interactions can further complicate effects on community dynamics as a consequence of intra-guild competition. The capacity for competitive mammalian carnivores to segregate their hunting and activity regimes is in major part a function of their similarity, in terms of body-size and dietary niche; termed the 'niche variation hypothesis'. In this study, we used camera-trapping to investigate intra-guild interactions and spatio-temporal partitioning among five mesocarnivores, the golden jackal (Canis aureus), European badger (Meles meles), red fox (Vulpes vulpes), European wildcat (Felis sylvestris) and stone marten (Martes foina), in the Stara Planina Mountains, Bulgaria. We collected a total of 444 images of golden jackals, 236 images of European badgers, 200 images of red foxes, 171 images of stone martens, and 145 images of European wildcats, from 6612 camera-days across fifteen camera-trapping stations. With respect to body size, the three smaller species (fox, wildcat and marten) were active in different time periods than the two larger competitors (jackal and badger) through both the warm and cold season. The more similar the trophic niche between species pairs (particularly relating to rodent consumption), the greater the spatio-temporal partitioning we observed within the pair; however, this adapted to seasonal dietary shifts. In conclusion, spatial and temporal (fine-scale and seasonal) niche partitioning appeared to reduce encounter probabilities and competition and may act to facilitate sympatric coexistence among this regional mesocarnivore guild.


An endocavitary rectal irradiation technique.
An alternative to standard short target-skin distance (TSD) rectal irradiation techniques is presented. The proctoscopic rectal cone is linked outside of the patient to a superficial X ray tube operated at 50 kVp and 25 mA. The cone is rigidly held in place by a hydraulic clamp and linked to the X ray tube with an electromagnetic lock. The X ray beam is filtered by 0.3 mm of aluminum, giving a half-value-layer of 0.27 mm of aluminum and an exposure rate of 720 R/min. (0.186 C/kg-min.) at the treatment TSD of 22 cm. The physical aspects of the technique are described in detail.


Effects of SpoIVA on the formation of spores and crystal protein in Bacillus thuringiensis.
In addition to forming spores, Bacillus thuringiensis (Bt) 4.0718 can produce toxins, insecticidal crystal protein (ICP) and vegetative insecticidal protein (Vip). The Bt spoIVA was successfully knocked out by gene recombination and was shown to inhibit sporulation. The mutant strain also exhibited significantly decreased growth and crystal formation, which inhibited spore formation and partially reduced the rate of crystal synthesis. The 50 % lethal concentrations (LC50) values of Bt 4.0718, replacement, complementation and multi-copy mutant strains against the fourth larval stage of H. armigera was determined as 5.422, 6.776, 6.223 and 5.018 μg/mL, respectively. A total of 1814 proteins were identified through isobaric tags for relative and absolute protein (iTRAQ), with 41 and 54 up and downregulated proteins observed. Gene ontology enrichment analysis showed that differentially expressed proteins were primarily involved in the biological process and molecular function. Quantitative real-time PCR analysis confirmed that 9 differential expressed genes exhibited a positive correlation between changes at transcriptional and translational levels. The results of this study provide a basis for further studies of the metabolic regulatory network of spores and crystal protein formation. Moreover, they can be used to ecologically safe insecticide of farmland production because the constructed Bt spoIVA mutants did not produce spores.Provides new ideas for the targeted improvement and application of environmentally friendly spore-free strains.


Characterizing liver sinusoidal endothelial cell fenestrae on soft substrates upon AFM imaging and deep learning.
Liver sinusoidal endothelial cells (LSECs) display unique fenestrated morphology. Alterations in the size and number of fenestrae play a crucial role in the progression of various liver diseases. While their features have been visualized using atomic force microscopy (AFM), the in situ imaging methods and off-line analyses are further required for fenestra quantification. Primary mouse LSECs were cultured on a collagen-I-coated culture dish, or a polydimethylsiloxane (PDMS) or polyacrylamide (PA) hydrogel substrate. An AFM contact mode was applied to visualize fenestrae on individual fixed LSECs. Collected images were analyzed using an in-house developed image recognition program based on fully convolutional networks (FCN). Key scanning parameters were first optimized for visualizing the fenestrae on LSECs on culture dish, which was also applicable for the LSECs cultured on various hydrogels. The intermediate-magnification morphology images of LSECs were used for developing the FCN-based, fenestra recognition program. This program enabled us to recognize the vast majority of fenestrae from AFM images after twice trainings at a typical accuracy of 81.6% on soft substrate and also quantify the statistics of porosity, number of fenestrae and distribution of fenestra diameter. Combining AFM imaging with FCN training is able to quantify the morphological distributions of LSEC fenestrae on various substrates. AFM images acquired and analyzed here provided the global information of surface ultramicroscopic structures over an entire cell, which is fundamental in understanding their regulatory mechanisms and pathophysiological relevance in fenestra-like evolution of individual cells on stiffness-varied substrates.


The effect of strontium ranelate on serum insulin like growth factor-1 and leptin levels in osteoporotic post-menopausal women: a prospective study.
The aim of this study was to investigate the effects of strontium ranelate on insulin like growth factor-1 (IGF-1), leptin and osteocalcin levels in osteoporotic post-menopausal women. Thirty-three women were given 2 g/day strontium ranelate for 6 months. Serum IGF-1, leptin and osteocalcin levels were measured before and after the treatment. Strontium ranelate treatment increased IGF-1 levels significantly (P = 0.02). Leptin and osteocalcin levels did not change significantly before and after the treatment. There was no correlation between basal and 6-months treatment levels of IGF-1 and leptin. Strontium ranelate increases serum IGF-1 levels in osteoporotic post-menopausal women.


Colon cancer: comprehensive evaluation with 64-section CT colonography using water enema as intraluminal contrast agent-a pictorial review.
Sixty-four-section CT colonography with water enema combines intracolonic neutral contrast agent with high-resolution CT images of the abdomen. Owing to submillimeter isotropic voxels, high-quality reformatted images are obtained. High-resolution images offer added value for the detection and localization of colonic lesions, evaluation of the local extent of the disease, and depiction, if any, of synchronous colorectal lesions and distant metastases. Sixty-four-section CT colonography with water enema has a major role in the evaluation of patients with colon cancer before planning therapy. It can be used to complement failed or incomplete colonoscopy and investigate the colon in elderly patients.


Central generation of bursting in the feeding system of the snail, Lymnaea stagnalis.
The central generation of burst activity was investigated in the buccal ganglia of Lymnaea. Eight different patterns of burst activity were generated by one or two consecutive phases of compound synaptic potentials resulting from activity of neurones. These inputs acted upon the different endogenous properties of buccal neurones such as post-inhibitory rebound and spike adaptation. Effects of synaptic inputs were reinforced by by electrotonic coupling of some buccal neurones of the same type.


Pediatric bilateral vocal cord granulomas presenting as airway foreign body following prolonged intubation due to COVID-19 related multisystem inflammatory syndrome.
This report is a case of bilateral vocal cord granulomas (VCG) in a 9-year-old child presenting as an airway foreign body and stridor following prolonged intubation due to COVID-19 pneumonia complicated by multisystem inflammatory syndrome in children (MIS-C). This case reports a 9-year-old male who presented to the emergency department with acute stridor concerning for airway obstruction. X-ray findings suggested an airway foreign body; however, on rigid bronchoscopy, ball-valving bilateral VCG obstructing 90% of the glottic airway were identified. After excision of the VCGs, the patient's symptoms resolved with no postoperative complications. With suspected foreign body aspiration in a pediatric patient with a history of prolonged intubation, it is important to consider a broad differential diagnosis. VCGs are rare complications of prolonged intubation that could be in children with prior history of intubation following COVID-19 pneumonia.


Multifocal tuberculous osteomyelitis.
A case of multifocal tuberculous osteomyelitis is reported. The lesions were found in the skull and ribs in a 10 year old girl with pulmonary tuberculosis and with tuberculous lymphadenitis. Complete healing was achieved under chemotherapy. Although radiologically the lesions appear "cystic", their true nature is that of granulomatous--caseous tuberculous osteomyelitis.


What information and the extent of information to be provided in an informed assent/consent form of pediatric drug trials.
This study aimed to determine the elements and the extent of information that child participants and their parents would like to read in an informed assent form (IAF)/informed consent form (ICF) of a pediatric drug trial. A descriptive survey was conducted to determine the perceived importance of each element of the ICF content from child participants and their parents who underwent informed assent/consent of a multi-center pediatric drug trial. The respondents were asked to indicate the level of importance of each item in a questionnaire, by giving a rating scale from 1 (not important) to 5 (very important). A total of 22 families, 17 child participants with the diagnosis of hematology or oncology diseases and 27 parents, were enrolled. Among 30 items, risk-benefit aspects (i.e., direct health benefit [mean: 4.71 for child respondents, 4.89 for parent respondents], indirect/societal benefit [mean: 4.65, 4.85], major foreseeable risk [mean: 4.47, 4.78], post-trial benefit/provision [mean: 4.59, 4.74], and all adverse effects of the drug including uncommon adverse effects [mean: 4.53, 4.74]) were perceived to be of most concerning items from both child participants' and parents' viewpoint. None of the items were considered 'slightly important' or lower by more than 20% of the respondents. For pediatric drug trials, risk-benefit information (including direct health benefit, indirect/societal benefit, and post-trial benefit/provision, as well as major foreseeable risk and adverse effects of the drug) should be made a salient feature of an IAF/ICF. This empirical data could help related stakeholders arrange essential information in order of importance and tailor an IAF/ICF to better suit child participants' and parents' needs, particularly for pediatric drug trials involving children with the diagnosis of hematology or oncology diseases.


Evaluation of corrosion on plasma sprayed and anodized titanium implants, both with and without bone cement.
The corrosion behavior of titanium with vacuum plasma sprayed titanium coatings and with anodized surfaces, both with and without polymeric bone cement were evaluated. Electrochemical extraction tests were carried out with subsequent analysis of the electrolyte by ICP-MS in order to verify our hypothesis of the ionic permeability of the polymer cement. The complexity of the situation resides in the existence of two interfaces: electrolyte-polymer and polymer-metal. The surface preparation (treatment of the surface) plays an important role in the corrosion resistance of titanium. The electrochemical magnitudes that were examined reveal that the plasma spray surfaces have the lowest corrosion resistance. The cement, in spite of having reduced electrical conductivity in comparison to metal, is an ionic transporter, and therefore capable of participating in the corrosion process. In the present study, we observed in fact crevice corrosion at the metal-cement interface. In the case of plasma spray surfaces, a process of diffusion of titanium particles in the electrolyte could accompany the crevice corrosion. In this study, we have shown that there is a corrosion process at the surface of the titanium through the cement which has as a consequence on the one hand the formation of titanium cations and on the other hand the growth of a passive layer on the titanium. In conclusion, we identified two principal factors that influence the corrosion process: [1] the type of surface treatment for the titanium, and [2] the ionic conductivity of the cement. There is indeed ionic transport through the cement; as evidenced by the presence of titanium in the electrolyte solution (ICP-MS analysis) and chloride at the surface of the titanium sample (EDX analysis). We show that the polymer cement is an ionic conductor and participates in the corrosion of the embedded titanium. We cannot deduce from our results, however, whether the polymer itself possesses corrosive properties. Long-term experiments will be necessary to study the degradation behavior of the polymer cement.


Immunohistological study of NM 23 and C-erbB-2 expression in primary tumor and metastases of colorectal adenocarcinoma.
Immunohistochemical study of expression of nm 23 and c-erbB-2 protein markers in primary tumor and metastases of colorectal adenocarcinoma showed that the incidence and expression of both protein markers in primary tumor tissue tended to increase after the appearance of liver metastases. Peculiarities of protein expression in tumor metastases in lymph nodes and liver and the possibility of using these markers as additional prognostic criteria in colorectal cancer are discussed.


Prevalence and characterization of hepatitis B and C virus infections in a needle-sharing population in Northern China.
The epidemiologies of hepatitis C virus (HCV) and hepatitis B virus (HBV) infections in specific populations in certain areas of China are poorly understood. A pilot survey of HCV/HBV infections was carried out in villages in Kuancheng County, Heben Province, where injection of sodium benzoate or amphetamines using shared needles has been a common practice. The aims of this study were to analyze the endemicity and characterize HCV/HBV infections in this population. Data on demographic characteristics and drug abuse were collected from individuals who signed informed consent forms. Serum HCV antibody (anti-HCV), hepatitis B surface antigen (HBsAg), and hepatitis B core antibody (anti-HBc) were measured in all participants. HCV RNA was measured in samples positive for anti-HCV using real-time polymerase chain reaction. Among 852 participants from 11 villages, 49.9% had used sodium benzoate or amphetamine at least once, by intravenous injection. The overall prevalence of anti-HCV, HCV RNA, anti-HBc, HBsAg, and HCV/HBV co-infection was 37.1%, 26.6%, 67.7%, 10.7%, and 30.0%, respectively. Two-hundred-twenty-three of 227 (98.2%) participants positive for HCV RNA were aged >40 years. Co-infection was related to sex, age, number of injections, and time from first injection. The rate of spontaneous HCV RNA clearance was 28.2% (89/316), and was related to the number of injections, time from first injection, and HBsAg positivity. However, HBsAg was related to the anti-HBc signal/cut-off ratio rather than to the above parameters. Trend tests demonstrated that the prevalence of anti-HCV, HCV RNA, and anti-HBc was related to the number of injections (P < 0.001), while HBsAg prevalence was not (P = 0.347). The prevalence of HCV and HBV infection is likely to be high among individuals older than 40 years in areas of needle sharing, and one-time screening for HCV infection should be offered to these populations.


Characterization of SepE and SepF for the N6-Glycosylated Adenine Structure Formation in Septacidin Biosynthesis.
Septacidin (1) represents a group of nucleoside antibiotics possessing a unique N6-glycosylated adenine core. They exhibit some fascinating bioactivities that are rare for other nucleoside antibiotics. Here we demonstrate that this unique structure in septacidin is formed by SepE and SepF. SepE is an unprecedented Fe(II)-dependent glycosyltransferase decorating the N6-position of AMP using ADP-l-glycero-β-d-manno-heptose (6) as a sugar donor. The Fe(II) may help SepE to bind AMP. SepF is an unusual glycosidase that detaches the N9-ribosyl-5-phosphate.


Age-dependent changes in in vivo ethanol metabolism and in the activity of hepatic enzymes involved in ethanol oxidation and microsomal functions.
The study of the influence of the age of the animals (13 to 53 weeks) on the rate of ethanol metabolism in vivo and the total activity of liver alcohol dehydrogenase and microsomal ethanol oxidizing system showed a progressive decline with age. These effects were observed concomitantly with a diminution in the content of cytochrome P-450 and microsomal functions related to oxidative and free-radical mediated reactions, namely, NADPH oxidase activity, NADPH-dependent oxygen uptake and NADPH-or t-butyl hydroperoxide-induced chemiluminescence. It is concluded that ageing is accompanied by a diminution in the total oxidative activity of the liver tissue, which would explain the depression in basal and ethanol-induced lipid peroxidation found in the oldest group of rats studied.


Outcome prognostic factors in inoperable malignant bowel obstruction.
Inoperable malignant bowel obstruction (MBO), a severe complication of peritoneal carcinomatosis, has a low desobstruction rate (30-40 %) and end-of-life decision-making is hampered by the lack of known prognostic factors. This study aimed to explore prognostic factors for desobstruction in MBO. All patients with inoperable MBO admitted in our large oncology hospital between 2010 and 2013 were treated following a clinical protocol based on antiemetics, steroids and two antisecretories, octreotide, and hyoscine butylbromide. Two prognostic factor analyses using logistic regressions were performed, one based on data from day 1 of admission and the other on data from day 8. Forty-five patients were included. Frequency of desobstruction was 48.9 %. In the analysis of prognostic factors on day 1, MBO episodes derived from functional physiopathologic mechanisms (vs. mechanic or mixed) were more prone to resolve (p < 0.001 corrected for multiple comparisons). Considering patients alive with persistent obstruction on day 8, a better clinical condition was the variable more associated with desobstruction, but without statistical significance after correction for multiple comparisons. A functional physiopathologic mechanism of MBO development may be an early prognostic factor for desobstruction. A high proportion of desobstruction was observed, suggesting that the combination of antisecretories with different mechanism of action warrants further investigation.


Pregnanolone, a metabolite of progesterone, stimulates LH-RH release: in vitro and in vivo studies.
The effect of 5 beta- and 5 alpha-reduced progestins on luteinizing hormone-releasing hormone (LH-RH) release was examined using either an in vitro superfusion or an in vivo push-pull perfusion (PPP) technique. Ovariectomized rats (at least 6 days post operation: OVX) were implanted subcutaneously in the neck region with a silastic capsule containing 17 beta-estradiol (235 micrograms/ml: E2). Two days afterwards, these OVX + E2 rats were subjected to experimental conditions. Pregnanolone(5 beta-pregnane-3 beta-ol-20-one) at a low concentration of 0.01 ng/ml and an apparent latency of 1 h stimulated in vitro LH-RH release from superfused hypothalamic fragments containing the preoptic-anterior hypothalamic-mediobasal hypothalamic area (POA-AHA-MBH). The stimulatory effect of pregnanolone required estrogen-priming and a pulsatile mode of administration. In addition, the effect appeared quite specific since other closely related steroids such as epipregnanolone, R-5020 and 5 alpha-DHP were ineffective. In OVX + E2 rats bearing a push-pull cannula (PPC) in the hypothalamic area, in vivo infusion of repetitive pulses of pregnanolone through the PPC was also effective, confirming in vivo the stimulatory effect of pregnanolone on LH-RH release obtained from in vitro preparations. In these experiments, the same OVX + E2 rats served both as an experimental animal receiving pulses of pregnanolone and as a control subject when perfused only with medium after a week interval. The basal release rate of LH-RH and the response to pulsatile pregnanolone were variable among animals apparently due to different locations of PPC in the hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS)


Increased affinity of choline acetyltransferase for choline in Alzheimer's disease: a steady-state kinetic study.
The steady-state kinetics of choline acetyltransferase (CAT) from autopsy samples of human caudate nucleus of aged controls and of patients with Alzheimer's disease was studied. In 10 samples from Alzheimer's disease-afflicted brains the affinity for the limiting substrate choline (Ch) was significantly higher: Michaelis constant KmCh was for these samples 1.93 +/- 0.72 mM while in the samples from 9 age-matched controls KmCh was 2.53 +/- 0.78 mM. The difference is statistically significant (P less than 0.05). Endogenous choline concentrations in the samples were 124 +/- 39 (n = 10) nmol/g wet wt. in the Alzheimer's disease-afflicted samples and 180 +/- 57 (n = 9) nmol/g wet weight (n = 9) in the control samples (P less than 0.05). The initial velocity at 70 microM acetyl co-enzyme (AcCoA) in Alzheimer's samples was 171.5 +/- 131.0 pmol [14C]acetyl choline [14C]ACh/mg protein/min as compared to the controls 422.1 +/- 231.0 pmol [14C]ACh/mg protein/min replicating many previous findings about decline of CAT activity in Alzheimer's disease. However, in the same samples the affinity for the other substrate acetyl-CoA (AcCoA) was significantly lower for the Alzheimer patients, KmAcCoA = 61 +/- 40 microM, than for the age-matched control patients, KmAcCoA = 28 +/- 8 microns (P less than 0.01). The data suggest some compensation of the loss of enzyme molecules via changed affinity for the limiting substrate, Ch.


The effects of leucotrofina or timolimfotropina on some parameters of carbohydrate metabolism in the Wistar rat.
A 10 days Leucotrofina or Timolimfotropina administration induced in Wistar rats a rise of glycemia and liver glycogen content, and a decrease of glycogen-phosphorylase a activity. These modifications lasted in time and may be due to an activation of the hepatic gluconcogenetic pathway, via a stimulated glucocorticoid production.


Expression of I-region-associated antigen (Ia) and interleukin 1 by subcultured human endothelial cells.
Activation of T cells requires three signals from an antigen-presenting cell: antigen, Ia determinants (HLA-D region determinants in man), and interleukin 1 (IL-1). Recent evidence has suggested that macrophages, dendritic cells, epidermal Langerhan's cells, and endothelial cells can each function as antigen-presenting cells (APC). If these cell types can independently function as APC, they should synthesize Ia determinants and secrete IL-1. To determine if endothelial cells fulfill these requirements, we have propagated human umbilical vein endothelial cells by serial subculture for extended periods of time and assessed Ia expression and IL-1 secretion. The endothelial cells were subcultured for 8 months (approximately 20 subcultures) and were found to display classic morphology and immunofluorescent staining for the endothelial cell-specific marker Factor VIII-related antigen. In a separate paper we have shown that these subcultured endothelial cells can present antigen to T cells in a HLA-D region-restricted fashion (C. R. Wagner, R. M. Vetto, and D. R. Burger, Subcultured human endothelial cells can independently function as fully competent antigen-presenting cells, accepted for publication, Hum. Immunol.). In this paper we present evidence demonstrating that extensively subcultured endothelial cells biosynthesize both HLA-DR and HLA-DS molecules after exposure to T cells and antigen or to a supernatant from antigen-activated T cells. Evidence is also presented that when endothelial cells are cultured in the presence of lipopolysaccharide they secrete a molecule(s) with IL-1 activity as assayed by LBRM-33-IA5 cell line production of interleukin 2.


Involvement of nigrotecto-reticulospinal pathways in the iminodipropionitrile (IDPN) model of spasmodic dyskinesias: a 2-deoxy-D-[1-14C]glucose study in the rat.
Chronic administration of iminodipropionitrile (IDPN) to rats causes a persistent behavioral syndrome characterized by lateral and vertical head twitches, random circling, and increased tactile and acoustic startle responses. In order to identify brain areas which are affected in rats manifesting this syndrome, we used the autoradiographic 2-deoxy-D-[1-14C]glucose ([14C]DG) method to map cerebral glucose utilization in IDPN-treated rats. One day after the development of the dyskinetic syndrome, there were significant decreases in local glucose utilization in the substantia nigra pars reticulata (SNr) and compacta (SNc), the dorsal raphe, the superficial and deep layers of the superior colliculus, the inferior colliculi, the interpeduncular nucleus, the medial and dorsolateral geniculate nuclei, and the superior and lateral vestibular nuclei. There were also significant decreases in layer 2 of the cingulate cortex and in the temporal and occipital cortices. In contrast, there were no changes in the motor cortex, the caudate-putamen, the nucleus accumbens, or the median raphe. These findings suggest that deleterious effects of IDPN on the nigrotectal pathways which affect head and neck movements and circling behaviors via the brainstem reticulospinal tracts may play an important role in the IDPN-induced persistent spasmodic dyskinetic syndrome in rats.


Role of biostimulants in mitigating the effects of climate change on crop performance.
Climate change is a critical yield-limiting factor that has threatened the entire global crop production system in the present scenario. The use of biostimulants in agriculture has shown tremendous potential in combating climate change-induced stresses such as drought, salinity, temperature stress, etc. Biostimulants are organic compounds, microbes, or amalgamation of both that could regulate plant growth behavior through molecular alteration and physiological, biochemical, and anatomical modulations. Their nature is diverse due to the varying composition of bioactive compounds, and they function through various modes of action. To generate a successful biostimulatory action on crops under different parameters, a multi-omics approach would be beneficial to identify or predict its outcome comprehensively. The 'omics' approach has greatly helped us to understand the mode of action of biostimulants on plants at cellular levels. Biostimulants acting as a messenger in signal transduction resembling phytohormones and other chemical compounds and their cross-talk in various abiotic stresses help us design future crop management under changing climate, thus, sustaining food security with finite natural resources. This review article elucidates the strategic potential and prospects of biostimulants in mitigating the adverse impacts of harsh environmental conditions on plants.


Mapping Dynamic Exposure: Constructing GIS Models of Spatiotemporal Heterogeneity in Artificial Stream Systems.
In flowing environments, the degree of turbulent flow determines the movement and distribution of chemicals. Variation in flow alters the patchiness of toxicant plumes within a stream ecosystem. This patchiness translates into variability in exposure pulses for organisms encountering the toxic plume. Throughout a stream, the processes that give rise to chemical plume structure will vary as a function of local flow characteristics. This research examines the influence of toxicant mode of entry and stream flow velocity on the spatiotemporal patterning of exposure. Two introduction treatments were evaluated: one mimicking groundwater and the other mimicking runoff. The influence of flow regime was examined through the comparison of models constructed under two stream flow velocities. Concentrations of a tracer molecule were recorded using an electrochemical monitoring system. From these localized, direct measurements, geographic information systems (GIS) were used to model exposure throughout the stream. Conceptualizing exposure as a series of toxicant pulses, exposure can be defined using a variety of chemical peak characteristics. Three-dimensional, layered maps were constructed defining exposure as the integrated area of toxicant peaks, the magnitude of peaks, and peak frequency. Differences in the spatial and temporal patterning of exposure were apparent both within treatments and between treatments. No two definitions of exposure yielded the same exposure distributions for any treatment. These models demonstrate that distribution of chemical exposure throughout a stream ecosystem is linked to both toxicant mode of introduction and stream hydrodynamics. Furthermore, these results demonstrate that optimal exposure modeling relies on first defining exposure.