A flexible two-photon fiberscope for fast activity imaging and precise optogenetic photostimulation of neurons in freely moving mice.
We developed a flexible two-photon microendoscope (2P-FENDO) capable of all-optical brain investigation at near cellular resolution in freely moving mice. The system performs fast two-photon (2P) functional imaging and 2P holographic photostimulation of single and multiple cells using axially confined extended spots. Proof-of-principle experiments were performed in freely moving mice co-expressing jGCaMP7s and the opsin ChRmine in the visual or barrel cortex. On a field of view of 250 μm in diameter, we demonstrated functional imaging at a frame rate of up to 50 Hz and precise photostimulation of selected groups of cells. With the capability to simultaneously image and control defined neuronal networks in freely moving animals, 2P-FENDO will enable a precise investigation of neuronal functions in the brain during naturalistic behaviors.


Association between copayment, medication adherence and outcomes in the management of patients with diabetes and heart failure.
To determine the association between copayment, medication adherence and outcomes in patients with Heart failure (HF) and Diabetes Mellitus (DM). PubMed, Scopus and Cochrane databases were searched using combinations of four sets of key words for: drug cost sharing; resource use, health and economic outcomes; medication adherence; and chronic disease. Thirty eight studies were included in the review. Concerning the direct effect of copayment changes on outcomes, the scarcity and diversity of data, does not allow us to reach a clear conclusion, although there is some evidence indicating that higher copayments may result in poorer health and economic outcomes. Seven and one studies evaluating the relationship between copayment and medication adherence in DM and HF population, respectively, demonstrated an inverse statistically significant association. All studies (29) examining the relationship between medication adherence and outcomes, revealed that increased adherence is associated with health benefits in both DM and HF patients. Finally, the majority of studies in both populations, showed that medication adherence was related to lower resource utilization which in turn may lead to lower total healthcare cost. The results of our systematic review imply that lower copayments may result in higher medication adherence, which in turn may lead to better health outcomes and lower total healthcare expenses. Future studies are recommended to reinforce these findings.


Psoralea corylifolia extract induces vasodilation in rat arteries through both endothelium-dependent and -independent mechanisms involving inhibition of TRPC3 channel activity and elaboration of prostaglandin.
Fructus Psoralea, Psoralea corylifolia L. (Leguminosae), has been widely used in traditional medicines for the treatment of dermatitis, leukoderma, asthma and osteoporosis. In this study, we sought to study mechanisms underlying the vasoactive properties of Psoralea corylifolia extract (PCE) and its active ingredients. To study mechanisms underlying the vasoactive properties of PCE prepared by extracting dried seeds of Psoralea corylifolia with 70% ethanol, isometric tension recordings of rat aortic rings and the ionic currents through TRPC3 (transient receptor potential canonical 3) channels were measured with the cumulative concentration (10-600 μg/mL) of PCE or its constituents. Cumulative treatment with PCE caused the relaxation of pre-contracted aortic rings in the presence and absence of endothelium with EC50 values of 61.27 ± 3.11 and 211.13 ± 18.74 μg/mL, respectively. Pretreatment with inhibitors of nitric oxide (NO) synthase, guanylate cyclase, or cyclooxygenase and pyrazole 3, a selective TRPC3 channel blocker, significantly decreased PCE-induced vasorelaxation (p < 0.01). The PCE constituents, bakuchiol, isobavachalcone, isopsoralen and psoralen, inhibited hTRPC3 currents (inhibited by 40.6 ± 2.7, 27.1 ± 7.9, 35.1 ± 4.8 and 47.4 ± 3.9%, respectively). Furthermore, these constituents significantly relaxed pre-contracted aortic rings (EC50 128.9, 4.5, 32.1 and 114.9 μg/mL, respectively). Taken together, our data indicate that the vasodilatory actions of PCE are dependent on endothelial NO/cGMP and also involved in prostaglandin production. PCE and its active constituents, bakuchiol, isobavachalcone, isopsoralen and psoralen, caused dose-dependent inhibition of TRPC3 channels, indicating that those ingredients attenuate Phe-induced vasoconstriction.


Spatial Confinement Modulates Macrophage Response in Microporous Annealed Particle (MAP) Scaffolds.
Macrophages are essential in the initiation, maintenance, and transition of inflammatory processes such as foreign body response and wound healing. Mounting evidence suggests that physical factors also modulate macrophage activation. 2D in vitro systems demonstrate that constraining macrophages to small areas or channels modulates their phenotypes and changes their responses to known inflammatory agents such as lipopolysaccharide. However, how dimensionality and pore size affect macrophage phenotype is less explored. In this work, the change in macrophage M1/M2 polarization when confined in microporous annealed particle (MAP) scaffolds is studied. Particles sizes (40, 70, and 130 µm) are selected using outputs from software LOVAMAP that analyzes the characteristics of 3D pores in MAP gels. As the size of building block particle correlates with pore size inside the scaffolds, the three types of scaffold allow us to study how the degree of spatial confinement modulates the behavior of embedded macrophages. Spatially confining macrophages in scaffolds with pore size on the scale of cells leads to a reduced level of the inflammatory response, which is correlated with a change in cell morphology and motility.


Apoptotic death of photoreceptors in the streptozotocin-induced diabetic rat retina.
Neurodegenerative changes in the diabetic retina occurring before diabetic retinopathy could be inevitable by the altered energy (glucose) metabolism, in the sense that dynamic image-processing activity of the retinal neurons is exclusively dependent on glucose. We therefore investigated the morphological changes in the neural retina, including neuronal cell death, of a streptozotocin-induced model of diabetes. Streptozotocin was intravenously injected. Rats were maintained hyperglycaemic without insulin treatment for 1 week and 4, 8, 12, and 24 weeks, respectively. Diabetic retinas were processed for histology, electron microscopy, and immunohistochemistry using the TUNEL method. A slight reduction in the thickness of the inner retina was observed throughout the diabetic retinas and a remarkable reduction was seen in the outer nuclear layer 24 weeks after the onset of diabetes. The post-synaptic processes of horizontal cells in the deep invaginations of the photoreceptors showed degeneration changes from 1 week onwards. A few necrotic ganglion cells were observed after 4 weeks. At 12 weeks, some amacrine cells and a few horizontal cells showed necrotic features. Three to seven cellular layers in the outer nuclear layer and nerve terminals, rolled by the fine processes of the Müller cells near the somata of the degenerated ganglion cells, were apparent at 24 weeks. Apoptosis appeared in a few photoreceptor cells at 4 weeks, and the number of apoptotic photoreceptors increased thereafter. These findings suggest that the visual loss associated with diabetic retinopathy could be attributed to an early phase of substantial photoreceptor loss, in addition to later microangiopathy.


Surgery improves breast cancer-specific survival in octogenarians with early-stage breast cancer.
No consensus exists on optimum therapy for older cancer patients. This singlecentre study was conducted to review the treatment and outcomes for octogenarian women treated for breast cancer. Data of all elderly breast cancer patients (≥80 years) with primary breast cancer treated at out institution between 1990 and 2009. Patients with carcinoma in-situ (stage 0) and advanced breast cancer (stage IV) were excluded. Breast cancer-specific survival and disease-free survival for the different patient groups were analysed according to the Kaplan-Meier method. The study population consisted of 259 patients (median age 84 years). There were 189 (73%) patients with early stage disease (I, IIA, IIB) and 70 (27%) with locally advanced disease (IIIA, IIIB, IIIC). A total of 175 (67.7%) patients underwent surgical treatment and 84 (32.4%) received primary endocrine treatment. Patients were followed for a median of 65 months. In patients with early stages, the mean breast cancer-specific survival was 108 months (95% CI 101-115) in the surgical group and 50 months (95% CI 39-61) in the non-surgical group (P < 0.01), whereas patients with locally advanced breast cancer breast cancer-specific survival was similar for the surgical and non-surgical groups. Breast cancer-specific survival and disease-free survival were significantly better among patients who underwent standard surgical treatment than among those with suboptimal surgery. In women ≥80 years with early-stage breast cancer, standard surgical treatment as compared with non-surgical therapy was associated with a better breast cancer-specific.


Aneurysms complicating coarctation of the aorta: anatomic aspects and evolution. Report of six successful surgical cases.
Six cases of coarctation of the aorta with aneurysms are reported. The 6 patients have been successfully operated with resection of the aneurysm and replacement of the aorta by a Dacron graft. The different anatomic characteristics are described. The authors emphasize the frequency of other aneurysms upstream of the coarctation, the risk of rupture (2 cases out of 6) and the technical difficulty of the operative procedure in comparison with a common coarctation. Aneurysm formation is a rare but severe sequel of coarctation in the adult and underlines the importance of surgical treatment of this lesion in childhood.


A radioimmunoassay with monoclonal antibodies for the detection of antigenic cell-free Fc receptor.
Making use of 125I-labelled monoclonal rat antibodies against mouse Fc receptor we have developed a solid-phase radioimmunoassay for cell-free antigenic mouse Fc receptor (FcR). An 80% acetone solution was used for fixation of relatively large amounts of soluble proteins on PVC microtiter plates. As a result of this treatment FcR practically lost its activity to bind the Fc portion of an IgG molecule but retained its antigenicity, the binding of specific anti-FcR (aFcR) antibody being increased following acetone fixation. Concentrations of cell-free FcR in NP-40 extracts of FcR-expressing cells were calculated from the linear part of a standard curve and expressed in units of antigenic activity, 1 unit being the amount of antigenic FcR capable of binding 1 microgram of 125I-aFcR. The method may be used for detecting cell-free FcR as a minor constituent in a mixture of proteins.


A review of measures of lymphedema.
Lymphedema usually is identified by patients, and rarely is it screened for routinely. Many assessments have been reported and have been used in evaluating a variety of treatments for lymphedema. A review of the literature was undertaken. Five frequently used measures of lymphedema include circumferential measures of limbs at various points (usually at bony landmarks); volumetric measures using limb submersion in water; skin tonometry, in which soft tissue compression is quantified; imaging techniques to describe tissue characteristics as well as to quantify soft tissue swelling (magnetic resonance imaging and computerized tomography; and ultrasound with and without Doppler flow studies for volumetric measures. Circumferential measures with calculations designed to compute limb volumes and volumetric measures are used most frequently, but these have some difficulty with reliability. No significant effort has been made to develop a patient based questionnaire that describes the size as well as the impact of lymphedema on an individual's functional level. Existing physical measures of lymphedema are available that are easy to use, inexpensive, have limited reliability, and do not address the issue of functional impact. Imaging techniques may provide valuable qualitative and quantitative information in selected populations.


Corrigendum: Potentiation of brain-derived neurotrophic factor-induced protection of spiral ganglion neurons by C3 exoenzyme/Rho inhibitor.
[This corrects the article DOI: 10.3389/fncel.2021.602897.].


An evaluation of plain roentgenograms prior to oral cholecystography in children.
Plain roentgenograms were taken prior to oral cholecystography in 100 children. Three children had their gallstones obscured by the oral cholecystographic agent. Consideration should be given to obtaining plain roentgenograms in children with a strong clinical suspicion of gallbladder disease and a normal oral cholecystogram.


Brachytherapy for lung cancer.
Brachytherapy (BT) is a minimally invasive anticancer radiotherapeutic modality where the tumor is directly irradiated via a radioactive source that is precisely implanted in or adjacent to the tumor. BT for lung cancer may be conducted in the form of endobronchial BT and radioactive seed implantation (RSI-BT), mainly for nonsmall cell lung cancer (NSCLC). For patients with early-stage lung cancer who are not suitable for surgery or external beam radiotherapy (EBRT), BT may be used as an alternative treatment, and curative results could be achieved in certain patients with cancer confined to the trachea lumen. For patients with locally advanced/metastatic lung cancer, BT could be selectively applied alone or as a boost to EBRT, which could improve the local tumor control and patient's survival. In addition, BT is also useful as a salvage treatment in select patients with locally recurrent/residual lung cancer that failed other treatments (e.g., surgery, chemotherapy, and EBRT). However, clinical outcomes are mainly obtained from retrospective studies. Prospective studies are limited and needed. In recent years, the introduction of modern image guidance, novel radioactive seeds, BT treatment planning systems (BT-TPS), after-loading technique, and three-dimensional printing template (3D-PT) assistance, among others, have potentially improved the clinical outcomes of BT. However, a comprehensive review of BT with newly published literature was lacking. This review is to discuss BT for NSCLC based on recent literature published in PubMed.


Esophageal pH assessment of gastroesophageal reflux in 18 patients and the effect of two prokinetic agents: cisapride and metoclopramide.
Foregut drug receptors permit inotropic manipulation of the dysmotility pattern associated with gastroesophageal reflux (GER). Two prokinetic agents, ie, Metoclopramide and Cisapride were assessed in 18 infants with severe GER (mean age 6.5 months) by means of 18-hour continuous intraesophageal pH monitoring. Six parameters were recorded, and the results compared before and during pharmacologic stimulation. Both agents improved the parameters measured, but Cisapride was found to be more effective in enhancing lower esophageal sphincter competence and esophageal motor function. Long-term assessment of both agents in the management of GER in infants is indicated.


Hepatocellular carcinoma presenting with multiple bone and soft tissue metastases and atypical cytomorphological features--a rare case report.
Hepatocellular carcinoma (HCC) with atypical cytomorphological features and presenting with bone and soft tissue metastasis is very rare. We report a 65-year-old male patient of HCC who presented with bone and soft tissue metastases and was clinically and radiologically suspected to have a soft tissue sarcoma. The patient presented with severe cervical pain with palpable masses in right scapular, nape of neck, and occiput area of scalp. Radiologically, these were large, bulky soft tissue masses expansile, destructive, and lytic in nature. Cytomorphologic studies revealed HCC with uncommon features of multinucleated osteoclast-like giant cell and very prominent intracytoplasmic hyaline bodies (IHBs). Cytology, immunohistochemistry on cell block preparation, rising serum α-fetoprotein (AFP) levels (1121.93-5000 ng/ml), and PIVKA II levels confirmed the diagnosis. The patient has been on follow-up on sorafinib for 2 months and is doing well. This case emphasizes the need for systematic approach in cases of HCC with atypical clinical presentation and unusual cytomorphology.


Nanoparticles decorated carbon nanotubes as novel matrix: A comparative study of influences of immobilization on the catalytic properties of Lensculinarisβ-galactosidase (Lcβ-gal).
In the present study, Multiwalled carbon nanotubes (MWCNT) decorated with two different nanoparticles namely tungsten disulfide (WS2) and tin oxide (SnO2), nanocomposites (NCs) were synthesized via hydrothermal method. Spectroscopic studies showed that both synthesized NCs possess nearly same functional groups but MWCNT-SnO2 NCs are rich in O-functional group. Microscopic studies revealed that both NCs have different morphological microstructure. Lens culinaris β-galactosidase (Lcβ-gal) was immobilized using glutaraldehyde cross-linker resulted in immobilization efficiency of 91.5% and 88% with MWCNT-WS2 and MWCNT-SnO2 NCs, respectively. Remarkable increase in rate of hydrolysis of whey lactose has been observed with both NCs i.e. Lcβ-gal immobilized MWCNT-WS2 hydrolyzes the 97% whey lactose in 1.5 h while MWCNT-SnO2 showed maximum 92% of whey hydrolysis in 2 h at optimum conditions. Both nanobiocatalyst could serve as a promising candidates for dairy industries and would offer a potential platform for enzyme based biosensor fabrication.


When donor funding leaves: an interrupted time-series analysis of the impact of integrating direct HIV care and treatment into public health services in a region of Johannesburg.
Early in South Africa's HIV response, donor-funded organizations directly provided HIV treatment through Comprehensive HIV Care, Management and Treatment sites (CCMTs), using their own and government staff. From 2012 to 2014 the donor-funded CCMT model was phased out, leaving nurses in South Africa's public clinics responsible for delivery of antiretroviral treatment (ART) services. We aimed to examine the impact on resources, staff workloads, and service delivery throughout this period of integration of HIV treatment into primary health clinics. We conducted an Interrupted Time-Series Analysis (ITSA) using data from three public clinics, including one former CCMT site, in one administrative region of Johannesburg. The ITSA was complemented by visual inspection of the data in Excel. We compared trends in expenditure, clinical staffing levels, patient headcounts, and services rendered at the clinics during four periods: pre-CCMT (2004-2007), CCMT operational (2007-2012), CCMT closure (2012-2014), and post-CCMT (2014-2016). Data were drawn from the country's District Health Information System, a national HIV treatment database, local budget and expenditure reports, National Health Laboratory Service charge records, and staff records. Closure of the CCMT differentially impacted the study clinics. As expected, ART services decreased at Clinic 1, where the CCMT was co-located, and increased at Clinics 2 and 3 possibly reflecting redistribution of patients. Despite a reduction in patient headcounts post-CCMT, Clinic 1 experienced a decrease in staff and a large increase in patients seen per clinical staff member per month. In contrast, Clinics 2 and 3 increased or maintained stable workforces, and staff workloads post closure were similar to pre-closure levels. Other primary care services-contraception and immunisations-seemed largely unaffected at Clinics 1 and 2. At Clinic 3, service delivery reduced, but this was accompanied by lowered patient headcounts generally, likely due to clinic renovations. In this study, integration of HIV treatment into primary healthcare services did not result in large-scale reductions in overall service delivery. One facility did experience increased staff workloads, but we were unable to assess service quality. To mitigate potential problems, monitoring systems should be introduced in advance and acknowledge the disparate and decentralised management of various data sources.


Performance of Pheromone-Baited Traps to Monitor the Seasonal Abundance of Tortrix Moths in Chestnut Groves.
(1) Background: Pammene fasciana (L.), Cydia fagiglandana (Zeller), and C. splendana (Hübner) (Lepidoptera: Tortricidae) are considered key moth pests of chestnut in Europe. (2) Methods: Investigations were performed in 2018-2019 in northern Italy. Sticky traps and commercially available pheromones were used for monitoring; moreover, two experimental pheromone blends were tested. All specimens were identified according to male genitalia and molecular analyses. Newly formed chestnut husks and fruits were randomly collected to evaluate the presence of larvae and/or feeding damage, by comparing it to trap catches. (3) Results: P. fasciana was present in all the sites, whereas Cydia species were recorded in three sites of six, with differences in abundance related to pheromone blends studied. Several non-target species, such as Oegoconia novimundi (Busck) and Cydia ilipulana (Walsingham), were present. Data about the seasonal flight activity are provided. (4) Conclusions: This research contributes to ascertaining the presence and abundance of tortrix moths in Italian chestnut groves, and the presence of non-target species highlights the risk of overestimating catches. Fruit damage recorded did not always reflect catches made by pheromone traps, suggesting that monitoring may underestimate the real size of moths' populations. All of the data acquired are important for planning specific control measures.


N-Methylanilide and N-methylbenzamide derivatives as phosphodiesterase 10A (PDE10A) inhibitors.
PDE10A is a recently identified phosphodiesterase with a quite remarkable localization since the protein is abundant only in brain tissue. Based on this unique localization, research has focused extensively on using PDE10A modulators as a novel therapeutic approach for dysfunction in the basal ganglia circuit including Parkinson's disease, Huntington's disease, schizophrenia, addiction and obsessive compulsive disorder. Medicinal chemistry efforts identified the N-methyl-N-[4-(quinolin-2-ylmethoxy)-phenyl]-isonicotinamide (8) as a nanomolar PDE10A inhibitor. A subsequent Lead-optimization program identified analogous N-methylanilides and their corresponding N-methylbenzamides (29) as potent PDE10A inhibitors, concurrently some interesting and unexpected binding modes were identified.


Activation of Erk1/2 and Akt following unilateral ureteral obstruction.
Chronic unilateral ureteral obstruction is a well characterized model of renal injury leading to tubulointerstitial fibrosis and distinct patterns of cell proliferation and apoptosis in the obstructed kidney. In this study we assessed the contribution of the mitogen activated protein kinase (MAPK)-ERK1/2 and the phosphatidylinositol 3 kinase (PI3K)-Akt pathways to early renal changes following unilateral obstruction. Increased activation of small Ras GTPase and its downstream effectors ERK1/2 and Akt was detected in ligated kidneys. The use of specific pharmacological inhibitors to either ERK1/2 or Akt activation led to decreased levels of fibroblast-myofibroblast markers in the interstitium while inhibition of PI3K reduced the number of proliferating cells and the amount of interstitial extracellular matrix deposition. Treatment with an ERK1/2 inhibitor diminished the number of apoptotic tubule and interstitial cells. Our results suggest a role for the MAPK-ERK1/2 and PI3K-Akt systems in early changes induced by ureteral obstruction and that inhibition of these signaling pathways may provide a novel approach to prevent progression of renal fibrosis.


Structure activity relationships of quinoline-containing c-Met inhibitors.
A series of quinoline-containing c-Met inhibitors were prepared and studied. Chemistry was developed to introduce a pyridyl moiety onto the 2-aryl ring present in a lead molecule which mitigated the potential for quinone formation relative to the original compound. The study also assessed the importance of an acylthiourea moiety present in the lead structure for effective binding to the c-Met protein ATP site.


Evidence for goal- and mixed evidence for false belief-based action prediction in 2- to 4-year-old children: A large-scale longitudinal anticipatory looking replication study.
Unsuccessful replication attempts of paradigms assessing children's implicit tracking of false beliefs have instigated the debate on whether or not children have an implicit understanding of false beliefs before the age of four. A novel multi-trial anticipatory looking false belief paradigm yielded evidence of implicit false belief reasoning in 3- to 4-year-old children using a combined score of two false belief conditions (Grosse Wiesmann, C., Friederici, A. D., Singer, T., & Steinbeis, N. [2017]. Developmental Science, 20(5), e12445). The present study is a large-scale replication attempt of this paradigm. The task was administered three times to the same sample of N = 185 children at 2, 3, and 4 years of age. Using the original stimuli, we did not replicate the original finding of above-chance belief-congruent looking in a combined score of two false belief conditions in either of the three age groups. Interestingly, the overall pattern of results was comparable to the original study. Post-hoc analyses revealed, however, that children performed above chance in one false belief condition (FB1) and below chance in the other false belief condition (FB2), thus yielding mixed evidence of children's false belief-based action predictions. Similar to the original study, participants' performance did not change with age and was not related to children's general language skills. This study demonstrates the importance of large-scaled replications and adds to the growing number of research questioning the validity and reliability of anticipatory looking false belief paradigms as a robust measure of children's implicit tracking of beliefs.


Inclusion of Interstate Migrant Workers in Kerala and Lessons for India.
An estimated 3.5 million interstate migrant workers have become an indispensable part of Kerala's economy. The state also offers the highest wages for migrant workers for jobs in the unorganised sector in the entire Indian subcontinent. Further, the state has evolved several measures for the inclusion of the workers and was able to effectively respond to their distress during the national lockdown. This paper examines labour migration to Kerala, key measures by the government to promote the social security of the workers and the state's response to the distress of migrant workers during lockdown, by synthesising the available secondary evidence. The welfare measures as well as interventions initiated by the state are exemplary and promising given the intent and provisions. However, some of them do not appear to have consideration of the grassroots requirements and implementation mechanisms to enhance access. As a result, the policy intent and substantial investments have not yielded the expected results. The state's effective response to the distress of workers during the lockdown emanates from its overall disaster preparedness and resilience achieved from confronting with two consecutive state-wide natural disasters and a public health emergency in the immediate past. While the government has played a strategic role through policy imperative and ensuring a synergistic response, the data presented by the state indicate a much larger but invisible role played by the employers and civil society in providing food and shelter to workers.


A subpopulation of germinal center B cells differentiate directly into antibody forming cells upon secondary immunization.
In the present study we sought to test the hypothesis that GC B cells may be stimulated to differentiate into AFCs during the induction of the secondary antibody response. To test this we gave HRP immune mice a booster immunization and looked for AFCs in the GCs making anti-HRP specific antibody. Within 3 days cells in the GCs were making anti-HRP. Most of the cells were just starting to make immunoglobulin in the rough endoplasmic reticulum but some were nearly mature plasma cells like those in the medullary cords. Furthermore, we isolated GC B cells after a boost with OVA and tested their ability to differentiate directly into AFCs. Data obtained using an OVA specific RIA indicated that purified GC B cells would differentiate and produce specific antibody in the absence of added T cells or antigen. Examination of the cells after 5 days in culture indicated that many of them had differentiated into large plasma cells. These results obtained both in vivo and in vitro demonstrate that shortly after booster immunization GC B cells receive the appropriate antigen and T cell signals in vivo to differentiate and become AFCs.


Arthroscopic fixation of humeral greater tuberosity fracture using a W-shaped suture.
Patients with greater tuberosity fractures of the humerus often require surgery. Therefore, there is a need to find a minimally invasive and effective surgical procedure with great patient outcomes. To evaluate the clinical outcomes of the W-shaped suture technique under shoulder arthroscopy in the treatment of greater tuberosity fractures of the humerus. In this retrospective study, a total of 17 patients were included. The fractures were closed, and there was no neurovascular injury. These patients underwent arthroscopically assisted reduction and internal fixation of the greater tuberosity fractures. Fixation was performed using sighting nails combined with a W-shaped suture. The imaging data of the patients were collected, and the ASES score, Constant-Murley score, and VAS score were used to evaluate the patient's outcome. At the last follow-up (at least 1 year), the range of motion in the affected shoulder was compared with that of the contralateral side. The operation was successful in all the patients. The average follow-up time was 13 months. There were no reported complications such as fracture displacement, nonunion, and internal fixation failure during the follow-up period. Post-operative X-ray examinations revealed good function recovery, with a healing time of between 10 and 12 weeks, and an average healing time of 11.5 weeks. Following the operation, patients reported reduced shoulder joint pain that no longer influenced their activity or caused discomfort in their daily life. The patient's VAS score ranged from 0 to 3, with an average of 0.52 ± 0.73, while at the last follow-up, the Constant-Murley score ranged from 83 to 97, with an average of 92.33 ± 7.55. The ASES score ranged from 81 to 98, with an average of 93.15 ± 6.93. At the last follow-up, there was no significant difference in the overall range of motion with the unaffected limb. This study demonstrates that the W-shaped suture can be used to effectively fix the fractures of the greater tuberosity of the humerus, by increasing the fixed area to promote healing.


Microenvironmental Control of Adipocyte Fate and Function.
The properties of tissue-specific microenvironments vary widely in the human body and demonstrably influence the structure and function of many cell types. Adipocytes are no exception, responding to cues in specialized niches to perform vital metabolic and endocrine functions. The adipose microenvironment is remodeled during tissue expansion to maintain the structural and functional integrity of the tissue and disrupted remodeling in obesity contributes to the progression of metabolic syndrome, breast cancer, and other malignancies. The increasing incidence of these obesity-related diseases and the recent focus on improved in vitro models of human tissue biology underscore growing interest in the regulatory role of adipocyte microenvironments in health and disease.


Consensus document on controversial issues in the diagnosis and treatment of bloodstream infections and endocarditis.
The treatment of severe bloodstream infections (sepsis, endocarditis, and infections of vascular prostheses) caused by Gram-positive microorganisms is made even more difficult by the emergence of resistant strains. The introduction of new antibiotics with activity against these strains has created new opportunities, but many controversial issues remain. The aim of this GISIG (Gruppo Italiano di Studio sulle Infezioni Gravi) working group - a panel of multidisciplinary experts - was to define recommendations for some controversial issues using an evidence-based and analytical approach. The controversial issues concerned the duration of therapy and role of aminoglycosides and teicoplanin in the treatment of Gram-positive bacterial endocarditis, the optimal use of the new antibiotics in the treatment of bloodstream infections caused by resistant Gram-positive strains, and the use of microbiological techniques (i.e., bactericidal serum testing and synergy testing) and of pharmacokinetic data (e.g., monitoring of plasma levels of antibiotics) in the treatment of difficult-to-treat Gram-positive bloodstream infections. A systematic literature search of randomized controlled trials and/or non-randomized studies was performed mainly using the MEDLINE database. A matrix was created to extract evidence from original studies using the CONSORT method to evaluate randomized clinical trials and the Newcastle-Ottawa Quality Assessment Scale for non-randomized studies. The GRADE method for grading the quality of evidence and strength of recommendation was applied.


Nanosecond time-resolved fluorescence measurements during protein denaturation.
A procedure is described by which the information available from nanosecond time-resolved fluorescence measurements can be used to study rates of reactions taking place on time scales of seconds to hours. A pulse fluorometer was modified so as to obtain a series of short sequential data collections which were rapidly stored on computer disk files. As an application of this new methodology, the unfolding of horse liver alcohol dehydrogenase under acid conditions was monitored by changes in the decay parameters of the intrinsic fluorescence. Although the individual decay curves each had relatively few counts (330 to 160 counts at the peak), a series of decay curves obtained as a function of time could be analyzed in terms of a biexponential function. It was found that the decrease in steady-state fluorescence could be explained most simply by a decrease in the amplitude associated with the longer of the two decay constants.


Characterization of two distinct Cl- conductances in fused human respiratory epithelial cells. I. Anion selectivities, stimulation and intermeshing signal transduction pathways.
With the aim of further elucidating the role of the epithelial Cl- conductance and its defect in cystic fibrosis (CF) patients we studied the properties and regulation of the Cl- conductance in primary cultures of human nasal polyp epithelia. To facilitate microelectrode punctures and to gain access to the cytoplasmic compartment for injection of antibodies, we prepared giant cells using a polyethylene-glycol fusion technique. The membrane potential (Vm) and resistance (Rm) and their responses to ionic substitutions in the bath were measured under control conditions and in the presence of different secretagogues. In non-CF cells Vm averaged-12.5 mV (SD +/- 6.6 mV, n = 69) and was independent of time after fusion, while Rm dropped from 12.4 +/- 7.3 M omega (n = 51) to 3.5 +/- 5.5 M omega (n = 24) in the 2nd week after fusion. The low Vm values reflected a vanishing K+ conductance in the presence of a dominating Cl- conductance that increased with time. In young cells, a Cl- conductance prevailed which could be stimulated by application of the Ca2+ ionophore, A23187, or of carbachol. As determined in CF cells, it had an outwardly rectifying current/voltage (ilV) relationship and exhibited the selectivity sequence I- > Br- > Cl- > F- > isethionate (ISE-) both in Vm and Rm measurements. With increasing age after fusion, a Cl- conductance prevailed in non-CF cells which could be stimulated by cyclic adenosine monophosphate (cAMP) or forskolin and which was downregulated by A23187. It had a linear ilV relationship and exhibited the selectivity sequence Br- > Cl- > I- > F- > ISE- if determined from Vm measurements, but a sequence of Cl- > Br- > F- = ISE- > I- if determined from Rm measurements. This points to multiple-ion pore behaviour of the respective Cl- channel. In agreement with observations described in the following publication, the results suggest that the cAMP-regulated Cl- conductance corresponds to the CF-gene product while the molecular nature of the Ca(2+)-regulated Cl conductance is not yet known.


Alphafetoprotein (AFP), concanavalin A non-reactive AFP and specific acetylcholinesterase in amniotic fluid from pathological pregnancies. Predictive values for open spina bifida.
Alphafetoprotein (AFP) and concanavalin A non-reactive alphafetoprotein determination and the acetylcholinesterase (AchE) qualitative test have been performed on amniotic fluid samples from 33 normal pregnancies, 44 pregnancies with fetal malformations and 8 normal pregnancies with elevated amniotic fluid alphafetoprotein (3 false positive AFP results, 5 contaminations with fetal blood). The validities of these three tests in detecting abnormal pregnancies are compared. The usefulness of the existing complementary tests in the detection of neural tube defects in a low neural tube defect incidence area is discussed. Risk figures for open spina bifida according to the prior risk situation and the results of maternal serum AFP, amniotic fluid AFP, AchE qualitative test and ultrasound examination have been calculated.


A systematic review and meta-analysis of diagnostic screening criteria for blunt cerebrovascular injuries.
Despite progress in diagnosing and managing blunt cerebrovascular injury (BCVI), controversy remains regarding the appropriate population to screen. A systematic review of published literature was conducted to summarize the overall incidence of BCVI and the various screening criteria used to detect BCVI. A meta-analysis was performed to evaluate which screening criteria may be associated with BCVI. Goals were to confirm inclusion of certain criteria in current screening protocols and possibly eliminate criteria not associated with BCVI. Studies published between January 1995 and April 2011 using digital subtraction angiography or CT angiography as a diagnostic modality and reporting overall BCVI incidence or prevalence of BCVI for specific screening criteria were examined. Screening criteria were analyzed using a random effects model to determine if an association with BCVI was present. The incidence range of BCVI was between 0.18% and 2.70% among approximately 122,176 blunt trauma admissions. The meta-analysis encompassed 418 BCVI and 22,568 non-BCVI patients. Of the 9 screening criteria analyzed, cervical spine (odds ratio [OR] 5.45; 95% CI 2.24 to 13.27; p < 0.0001) and thoracic (OR 1.98; 95% CI 1.35 to 2.92; p = 0.001) injuries demonstrated a significant association with BCVI. Patients with cervical spine and thoracic injuries had significantly greater likelihoods of BCVI compared with patients without these injuries. All patients with either injury should be screened for BCVI. Multivariate logistic regression analysis is needed to elucidate the possible impact of the combined presence of screening criteria, but it was not possible in our study due to limitations in data presentation. Standardized reporting of BCVI data is not established and is recommended to permit future collaboration.


GSK-3β manipulates ferroptosis sensitivity by dominating iron homeostasis.
Ferroptosis is a newly characterized form of non-apoptotic-programmed cell death, which is driven by the lethal accumulation of iron-catalyzed lipid peroxides. Uncontrolled ferroptosis is implicated in the pathogenesis of a group of human diseases, while targeted induction of ferroptosis provides a potent therapeutic design for cancers. During the past decade, the fundamental regulatory circuits of ferroptosis have been identified. In this study, we show that the multifaceted Ser/Thr protein kinase GSK-3β acts as a positive modulator of the ferroptosis program. Pharmacological inhibition of GSK-3β by selective inhibitor LY2090314 or genetic KD of GSK-3β by shRNA potently promotes ferroptotic resistance. GSK-3β KD antagonizes the expression of iron metabolic components including DMT1, FTH1, and FTL, leading to the disruption of iron homeostasis and decline in intracellular labile free iron level. Taken together, our findings elaborate an indispensable role of GSK-3β in determining ferroptotic sensitivity by dominating cellular iron metabolism, which provides further insight into GSK-3β as a target for cancer chemotherapy.


Evolution of residual stress and crack morphologies during 3D FIB tomographic analysis of alumina.
Three-dimensional focused ion beam (FIB) tomography is increasingly being used for 3D characterization of microstructures in the 50 nm-20 microm range. FIB tomography is a destructive, invasive process, and microstructural changes may potentially occur during the analysis process. Here residual stress and crack morphologies in single-crystal sapphire samples have been concurrently analyzed using Cr3+ fluorescence spectroscopy and FIB tomography. Specifically, maps of surface residual stress have been obtained from optically polished single-crystal alumina [surface orientation (1 ī 0 2)], from FIB milled surface trenches, from Vickers micro-indentation sites (loads 50 g-300 g), and from Vickers micro-indentation sites during FIB serial sectioning. The residual stress maps clearly show that FIB sputtering generates residual stress changes. For the case of the Vickers micro-indentations, FIB sputtering causes significant changes in residual stress during the FIB tomographic serial sectioning. 3D reconstruction of the crack distribution around micro-indentation sites shows that the cracks observed are influenced by the location of the FIB milled surface trenches due to localized stress changes.


A State-of-Art Review of Digital Technologies for the Next Generation of Tinnitus Therapeutics.
Background: Digital processing has enabled the development of several generations of technology for tinnitus therapy. The first digital generation was comprised of digital Hearing Aids (HAs) and personal digital music players implementing already established sound-based therapies, as well as text based information on the internet. In the second generation Smart-phone applications (apps) alone or in conjunction with HAs resulted in more therapy options for users to select from. The 3rd generation of digital tinnitus technologies began with the emergence of many novel, largely neurophysiologically-inspired, treatment theories that drove development of processing; enabled through HAs, apps, the internet and stand-alone devices. We are now of the cusp of a 4th generation that will incorporate physiological sensors, multiple transducers and AI to personalize therapies. Aim: To review technologies that will enable the next generations of digital therapies for tinnitus. Methods: A "state-of-the-art" review was undertaken to answer the question: what digital technology could be applied to tinnitus therapy in the next 10 years? Google Scholar and PubMed were searched for the 10-year period 2011-2021. The search strategy used the following key words: "tinnitus" and ["HA," "personalized therapy," "AI" (and "methods" or "applications"), "Virtual reality," "Games," "Sensors" and "Transducers"], and "Hearables." Snowballing was used to expand the search from the identified papers. The results of the review were cataloged and organized into themes. Results: This paper identified digital technologies and research on the development of smart therapies for tinnitus. AI methods that could have tinnitus applications are identified and discussed. The potential of personalized treatments and the benefits of being able to gather data in ecologically valid settings are outlined. Conclusions: There is a huge scope for the application of digital technology to tinnitus therapy, but the uncertain mechanisms underpinning tinnitus present a challenge and many posited therapeutic approaches may not be successful. Personalized AI modeling based on biometric measures obtained through various sensor types, and assessments of individual psychology and lifestyles should result in the development of smart therapy platforms for tinnitus.


Resolvin E1 attenuates doxorubicin-induced cardiac fibroblast senescence: A key role for IL-1β.
Cardiac fibroblasts (CFs) undergo senescence in reaction to different stressors, leading to a poor prognosis of cardiac disease. Doxorubicin (Doxo) is an antineoplastic drug with strong cardiotoxic effects, which induces IL-1β secretion and thus, triggers a potent pro-inflammatory response. Doxo induces CFs senescence; however, the mechanisms are not fully understood. Different pharmacological strategies have been used to eliminate senescent cells by inducing their apoptosis or modifying their secretome. However, Resolvin E1 (RvE1), a lipid derivative resolutive mediator with potent anti-inflammatory effects has not been used before to prevent CFs senescence. CFs were isolated from adult male C57BL/6J mice and subsequently stimulated with Doxo, in the presence or absence of RvE1. Senescence-associated β-galactosidase activity (SA-β-gal), γ-H2A.X, p53, p21, and senescence-associated secretory phenotype (SASP) were evaluated. The involvement of the NLRP3 inflammasome/interleukin-1 receptor (IL-1R) signaling pathway on CFs senescence was studied using an NLRP3 inhibitor (MCC950) and an endogenous IL-1R antagonist (IR1A). Doxo is able to trigger CFs senescence, as evidenced by an increase of γ-H2A.X, p53, p21, and SA-β-gal, and changes in the SASP profile. These Doxo effects were prevented by RvE1. Doxo triggers IL-1β secretion, which was dependent on NLRP3 activation. Doxo-induced CFs senescence was partially blocked by MCC950 and IR1A. In addition, IL-1β also triggered CFs senescence, as evidenced by the increase of γ-H2A.X, p53, p21, SA-β-gal activity, and SASP. All these effects were also prevented by RvE1 treatment. CONCLUSION: These data show the anti-senescent role of RvE1 in Doxo-induced CFs senescence, which could be mediated by reducing IL-1β secretion.


The effects of hyperbaric oxygenation on bone-titanium implant interface strength with and without preceding irradiation.
This study investigated the influence of a single 15-Gy dose of irradiation on the capacity of titanium screws to integrate in irradiated bone tissue. The biomechanical force necessary to unscrew the titanium implants 8 weeks after placement was 54% lower for implants in irradiated bone tissue compared to implants in nonirradiated bone tissue. Postirradiation use of hyperbaric oxygen treatment at 2.8 ATA (2-hour daily treatments for 21 days) increased the biomechanical force necessary to unscrew the titanium implants by 44% in irradiated bone and by 22% in nonirradiated bone.


Characterization of immunoregulatory cytokines produced by epidermal cells.
The epidermis has been identified as an important site for the initiation of immunological events. In addition to the macrophage-like Langerhans cells, keratinocytes within the epidermal cells have been shown to act as immunoregulatory cells through the secretion of cytokines such as epidermal cell-derived thymocyte-activating factor (ETAF) and interleukin 3. Epidermal cell-derived interleukin 3 (EC IL-3), like lymphocyte-derived IL-3, induced the proliferation of IL-3-dependent mast cell-like cell lines. Biochemically, EC IL-3 was a heat-stable protein with a molecular weight of approximately 30 kD. Upon chromatofocusing, EC IL-3 exhibited three isoelectric points, at pI 7.8, 7.4, and 7.1. Furthermore, an antiserum against IL-3 neutralized EC IL-3 activity, suggesting that the molecules are closely related and share similar epitopes. ETAF-like macrophage-derived interleukin 1 (IL-1) is a low molecular weight protein with a multiplicity of amplifying effects on immunological and inflammatory reactions. Thus BALB/c mice were immunized with partially purified IL-1, and immune spleen cells were hybridized with plasmocytoma cells. Supernatants of the hybridoma cultures were screened for their capacity to inhibit IL-1-induced thymocyte proliferation. After expansion and cloning, one clone was selected for ascitic antibody production. The monoclonal anti-IL-1 antibody inhibited both the IL-1-dependent thymocyte and the fibroblast proliferation. Furthermore, the antibody blocked murine and human ETAF activity, suggesting that ETAF and IL-1 share antigenically similar domains. Moreover, by using the monoclonal antibody bound to Staphylococcus aureus cells, it was possible to immunoprecipitate IL-1. In contrast, anti-IL-1 antibody did not inhibit IL-2 or IL-3 activity. These findings demonstrate that the production of immunoregulatory cytokines is not confined to cells of the immune system and that keratinocytes through the production of ETAF and EC IL-3 may mediate inflammatory and hypersensitivity reactions. Furthermore, the monoclonal anti-IL-1 antibody may provide a useful tool for the development of new immunoassays to detect IL-1/ETAF and thereby facilitate the investigation of the role of these cytokines during the pathogenesis of inflammatory diseases.


Gene therapy for cardiovascular disease mediated by ultrasound and microbubbles.
Gene therapy provides an efficient approach for treatment of cardiovascular disease. To realize the therapeutic effect, both efficient delivery to the target cells and sustained expression of transgenes are required. Ultrasound targeted microbubble destruction (UTMD) technique has become a potential strategy for target-specific gene and drug delivery. When gene-loaded microbubble is injected, the ultrasound-mediated microbubble destruction may spew the transported gene to the targeted cells or organ. Meanwhile, high amplitude oscillations of microbubbles increase the permeability of capillary and cell membrane, facilitating uptake of the released gene into tissue and cell. Therefore, efficiency of gene therapy can be significantly improved. To date, UTMD has been successfully investigated in many diseases, and it has achieved outstanding progress in the last two decades. Herein, we discuss the current status of gene therapy of cardiovascular diseases, and reviewed the progress of the delivery of genes to cardiovascular system by UTMD.


Hydrophilic cobalt sulfide nanosheets as a bifunctional catalyst for oxygen and hydrogen evolution in electrolysis of alkaline aqueous solution.
Hydrophilic medium and precursors were used to synthesize a hydrophilic electro-catalyst for overall water splitting. The cobalt sulfide (Co3S4) catalyst exhibits a layered nanosheet structure with a hydrophilic surface, which can facilitate the diffusion of aqueous substrates into the electrode pores and towards the active sites. The Co3S4 catalyst shows excellent bifunctional catalytic activity for both the oxygen evolution reaction (OER) and hydrogen evolution reaction (HER) in alkaline solution. The assembled water electrolyzer based on Co3S4 exhibits better performance and stability than that of Pt/C-RuO2 catalyst. Thereforce the hydrophilic Co3S4 is a highly promising bifunctional catalyst for the overall water splitting reaction.


Involvement of fibronectin in in vitro regeneration of retinal pigment epithelium.
Argon laser photocoagulation was placed on the confluent monolayer of cultured chick retinal pigment epithelial cells as a model of the regeneration process of retinal pigment epithelium after laser burn. The intense fibrillar net immunofluorescent pattern of fibronectin appeared on the burnt area from 2 h after the laser application, before the beginning of tissue reconstruction. Fibronectin was observed for several days, then became undetectable before the complete regeneration of retinal pigment epithelial cells. This indicates that fibronectin is involved in the early regeneration process of retinal pigment epithelium.


Distinguishing histopathologic features of acantholytic dermatoses and the pattern of acantholytic hypergranulosis.
Acantholysis can be seen in multiple skin diseases. Adnexal acantholysis has been regarded as a feature distinguishing pemphigus vulgaris (PV) from acantholytic conditions. A retrospective review of the histopathologic features of diseases with acantholysis including PV, pemphigus foliaceus (PF), Hailey-Hailey disease (HHD), Darier disease (DD), Grover disease, and pityriasis rubra pilaris (PRP) was performed. Biopsies of PV (n = 49), HHD (n = 27), DD (n = 25), Grover disease (n = 65), and PRP (n = 33) showed suprabasilar acantholysis. Acantholysis was limited to the lower epidermis in PV and PRP, and involved all epidermal layers in HHD, DD, and Grover disease. Acantholysis in PF (n = 38) mainly involved the upper epidermis. Follicular acantholysis occurred more frequently in PV and PF (P < 0.0001). Eccrine acantholysis was found in PV (42%), HHD (18%), PF (13%), and DD (4%). Grover disease, DD, and HHD had greater dyskeratosis (P < 0.0001). Neutrophils were more common in PV, PF, and HHD, while eosinophils were more common in Grover disease and DD. A pattern termed acantholytic hypergranulosis occurred predominantly in PF. Adnexal acantholysis does not reliably distinguish PV from PF. The level of acantholysis, degree of dyskeratosis, and acantholytic hypergranulosis are distinguishing features between the two types of pemphigus and other acantholytic disorders.


Cytotoxicity of human umbilical cord blood-derived mesenchymal stem cells against human malignant glioma cells.
Mesenchymal stem cells (MSCs) represent a potential useful source for cell-based glioma therapies because these cells evidence both orthodox and unorthodox plasticity and also show tropism for cancer. In this study, the authors attempted to access the cytotoxicity of human umbilical cord blood (hUCB)-derived MSCs, with or without cytokine activations against malignant glioma cells. hUCB-derived MSCs were activated by interleukin-2, interleukin-15, granulocyte macrophage colony-stimulating factor, and combinations. The hUCB-derived MSCs and activated hUCB-derived MSCs were effector cells. The cytotoxicity of the unactivated hUCB-derived MSCs and activated hUCB-derived MSCs against the target cells (human malignant glioma cells) was estimated via visual survival cell assays and transwell inserts. Phenotypic changes occurring in these hUCB-derived MSCs before and after cytokine activation were determined via flow cytometry. The secreted proteins from these effector cells were estimated via enzyme-linked immunosorbent assays. We noted a significant cytotoxicity of hUCB-derived MSCs against malignant glioma cells. In addition, the hUCB-derived MSCs activated with cytokines evidenced significantly higher cytotoxicity than that observed with unactivated hUCB-derived MSCs. Differentiated immune effectors cells from the hUCB-derived MSCs after cytokine activation were not shown to have increased in number. However, the activated hUCB-derived MSCs secreted more immune response-related proteins (interleukin 4, interferon-gamma) than did the unactivated hUCB-derived MSCs. The data collected herein confirm for the first time that hUCB-derived MSCs, with or without activation, evidence significant cytotoxicity against human malignant glioma cells, and the immune response-related proteins secreted in this process may perform relevant functions.


Atmospheric deposition of organochlorines (PCBs and pesticides) in northern France.
Bulk deposition measurements were made in northern France for a number of organochlorines (PCBs, HCB, pp'DDE, alpha-HCH and gamma-HCH) over a 1-y period, at urban, semi-rural, rural and forest sites located in accordance with prevailing wind direction. The west-east rise of PCB bulk deposition (average annual values as sigma 7) ranged from 12.2 to 46.8 ng l(-1) and showed the anthropogenic influence arising from towns, industries, storage areas and landfills over continental areas. The values were maximal at the urban site 3, Paris (122 ng l(-1)) and were still high at the eastern site 6, Abreschviller near landfills (62 ng l(-1)). Also, the highest annual deposits were found at sites 3 and 6 (Paris and Abreschviller), 38.6 and 47.3 microgm-2, respectively, i.e. 3.6 and 4 times higher than the western site value: Pleumeur. A temporal trend was observed, at the urban site where a rise occurred (up to 441 ng l(-1)) in March and April. PCB distribution according to the chlorination degree displayed high proportions of 3 Cl and 4 Cl congeners, particularly in the forest area. Annual gamma-hexachlorocyclohexane (gamma-HCH) concentration values at sites 1 (Ouessant) and 2 (Pleumeur) were close to the background noise (1.7 ng l(-1)). At the agricultural (4) and the urban (3) sites, values were maximal (19.2 and 15.9 ng l(-1)) with peaks in spring and autumn. At Pleumeur, without any local input, negative correlations were found between PCB/temperature (r = -0.503, p < 0.05), HCB/temperature ( r = -0.549, p < 0.01) and gamma-HCH/temperature ( r = -0.675, p < 0.01). A clear influence of south-west winds upon the magnitude of PCB fluxes throughout the sites was noticed. Whereas there was no global decrease of PCB contamination since 1986, the general trend of gamma-HCH total deposits was a 10-time fall, as a result of the restricting legislation in use.


Embryonic stem cell-like subpopulations are present within Schwannoma.
There is accumulating evidence of the presence of embryonic stem cell (ESC)-like cells in benign tumors. This study aimed to identify ESC-like cells in Schwannoma using the induced-pluripotent stem cell (iPSC) markers OCT4, SOX2, NANOG, KLF4 and c-MYC. Immunohistochemical (IHC) staining (n = 20) and RT-qPCR (n = 6) were performed on Schwannoma tissue samples (STS) to investigate protein and mRNA expression of these iPSC markers, respectively. Immunofluorescence (IF) staining was performed to investigate co-localization of the iPSC markers with CD34, α-SMA and CD133. IHC staining and RT-qPCR demonstrated protein and mRNA expression of all five iPSC markers, respectively. IF staining showed expression of SOX2, KLF4 and c-MYC on the tumor cells and the endothelium of the tumor microvessels which also expressed OCT4, while NANOG was exclusively expressed on the endothelium of the tumor microvessels. The OCT4+/CD34+ endothelium expressed CD133. We have identified a putative OCT4+/SOX2+/NANOG+/KLF4+/c-MYC+/CD133+ ESC-like subpopulation on the endothelium of tumor microvessels and an OCT4-/SOX2+/NANOG-/KLF4+/c-MYC+/CD133+ ESC-like subpopulation, within Schwannoma.


Purification and characterization of human liver cytochrome P-450-ALC.
Cytochrome P-450-ALC, an ethanol-oxidizing form of microsomal cytochrome P-450 (P-450), has been purified from human liver. P-450-ALC (Mr = 54,000 daltons) is a low-spin ferric hemeprotein with a CO-reduced Soret maximum at 452 nm, and has an NH2-terminal amino acid sequence nearly identical to that deduced from a human P-450-ALC cDNA clone. In a reconstituted system, P-450-ALC oxidizes ethanol and aniline at turnover rates (12.2 and 7.3 nmol min-1, respectively) 10-fold greater than two other human P-450 isozymes (termed P-450-B and P-450-C) purified from the same liver. Both P-450-ALC and P-450-C effectively demethylate N-nitrosodimethylamine (NDMA) at low substrate concentrations (0.5 mM), especially in the presence of cytochrome b5. Our results provide direct evidence for a liver P-450 isozyme in humans with catalytic properties similar to the related alcohol-inducible rodent P-450s and also reveal a new human NDMA demethylase.


Dietary glutamine supplementation affects macrophage function, hematopoiesis and nutritional status in early weaned mice.
To investigate the effect that early weaning associated with the ingestion of either a glutamine-free or supplemented diet has on the functioning of peritoneal macrophages, hematopoiesis and nutritional status of mice. Swiss Webster mice were early weaned on their 14th day of life and distributed to two groups, being fed either a glutamine-free diet (-GLN) or a glutamine-supplemented diet (+GLN). Animals belonging to a control group (CON) were weaned on their 21st day of life. The -GLN and +GLN groups had a lower lean body mass, carcass protein and ash content, plasma glutamine concentration and lymphocyte counts both in the peripheral blood and bone marrow when compared to the CON group (P<0.05). Dietary supplementation with glutamine reversed both the lower concentrations of protein and DNA in the muscle and liver, as well as the reduced capacity of spreading and synthesizing nitric oxide, hydrogen peroxide, TNF-alpha, IL-1 beta and IL-6 in cultures of peritoneal macrophages obtained from the -GLN group (P<0.05). These data indicate that the ingestion of glutamine modulates the function of peritoneal macrophages in early weaned mice. However, a glutamine-supplemented diet cannot substitute maternal milk in respect to immunological and metabolic parameters.


Progression to overt proteinuria in microalbuminuric Koreans with non-insulin-dependent diabetes mellitus.
Long-term data concerning the progression of microalbuminuria are not available in Koreans with non-insulin-dependent diabetes mellitus (NIDDM). To elucidate potential risk factors of the development of overt proteinuria in microalbuminuric Koreans with NIDDM, we studied retrospectively 46 patients with NIDDM. Between 1989 and 1990, they were found to have persistent microalbuminuria, and then were followed up regularly. Urinary albumin excretion rates (UAEs) were measured on a 24-h urine sample. Microalbuminuria was defined as UAE between 20 and 200 microg/min, and overt proteinuria as UAE > 200 microg/min on two consecutive occasions. After a mean of 4.5 years (range 3-6), 23 patients progressed to overt proteinuria (progressors), and others remained microalbuminuric (nonprogressors). Duration of diabetes was significantly longer in progressors than in nonprogressors. Mean fasting plasma glucose and HbA1c levels during the follow-up were significantly higher in progressors compared with nonprogressors (11.5+/-3.6 vs. 8.7+/-2.5 mmol/l, P=0.006, and 8.9+/-1.5 vs. 7.5+/-1.4%, P=0.005, respectively). In addition, frequencies of overt proteinuria were significantly higher in patients with their mean HbA1c > 8% during follow-up than in patients with their mean HbA1c < or=80% during follow-up (65.2 vs. 30.4%, P=0.015). Mean systolic blood pressure and mean diastolic blood pressure during follow-up tended to be higher in progressors compared with nonprogressors. Multiple logistic regression analysis revealed that mean HbA1c levels and mean systolic blood pressure during the follow-up were the most significant predictors for the incidence of overt proteinuria at 4.5-year follow-up, when adjusted for various factors (P=0.023, P=0.038, respectively). We conclude that poor glycemic control, along with elevated systolic blood pressure, were powerful predictors for the development of overt proteinuria in microalbuminuric Koreans with NIDDM.


1,4-Dihydropyridine derivatives without Ca2+-antagonist activity up-regulate Psma6 mRNA expression in kidneys of intact and diabetic rats.
Impaired degradation of proteins by the ubiquitin-proteasome system (UPS) is observed in numerous pathologies including diabetes mellitus (DM) and its complications. Dysregulation of proteasomal degradation might be because of altered expression of genes and proteins involved in the UPS. The search for novel compounds able to normalize expression of the UPS appears to be a topical problem. A novel group of 1,4-dihydropyridine (1,4-DHP) derivatives lacking Ca2+-antagonists activities, but capable to produce antidiabetic, antioxidant and DNA repair enhancing effects, were tested for ability to modify Psma6 mRNA expression levels in rat kidneys and blood in healthy animals and in rats with streptozotocin (STZ) induced DM. Psma6 gene was chosen for the study, as polymorphisms of its human analogue are associated with DM and cardiovascular diseases. 1,4-DHP derivatives (metcarbatone, etcarbatone, glutapyrone, J-9-125 and AV-153-Na) were administered per os for three days (0.05 mg/kg and/or 0.5 mg/kg). Psma6 gene expression levels were evaluated by quantitative PCR. Psma6 expression was higher in kidneys compared to blood. Induction of diabetes caused increase of Psma6 expression in kidneys, although it was not changed in blood. Several 1,4-DHP derivatives increased expression of the gene both in kidneys and blood of control and model animals, but greater impact was observed in kidneys. The observed effect might reflect coupling of antioxidant and proteolysis-promoting activities of the compounds.


Challenge for 3D culture technology: Application in carcinogenesis studies with human airway epithelial cells.
Lung cancer is still one of the major intractable diseases and we urgently need more efficient preventive and curative measures. Recent molecular studies have provided strong evidence that allows us to believe that classically well-known early airway lesions such as hyperplasia, metaplasia, dysplasia and carcinoma in situ are really precancerous lesions progressing toward cancer but not necessarily transient and reversible alteration. This suggests that adequate early control of the precancerous lesions may lead to improved prevention of lung cancer. This knowledge is encouraging in view of the imminent necessity for additional experimental systems to investigate the causal mechanisms of cancers directly in human cells and tissues. There are many questions with regard to various precancerous lesions of the airways. For example, should cells, before reaching a stage of invasive carcinoma, undergo all precancerous stages such as hyperplasia or metaplasia and dysplasia, or is there any shortcut to bypass one or more of the precancerous stages? For the study of such questions, the emerging 3-dimensional (3D) cell culture technology appears to provide an effective and valuable tool. Though a great challenge, it is expected that this in vitro technology will be rapidly and reliably improved to enable the cultures to be maintained in an in vivo-mimicking state of differentiation for much longer than a period of at best a few months, as is currently the case. With the help of a "causes recombination-Lox" (Cre-lox) technology, it has been possible to trace cells giving rise to specific lung tumor types. In this short review we have attempted to assess the future role of 3D technology in the study of lung carcinogenesis.


Sentinel Listeriosis Surveillance in Selected Hospitals, China, 2013-2017.
During 2013-2017, a total of 211 cases of listeriosis were reported by 64 sentinel hospitals in China to a national foodborne disease surveillance network. The average case-fatality rate was 31.2% for perinatal cases and 16.4% for nonperinatal cases. Sequence types 87 and 8 were the most prevalent types.


[Esophagopleural fistula; a rare complication after pneumonectomy (author's transl)].
This work deals with 8 own cases of esophagopleural fistula observed in 807 pneumonectomies with mortality rate of 37%. The pathogenesis as well as the treatment of esophagopleural fistulae based on world literature and own experience are discussed. The esophagopleural fistula is serious complication usually connected with pneumonectomy. Esophagography which should be done in every case after pneumonectomy ensures the diagnosis and before the operation shows any pathology of esophagus in its course and position. Small esophagopleural fistulae heal best after gastrostomy but larger ones require beside a double row suture of esophageal wall and additional covering with muscle flap.


Slower activation of insulin action in hypertension associated with obesity.
To determine whether kinetic abnormalities in the onset of insulin action contribute to the insulin resistance in obesity-associated hypertension. We monitored the rate of increase in glucose infusion during 6 h of hyperinsulinemic (40 mU/m2 per min) euglycemic clamps in hypertensive and normotensive obese subjects. The two groups of hypertensive (n=9) and normotensive (n=9) subjects were matched for age (48+/-2 versus 45+/-5 years), sex (five males and four females versus four males and five females) and body mass index (42+/-3 versus 40+/-2 kg/m2). In all subjects, the glucose infusion rate required to maintain euglycemia increased progressively during the clamp studies to achieve maximal, steady-state values within the fifth hour. During the first 2 h of the clamp, mean glucose infusion rate, the traditional approach to assessing insulin sensitivity, was lower in the hypertensive than in the normotensive obese patients (2.04+/-0.13 versus 3.29+/-0.41 mg/kg per min, respectively; P < 0.05). In contrast, the maximal steady-state glucose infusion rate, calculated as the mean value during the sixth hour of clamping, was similar in the hypertensive and in the normotensive obese patients (4.48+/-0.43 versus 4.81+/-0.45 mg/kg per min, respectively; NS). The time required to reach the half-maximal glucose infusion rate was greater in the hypertensive than normotensive obese patients (91+/-12 versus 38+/-5 min, respectively; P< 0.05). In obesity, hypertension was associated with a slower rate of activation of the insulin effect on glucose metabolism, whereas the maximal steady-state insulin effects were not altered by elevated blood pressure. Thus, the link between obesity and hypertension may be associated with the kinetics of onset of insulin action.


Changes in household food security, access to health services and income in northern Lao PDR during the COVID-19 pandemic: a cross-sectional survey.
We assessed the relative difficulty in meeting food needs during the COVID-19 pandemic compared with before; determined the relationship between pandemic-associated difficulties in food access and household, maternal and child food security; and identified resiliency-promoting strategies. A cross-sectional survey of households undertaken in November 2020. Rural districts of Luang Prabang Province, Lao People's Democratic Republic. Households (N=1122) with children under 5 years. Survey respondents reported the relative ease of access of food and healthcare as well as changes in income and expenditures compared with before March 2020. We determined indicators of food security and source of foods consumed for households, women and children, as well as prevalence of malnutrition in children under 5. Nearly four-fifths (78.5%) found it harder to meet household food needs during the pandemic. The most common reasons were increased food prices (51.2%), loss of income (45.3%) and decreased food availability (36.6%). Adjusting for demographics, households with increased difficulty meeting food needs had lower food consumption scores and child dietary diversity. Over 85% of households lost income during the pandemic. Decreased expenditures was associated with reliance on more extreme coping strategies to meet food needs. The households who experienced no change in meeting food needs produced a greater percentage of their food from homegrown methods (4.22% more, 95% CI 1.28 to 7.15), than households who found it more difficult. Pandemic-associated shocks may have large effects on food insecurity. Action is needed to mitigate consequences of the pandemic on nutrition. Local food production and safety net programmes that offset income losses may help.


A qualitative evaluation of the revised amyotrophic lateral sclerosis functional rating scale (ALSFRS-R) by the patient community: a web-based cross-sectional survey.
The revised Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS-R) is the most commonly used outcome measure in ALS studies. The aim of this study was to identify potential limitations of the ALSFRS-R from the perspective of people living with ALS and their caregivers. A web-based survey was developed by investigators, people living with ALS, and their caregivers, and shared across social media. For each item, participants were asked, "Can you think of a situation where you might not be able to answer this item accurately or that your answer might not reflect your abilities?" Responses were divided into two categories: criticisms that could be addressed in a manual or issues with the items/responses that would require measure modification. 57 participants (72% participants with ALS, 28% caregivers) responded to at least one item question, of which 71.9% expressed concern about at least one item. The most frequently identified items were speech, walking, and cutting food. Common criticisms were: language used is of a medical literacy level too high; item is situational; difficult to distinguish the difference between response choices; and the structure and/or underlying assumptions of the item makes it difficult to answer. Several items of the ALSFRS-R were considered to inaccurately reflect the abilities of patients with ALS. The ALSFRS-R may need a revision to address these issues, preferably in co-development with people living with ALS and their caregivers, and/or alternate outcome measures should be considered for patients with ALS.


Groundwater level prediction in Apulia region (Southern Italy) using NARX neural network.
In the Mediterranean area, the high water demand frequently leads to an excessive exploitation of the water resource, which involves a qualitative degradation of the freshwaters stored in coastal karst aquifers, as a result of phenomena such as sea saltwater intrusion. In this study, the NARX network was used to predict the daily groundwater level fluctuation for 76 monitored wells located on the Apulian territory. A preliminary analysis on reference wells was performed in order to assess the impact on the groundwater level prediction of two input parameters, rainfall and evapotranspiration, and the sensitivity to changes of training algorithm and input time delay. Based on the findings of the preliminary analysis, a comprehensive regional analysis and extensive sub-regional analyses were performed, proving the reliability of the NARX-BR network for the groundwater level prediction in wells located on different hydrogeological structures. The accurate results obtained for the Apulia region suggest the NARX network application for groundwater level prediction in other areas affected by groundwater resource management issues.


The primary study of low-dose pancreas perfusion by 640- slice helical CT: a whole-organ perfusion.
To discuss the feasibility of low-dose whole-pancreas imaging utilizing 640-slice dynamic volume CT.80 patients (40 cases of normal pancreas and 40 patients supposed of having pancreatic carcinoma or focal pancreatic space-occupying lesions were mainly refered) referred for CT pancreas perfusion were enrolled in the study. 80 patients randomly assigned to 3 groups: Group ① (whole sequence). Group ② (odd number sequence). Group ③ (even number group)(Compared to ①, the scanning times and effective radiate dose of ② and ③ decreased about 50% respectively). The head, body, tail of each normal pancreas without any pancreatic disease, lesion and lesion-surrounding areas of each pancreatic cancer were selected as ROI, and tissue peak, blood flow are measured.According to pathology and clinical materials, 27 patients were diagnosed as pancreatic cancer; 40 patients were diagnosed as normal pancreas. The tissue peak and blood flow of the head, body, tail of normal pancreas without any pancreatic disease are 109.63 ± 16.60 and 131.90 ± 41.61, 104.38 ± 19.39 and 127.78 ± 42.52, 104.55 ± 15. 44 and 123.50 ± 33.44 respectively. The tissue peak and blood flow of pancreatic cancer is 59.59 ± 18.20 and 60.00 ± 15.36. For and between each group, there is no significant statistical difference for the tissue peak and blood flow of normal areas of the head, body, tail of normal pancreas. There is statistical difference for the tissue peak and blood flow of lesion and lesion-surrounding areas of pancreatic cancer in each group. However, there is no statistical difference for the tissue peak and blood flow of normal and diseasing areas between 3 groups.Low-dose whole-pancreas perfusion with 640-slice dynamic volume CT is feasible.


New highly irritant euphorbia factors from latex of Euphorbia tirucalli L.
From the latex of Euphorbia tirucalli L. growing in Madagascar, 5 new euphorbia factors were isolated. They were characterized as 13-O-acetyl-12-O-acylphorbol- and 12-O-acetyl-13-O-acylphorbol derivatives carrying homologous conjugated unsaturated fatty acids as acyl groups. Furthermore, 2 mixtures of homologous 3-O-acylingenol derivatives are obtained carrying the same type of unsaturated fatty acids. Due to their highly unsaturated acyl groups all Euphorbia factors or factor groups isolated are highly sensitive to autoxidation.


Midwall fibrosis is an independent predictor of mortality in patients with aortic stenosis.
The goal of this study was to assess the prognostic significance of midwall and infarct patterns of late gadolinium enhancement (LGE) in aortic stenosis. Myocardial fibrosis occurs in aortic stenosis as part of the hypertrophic response. It can be detected by LGE, which is associated with an adverse prognosis in a range of other cardiac conditions. Between January 2003 and October 2008, consecutive patients with moderate or severe aortic stenosis undergoing cardiovascular magnetic resonance with administration of gadolinium contrast were enrolled into a registry. Patients were categorized into absent, midwall, or infarct patterns of LGE by blinded independent observers. Patient follow-up was completed using patient questionnaires, source record data, and the National Strategic Tracing Service. A total of 143 patients (age 68 ± 14 years; 97 male) were followed up for 2.0 ± 1.4 years. Seventy-two underwent aortic valve replacement, and 27 died (24 cardiac, 3 sudden cardiac deaths). Compared with those with no LGE (n = 49), univariate analysis revealed that patients with midwall fibrosis (n = 54) had an 8-fold increase in all-cause mortality despite similar aortic stenosis severity and coronary artery disease burden. Patients with an infarct pattern (n = 40) had a 6-fold increase. Midwall fibrosis (hazard ratio: 5.35; 95% confidence interval: 1.16 to 24.56; p = 0.03) and ejection fraction (hazard ratio: 0.96; 95% confidence interval: 0.94 to 0.99; p = 0.01) were independent predictors of all-cause mortality by multivariate analysis. Midwall fibrosis was an independent predictor of mortality in patients with moderate and severe aortic stenosis. It has incremental prognostic value to ejection fraction and may provide a useful method of risk stratification.


Characterizing the skull base in craniofacial microsomia using principal component analysis.
The aim of this study was to compare the anatomical differences in the skull base between the affected and non-affected side in patients with craniofacial microsomia (CFM), and to compare the affected and non-affected sides with measurements from a normal population. Three-dimensional computed tomography scans of 13 patients with unilateral CFM and 19 normal patients (age range 7-12 years) were marked manually with reliable homologous landmarks. Principal component analysis (PCA), as part of a point distribution model (PDM), was used to analyse the variability within the normal and preoperative CFM patient groups. Through analysis of the differences in the principal components calculated for the two groups, a model was created to describe the differences between CFM patients and normal age-matched controls. The PDMs were also used to describe the shape changes in the skull base between the cohorts and validated this model. Using thin-plate splines as a means of interpolation, videos were created to visualize the transformation from CFM skull to normal skull, and to display the variability in shape changes within the groups themselves. In CFM cases, the skull base showed significant asymmetry. Anatomical areas around the glenoid fossa and mastoid process showed the most asymmetry and restriction of growth, suggesting a pathology involving the first and second pharyngeal arches.


A retinal smear technique for varying grating contrast.
We present here a technique for modulating contrast in gratings seen in Maxwellian-view or on a projection screen. The method can be used with achromatic or chromatic grating stimuli. It is based on a high frequency oscillatory displacement of the grating image, which in turn produces a smearing of the image on the observer's retina. The waveform of the displacement and its amplitude determine the appearance and contrast of the image produced on the retina.


Effects of naftidrofuryl in patients with intermittent claudication.
In a randomized, double-blind, placebo-controlled study in patients with peripheral arterial occlusive disease stage IIb of Fontaine's classification, the efficacy of naftidrofuryl, a vasoactive substance, was investigated. Forty patients--31 men and 9 women with an average age of 62.98 +/- 10.65 years--were admitted to the study. All had a history of claudication for at least six months. The duration of the trial was eighteen weeks. After a washout period of two weeks the patients received either two times 400 mg naftidrofuryl or two identical placebo tablets daily for eight weeks. Subsequently a crossover from verum to placebo and vice versa was carried out and the alternative medication was administered for another eight weeks. Clinical results were evaluated by measuring painfree and maximal walking distance by treadmill test at a speed of 3.2 km/h and an inclination of 12 degrees. Systolic ankle pressure and brachial pressure were measured by Doppler ultrasound, and ankle/arm pressure ratio was calculated. Treadmill test, Doppler ultrasound examination, and laboratory data analysis were performed at the beginning of every study period, every four weeks during and at the end of the study. Results. The patients in both groups showed an initial homogeneity of age, risk factors, concomitant diseases, walking distance, and Doppler indices. After eight weeks naftidrofuryl resulted in a statistically significant increase in painfree (p less than 0.02) and maximal walking distance (p less than 0.05). The placebo-treated group showed only a slight, statistically nonsignificant increase in both painfree and maximal walking distance.(ABSTRACT TRUNCATED AT 250 WORDS)


Restricted quasispecies variation following infection with the GB virus B.
The extent of genetic variability following acute infection of tamarins with GB virus B (GBV-B) is not known. In this study we attempted to define the quasispecies variation of GBV-B 17 days post-infection, by PCR amplification of GBV-B RNA extracted from serum and liver. Cloning followed by sequencing revealed a small number of changes in the three regions studied, namely the 5' untranslated region, E2 and NS3. Moreover, there was no region of high amino acid variability in E2, akin to hypervariable region 1 of hepatitis C virus. This was further confirmed by analysing sequences from two additional animals obtained at a similar time point post-infection. Nevertheless, it was apparent that different variants with one or two amino acid substitutions in the region studied had been selected when comparing the sequences from the three animals. This restricted sequence variation of GBV-B during acute hepatitis may explain the infrequent progression of the infection to a chronic stage.


Profile of von Willebrand factor antigen and von Willebrand factor propeptide in an overall TIA and ischaemic stroke population and amongst subtypes.
Von Willebrand factor propeptide (VWF:Ag II) is proposed to be a more sensitive marker of acute endothelial activation than von Willebrand factor antigen (VWF:Ag). Simultaneous data on VWF:Ag and VWF:Ag II profiles are very limited following TIA and ischaemic stroke. In this prospective, observational, case-control study, plasma VWF:Ag and VWF:Ag II levels were quantified in 164 patients≤4weeks of TIA or ischaemic stroke (baseline), and then ≥14days (14d) and ≥90days (90d) later, and compared with those from 27 healthy controls. TIA and stroke subtyping was performed according to the TOAST classification. The relationship between VWF:Ag and VWF:Ag II levels and platelet activation status was assessed. 'Unadjusted' VWF:Ag and VWF:Ag II levels were higher in patients at baseline, 14d and 90d than in controls (p≤0.03). VWF:Ag levels remained higher in patients than controls at baseline (p≤0.03), but not at 14d or 90d after controlling for differences in age or hypertension, and were higher in patients at baseline and 90d after controlling for smoking status (p≤0.04). 'Adjusted' VWF:Ag II levels were not higher in patients than controls after controlling for age, hypertension or smoking (p≥0.1). Patients with symptomatic carotid stenosis (N=46) had higher VWF:Ag and VWF:Ag II levels than controls at all time-points (p≤0.002). There was no significant correlation between platelet activation status and VWF:Ag or VWF:Ag II levels. VWF:Ag and VWF:Ag II levels are increased in an overall TIA and ischaemic stroke population, especially in patients with recently symptomatic carotid stenosis. VWF:Ag II was not superior to VWF:Ag at detecting acute endothelial activation in this cohort and might reflect timing of blood sampling in our study.


The structure and development of avian lumbosacral specializations of the vertebral canal and the spinal cord with special reference to a possible function as a sense organ of equilibrium.
The avian lumbosacral vertebral column and spinal cord show a number of specializations which have recently been interpreted as a sense organ of equilibrium. This sense organ is thought to support balanced walking on the ground. Although most of the peculiar structures have been described previously, there was a need to reevaluate the specializations with regard to the possible function as a sense organ. Specializations were studied in detail in the adult pigeon. The development of the system was studied both in the pigeon (semiprecocial at hatching) and in the chicken (precocial). Specializations in the vertebral canal consist of a considerable enlargement, which is not due to an increase in the size of the spinal nervous tissue, but to a large glycogen body embedded in a dorsal rhomboid sinus. The dorsal wall of the vertebral canal shows segmented bilateral dorsal grooves, which are covered by the meninges towards the lumen of the vertebral canal leaving openings in the midline and laterally. This results in a system of lumbosacral canals which look and may function similar to the semicircular canals in the inner ear. Laterally these canals open above ventrolateral protrusions or accessory lobes of the spinal cord which contain neurons. There are large subarachnoidal cerebrospinal fluid spaces, lateral and ventral to the accessory lobes. Movement of this fluid is thought to stimulate the lobes mechanically. As to the development of avian lumbosacral specializations, main attention was given to the organization of the lobes and the adjacent fluid spaces including the dorsal canals. In the pigeon the system is far from being adult-like at hatching but maturates rapidly after hatching. In the chicken the system looks already adult-like at hatching. The implications derived from the structural findings are discussed with regard to a possible function of the lumbosacral specializations as a sense organ of equilibrium. The adult-like organization in the newly hatched chickens, which walk around immediately after hatching, supports the assumed function as a sense organ involved in the control of locomotion on the ground.


Anxiogenic effects of a beta-carboline on tonic immobility and open field behavior in chickens (Gallus gallus).
Tonic immobility (TI) is an innate form of active motor inhibition displayed by many species in response to restraint. It is strongly influenced by manipulations that affect fear, and is thought to be the last in a series of responses to attack by a predator. The suggestion that GABA systems may be involved in TI was investigated by assessing the effects of the alpha-GABAA inverse agonist beta-CCM (beta-carboline-3-carboxylic acid-N-methylamide), which is thought to have anxiogenic properties, on the immobility response and open field behavior in chickens (Gallus gallus). Birds given beta-CCM displayed longer durations of TI than those given control injections. Although there was a tendency towards increased susceptibility in the groups given the drug, it was not significant. In the open field test, the drug reduced activity and vocalization relative to control levels. The results are discussed in terms of the involvement of GABA systems in TI and related behaviors, and the suggestion that alpha-GABAA inverse agonists are anxiogenic agents.


A specific diagnostic antigen of Echinococcus granulosus with an apparent molecular weight of 8 kDA.
An echinococcus antigen with an apparent molecular weight of 8 kDa was identified as diagnostically important. An immunoblot assay using this antigen was 91% sensitive for surgically confirmed Echinococcus granulosus hydatid disease of the liver. Specificity was 100% for echinococcosis. Marked cross-reactivity was observed with serum specimens from patients with E. multilocularis and E. vogeli infections. The 8 kDa component was not related to the widely recognized echinococcus antigen 5.


Inflammatory cytokines regulate the expression of glycosyltransferases involved in the biosynthesis of tumor-associated sialylated glycans in pancreatic cancer cell lines.
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant stroma containing several pro-inflammatory cytokines, which are described to modulate the expression of important genes related to tumor promotion and progression. In the present work we have investigated the potential role of these cytokines in the biosynthesis of tumor-associated carbohydrate antigens such as sialyl-Lewis(x) (SLe(x)) through the regulation of specific glycosyltransferase genes. Two human PDAC cell lines MDAPanc-3 and MDAPanc-28 were treated with pro-inflammatory cytokines IL-1β, TNFα, IL-6 or IL-8, and the content of tumor-associated carbohydrate antigens at the cell membrane was analyzed by flow cytometry. In addition, variation in the mRNA expression of sialyltransferase (ST) and fucosyltransferase (FUT) genes, which codify for the ST and FucT enzymes involved in the carbohydrate antigens' biosynthesis, was determined. The inflammatory microenvironment of PDAC tissues and the expression of Lewis-type antigens were analyzed by immunohistochemistry to find a possible correlation between inflammation status and the presence of tumor-associated carbohydrate antigens. IL-1β stimuli increased SLe(x) and α2,6-sialic acid levels in MDAPanc-28 cells and enhanced the mRNA levels of ST3GAL3-4 and FUT5-7, which codify for ST and FucT enzymes related to SLe(x) biosynthesis, and of ST6GAL1. IL-6 and TNFα treatments increased the levels of SLe(x) and Le(y) antigens in MDPanc-3 cells and, similarly, the mRNA expression of ST3GAL3-4, FUT1-2 and FUT6, related to these Lewis-type antigens' biosynthesis, were increased. Most PDAC tissues stained for SLe(x) and SLe(a) and tended to be expressed in the tumor samples with a higher presence of inflammatory immune cells. The inflammatory microenvironment can modulate the glycosylation pattern of PDAC cells, increasing the expression of tumor-associated sialylated antigens such as SLe(x), which contributes to pancreatic tumor malignancy.


LH receptor gene expression in cumulus cells in women entering an ART program.
Luteinizing hormone (LH) exerts its actions through its receptor (LHR), which is mainly expressed in theca cells and to a lesser extent in oocytes, granulosa and cumulus cells. The aim of the present study was the investigation of a possible correlation between LHR gene and LHR splice variants expression in cumulus cells and ovarian response as well as ART outcome. Forty patients undergoing ICSI treatment for male factor infertility underwent a long luteal GnRH-agonist downregulation protocol with a fixed 5-day rLH pre-treatment prior to rFSH stimulation and samples of cumulus cells were collected on the day of egg collection. RNA extraction and cDNA preparation was followed by LHR gene expression investigation through real-time PCR. Furthermore, cumulus cells were investigated for the detection of LHR splice variants using reverse transcription PCR. Concerning LHR expression in cumulus cells, a statistically significant negative association was observed with the duration of ovarian stimulation (odds ratio = 0.23, p = 0.012). Interestingly, 6 over 7 women who fell pregnant expressed at least two specific types of LHR splice variants (735 bp, 621 bp), while only 1 out of 19 women that did not express any splice variant achieved a pregnancy. Consequently, the present study provide a step towards a new role of LHR gene expression profiling as a biomarker in the prediction of ovarian response at least in terms of duration of stimulation and also a tentative role of LHR splice variants expression in the prediction of pregnancy success.


Renin gene expression in rat tissues: a new quantitative assay method for rat renin mRNA using synthetic cRNA.
A genomic renin exon 9 fragment was subcloned into vector pSPT18 and used for in vitro transcription to obtain 32P-labeled rat renin cRNA. Using this cRNA, we established quantitative solution hybridization and specific Northern blotting assays for renin mRNA. We were able to detect renin mRNA in the kidney, heart ventricle and atrium, brain, testis, and adrenal gland of male rats in the concentrations of 430 +/- 8.1, 110 +/- 1.9, 43 +/- 0.9, 64 +/- 1.1, 47 +/- 0.9 and 11 +/- 0.2 pg mRNA/mg of total RNA, respectively.


Syngrafting skin among mice of similar and different ages increases the number of Langerhans cells and decreases responsiveness to 1,4-dinitrofluorobenzene.
Counting on suction blister skin biopsies, the number of Langerhans cells per mm2 in BALB/c mouse epidermis was found identical in 1-, 11-, and 22-month-old animals, but consistently elevated in syngrafted skin 10 months after transplantation. This effect of grafting was independent of whether young or old skin was transferred to young or old recipients. Skin swelling induced by dinitrofluorobenzene (DNFB) sensitization was reduced in 22-month-old non-grafted mice and in grafted skin irrespective of donor and recipient age.


Thecal plate morphology, molecular phylogeny, and toxin analyses reveal two novel species of Alexandrium (Dinophyceae) and their potential for toxin production.
This study describes two novel species of marine dinophytes in the genus Alexandrium. Morphological characteristics and phylogenetic analyses support the placement of the new taxa, herein designated as Alexandrium limii sp. nov. and A. ogatae sp. nov. Alexandrium limii, a species closely related to A. taylorii, is distinguished by having a shorter 2'/4' suture length, narrower plates 1' and 6'', with larger length: width ratios, and by the position of the ventral pore (Vp). Alexandrium ogatae is distinguishable with its metasert plate 1' having almost parallel lateral margins, and by lacking a Vp. Production of paralytic shellfish toxins (PSTs), cycloimines, and goniodomins (GDs) in clonal cultures of A. ogatae, A. limii, and A. taylorii were examined analytically and the results showed that all strains contained GDs, with GDA as major variants (6-14 pg cell-1) for all strains except the Japanese strain of A. limii, which exclusively had a desmethyl variant of GDA (1.4-7.3 pg cell-1). None of the strains contained detectable levels of PSTs and cycloimines.


Development of a risk assessment tool for women with a family history of breast cancer.
Innovative technologies that enable the collection of family history information and the assessment of breast cancer risk have a potential to enhance the quality of preventive care. We developed a computerized tool that supports stratification of breast cancer risk, genetic risk assessment in the clinical environment (GRACE). In a preliminary evaluation of the tool's impact, we randomized women with a family history of breast cancer (n=72) to either use the GRACE tool or undergo risk counseling by a nurse specialist. There was no statistically significant differences between the GRACE and nurse counselling groups in risk perceptions (F=.03, P>.05) and cancer-related worries (F=.80, P>.05). However, patients reported more positive attitudes toward working with the nurse. It was feasible to use GRACE in a Clinic. Additional research is required to identify solutions for providing emotional support in conjunction with the tool.