Approaching Combined Rectal and Vaginal Prolapse.
Combined rectal prolapse and pelvic organ prolapse surgery provides significant quality-of-life benefits with improvements in bothersome symptoms of pain, bulge, constipation, urinary retention, as well as bowel and bladder incontinence. Robotic surgery is the ideal tool for a combined surgical repair. It allows enhanced suturing in the deep pelvis, three-dimensional (3D) visualization of the presacral space and easy mobilization of the rectum and dissection of the vagina. Combined procedures can be offered to patients with the advantages of a single operation and concurrent recovery period without increasing complications. In this article, we highlight our approach to combined prolapse repair.


Precise control of the degree and regioselectivity of functionalization in nitro- and amino-functionalized di(trispyrazolylborato)iron(II) spin crossover complexes.
Di(trispyrazolylborato)iron(II) ([Tp2Fe]) complexes represent one of the most robust classes of spin-crossover complexes. Their stability renders them particularly suitable for integration in nanoscale devices, e.g. as sensors or information storage units. While prior studies of the functionalization of those derivatives have been focused on the electronic and steric effects of alkyl and -CF3 groups in position 3, a pyrazole exchange reaction between nitropyrazole and either trispyrazolylborate or its iron complex allows the regioselective installation of nitro substituents in positions 3, 4 and 5 of the [Tp2Fe] complexes. The degree of substitution can be varied from 1 to 4 functionalized pyrazoles per complex. The amine-functionalized analogues are accessed by reduction of the nitro analogues under hydrogen transfer conditions. With the exception of di- and tetra-3-NO2 substituted complexes, all derivatives display spin crossover properties in the solid state, with transition temperatures ranging from 180 to 380 K and showing different degrees of abruptness but no hysteresis. The Slichter-Drickamer model was used to extract the empirical thermodynamic transition parameters, allowing a systematic investigation of the influence of stoichiometry, position, and electronic nature of the substitution on the magnetic properties of the complexes. The steric effects dominate for substitution in position 3 but the electronic effects are significant for the other positions.


Imprinting of lymphocytes with melanoma antigens acquired by trogocytosis facilitates identification of tumor-reactive T cells.
Trogocytosis is a contact-dependent intercellular transfer of membrane fragments and associated molecules from APCs to effector lymphocytes. We previously demonstrated that trogocytosis also occurs between tumor target and cognate melanoma Ag-specific cytotoxic T cells (CTL). In this study, we show that, following trogocytosis, immune effector cells acquire molecular components of the tumor, including surface Ags, which are detectable by specific mAbs. We demonstrate that CD8(+) and CD4(+) T cells from melanoma patients' PBMC and tumor-infiltrating lymphocytes (TIL) capture melanoma Ags, enabling identification of trogocytosing lymphocytes by staining with Ag-specific Abs. This finding circumvents the necessity of tumor prelabeling, which in the past was mandatory to detect membrane-capturing T cells. Through the detection of melanoma Ags on TIL, we sorted trogocytosing T cells and verified their preferential reactivity and cytotoxicity. Furthermore, tumor Ag-imprinted T cells were detected at low frequency in fresh TIL cultures shortly after extraction from the tumor. Thus, T cell imprinting by tumor Ags may allow the enrichment of melanoma Ag-specific T cells for research and potentially even for the adoptive immunotherapy of patients with cancer.


[Treatment of anti-neutrophil cytoplasmic antibody related vasculitis].
ANCA-associated vasculitides are a well-known clinico-pathological group of systemic diseases comprising microscopic poliangiitis, granulomatosis with poliangiitis and eosinophilic granulomatosis with poliangiitis. This article shows contemporary treatment of this diseases with extensive literature review. Stepwise treatment of ANCA-associated vasculitides is divided into induction therapy and remission maintenance therapy. Standard induction therapy is a combination of glucocorticoids and cyclophosphamide, and in maintenance therapy, combination of low-dose glucocorticoids and azathioprine or methotrexate is used. Leading rheumatology and nephrology associations developed treatment guidelines. Since ANCA-associated vasculitides are relatively rare diseases, there are only few randomized controlled studies to provide high level of evidence and treatment recommendations. Most patients achieve remission, but relapses often occur. The main treatment considerations, apart from frequently relapsing disease, are disease refractory to treatment and potentially harmful effects of immunosuppressants, especially cyclophosphamide. Future studies are needed to determine the effects of less toxic immunosuppressants, mainly biological agents.


[Influence of the work environment on burn-out and depression in nurses].
Burn out is defined as a state of psychological exhaustion in the face of an unfavourable work environment. In the health sector, this includes, among other things, an excessive care burden, the lack of autonomy or control over work, the vicious circle of absenteeism, the lack of support, moral and sexual harassment, discrimination or the Covid-19 health crisis. Burn-out also increases the risk of depression among care workers.


Recombinant aprotinin homologue with new inhibitory specificity for cathepsin G.
The substitution of amino acids in the reactive site of aprotinin, a bovine serine proteinase inhibitor with potent activity against trypsin, plasmin and tissue kallikrein, led to a change in specificity of the inhibitor. Twelve new aprotinin variants prepared by recombinant DNA technology and expressed in Escherichia coli clearly demonstrated that the neighbouring groups of the P1 residue, in particular P'2, contribute to the specificity of the inhibitor, while earlier investigations on semisynthetically prepared variants revealed the importance of the P1 residue in dominating the inhibitory specificity. Recombinant aprotinin variants which act specifically against chymotrypsin-like proteinases, were obtained by substitution of the amino acids in position P1 and P'2 by hydrophobic amino acids like phenylalanine, tyrosine and leucine. Some of these variants, particularly those with phenylalanine or leucine substitutions, were also found to exhibit inhibitory activity against cathepsin G with an equilibrium constant of dissociation Ki of 10(-8) M. Inhibitory specificity against cathepsin G was not found in any semisynthetic variant prepared earlier.


Assessment of correlation between CT angiographic clot load score, pulmonary perfusion defect score and global right ventricular function with dual-source CT for acute pulmonary embolism.
The purpose of this study was to prospectively investigate the correlation between CT angiographic clot load (CTACL) score, pulmonary perfusion defect (PPD) score and the global right ventricular function in the assessment of pulmonary embolism (PE) severity. 49 patients with acute PE, who underwent dual-source CT scan, were included in the study. CT angiography and perfusion imaging were performed. Data from electrocardiogram-gated coronary angiography scanning protocol were used for right ventricular function analysis. Two readers evaluated the CTACL and PPD scores using the Qanadli and Chae methods, respectively. The PPD score had a strong positive correlation with the CTACL score (r=0.72, p<0.001) and both scores in turn had a strong positive correlation with the right ventricular/left ventricular (RV/LV) diameter ratio (r=0.60, r=0.62, p<0.001). However, the PPD score had a strong negative correlation with ejection fraction (EF) (r=-0.63, p<0.001) while the CTACL score had a low negative correlation with EF (r=-0.33, p=0.02). Between the RV/LV<1 group (n=35) and the RV/LV >1 group (n=14), the PPD score, CTACL score, pulmonary artery trunk diameter, EF and reflux of inferior vena cava were significantly different, all with p<0.001. The end-systolic volume (p=0.01) was significantly different but the end-diastolic volume (p=0.11) and stroke volume (p=0.08) showed no statistically significant difference between the two groups. Therefore, considering PPD scores, CTACL scores and cardiovascular manifestations together may be helpful in the evaluation of PE severity.


The effect of psychological distress on medication adherence in persons with HIV infection in Nigeria.
A high level of adherence to prescribed antiretroviral (ARV) regimens is required to achieve and maintain suppression of human immunodeficiency virus (HIV) replication and prevent drug resistance. This study aimed to determine the possible relationship between psychopathology and ARV medication adherence in Nigeria. Persons with HIV infection (N=182) completed various questionnaires on sociodemographic and clinical details, general psychopathology, self-esteem, and medication adherence. Low medication adherence was reported in 26.9% of the participants; significant correlates included presence of psychopathology and perceived poor social support. The success of any intervention policy for HIV-infected persons in sub-Saharan Africa must consider both low level of medication adherence and its associated factors.


Children as messengers of health knowledge? Impact of health promotion and water infrastructure in schools on facial cleanliness and trachoma in the community.
Health promotion is essential to the SAFE strategy for trachoma elimination. Schools are a valuable venue for health promotion. However, there is little literature about the impact of health education and water infrastructure in schools on facial cleanliness and trachoma in the community. Our study aimed to describe the current state of school health promotion in Kongwa, Tanzania, and to examine the transferability of health messages from schools to the community at large. A cross-sectional survey was carried out in all 92 villages in Kongwa district, which included 85 primary schools. Data were collected on health messages and water infrastructure in the schools. A random sample of 3084 children aged 0-5 were examined for facial cleanliness in all villages. In 50 villages, a random sample of 50 children aged 1-9 per village were examined for follicular trachoma (TF). Thirty-seven (44.6%) schools had educational materials on face-washing. Fifty (60.2%) schools had a washing station. The presence of a health teacher was correlated with having posters on face washing in classrooms. The presence of face-washing materials was correlated with the availability of washing stations. Neither teachers mentioning face-washing in health curricula nor educational materials in classrooms were associated with clean faces or trachoma in the community. Having a washing station in the school was associated with lower community rates of trachoma. Primary school health messages and materials on trachoma were not associated with clean faces or lower rates of trachoma in the community. The target audience for primary school health promotion is likely the students themselves, without immediate rippling effects in the community. A long-term perspective should be considered during the implementation of health promotion in schools. The goal of school health promotion should be training the next generation of parents and community health leaders in combatting trachoma.


Update on Paraneoplastic Cerebellar Degeneration.
Purpose of review: To provide an update on paraneoplastic cerebellar degeneration (PCD), the involved antibodies and tumors, as well as management strategies. Recent findings: PCD represents the second most common presentation of the recently established class of immune mediated cerebellar ataxias (IMCAs). Although rare in general, PCD is one of the most frequent paraneoplastic presentations and characterized clinically by a rapidly progressive cerebellar syndrome. In recent years, several antibodies have been described in association with the clinical syndrome related to PCD; their clinical significance, however, has yet to be determined. The 2021 updated diagnostic criteria for paraneoplastic neurologic symptoms help to establish the diagnosis of PCD, direct cancer screening, and to evaluate the presence of these newly identified antibodies. Recognition of the clinical syndrome and prompt identification of a specific antibody are essential for early detection of an underlying malignancy and initiation of an appropriate treatment, which represents the best opportunity to modulate the course of the disease. As clinical symptoms can precede tumor diagnosis by years, co-occurrence of specific symptoms and antibodies should prompt continuous surveillance of the patient. Summary: We provide an in-depth overview on PCD, summarize recent findings related to PCD, and highlight the transformed diagnostic approach.


Syntaxin-1A inhibition of P-1075, cromakalim, and diazoxide actions on mouse cardiac ATP-sensitive potassium channel.
Syntaxin (Syn)-1A binds sulfonylurea receptor (SUR) nucleotide binding folds of cardiac myocyte (SUR2A) and islet beta-cells (SUR1) to inhibit ATP-sensitive potassium (K(ATP)) channels. We further reported that Syn-1A reduced the potency and efficacy of beta-cell-specific K(ATP) channel openers (KCOs). Here, we examined whether Syn-1A would influence non-specific (diazoxide) and SUR2-specific KCOs [N-cyano-N'-(1,1-dimethylpropyl)-N''-3-pyridylguanidine (P-1075) and cromakalim] on cardiac myocyte K(ATP) channels activation. Confocal microscopy and Western blotting verified the presence of both Syn-1A and -1B expressions on rodent cardiac ventricular myocytes. Inside-out patch-clamp electrophysiology was utilized to examine the effects of these syntaxins on K(ATP) macroscopic currents activated by various KCOs from a stable cell line expressing the potassium inward rectifier 6.2 (Kir6.2)/SUR2A and from C57BL/6 male mouse ventricular myocytes. Syn-1A inhibited the current amplitude activated by P-1075, cromakalim and diazoxide via its H3 but not Habc domain. Syn-1B exhibited similar inhibitory effects on P-1075 activation of K(ATP) currents. In examining for direct effects of Syn-1A on the KCO binding to cardiac SUR2 receptors, we found that Syn-1A did not directly affect [(3)H]-P-1075 binding to rat cardiac membrane SUR2A at maximum binding capacity, but was able to mildly reduce the affinity of cold P-1075 and cromakalim to displace [(3)H]-P-1075 binding. In conclusion, Syn-1A (and Syn-1B) could inhibit K(ATP) currents activated by SUR2A-acting KCOs. Potential fluctuations in the levels of these syntaxins in the myocardium may affect the therapeutic effectiveness of cardiac KCOs.


[The role of gamma delta T cells in infectious disease].
There have been several lines of evidence that at least significant fraction of gamma delta--T cells are specialized to recognize epitopes on mycobacterial antigens including 65 KD heat shock protein(HSP). Since HSP is known to be highly conserved in amino acid sequences from prokaryotes to eukaryotes, it is possible that the HSP--specific gamma delta--T cells may recognize the endogenous HSP expressed by autologous cells. The broad--reactive gamma delta--T cells may be responsible for protection during the period between an early stage covered mainly by phagocytes and a late stage covered by typical immunities in terms of the time sequence after microbial invasions.


[Headache in Horton's disease: the clinical picture and physiopathogenetic mechanisms].
Headache is no doubt one of the most frequent symptom in Horton's disease. Nevertheless it has neither received a clear definition nor it has been studied in its pathogenetic aspects. The authors report 32 cases of Horton's disease in acute phase with arteritic localizations in different areas. Among them 24 (75%) had headache which appeared with different features from case to case. The authors identify 3 different kinds of headache, all recognizable in the secondary medical forms: epicritic headache (the most frequent), deep headache and generic headache (the rarest). The characters and the pathogenetic mechanisms of each form are also described.


Consequences of introducing a disulfide bond into an antibacterial and hemolytic peptide.
The effect of introducing a disulfide bridge between the N- and C-terminal ends on the structure and biological activities of the 13-residue linear peptide PKLLKTFLSKWIG(SPFK), which has both antibacterial and hemolytic activity, have been investigated. The terminal amino acids P and G in SPFK were replaced by cysteines to form a disulfide bridge. The linear peptides C(Acm)KLLKTFLSKWIC(Acm) and C(Acm) KLLKTFLSKWIC(Acm)-amide, where Acm is acetamidomethyl group, showed antibacterial activity but did not possess hemolytic activity unlike SPFK. Introduction of an S-S bridge resulted in enhanced hemolytic activity compared with SPFK. The hemolytic activity was particularly pronounced in the cyclic peptide CKLLKTFLSKWIC-amide. Circular dichroism studies indicate that the cyclic peptides tend to adopt distorted helical structures. The cyclic peptides also have a greater affinity for lipid vesicles, which could be the reason for the effective perturbation of the erythrocyte membrane.


Patients with fibromyalgia show increased beta connectivity across distant networks and microstates alterations in resting-state electroencephalogram.
Fibromyalgia (FM) is a chronic condition characterized by widespread pain of unknown etiology associated with alterations in the central nervous system. Although previous studies demonstrated altered patterns of brain activity during pain processing in patients with FM, alterations in spontaneous brain oscillations, in terms of functional connectivity or microstates, have been barely explored so far. Here we recorded the EEG from 43 patients with FM and 51 healthy controls during open-eyes resting-state. We analyzed the functional connectivity between different brain networks computing the phase lag index after group Independent Component Analysis, and also performed an EEG microstates analysis. Patients with FM showed increased beta band connectivity between different brain networks and alterations in some microstates parameters (specifically lower occurrence and coverage of microstate class C). We speculate that the observed alterations in spontaneous EEG may suggest the dominance of endogenous top-down influences; this could be related to limited processing of novel external events and the deterioration of flexible behavior and cognitive control frequently reported for FM. These findings provide the first evidence of alterations in long-distance phase connectivity and microstate indices at rest, and represent progress towards the understanding of the pathophysiology of fibromyalgia and the identification of novel biomarkers for its diagnosis.


A Game Theoretic Analysis of the Dual Function of Antibiotics.
There are two major views toward the role of antibiotics in microbial social interactions. The classical view is that antibiotics serve as weapons, produced by a bacterial species, at a significant cost, to inhibit the growth of its competitors. This view is supported by observations that antibiotics are usually upregulated by stress responses that infer the intensity of ecological competition, such as nutrient limitation and cellular damage, which point out to a competitive role for antibiotics. The other ecological function frequently assigned to antibiotics is that they serve as signaling molecules which regulate the collective behavior of a microbial community. Here, we investigate the conditions at which a weapon can serve as a signal in the context of microbial competition. We propose that an antibiotic will serve as a signal whenever a potential alteration of the growth behavior of the signal receiver, in response to a subinhibitory concentration (SIC) of the antibiotic, reduces the competitive pressure on the signal producer. This in turn would lead to avoiding triggering the stress mechanisms of the signal producer responsible for further antibiotics production. We show using individual-based modeling that this reduction of competitive pressure on the signal producer can happen through two main classes of responses by the signal recipient: competition tolerance, where the recipient reduces its competitive impact on the signal producer by switching to a low growth rate/ high yield strategy, and niche segregation, where the recipient reduces the competitive pressure on the signal producer by reducing their niche overlap. Our hypothesis proposes that antibiotics serve as signals out of their original function as weapons in order to reduce the chances of engaging in fights that would be costly to both the antibiotic producer as well as to its competitors.


Trends in Polyethylene Design and Manufacturing Characteristics for Total Knee Arthroplasty: An Analysis From the American Joint Replacement Registry.
Advances in polyethylene bearing characteristics have led to various options for total knee arthroplasty. We examine trends in use of polyethylene in the United States. An analysis of American Joint Replacement Registry data from 2012 to 2019 for primary total knee arthroplasty procedures was conducted. Design and manufacturing characteristics were obtained and validated in the American Joint Replacement Registry dataset. Polyethylene bearing design and manufacturing use characteristics were compared by year, gender, age group, and geographic region of the procedure. Descriptive analysis was performed. We identified 993,292 cases over the study period. In total, 791,233 (80%) cases had complete device data available and were included in analysis. In addition, 366,280 (49.4%) cases utilized minimally stabilized bearings compared to 374,809 (50.6%) for posterior-stabilized bearings. Posterior-stabilized use was stable over the study period and highest in the Northeast (64.5%; P < .0001). Cruciate-retaining use decreased from 29.7% to 20.2% and anterior-stabilized use increased from 17.4% to 25.2%. Conventional polyethylene use decreased from 42.8% to 21.9%, and was highest in the 81+ age group (37.8%; P < .0001) and Northeast (43.0%; P < .0001). Highly cross-linked polyethylene (HXLPE) with antioxidant use increased from 9.08% to 31.8%, while HXLPE without antioxidant use remained constant. There was an increase in use of anterior-stabilized and pivot bearings and an increase in use of HXLPE with antioxidant. Additionally, there were noted age group and regional differences in the use of various bearings. Further study should evaluate if these changing utilization trends are related to clinical outcomes, or due to regional training, preferences, or manufacturer pressures. Level III, retrospective.


Pharmacokinetics of pipemidic acid in healthy middle-aged volunteers and elderly patients with renal insufficiency.
Pipemidic acid in bacteriological screening was more potent than nalidixinic acid, notably against Pseudomonas. In a study in middle-aged female volunteers, the relative bioavailability of a single 500 mg tablet of pipemidic acid was good. The main pharmacokinetic parameters were: tmax, 2.3 h; Cmax, 3.3 micrograms/ml; t1/2, 4.6 h; Varea, 2.04 l/kg; Cltot, 5.48 ml/min per kg; Clren, 3.05 ml/min per kg; Ae(0-24), 55.7% of dose. In elderly inpatients with normal renal function, the main alterations from middle-aged volunteers were: slightly elevated Cmax (+28%) and AUC (+13%) and significantly decreased Varea (-51%), Clren (-34%) and Ae(0-24) (-31%). In elderly inpatients with renal insufficiency, Clren (-58%) was decreased compared with elderly inpatients with normal renal function. Concn. of pipemidic acid in urine still exceeded several-fold the MIC values of the most pathogenic bacteria. Among all the subjects, there was a negative correlation between Clcreat and AUC0-infinity, and a positive correlation between Clcreat and Cltot, Clren and Ae(0-24). In the treatment of urinary-tract infections with pipemidic acid, a mild renal insufficiency (Clcreat 30 ml/min or more) does not necessitate dose adjustments.


Potential role of cyclin-dependent kinase 4/6 inhibitors in the treatment of squamous cell carcinoma of the head and neck.
Human papillomavirus (HPV)-negative squamous cell carcinoma of the head and neck (SCCHN) is mainly driven by genetic aberrations involved in the cell cycle pathway resulting in cyclin-dependent kinase (CDK) 4 and 6 activation. This supports the investigation of the activity of CDK4/6 inhibitors in this disease. We review the therapeutic potential of CDK4/6 inhibitors in SCCHN. CDK4/6 inhibitors in monotherapy have demonstrated cytostatic activity in HPV-negative SCCHN. Combination with epidermal growth factor inhibitors, with phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin pathways inhibitors or with immunotherapy, have shown promising preclinical efficacy. No strong predictive biomarkers of response or resistance have been firmly identified.Phase I clinical trials have demonstrated that palbociclib or ribociclib in combination with cetuximab is well tolerated. A phase II single-arm trial combining palbociclib/cetuximab has shown promising results. Inhibition of CDK4/6 represents a new potential treatment for HPV-negative SCCHN patients. Randomized clinical trials that investigate these compounds in an unbiased manner are needed to fully evaluate their efficacy. However, it is unlikely that all the patients will benefit from this new approach. To determine a molecular profile/phenotype that will predict CDK4/6 inhibitor activity, researchers will have to take into account simultaneously occurring events in the cyclin-D/CDK4/CDK6/retinoblastoma and associated pathways.


EU health co-design policies to counteract the COVID-19 pandemic effect promoting physical activity.
The research is placed in the context of interdisciplinary medical-legal studies on the importance of promoting physical activity as a public health tool. The aim was to highlight the tools that can be used by EU members for planning interventions aimed at overcoming the consequences of the COVID-19 pandemic and for responding to a future crisis. First, the medical resources relating to the indirect and direct effects of the COVID-19 pandemic are analysed. Then, the results are compared with the measures of the EU bodies to verify the correspondence of the scientific arrests, with the political-regulatory interventions. It was found that the prolonged closure of sports centres and the contagion from COVID-19 produce affects the body in a way that can only be recovered by motor activity. However, in the EU, there does not exist a regulatory harmonization about health issues that can directly impose the Members to implement their legislation to promote motor activity. The signing of the Rome Declaration at the Global Health Summit on 21 May 2021 constitutes an important and concrete commitment for the exchange in the medical-scientific field, and for an effective co-design of intervention strategies for the relaunch of physical activity within projects such as EU4Health and the two-year HealthyLifestyle4All campaign.


Docosahexaenoic acid enhances the susceptibility of human colorectal cancer cells to 5-fluorouracil.
Powerful growth-inhibitory action has been shown for n-3 polyunsaturated fatty acids against colon cancer cells. We have previously described their ability to inhibit proliferation of colon epithelial cells in patients at high risk of colon cancer. In the work reported here we investigated the ability of docosahexaenoic acid (DHA) to potentiate the antineoplastic activity of 5-fluorouracil (5-FU) in p53-wildtype (LS-174 and Colo 320) and p53-mutant (HT-29 and Colo 205) human colon cancer cells. When in combination with DHA, 5-FU was used at concentrations ranging from 0.1 to 1.0 microM, much lower than those currently found in plasma patients after infusion of this drug. Similarly, the DHA concentrations (< or =10 microM) used in combination with 5-FU were lower than those widely used in vitro and known to cause peroxidative effects in vivo. Whereas the cells showed different sensitivity to the growth-inhibitory action of 5-FU, DHA reduced cell growth independently of p53 cellular status. DHA synergized with 5-FU in reducing colon cancer cell growth. The potentiating effect of DHA was attributable to the enhancement of the proapoptotic effect of 5-FU. DHA markedly increased the inhibitory effect of 5-FU on the expression of the antiapoptotic proteins BCL-2 and BCL-XL, and induced overexpression of c-MYC which has recently been shown to drive apoptosis and, when overexpressed, to sensitize cancer cells to the action of proapoptotic agents, including 5-FU. Our results indicate that DHA strongly increases the antineoplastic effects of low concentrations of 5-FU. Overall, the results suggest that combinations of low doses of the two compounds could represent a chemotherapeutic approach with low toxicity.


Whole-cell patch recording from Drosophila larval neurons.
The fruit fly Drosophila melanogaster has been instrumental in expanding our understanding of early aspects of neural development. The use of this model system has greatly added to our knowledge of neural cell-fate determination, axon guidance, and synapse formation. It has also become possible to access and make electrophysiological recordings directly from neurons in situ in an intact central nervous system (CNS), which has facilitated studies of the development and regulation of neuronal signaling. This protocol describes a procedure for revealing larval motor neurons and applying whole-cell patch recording techniques to these cells. The useful lifetime of first-instar larval preparations is ∼30 min, and that of third-instar CNS preparations is up to 1 h. It is therefore recommended that fresh preparations are used and that no breaks are taken during the procedure, although there may be time to pull and polish a patch pipette.


Caudal Intraparietal Sulcus and three-dimensional vision: A combined functional magnetic resonance imaging and single-cell study.
The cortical network processing three-dimensional (3D) object structure defined by binocular disparity spans both the ventral and dorsal visual streams. However, very little is known about the neural representation of 3D structure at intermediate levels of the visual hierarchy. Here, we investigated the neural selectivity for 3D surfaces in the macaque Posterior Intraparietal area (PIP) in the medial bank of the caudal intraparietal sulcus (IPS). We first identified a region sensitive to depth-structure information in the medial bank of the caudal IPS using functional Magnetic Resonance Imaging (fMRI), and then recorded single-cell activity within this fMRI activation in the same animals. Most PIP neurons were selective for the 3D orientation of planar surfaces (first-order disparity) at very short latencies, whereas a very small fraction of PIP neurons were selective for curved surfaces (second-order disparity). A linear support vector machine classifier could reliably identify the direction of the disparity gradient in planar and curved surfaces based on the responses of a population of disparity-selective PIP neurons. These results provide the first detailed account of the neuronal properties in area PIP, which occupies an intermediate position in the hierarchy of visual areas involved in processing depth structure from disparity.


[The role of phosphoinositides, protein kinase C and protein kinase A in the K+ regulatory signal transduction in human adrenocortical cells].
The messenger mechanisms mediating K+ regulatory signals in human adrenocorticocytes were studied. It was shown that potassium ions initiated decay of polyphosphoinositides to inositolphosphates and obviously diacylglycerol. The latter compounds activate protein kinase C as affected by different agonists. Using western blotting method we showed translocation of PKCalpha from cytosol to membranes after adrenal tissue preincubation in the medium with increased K+ content (8.5 mM). Translocation means activation of the enzyme. Activity of PKC increased in the microsomal fraction and did not change in cytosol. Increased concentration of K+ in the incubation medium also activates protein kinase A, although to a lesser extent compared to PKC. Unlike PKC activity of PKA was changed in cytosol as well. The possibility of involvement of several messenger systems in K+ signal transduction in human adrenocortical cells as well as the hypothesis on cross-talk between messenger mechanisms for main physiological agonists controlling aldosterone biosynthesis in the adrenals are discussed.


Contributions of the Education through Work for Health Program to the preceptors of Primary Healthcare.
To analyze the contributions of the Education through Work for Health Program to the preceptors of Primary Healthcare working in family health units in Feira de Santana, Bahia. A qualitative study conducted with preceptors in the months of March and April 2016, adopting preceptors who had worked in family health units linked to the Program for a minimum period of two years as inclusion criterion. Preceptors on vacation, maternity leave or on leave for different reasons were excluded. The collected data were interpreted based on the content analysis technique. A total of 16 preceptors were interviewed. The Program's contributions to the preceptors were as follows: expanding the perspective on health work, gaining experience in the teaching-learning process and the motivation to continue and search for new studies. The Program enabled preceptors to take a broader look at health work in the context of Primary Healthcare, to gain experience in training processes and motivation to develop new studies, in addition to contributing in a responsible way to the consolidation of the Unified Health System.


The community periodontal index of treatment needs (CPITN) procedure for population groups and individuals.
The FDI-WHO Joint Working Group 1 on periodontal diseases supports the use of the CPITN as an epidemiological screening procedure for periodontal treatment needs in populations and also, in a modified form for screening and monitoring of individuals by dental practitioners. The advantages of the CPITN procedure have become more evident following 7 years of development and field experience. This report reviews the CPITN procedure and describes in detail the use of the method for both purposes. Methods, clinical criteria and evaluation of the CPITN data are discussed. The CPITN is primarily a screening procedure which requires clinical assessment for the presence or absence of periodontal pockets, calculus and gingival bleeding. Use of a special CPITN periodontal probe (or its equivalent) is recommended. For epidemiological purposes in adult populations, 10 specified index teeth are examined; for persons under 20 years of age only six index teeth are specified. In dental practice all teeth are examined and the highest score for each sextant noted. Only 6 scores are recorded. Measures of gingival recession, tooth mobility, intensity of inflammation, precise identification of pocket depths or differentiation between supra- and subgingival calculus are not included in the CPITN. Individuals are assigned to one of four treatment need categories determined from their CPITN scores. Detailed explanations are given for the evaluation of epidemiological data. Four tabulations provide an insight into the overall pattern of prevalence, severity and categories of treatment need. For health services planning, the data provide a basis for estimating overall population needs in terms of treatment categories and the clinical personnel required for periodontal care. In the clinical care situation the procedure offers a simple screening method for determining the level of intervention required and also a tool for the longitudinal monitoring of disease changes. Used with commonsense and an understanding of periodontal disease, the CPITN procedure provides the epidemiologist and the practitioner with a practical means of assessing periodontal treatment needs.


Hematological parameters in largemouth bass (Micropterus salmoides) with formalin-preservation: comparison between wild tournament-caught and captive-raised fish.
Largemouth bass (Micropterus salmoides) are an economically important freshwater fish species that have been investigated for both the short and long-term effects of stress, secondary to angling. Limited data has been published on the hematological parameters of this species and blood sample stability is a notable limitation of hematologic field studies. A relatively novel technique using 10% neutral buffered formalin preserves heparinized whole blood and maintains blood cell stability beyond one month in striped bass. The objective of this study was to evaluate the differences in hematological parameters between tournament-caught and captive-raised largemouth bass using whole blood preservation with neutral buffered formalin. Two populations of largemouth bass (n = 26 wild; n = 29 captive) underwent coccygeal venipuncture to collect heparinized whole blood for packed cell volume, total solids, and manual differential. Formalin preservation of heparinized whole blood facilitated manual hemocytometer analysis. Results were compared between the populations (tournament-caught, and captive-raised) with Wilcoxon rank sum test, a Hotelling's T 2 test, and Bonferroni simultaneous 95% confidence intervals to determine significance. The mean packed cell volume (44.9 ± 5.4%) and total solids (7.2 ± 1.1 g/dL) were significantly higher, while the total leukocyte count (7.08 ± 1.86 × 103/µL) was significantly lower in the wild tournament-caught population of largemouth bass, as compared to the captive-raised counterparts (PCV 34.4 ± 7.2%; TS 5.2 ± 1.0 g/dL; WBC 16.43 ± 8.37 × 103/µL). The wild population demonstrated a significantly distinct leukogram characterized by a neutropenia (24.1 ± 12.7%), lymphocytosis (67.7 ± 13.0%), and monocytopenia (8.3 ± 2.9%), while the erythrocyte and thrombocyte counts were not significantly different between populations. Numerous factors have been demonstrated to influence hematologic parameters in fish including age, size, sex, temperature, environmental oxygen level, population density, and infection. The wild population endured stress during angling capture, live-well hypoxia, transport, and extended air exposures at weigh in, which may have caused a stress leukopenia as well as osmoregulatory dysfunction and subsequent hemoconcentration. Further evaluation of seasonal impact as well as increased sample size is warranted to enhance our understanding of largemouth bass hematology. This study concluded that wild largemouth bass captured via tournament angling have higher packed cell volume and total solids, and lower total leukocyte counts, compared to captive-reared individuals. Through the completion of this study, we demonstrated the successful use of 10% neutral buffered formalin to preserve heparinized whole blood for precise hemocytometer cell counts in a new teleost species, the largemouth bass.


Implantation of a synthetic cornea: design, development and biological response.
Our goal was to evaluate 3 different designs of synthetic corneas in vivo. All devices had a transparent hydrogel center molded to a porous peripheral skirt. Over 30 devices were implanted into rabbits and followed for up to 6 months. The devices were preseeded with rabbit stromal fibroblasts, which enhanced the rate of fibroplasia. The anterior surface of the hydrogel was modified using argon rf plasma treatments. Clinical examinations were performed, and histological analyses were conducted on tissue throughout the time course. Our optimal model ranged from 4.5 to 6 mm and had an extended porous skirt increasing the surface area for fibroplasia and ultimate anchorage of the device. Fibroplasia occurred in this model, and collagen was detected by 28 days. The anterior chamber was normal with no detectable leakage of aqueous humor. Glycosaminoglycans were detected and followed the time course outlined previously when porous material itself was inserted into the stroma. We present the first demonstration that rabbit limbal epithelial cells can migrate onto the synthetic cornea in vivo.


An evaluation of the referral transport system of Patna, Bihar.
The key objective of the study was to evaluate the coverage and functioning of the referral transport system under NRHM in block PHCs of district Patna in Bihar. A cross-sectional descriptive study was conducted during October-November, 2008 in 16 block PHCs in Patna. In-depth interviews were conducted at 811 households where there was an occasion to transfer a patient to a hospital in the previous two months time. Medical officer in-charge and civil surgeon of the district were also interviewed. Besides, focus group discussions were conducted with the community members and leaders regarding the functioning of referral transport. Availability of the referral transport services was irregular mainly due to deputation of the vehicles for flood relief activities or other purposes. 93 (11.5%) of respondents used the PHC transport facilities, of which 52.7% got it instantaneously. 'Dial an ambulance 102' services were mainly used by urban clients. The system was following an arbitrary cost structure. 84.2% of the clients belonging to below poverty line had to pay for the service and are afraid of availing the services. Inadequate number of ambulances in PHCs, unequipped ambulances, lack of life saving equipments, 24 hours duty by single driver, arbitrary cost frame work and urban preference for services were some of the factors leading to unpopularity of the scheme.


Angiotensinase and vasopressinase activities in hypothalamus, plasma, and kidney after inhibition of angiotensin-converting enzyme: basis for a new working hypothesis.
Reducing angiotensin II (Ang II) production via angiotensin-converting enzyme (ACE) inhibitors is a key approach for the treatment of hypertension. However, these inhibitors may also affect other enzymes, such as angiotensinases and vasopressinase, responsible for the metabolism of other peptides also involved in blood pressure control, such as Ang 2-10, Ang III, Ang IV, and vasopressin. We analyzed the activity of these enzymes in the hypothalamus, plasma, and kidney of normotensive adult male rats after inhibition of ACE with captopril. Aspartyl- (AspAP), glutamyl- (GluAP), alanyl- (AlaAP) and cystinyl-aminopeptidase (CysAP) activities were measured fluorimetrically using arylamides as substrates. Systolic blood pressure (SBP), water intake, and urine flow were also measured. Captopril reduced SBP and increased urine flow. In the hypothalamus, GluAP and AspAP increased, without significant changes in either AlaAP or CysAP. In contrast with effects in plasma, GluAP was unaffected, AspAP decreased, while AlaAP and CysAP increased. In the kidney, enzymatic activities did not change in the cortex, but decreased in the medulla. These data suggest that after ACE inhibition, the metabolism of Ang I in hypothalamus may lead mainly to Ang 2-10 formation. In plasma, the results suggest an increased formation of Ang IV together with increased vasopressinase activity. In the kidney, there is a reduction of vasopressinase activity in the medulla, suggesting a functional reduction of vasopressin in this location. The present data suggest the existence of alternative pathways in addition to ACE inhibition that might be involved in reducing BP after captopril treatment.


Cyclosporine for pyoderma gangrenosum.
A case of pyoderma gangrenosum that occurred in the absence of underlying illness and was initially misdiagnosed is described. Hemorrhagic pustules and ulcers appeared over vein-graft sites after open heart surgery and were treated with cyclosporine. The result was rapid and complete resolution. It is the purpose of this report to emphasize pyoderma gangrenosum as a cause of ulceration in healthy individuals and to highlight the addition of cyclosporine to the therapeutic armamentarium.


A Systematic Investigation Unveils High Coinfection Status of Porcine Parvovirus Types 1 through 7 in China from 2016 to 2020.
Porcine parvovirus genotype 1 (PPV1) causes reproductive disorder in swine and is prevalent in China. Recently, six new genotypes of PPVs (PPV2 through PPV7) have also been detected in Chinese swine herds. However, the coinfection status of all these seven genotypes of PPVs (PPV1-7) in China was not clarified yet. In this study, we developed a panel of PPV1-7 PCR assays with satisfied specificity, sensitivity and reproducibility and then applied to the detection of PPV1-7 in 435 clinical samples collected from eight provinces of China in 2016-2020. A total of 55.40% samples (241 out of 435) were PPV positive, while PPV2 and PPV3 (both 22.53%) belonging to the genus of Tetraparvovirus were the most prevalent genotypes. Noticeably, PPV1-7 strains were more prevalent in nursery and finishing pigs than in suckling pigs. In addition, coinfection could be detected in all eight provinces and 27.36% (119/435) samples were coinfected with two to five genotypes of PPVs. Meanwhile, the coinfection of PPVs with PCV2 was 22.30% (97/435). Twenty complete genomes of representative PPV1-7 were determined, and phylogenetic analysis confirmed the genotyping results by sequence comparisons and PCR assays. Remarkably, the PPV7 HBTZ20180519-152 strain from domestic pig was recombined from parental JX15-like and JX38-like isolates from wild boars. Selective pressure analysis based on VP2 sequences of PPV1-7 showed that they were predominantly under negative selection, while few positive selection sites could be detected in VP2 of PPV7. Overall, this systematic investigation unveils high prevalence and coinfection of PPV1-7 in China from 2016 to 2020. IMPORTANCE Porcine parvoviruses (PPVs) are prevalent in China associating with reproductive failure in swine. The coinfection of seven genotypes of PPVs (PPV1-7) might have synergistic effects on PPV1 associated SMEDI syndrome. However, the coinfection status of PPV1-7 in China is not clear yet. This study showed that PPV1-7 strains are highly prevalent (55.40%) in China and mainly in nursery and finishing pigs in recent years. In addition, the coinfections of different genotypes of PPVs (27.36%) and PPVs with PCV2 (22.30%) are common. Geographic analysis indicated that different genotypes of PPVs are widely cocirculating in China. Intriguingly, a PPV7 strain from the domestic pig was detected as a recombinant from two wild boar isolates. Selective pressure analyses showed that PPV1-7 are mainly under purifying selection. Our findings provide the first systematic investigation on the prevalence, coinfection, and evolution of PPV1 through PPV7 in Chinese swineherds from 2016 to 2020.


Spatial cognition in the context of foraging styles and information transfer in ants.
Ants are central-place foragers: they always return to the nest, and this requires the ability to remember relationships between features of the environment, or an individual's path through the landscape. The distribution of these cognitive responsibilities within a colony depends on a species' foraging style. Solitary foraging as well as leader-scouting, which is based on information transmission about a distant targets from scouts to foragers, can be considered the most challenging tasks in the context of ants' spatial cognition. Solitary foraging is found in species of almost all subfamilies of ants, whereas leader-scouting has been discovered as yet only in the Formica rufa group of species (red wood ants). Solitary foraging and leader-scouting ant species, although enormously different in their levels of sociality and ecological specificities, have many common traits of individual cognitive navigation, such as the primary use of visual navigation, excellent visual landmark memories, and the subordinate role of odour orientation. In leader-scouting species, spatial cognition and the ability to transfer information about a distant target dramatically differ among scouts and foragers, suggesting individual cognitive specialization. I suggest that the leader-scouting style of recruitment is closely connected with the ecological niche of a defined group of species, in particular, their searching patterns within the tree crown. There is much work to be done to understand what cognitive mechanisms underpin route planning and communication about locations in ants.


Human brain tumors: assessment with in vivo proton MR spectroscopy.
To better understand variations in spectra of brain tumors, 122 in vivo proton spectra of brain tumors in 82 patients were analyzed. The changes in relative metabolite concentrations compared with those in normal spectra and the presence of any new metabolite were assessed. To evaluate the clinical usefulness of in vivo hydrogen-1 magnetic resonance (MR) spectroscopy in brain tumors, the authors looked for specific spectral changes on the basis of tumor grade. All tumor spectra showed differences from normal reference spectra. The differential diagnosis of the spectra was limited because intraindividual differences between spectra of one tumor at different locations were often larger than differences between spectra of tumors with different histologic characteristics. However, the variations in metabolite concentrations, and especially the presence or absence of aliphatic signals, were proved to be indicators of the histologic grade of tumor. The observed spectral patterns conformed to a four-compartment model, described herein, which is proposed to improve the interpretation of brain spectra.


Restricted exchange microenvironments for cell culture.
Metabolite diffusion in tissues produces gradients and heterogeneous microenvironments that are not captured in standard 2D cell culture models. Here we describe restricted exchange environment chambers (REECs) in which diffusive gradients are formed and manipulated on length scales approximating those found in vivo. In REECs, cells are grown in 2D in an asymmetric chamber (<50 μL) formed between a coverglass and a glass bottom cell culture dish separated by a thin (~100 μm) gasket. Diffusive metabolite exchange between the chamber and bulk media occurs through one or more openings micromachined into the coverglass. Cell-generated concentration gradients form radially in REECs with a single round opening (~200 μm diameter). At steady state only cells within several hundred micrometers of the opening experience metabolite concentrations that permit survival which is analogous to diffusive exchange near a capillary in tissue. The chamber dimensions, the openings' shape, size, and number, and the cellular density and metabolic activity define the gradient structure. For example, two parallel slots above confluent cells produce the 1D equivalent of a spheroid. Using REECs, we found that fibroblasts align along the axis of diffusion while MDCK cells do not. MDCK cells do, however, exhibit significant morphological variations along the diffusive gradient.


The human tumour suppressor gene p53 is alternatively spliced in normal cells.
Alternative splicing affecting the p53 carboxy-terminus has previously been described in mouse but not in normal human cells. We report here the detection in normal human lymphocytes of an alternatively spliced form of human p53 mRNA containing an additional 133 bp exon derived from intron 9. This splice variant encodes a truncated protein of 341 amino-acids including 10 new amino-acids derived from the novel exon. The truncated protein, which lacks part of the p53 tetramerization domain, fails to bind DNA in vitro and has a transcriptional defect in vivo in both yeast and mammalian cells. Quantitative RT-PCR experiments suggest that the alternatively spliced form is only present in significant amounts in quiescent cells. Considering the numerous functions ascribed to the carboxy-terminus of the p53 protein, this splice variant may have important implications for the biological role of p53 in normal cells.


Role of Pre-operative Inflammatory Markers as Predictors of Lymph Node Positivity and Disease Recurrence in Well-Differentiated Pancreatic Neuroendocrine Tumours: Pancreas2000 Research and Educational Program (Course 9).
Pancreatic neuroendocrine tumours (PNET) is a rare disease and in the absence of metastases, surgical resection is recommended. Key factors affecting survival in PNETs are the stage and grade of the disease, but there is increasing evidence suggesting lymph node involvement is associated with shorter disease-free and overall survival. Ability to predict the likelihood of lymph node involvement at the time of diagnosis would affect surgical decision making in these patients. A systemic inflammatory index such as neutrophil to lymphocyte ratio or platelet to lymphocyte ratio has been associated with poor prognosis in several cancers. Method: This study is a retrospective multi-centre study. The data including pre-operative inflammatory markers such as haemoglobin, neutrophil, lymphocyte counts and pathological data including number of positive lymph nodes, tumour grade and size, are collected to assess the association between inflammatory index and lymph node involvement. Conclusion: This study aims to assess the value of routinely available pre-operative haematological markers in predicting lymph node involvement in non-functioning PNETs.


Response surface optimization for determination of volatile organic compounds in water samples by headspace-gas chromatography-mass spectrometry method.
The determination of benzene, toluene, p-xylene, tetrachloroethene, chlorobenzene, and dibromomethane in water by headspace analysis combined with gas chromatography-mass spectrometry (HS-GC-MS) has been investigated. An optimization strategy for the analysis of the six volatile organic compounds (VOCs) is presented with the experimental design and response surface methodology aid. Thermostatting time, temperature, and salt quantity were optimized by using a central composite design, and quadratic models relating peak areas to the three factors were built. The mathematical models were tested on a number of simulated data set and had a coefficient of R2>0.98. The factor effects were visualized as three-dimensional response surfaces and contour plots. The optimal conditions were achieved in 36.8 min, at 90 degrees C, and with 6 g of NaCl. The method showed a good agreement between the experimental data and predictive values throughout the studied parameter space, and was suitable for optimization studies of the VOCs in water by the HS-GC-MS method.


Questioning the validity of the 4+/5+ binge or heavy drinking criterion in college and clinical populations.
The terms 'binge drinking' and 'heavy drinking' are both operationalized typically as 4+/5+ standard drinks per occasion for women/men, and are used commonly as a proxy for non-problematic (<4/<5) versus problematic (4+/5+) drinking in multiple research contexts. The Food and Drug Administration in the United States recently proposed the 4+/5+ criterion as a primary efficacy end-point in their guidance for trials examining new medications for alcohol use disorders (AUDs). Internationally, similar cut-offs have been proposed, with the European Medicines Agency having identified reductions in the number of heavy drinking days (defined as 40/60 g pure alcohol in women/men) as a primary end-point for efficacy trials with a harm reduction goal. We question the validity of the 4+/5+ cut-off (and other similar cut-offs) on multiple accounts. The 4+/5+ cut-off has not been shown to have unique predictive validity or clinical utility. The cut-off has been created based on retrospective self-reports and its use demonstrates ecological bias. Given strong evidence that the relationship between alcohol consumption and problems related to drinking is at least monotonic, if not linear, there is little existing evidence to support the 4+/5+ cut-off as a valid marker of problematic alcohol use. There is little empirical evidence for the 4+/5+ standard drinks per occasion threshold for 'binge' or 'heavy' drinking in indexing treatment efficacy. Further consideration of an appropriate threshold seems to be warranted.


1,1'-[m-Phenyl-enebis(nitrilo-methanylyl-idene)]dinaphthalen-2-ol-1,1'-[m-phenyl-enebis(imino-methanylyl-idene)]dinaphthalen-2(1H)-one (0.58/0.42).
In the solid state the title Schiff base, 0.58C(28)H(20)N(2)O(2)·0.42C(28)H(20)N(2)O(2), exists both as the keto-imino and as the enol-amino tautomer, which is manifested in the disorder of the H atom in the intra-molecular hydrogen-bonded ring. The naphthalene ring systems show some distortion, which is consistent with the quinoid effect. The ratio of the enol form refined to 58 (5)%. The mol-ecule has crystallographically imposed symmetry: a twofold axis passes through the central benzene ring. Crystals are built up of layers parallel to (010). Stacking interactions between the layers involve only standard van der Waals attraction forces between apolar groups. The alignment of the aromatic rings in neighbouring layers shows a herringbone motif. A weak C-H⋯O inter-action is observed.


Metnase and EEPD1: DNA Repair Functions and Potential Targets in Cancer Therapy.
Cells respond to DNA damage by activating signaling and DNA repair systems, described as the DNA damage response (DDR). Clarifying DDR pathways and their dysregulation in cancer are important for understanding cancer etiology, how cancer cells exploit the DDR to survive endogenous and treatment-related stress, and to identify DDR targets as therapeutic targets. Cancer is often treated with genotoxic chemicals and/or ionizing radiation. These agents are cytotoxic because they induce DNA double-strand breaks (DSBs) directly, or indirectly by inducing replication stress which causes replication fork collapse to DSBs. EEPD1 and Metnase are structure-specific nucleases, and Metnase is also a protein methyl transferase that methylates histone H3 and itself. EEPD1 and Metnase promote repair of frank, two-ended DSBs, and both promote the timely and accurate restart of replication forks that have collapsed to single-ended DSBs. In addition to its roles in HR, Metnase also promotes DSB repair by classical non-homologous recombination, and chromosome decatenation mediated by TopoIIα. Although mutations in Metnase and EEPD1 are not common in cancer, both proteins are frequently overexpressed, which may help tumor cells manage oncogenic stress or confer resistance to therapeutics. Here we focus on Metnase and EEPD1 DNA repair pathways, and discuss opportunities for targeting these pathways to enhance cancer therapy.


GATA Binding Protein 3 (GATA-3) expression evaluation as prognostic factor in breast cancer and its relationship with other immunemarkers.
Breast cancer is the most frequent cancer in which the mortality rate could be decreased by proper management. The GATA3 transcription factor is one of the most frequently mutated genes in breast cancer. We studied the immunohistochemical (IHC) expression of estrogen and progesterone receptor, human epidermal growth factor receptor 2, and GATA-3 in 166 radical/partial mastectomy specimens having different histologic grades and stages of breast carcinoma. All samples were obtained from the pathology department of Sina hospital in Tehran-Iran from 2010 to 2016. There was a direct relationship between the luminal subtype carcinoma and higher GATA-3 expression (P-value: 0.001) and between triple-negative carcinoma and lower GATA-3 expression (P-value: 0.001). Moreover, there was a direct relationship between the metastasis rate and the tumor's grade with GATA-3 staining (P-value: 0.000 and 0.001, respectively). GATA-3 expression is related to the histopathologic and prognostic factors. GATA3 can be introduced as an important predictor in breast cancer patients.


Hybridization Networks of mRNA and Branched RNA Hybrids.
Messenger RNA (mRNA) is emerging as an attractive biopolymer for therapy and vaccination. To become suitable for vaccination, mRNA is usually converted to a biomaterial, using cationic peptides, polymers or lipids. An alternative form of converting mRNA into a material is demonstrated that uses branched oligoribonucleotide hybrids with the ability to hybridize with one or more regions of the mRNA sequence. Two such hybrids with hexamer arms and adamantane tetraol as branching element were prepared by solution-phase synthesis. When a rabies mRNA was treated with the branched hybrids at 1 M NaCl concentration, biomaterials formed that contained both of the nucleic acids. These results show that branched oligoribonucleotides are an alternative to the often toxic reagents commonly used to formulate mRNA for medical applications.


Re-exploring the pelvic neuroanatomy from a new perspective and a potential guidance for TaTME: a "bottom-up" approach.
Neuro-anatomy of the perineum has gained renewed attention due to its significance in the transanal procedures for rectal cancer (eg TaTME). Surgeons embarking on this technique must have sophisticated knowledge and a precise anatomical understanding of the perineum before proceeding with this reversed rectal approach. We report anatomical observations deriving from a relevant experience in the colorectal surgery field. The collective multicenter experience of the present study is clinically relevant and based on the rectal and transanal resections performed in colorectal centers of excellence from Greece, UK, and Italy over the last 10 years (2011-2020). From the original anatomical and intraoperative observations derived from collective cases operated by this multicenter group of colorectal surgical centers in three European countries, data were retrieved and analyzed in collaboration with specialist researchers of human anatomy and interpreted for their clinical significance and potential use for preoperative planning and intraoperative guidance during TaTME. This descriptive article demonstrates in detail the neurogenic pathways encountered in the perineum and pelvic cavity during transanal procedures. Specific anatomical and topographic implications are also included serving as a guide for colorectal surgeons to perform a nerve-sparing procedure. transanal approach for rectal excision offers new insights into the complex pelvic and perineal neuroanatomy while the procedure itself remains a challenge for surgeons. Preoperative anatomical planning and 3D reconstruction may help in anticipating technical difficulties, resulting in more precise surgical dissections and decreased postoperative complications.


Lack of association of endothelial nitric oxide synthase polymorphisms and migraine.
The aim of this study was to evaluate if 2 functional endothelial nitric oxide synthase (eNOS) gene polymorphisms might be risk factors for migraine. Nitric oxide synthase promotes the synthesis of nitric oxide (NO). NO is a potent vasodilator and mediates several processes involved in migraine pathophysiology. Only one study has suggested an association with migraine with aura. We performed a sex- and age-matched case-control study using 2 eNOS polymorphisms (rs1800779 and rs1799983), which are in linkage disequilibrium. Genotypes were obtained with allele-specific probes in a real-time polymerase chain reaction assay. Genotypic and allelic distributions were compared with chi(2) method. We also estimated the reconstructed haplotypes and calculated ORs for individual haplotypes. A total of 337 migraine patients (188 with aura) and 341 healthy controls were recruited. We found no significant differences in the distribution of genotypes and alleles for either polymorphism among clinical subgroups. Neither rs1800779 nor rs1799983 polymorphisms increased the risk for suffering from migraine aura. As others have previously reported, we failed to demonstrate genetic association of the eNOS gene with migraine.


Rhodium-catalyzed intramolecular, anti-Markovnikov hydroamination. Synthesis of 3-arylpiperidines.
The intramolecular anti-Markovnikov hydroamination of 1-(3-aminopropyl)vinylarenes in the presence of a readily available rhodium catalyst to form 3-arylpiperidines is reported. In contrast to intermolecular hydroamination of vinylarenes, which occurred in high yields in the presence of rhodium catalysts containing DPEphos, the intramolecular reaction occurred in high yield in the presence of [Rh(COD)(DPPB)]BF4 as catalyst. Reactants with substituents beta to the nitrogen occurred in high yield, and these reactions formed 3,5-disubstituted piperidines with high diastereomeric excess. The regiochemistry of these cyclizations contrasts with the regiochemistry of intramolecular hydroaminations catalyzed by lanthanide complexes, group III metal complexes, and platinum complexes, all of which have been reported to form cyclization products from Markovnikov addition.


Hyperimmune globulin in pregnancy for the prevention of congenital cytomegalovirus disease.
Cytomegalovirus (CMV) is the most common and serious cause of congenital infections in developed countries since it is capable of infecting the fetus after both primary and recurrent maternal infection, and can be spread for years by infected children. Areas covered: Animal and human pregnancy studies about the prevention of congenital CMV infection and disease by CMV-specific hyperimmune globulin (HIG). Commercial HIG is manufactured from the plasma of selected donors with high anti-CMV antibody avidity and titers. Expert commentary: Currently available experimental and clinical studies and case reports support the possible effectiveness and safety of HIG infusions in pregnancy for the prevention of congenital CMV disease. The knowledge about the potential efficacy of preventive or therapeutic HIG administration should be enlarged by multi-center randomized studies, which may be favored by the implementation of CMV screening. Meanwhile, if ultrasound examinations show signs of fetal injury, or CMV is detected in the amniotic fluid, the patients should be advised about the possible option of HIG therapy.


Electrotransfer of cDNA coding for a heterologous prion protein generates autoantibodies against native murine prion protein in wild-type mice.
Prion diseases (e.g., Creutzfeldt-Jakob disease in humans) are always fatal neurodegenerative disorders characterized by conversion of the ubiquitous cellular prion protein (PrP(c)) into a pathological conformer. Immunological strategies are considered as promising prophylactic or therapeutic approaches but, unfortunately, vaccination attempts until now have been very disappointing in wild-type animals because of immune tolerance to self PrP(c). Encouraging results have come from recent experiments carried out through genetic immunization (i.e., injection in mice of cDNA coding for murine prion protein [PrP]) or heterologous protein immunization (i.e., injection in mice of PrP from another species), albeit the levels of autoantibodies in wild-type animals remained generally low. Here we investigated whether combining the potential benefits of these two last approaches, namely using genetic immunization with the cDNA coding for a heterologous PrP, could more efficiently break immune tolerance. Wild-type mice were thus vaccinated with cDNA coding for human PrP(c), fused or unfused to a stimulatory T-cell epitope, using or not using electrotransfer of DNA. After three DNA injections, mice receiving electrotransferred DNA developed a strong immune response, oriented toward the humoral Th2 type, characterized not only by high IgG1 and IgG2a antibody titers against the heterologous human PrP(c), but also, as expected, by significant amounts of autoantibodies recognizing the native conformation of murine PrP(c) expressed on cell membranes as revealed by flow cytometry and immunofluorescence. These results hence open the way for investigation of the possible protective effects of anti-PrP(c) autoantibodies in infected mouse models. More generally, our results suggest that this original immunization strategy could be of value for circumventing tolerance to poorly immunogenic proteins.


Herbal products: behaviors and beliefs among Italian women.
The use of phytotherapy is growing worldwide, but the popular perception is that this kind of approach is natural and therefore safer than traditional medicine; for this reason the use is frequently not communicated to the doctor. Instead, even if many herbal remedies are benign in nature, some of these therapies have potentially harmful side effects or adverse interactions with other medications. So, the purpose of our study was to analyze the behavior patterns and decision-making modalities about herbal remedy use among a sample of Italian women. During a 5-month period, interviews to women attending the outpatient ambulatory of an urban university general hospital were made on the basis of a pre-structured 25-item questionnaire. Among a random study population of 1,044 subsequent patients, 491 women (47.03%) reported to have been taking one or more herbal products in the last year, sometimes used during pregnancy or given to their children (35.23%). The 10 most frequently used herbal products reported were propolis, aloe, valerian root, blueberry, fennel, dandelion, mallow, arnica, thyme, and Echinacea. The major purposes for using these products were to stimulate the immune system and to cure respiratory problems. 47/491 (9.57%) women reported side effects, but only 36% referred to the doctor. In most of the cases, herbal products were taken in combination with drugs (44.61%) or homeopathic treatments (11.81%). The majority of our women did not obtain information about this kind of therapy from a health care provider (72.71%). The present survey highlights the general use of phytomedicines by a sample of Italian women, the potential risk of their confidence with the 'natural world,' and the lack of discussion on this argument with doctors and pharmacists. This suggests the importance of training for health care providers and the need of informational programs for consumers.


Complete mitochondrial genome of a Neotropical dobsonfly Chloronia mirifica Navás, 1925 (Megaloptera: Corydalidae), with phylogenetic implications for the genus Chloronia Banks, 1908.
The complete mitochondrial (mt) genome of Chloronia mirifica Navás, 1925 was determined and analyzed. This represents the first mt genome of the Neotropical endemic dobsonfly genus Chloronia Banks, 1908. The mt genome of C. mirifica is a typical circular DNA of 15877 base pairs with an A+T 77.1%. It has an ancestral gene arrangement of the insect mt genome composed of 37 genes. Eleven of the 13 protein coding genes (PCGs) start with ATT and ATG, while nad6 and nad1 start with ATC and TTG respectively. Five PCGs end with a single T, while the others have a complete termination codon TAG or TAA. Most tRNAs are folded into the typical clover-leaf structure except for the trnS1 whose dihydrouridine arm is a simple loop. The secondary structure of rrnL consists of five structural domains and 50 helices, while the rrnS includes three domains and 34 helices. The phylogenetic analysis based on all 13 PCGs and two rRNA genes of the mt genome recovers Chloronia to be the sister group of Corydalus Latreille, 1802.


Whole-exome sequencing identifies a novel INS mutation causative of maturity-onset diabetes of the young 10.
Monogenic diabetes is often misdiagnosed with type 2 diabetes due to overlapping characteristics. This study aimed to discover novel causative mutations of monogenic diabetes in patients with clinically diagnosed type 2 diabetes and to explore potential molecular mechanisms. Whole-exome sequencing was performed on 31 individuals clinically diagnosed with type 2 diabetes. One novel heterozygous mutation (p.Ala2Thr) in INS was identified. It was further genotyped in an additional case-control population (6523 cases and 4635 controls), and this variant was observed in 0.09% of cases. Intracellular trafficking of insulin proteins was assessed in INS1-E and HEK293T cells. p.Ala2Thr preproinsulin-GFP was markedly retained in the endoplasmic reticulum (ER) in INS1-E cells. Activation of the PERK-eIF2α-ATF4, IRE1α-XBP1, and ATF6 pathways as well as upregulated ER chaperones were detected in INS1-E cells transfected with the p.Ala2Thr mutant. In conclusion, we identified a causative mutation in INS responsible for maturity-onset diabetes of the young 10 (MODY10) in a Chinese population and demonstrated that this mutation affected β cell function by inducing ER stress.


Avulsion fractures of the proximal tibial epiphysis.
Fractures of the tibia through the proximal epiphysis are rare. This injury usually results from severe direct or indirect force about the knee, and has not been described as resulting from a patellar tendon avulsion injury. Four patients presented with five avulsion fractures of the proximal tibial epiphysis. All were older adolescent males who had been engaged in jumping sports when the injury occurred; one had bilateral injury. All the patients were treated by closed reduction and plaster cast immobilization for 4-8 weeks, with satisfactory results. On the basis of our cases and five cases previously reported, the authors would agree with Ryu and Debenham's suggestion that the Watson-Jones classification, which divides avulsion fractures of the tibial tubercle into three types, should be expanded to include this fourth type - avulsion fracture of the proximal tibial epiphysis.


Resveratrol in prevention and treatment of common clinical conditions of aging.
Resveratrol is a potent member of the class of natural, plant-derived chemicals known as polyphenols. These help explain in part why a diet high in fruit and vegetables confers health benefits and are associated with reduced risk of common complex conditions such as cardiovascular disease, cancer, diabetes, and Alzheimer's disease. We present the latest molecular findings that account for the beneficial actions of resveratrol. The intracellular pathways activated are crucial for anti-oxidant defence, regulation of the cell cycle, mitochondrial energy production, vascular tone, oncogene suppression, and many other phenomena which if unchecked lead to morbidity and mortality from onset and progression of these various diseases. While a healthy diet and lifestyle is strongly recommended in prevention of such conditions, the future bodes well for the use of resveratrol and analogues of higher potency than the natural form for treatment of diseases that afflict humans, particularly as they age.


Advanced backcross QTL analysis: a method for the simultaneous discovery and transfer of valuable QTLs from unadapted germplasm into elite breeding lines.
Advanced backcross QTL analysis is proposed as a method of combining QTL analysis with variety development. It is tailored for the discovery and transfer of valuable QTL alleles from unadapted donor lines (e.g., land races, wild species) into established elite inbred lines. Following this strategy, QTL analysis is delayed until the BC2 or BC3 generation and, during the development of these populations, negative selection is exercised to reduce the frequency of deleterious donor alleles. Simulations suggest that advanced backcross QTL analysis will be effective in detecting additive, dominant, partially dominant, or overdominant QTLs. Epistatic QTLs or QTLs with gene actions ranging from recessive to additive will be detected with less power than in selfing generations. QTL-NILs can be derived from advanced backcross populations in one or two additional generations and utilized to verify QTL activity. These same QTL-NILs also represent commercial inbreds improved (over the original recurrent inbred line) for one or more quantitative traits. The time lapse from QTL discovery to construction and testing of improved QTL-NILs is minimal (1-2 years). If successfully employed, advanced backcross QTL analysis can open the door to exploiting unadapted and exotic germplasm for the quantitative trait improvement of a number of crop plants.


Ratio of paricalcitol dosage to serum parathyroid hormone level and survival in maintenance hemodialysis patients.
Several observational studies have indicated that vitamin D receptor activators (VDRA), including paricalcitol, are associated with greater survival in maintenance hemodialysis (MHD) patients; however, patients with higher serum parathyroid hormone (PTH), indicative of a more severe secondary hyperparathyroidism and higher death risk, are usually given higher VDRA dosages, which can lead to confounding by medical indication and attenuated survival advantage of high VDRA dosages. It was hypothesized that the ratio of the administered paricalcitol dosage to serum PTH level discloses better the underlying dosage-survival association. The 3-yr mortality predictability of the administered paricalcitol during the first 3 mo of the cohort divided by averaged serum intact PTH during the same period was examined in 34,307 MHD patients from all DaVita dialysis clinics across the United States using Cox regression. MHD patients were 60.8 +/- 15.4 yr of age and included 47% women, 34% black patients, and 47% patients with diabetes. Initially, the ratio of paricalcitol (mircrog/wk) to PTH (pg/ml) was divided into four groups: 0 (reference), 1 to <30, 30 to <60, and >60 x 10(-3). Unadjusted, case mix-adjusted (demographics, comorbidity, and Kt/V), and malnutrition-inflammation complex syndrome-adjusted models, the death rate ratio for the paricalcitol/PTH index groups, were 0.99, 0.95, and 0.92. Restricted cubic splines analyses were consistent with a linear relation. Higher weekly paricalcitol dosage per each unit of serum PTH seems to have an incremental association with greater survival in MHD patients. The observed dosage-response phenomenon needs to be confirmed in clinical trials.


PPAR-gamma activation as an anti-inflammatory therapy for respiratory virus infections.
Newly emerged influenza viruses have attracted extensive attention due to their high infectivity, proinflammatory actions, and potential to induce worldwide pandemics. Frequent mutations and gene reassortments between viruses complicate the development of protective vaccines and antiviral therapeutics. In contrast, targeting the host response for the development of novel cost-effective, broad-based prophylactic or therapeutic agents holds greater promise. Since inflammation often parallels the development of productive immune responses, virus-induced pulmonary inflammation and injury represents an additional challenge to the development of broad-based immunotherapeutics. Excessive inflammatory responses to respiratory viruses, also known as "cytokine storm," are largely due to immune dysregulation that manifests as proinflammatory cytokine secretion. In addition to modulating lipid and glucose metabolism, peroxisome proliferator-activated receptors (PPAR) play important roles in antagonizing core inflammatory pathways such as NF-kappaB, AP1, and STAT. Their role in regulating inflammatory responses caused by pulmonary pathogens is receiving increasing attention, setting the stage for the discovery of novel applications for anti-diabetic and lipid-lowering drugs. This review focuses on the potential use of PPAR-gamma agonists to downregulate the inflammatory responses to respiratory virus-related pulmonary inflammation.


Skeletal growth of children from the Iron Age site at K2 (South Africa).
Cross-sectional growth data were obtained from the skeletal remains of children from the Iron Age site of K2 near the Limpopo River. Standard measurements of the diaphyseal lengths of the long bones from both limbs were recorded and compared to published skeletal data. For this purpose, data on Eskimo and Aleut skeletons, Libben skeletons, and skeletons from Indian Knoll and Altenerding were used. An attempt to study growth allometrically was made. K2 children were growing as well as children from these other groups. Comparison of data for K2 children with those on living South African "Cape Coloured" rural children, studied during the late 1980s, shows the similarity of growth of both groups.


Synchronous primary endometrial and ovarian cancers: a multicenter review of 63 cases.
To investigate clinicopathologic characteristics, therapeutic methods, and prognostic factors in women with synchronous primary endometrial and ovarian cancers (SEOCs). A retrospective review of 2 cancer registry databases in Turkey was conducted to identify patients diagnosed with SEOCs between January 1995 and December 2012. Patients with recurrent, metastatic, and metachronously occurring tumors were excluded. Multivariate logistic regression models were used to identify prognostic predictors for progression-free survival (PFS) and overall survival (OS). The analysis included 63 women with SEOCs. Seventy-six percent of the patients had stage I endometrial cancer, and 60% of the patients had stage I ovarian cancer. Thirty-seven patients (58.7%) had endometrioid/endometrioid histology. Optimal cytoreduction was obtained in 47 (74.6%) patients. Recurrence developed in 17 patients (27%). Multivariate analysis confirmed lymphovascular space invasion (LVSI) as an independent poor prognostic factor for OS (odds ratio [OR] 3.1, p = 0.045), whereas early-stage disease and optimal cytoreduction were found to be independent good prognostic factors for both PFS (OR 12.85, p<0.001 and OR 4.58, p = 0.004, respectively) and OS (OR 7.31, p = 0.002 and OR 2.95, p = 0.028, respectively). The 3- and 5-year OS rates were 74% and 69%, respectively. Our study demonstrated that optimal cytoreduction, early-stage disease, and LVSI are the most significant factors affecting survival in women with SEOC.


Is bronchoscopic lung biopsy helpful in the management of patients with diffuse lung disease?
Few studies have evaluated how the results of bronchoscopic lung biopsy (BLB) affect clinical management. The objective of the present study was to evaluate the clinical usefulness of BLB in the overall management of patients with diffuse pulmonary diseases. The study was a retrospective analysis of patients who underwent fluoroscopy-guided BLB over a 2-yr period. Patients whose biopsy was of a lung mass or single dominant lung nodule were excluded. The usefulness of BLB was assessed to determine whether the results affected the clinical management of these patients. During the study period, 603 patients underwent 651 BLB procedures. The results of BLB were clinically helpful in 494 (75.9%) out of 651 procedures. No diagnostic abnormality was identified in 107 (16.4%) out of the 651 biopsy procedures. This finding was clinically helpful in 59 (55.1%) out of 107 procedures, as the results excluded specific processes suspected before BLB. In 52 procedures (8% of all BLB), no lung parenchyma was obtained. In conclusion, bronchoscopic lung biopsy is a clinically useful test in approximately 75% of procedures. In the 25% of bronchoscopic lung biopsies that were clinically unhelpful, the reason for failure in approximately one-third of patients was the failure of the procedure to obtain an adequate quantity of lung parenchyma for a meaningful histological analysis.


Efficacy and safety of rifampicin in patients with persistent hepatocellular secretory failure.
Persistent hepatocellular secretory failure (PHSF) is a rare condition of severe cholestasis caused by drugs, toxins, infection, or temporary biliary obstruction. Real-world data on rifampicin in cholestasis, particularly among patients with deep jaundice, are scarce. We aimed to expand the knowledge on the efficacy and safety of rifampicin treatment in PHSF patients. Sixteen patients with PHSF who had received rifampicin treatment (150-300 mg/d) at our institution from September 2016 to July 2020 were included. Treatment efficacy was assessed by 20% improvement in serum total bilirubin (TBIL) concentration at 4 weeks. Follow-up was continued until the concentration of TBIL returned to normal. Among the 16 enrolled patients, 12 had predisposing factors (drugs, infection, or transient biliary obstruction). ATP8B1 or ABCB11 mutations were detected in the other four patients without trigger events. UGT1A1 mutations were found in 7/10 patients. Before rifampicin treatment, the median TBIL level was 352 μmol/L (range 171-591 μmol/L). TBIL > 20% improvement was observed in 14 patients at 4 weeks. TBIL levels of 14 patients eventually returned to normal after 6-12 weeks of rifampicin treatment. The remaining two patients who did not respond to rifampicin finally recovered after nasobiliary drainage. Except for one patient with transient drug-induced hepatitis, no other serious adverse events were observed. Rifampicin could be a promising option for most PHSF patients. Most PHSF patients have UGT1A1 deficiency, which may be the target of rifampicin.


Binding sites for endotoxins (lipopolysaccharides) on human monocytes.
The nature of the binding sites for LPS on human monocytes was investigated using [3H] labeled intact LPS from Neisseria meningitidis and from Salmonella minnesota R7, and the [3H] labeled purified inner core region (PS-OMe) of S.m. R7 LPS. In the presence of serum, intact LPS from enterobacterial and nonenterobacterial strains bound to monocytes in a dose-dependent, saturable, and displaceable fashion. N.m. LPS and LPS from the enterobacterial strain of Escherichia coli 0111-B4 bound to the same sites on monocytes as assessed in competitive binding experiments. Specific binding of intact LPS to monocytes occurred through the CD14 molecule as shown by the ability of mAb and of F(ab')2 fragments of mAb directed against specific epitopes of CD14 to inhibit the binding of [3H]-LPS to cells and by the lack of binding of intact LPS to CD14-deficient cells from patients with paroxysmal nocturnal hemoglobinuria. Specific binding of LPS to monocytes was not mediated by the CD11/CD18 complex because mAb to the alpha and beta chains of the Leu-CAM molecules did not alter the binding of LPS to cells and because LPS did not inhibit the binding of labeled mAb to monocytes. [3H]-PS-OMe also bound in a dose-dependent and displaceable fashion to monocytes involving an unidentified, non-CD14, binding site on the cells. Binding of LPS to monocytes also involved nonsaturable binding sites for hydrophobic structures of LPS as evidenced in binding experiments performed in the absence of serum. These observations indicate that intact LPS may interact with the monocyte membrane in at least three ways including serum-dependent binding to CD14 and to a lectin-like receptor, and serum-independent hydrophobic interactions.


Joint loads and muscle force distribution during classical and jazz pirouettes.
The objective of this study was to analyze the muscle force distribution and lower limb joint loading during two types of pirouettes and check which muscle in which pirouette generates the highest force and which joint is the most loaded. Skilled dancers (n = 16) performed single-turn pirouettes in jazz and classic styles. Kinematic and kinetic data were collected using the Vicon system and Kistler plates. The joint reaction forces and muscle forces were calculated using a musculoskeletal model in the AnyBody Modeling System. No statistically significant differences were found for the duration of the turn in both pirouettes. The range of motion in all joints of supporting leg in sagittal plane and in hip joint in frontal plane for non-supporting leg were significantly higher in classic pirouette. The ankle joint was the most loaded joint in both pirouettes and its maximal value was significantly higher in classic pirouette. The force generated by ankle plantar flexors muscles was significantly greater in the jazz pirouette in turn phase. For the nonsupport limb, external hip rotators generated significantly greater force when performing the classic pirouette. It seems that early stage dancers may start their lessons with jazz pirouettes, where necessary joint mobility is lesser. They also are supposed to increase muscle strength and body awareness with such proceedings. A better awareness of the mechanical loads on the musculoskeletal system which a dancer performing pirouettes faces should have an impact on the way dance classes are conducted and choreographic elements are sequenced.


Liver Steatosis: Better Predictor of CKD in MAFLD Than Liver Fibrosis as Determined by Transient Elastography With Controlled Attenuation Parameter.
Background: Changing the term/concept of the non-alcoholic fatty liver disease (NAFLD) to metabolic dysfunction associated fatty liver disease (MAFLD) may broaden the pathological definition that can include chronic renal involvement, and, possibly, changes chronic kidney disease's (CKD's) epidemiological association with liver disease, because CKD is associated with metabolic disorders and almost all patients with CKD present some form of an atherogenic dyslipidemia. Our study explores the relationship between MAFLD and CKD using Transient Elastography (TE) with a Controlled Attenuated Parameter (CAP). Methods: We evaluated 335 patients with diabetes with MAFLD and with high CKD risk using TE with CAP (FibroScan®). The CKD was defined according to Kidney Disease Improving Global Outcomes (KDIGO) 2012 guidelines. Logistic regression and stepwise multiple logistic regression were used to evaluate the factors associated with CKD. In addition, a receiver operating characteristic curve (ROC) analysis was used to assess the performance of CAP and TE in predicting CKD and its optimal threshold. Results: The prevalence of CKD in our group was 60.8%. Patients with CKD had higher mean liver stiffness measurements (LSM) and CAP values than those without CKD. We found that hepatic steatosis was a better predictor of CKD than fibrosis. Univariate regression showed that CAP values >353 dB/m were predictive of CKD; while the multivariate regression analysis (after adjustment according to sex, body mass index (BMI), low-density lipoprotein cholesterol (LDLc), and high-density lipoprotein cholesterol (HDLc), and fasting glucose) showed that CAP values >353 dB/m were more strongly associated with the presence of CKD compared to the LSM (fibrosis) values. Conclusion: In patients with MAFLD, CAP-assessed steatosis appears to be a better predictor of CKD compared to LSM-assessed hepatic fibrosis.


In vitro characteristics of pulmonary neuroendocrine cells isolated from rabbit fetal lung. I. Effects of culture media and nerve growth factor.
Pulmonary neuroendocrine (NE) cells, dispersed throughout the airway mucosa as single cells and as innervated clusters (neuroepithelial bodies), were isolated from rabbit fetal lung and studied in short-term culture. The effects of culture media and nerve growth factor (NGF) on in vitro maintenance, differentation, and cell kinetics of isolated NE cells were examined. For demonstration of NE cells in intact lung, during cell separation and after culture, immunostaining for serotonin, formaldehyde-induced fluorescence method, histochemical reaction for acetylcholinesterase, and electron microscopy were used. The isolation procedure consisted of mechanical and enzymatic dissociation of lung tissue followed by separation of isolated cells on a discontinuous gradient of Percoll, resulting in 5- to 10-fold enrichment in NE cells. Cell fractions enriched in NE cells were cultured up to 7 days either in supplemented alpha-minimal essential medium with fetal bovine serum or in defined, hormone-supplemented, serum-free medium. NGF (2.5 S 5 to 50 ng/ml) was added to both serum-supplemented and serum-free media; cultures without NGF served as control. The number of serotonin-immunoreactive NE cells maintained in serum-supplemented medium (0.5% fetal bovine serum) increased significantly (p less than 0.05) on days 4 and 7 compared with cultures grown in serum-free medium. NE cells maintained in serum-supplemented medium incorporated [3H]thymidine and their labeling index was significantly increased (p less than 0.01) on day 7, whereas few or no NE cells were labeled in cultures grown in serum-free medium. NGF had no effect on the maintenance or kinetics of NE cells. Cultured NE cells formed elongated (unipolar or bipolar) neurite-like cytoplasmic processes with a button-like ending, regardless of the presence of NGF. Amine accumulated in perinuclear cytoplasm and in button-like endings. Staining for acetylcholinesterase (strongly positive in intact neuroepithelial bodies) was not detectable after separation into single cells, culture, or exposure to NGF. This study demonstrates that NE cells can be isolated from rabbit fetal lung and maintained in short-term culture. Low concentrations of fetal bovine serum enhanced the in vitro maintenance of NE cells, whereas NGF had no such effect. A feeder layer may be also important, since NE cells were closely associated with lung epithelial or fibroblast-like cells. The formation of neurite-like processes appears to be an expression of paracrine/paraneuron-type cell differentiation not mediated by NGF.(ABSTRACT TRUNCATED AT 400 WORDS)


Complexity of ballooned hepatocyte feature recognition: Defining a training atlas for artificial intelligence-based imaging in NAFLD.
Histologically assessed hepatocyte ballooning is a key feature discriminating non-alcoholic steatohepatitis (NASH) from steatosis (NAFL). Reliable identification underpins patient inclusion in clinical trials and serves as a key regulatory-approved surrogate endpoint for drug efficacy. High inter/intra-observer variation in ballooning measured using the NASH CRN semi-quantitative score has been reported yet no actionable solutions have been proposed. A focused evaluation of hepatocyte ballooning recognition was conducted. Digitized slides were evaluated by 9 internationally recognized expert liver pathologists on 2 separate occasions: each pathologist independently marked every ballooned hepatocyte and later provided an overall non-NASH NAFL/NASH assessment. Interobserver variation was assessed and a 'concordance atlas' of ballooned hepatocytes generated to train second harmonic generation/two-photon excitation fluorescence imaging-based artificial intelligence (AI). The Fleiss kappa statistic for overall interobserver agreement for presence/absence of ballooning was 0.197 (95% CI 0.094-0.300), rising to 0.362 (0.258-0.465) with a ≥5-cell threshold. However, the intraclass correlation coefficient for consistency was higher (0.718 [0.511-0.900]), indicating 'moderate' agreement on ballooning burden. 133 ballooned cells were identified using a ≥5/9 majority to train AI ballooning detection (AI-pathologist pairwise concordance 19-42%, comparable to inter-pathologist pairwise concordance of between 8-75%). AI quantified change in ballooned cell burden in response to therapy in a separate slide set. The substantial divergence in hepatocyte ballooning identified amongst expert hepatopathologists suggests that ballooning is a spectrum, too subjective for its presence or complete absence to be unequivocally determined as a trial endpoint. A concordance atlas may be used to train AI assistive technologies to reproducibly quantify ballooned hepatocytes that standardize assessment of therapeutic efficacy. This atlas serves as a reference standard for ongoing work to refine how ballooning is classified by both pathologists and AI. For the first time, we show that, even amongst expert hepatopathologists, there is poor agreement regarding the number of ballooned hepatocytes seen on the same digitized histology images. This has important implications as the presence of ballooning is needed to establish the diagnosis of non-alcoholic steatohepatitis (NASH), and its unequivocal absence is one of the key requirements to show 'NASH resolution' to support drug efficacy in clinical trials. Artificial intelligence-based approaches may provide a more reliable way to assess the range of injury recorded as "hepatocyte ballooning".


Gliomatosis cerebri diagnostic challenge: two case reports.
Gliomatosis cerebri is a specific entity defined as a diffuse neoplastic glial cell infiltration of the brain, preserving the architecture of the normal surrounding tissues, involving more than 2 cerebral lobes. Clinical symptoms or radiologic features are nonspecific, and patients are often misdiagnosed with other neurologic diseases. Here, we report the diagnostic workup of 2 patients with gliomatosis cerebri, discussing the clinical, radiologic, and pathologic findings. Case 1: a 64-year-old woman who presented with an intracranial hypertension syndrome and had symmetrical white matter T2-weighted and fluid-attenuated inversion recovery hyperintensities pattern on magnetic resonance imaging; and case 2: a 54-year-old man with the diagnosis of multiple sclerosis for 8 years who presented with de novo cognitive impairment and focal deficits. This report highlights the difficulty of this differential diagnosis and the need of considering it also in the presence of a symmetrical pattern of white matter involvement. Cerebral biopsy remains crucial for the correct diagnosis and treatment approach.