Genetic characterization of Toxoplasma gondii from cats in Yunnan Province, Southwestern China.
Cats are the definitive hosts of Toxoplasma gondii. The distribution of genetic diversity of T. gondii in cats is of importance to understand the transmission of this parasite. The objective of this study was to genetically characterize T. gondii isolates from cats in Yunnan province, southwestern China. Genomic DNA was extracted from 5-10 g cat tissue samples (brain, tongue, heart, and liver). Using multilocous polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology, we determined genetic diversity of T. gondii isolates from cats in Yunnan province. In total, 175 stray cats were tested for T. gondii DNA, respectively, 44 (25.14%) of which were found to be positive for the T. gondii B1 gene by PCR amplification. The positive DNA samples were typed at 11 genetic markers, including 10 nuclear markers, namely, SAG1, 5'-3'SAG2, alternative SAG2, SAG3, GRA6, L358, PK1, BTUB, c22-8, c29-2 and an apicoplast locus Apico. Of these, 16 isolates from cats were genotyped with data for more than 9 loci, revealed 5 genotypes in total, of which 11 of 16 samples were identified as ToxoDB#9, two samples may belong to genotye #225, one was Type II, one was ToxoDB#3, and one was ToxoDB#20 (http://toxodb.org/toxo/). The results of the present study indicated a wide distribution of T. gondii infection in cats in Yunnan province, which may pose significant public health concerns. To our knowledge, the present study is the first report of T. gondii prevalence and genotypes in cats in southwestern China, and the first report of Type II T. gondii from cats in China.


IL-15Ralpha chaperones IL-15 to stable dendritic cell membrane complexes that activate NK cells via trans presentation.
Natural killer (NK) cells are innate immune effectors that mediate rapid responses to viral antigens. Interleukin (IL)-15 and its high affinity IL-15 receptor, IL-15Ralpha, support NK cell homeostasis in resting animals via a novel trans presentation mechanism. To better understand how IL-15 and IL-15Ralpha support NK cell activation during immune responses, we have used sensitive assays for detecting native IL-15 and IL-15Ralpha proteins and developed an assay for detecting complexes of these proteins. We find that IL-15 and IL-15Ralpha are preassembled in complexes within the endoplasmic reticulum/Golgi of stimulated dendritic cells (DCs) before being released from cells. IL-15Ralpha is required for IL-15 production by DCs, and IL-15 that emerges onto the cell surface of matured DCs does not bind to neighboring cells expressing IL-15Ralpha. We also find that soluble IL-15-IL-15Ralpha complexes are induced during inflammation, but membrane-bound IL-15-IL-15Ralpha complexes, rather than soluble complexes, support NK cell activation in vitro and in vivo. Finally, we provide in vivo evidence that expression of IL-15Ralpha specifically on DCs is critical for trans presenting IL-15 and activating NK cells. These studies define an unprecedented cytokine-receptor biosynthetic pathway in which IL-15Ralpha serves as a chaperone for IL-15, after which membrane-bound IL-15Ralpha-IL-15 complexes activate NK cells via direct cell-cell contact.


Acute perioperative normovolaemic haemodilution in major maxillofacial surgery.
Extensive blood loss can occur when patients have major operations for head and neck neoplasia. The risks and costs of allogeneic blood transfusion have been well documented. Acute perioperative normovolaemic haemodilution (APNH) is a method of saving blood and autologous transfusion. The purpose of this study was to compare a group of patients having major head and neck operations who had APNH, with a historical group, to assess the need for allogeneic transfusion. Of the 21 patients who had APNH only one patient required allogeneic blood. In the historical group of 24 patients, 15 patients had required allogeneic blood (p=0.0001). APNH is an economic, safe and practical way of saving blood.


Neurocognitive effects of chemotherapy and endocrine therapies in the treatment of breast cancer: recent perspectives.
With an estimated 207,090 patients diagnosed with breast cancer in 2010, the role of chemotherapy-induced cognitive impairment is of growing importance. Studies to determine the impact of chemotherapy-induced cognitive impairment have been hindered by difficulties in study-design, in particular, study methodology. Here, we present a review of existing studies and discuss several mechanisms for chemotherapy-induced neurocognitive impairment in breast cancer patients, such as direct neurotoxic injury, telomere shortening, oxidative stress, cytokine dysregulation, estrogen-mediated effects, and the role of certain genetic polymorphisms. Decreased estrogen levels may serve as a link between multiple mechanisms potentiating the effects of the chemotherapy-induced cognitive impairment.


Association of depressive symptoms with coagulation factors in young healthy individuals.
Depression has been reported to be an independent risk factor for coronary heart disease (CHD). We investigated the association of depressive symptoms with lipids and coagulation factors in young individuals free of CHD. We recruited 1073 young healthy individuals candidates for military academies (mean age=18.4+/-0.8 years, males 762) in whom the presence of depressive symptoms was assessed by using the depression scale of Minnesota Multiphasic Personality Inventory test. Total cholesterol, triglycerides, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, lipoprotein (a), fibrinogen, factors VII, VIII and X were measured. The depression score ranged from 22 to 90. The participants were divided into quartiles according to the depression score. Three hundred twenty-two subjects were classified in the upper quartile (score>48) and 269 in the lower quartile (score<37) of the depression score. Factor VII (102.95+/-24 versus 98.5+/-20%) and X levels (92+/-11 versus 89.7+/-10%) were significantly higher in individuals in the upper quartile compared to the lower quartile of the depression score. In a logistic regression model with factor VII as dependent variable (upper versus lower quartile) and depression score, age, gender, body mass index, exercise and smoking as predictor variables, depression was an independent predictor of factor VII levels with an adjusted odds ratio for high levels of factor VII of 1.05 (95% confidence interval 1.008-1.09, p=0.01). Factor VII levels were associated with triglycerides (r=0.21, p=0.001) while factor X with triglycerides (r=0.22, p<0.001) and cholesterol levels (r=0.12, p<0.001). Depressed mood is associated with a hypercoagulant profile as it is expressed by the higher levels of coagulation factors VII and X. This might partially explain the higher propensity for CHD of people with depressive symptoms.


The efficacy and safety of second-line treatments of refractory and/or high risk pregnant antiphospholipid syndrome patients. A systematic literature review analyzing 313 pregnancies.
The most efficacious strategy to manage pregnant patients with antiphospholipid syndrome (APS) refractory to conventional heparin/low-dose aspirin treatment or at high risk of adverse pregnancy outcomes has not been determined with any degree of certainty. The study set out to evaluate the efficacy and safety of the second-line treatments most frequently used in addition to conventional therapy, and the data were analyzed to identify which is/are associated to the best pregnancy outcomes. A systematic review of the literature on studies concerning second-line treatments for refractory and/or high risk pregnant APS women published between February 2006 and February 2020 was conducted. The records were retrieved by searching Medline via Pubmed, the Web of Science platform, the Cochrane library database and clinicaltrials.gov. Fourteen studies met the eligibility criteria of the review: six retrospective cohort studies, one case-control, one case-series and six case reports. The results of single treatment protocols based upon hydroxychloroquine (HCQ), low-dose steroids (LDS), intravenous immunoglobulins (IVIG), plasma exchange (PE) or pravastatin and of combination protocols based upon HCQ+LDS, IVIG+LDS, PE+LDS and PE+IVIG used during 313 pregnancies in 303 APS women were analyzed and compared. The second-line treatments produced 261/313 (83.4%) live births; severe pregnancy complications were registered in 75/313 (24%) pregnancies. Drug side-effects were observed in 3/313 (0.9%) pregnancies. Statistical analysis identified a significantly higher live birth rate and/or a significantly lower number of severe complications in the pregnancies treated with IVIG, HCQ, pravastatin, PE+IVIG and PE+LDS. Our results suggest using low-dose IVIG (< 2 g/Kg/month) or HCQ 400 mg/day starting before pregnancy in women with APS refractory to conventional therapy, while high-dose IVIG (2 g/Kg/month) associated with PE or alone in those with high risk±refractory APS.


The relationship of amniotic fluid fluorescence polarization to neonatal lung function.
Steady-state fluorescence polarization (FP) of 1,6-diphenyl-1,3,5-hexatriene (DPH) in monkey and human amniotic fluid was studied over a wide range of gestational ages. In both of these systems, the fluorescence polarization decreased with advancing gestational age. In the monkey, these measurements were correlated with both biochemical and physiologic parameters of lung function, including maximal lung volume, alveolar stability, percentage of disaturated phosphatidylcholine in lung homogenate, and lecithin/sphingomyelin ratio of amniotic fluid. Fluorescence polarization values correlated well with the lung disaturated phosphatidylcholine content expressed as a percentage of phosphatidylcholine, thus suggesting that the fluorescent probe interacts with a fraction of the amniotic fluid which is closely related to development of the pulmonary surfactant. system. Comparison of monkey and human amniotic fluid fluorescence polarizations showed a greater anisotropy of DPH in the monkey fluid at all stages of gestation, thereby indicating a greater microviscosity in monkey pulmonary surfactant.


Heterogeneity of human liver, muscle, and adipose tissue insulin receptor.
We have studied the structure and function of the human insulin receptor in liver, skeletal muscle and adipose tissue. The alpha-subunit of the insulin receptor for liver, muscle and adipose tissue migrated on SDS-PAGE with Mrs 137632 +/- 216, 134034 +/- 1080, and 133575 +/- 165, respectively (p less than 0.05). Treatment of these receptors with neuraminidase decreased their molecule sizes and eliminated the relative size differences between the receptors. Three monoclonal antibodies (5A1, 10D9, and 20H3), directed towards different epitopes of the human insulin receptor alpha-subunit were used to probe immunological differences among the receptors. Antibodies 5A1 and 20H3 recognized all the receptors, whereas 10D9 recognized muscle and adipose tissue receptors but not liver receptors. The mobility of insulin receptor beta-subunit in the absence of insulin was the same in all tissues with a similar phosphorylation-induced decrease in mobility in SDS-PAGE in the presence of insulin. However, insulin stimulated autophosphorylation per receptor was different being greatest (p less than 0.05) in muscle (334 +/- 104 32P cpm) and similar in adipose tissue (114 +/- 10) and liver (183 +/- 68). These studies indicate, therefore, that the human insulin receptor is heterogeneous among the major target tissues for insulin, and raise the possibility that this heterogeneity may account for tissues' specific differences in insulin's biological messages.


IGF-I regulates caveolin 1 and IRS1 interaction in caveolae.
Caveolae are hot spots in IGF-I signalling as suggested by the facts that IGF-I receptors localize in caveolae, directly interact with and tyrosine phosphorylate caveolin 1, the major caveolar protein. Also a number of IGF-IR substrates reside in caveolae, supporting a role of these organelles in the regulation of IGF-I action. Recently, we have demonstrated that IGF-I could specifically regulate Shc phosphorylation in caveolae. Here we show that also IRS1 localizes in this region where it is tyrosine phosphorylated in the presence of IGF-I. Moreover, IRS1 co-immunoprecipitates with caveolin 1 and the specific phosphocaveolin 1-IRS1 interaction is increased by IGF-I.


Psychological aspects of traumatic injury in children and adolescents.
Each year millions of children are exposed to some form of extreme traumatic stressor. These traumatic events include natural disasters (e.g., tornadoes, floods, hurricanes), motor vehicle accidents, life-threatening illnesses and associated painful medical procedures (e.g., severe burns, cancer, limb amputations), physical abuse, sexual assault, witnessing domestic or community violence, kidnapping, and sudden death of a parent. During times of war, violent and nonviolent trauma (e.g., lack of fuel and food) may have terrible effects on children's adjustment. The events of September 11, 2001 and the unceasing suicidal attacks in the Middle East underscore the importance of understanding how children and adolescents react to disasters and terrorism. The body of literature related to children and their responses to disasters and trauma is growing. Mental health professionals are increasing their understanding about what factors are associated with increased risk (vulnerability) and affect how children cope with traumatic events. Researchers recognize that children's responses to major stress are similar to adults' (reexperiencing the event, avoidance, and arousal) and that these responses are not transient. A review of the literature indicates that PTSD is the most common psychiatric disorder after traumatic experiences, including physical injuries. There is also evidence for other comorbid conditions, including mood, anxiety, sleep, conduct, learning, and attention problems. In terms of providing treatment, CBT emerges as the best validated therapeutic approach for children and adolescents who experienced trauma-related symptoms, particularly symptoms associated with anxiety or mood disorders. The best approach to the injured child requires injury and pain assessment followed by specific interventions, such as pain management, brief consultation, and crisis intervention immediately after the specific traumatic event. Family support also may be necessary to help the family through this difficult period. The main conclusion that arises from the research on resilience in development is that extraordinary resilience and recovery power of children depend on basic human protective systems operating in their favor. This finding has produced a fundamental change in the framework for understanding and helping children at high risk or already in trouble. This shift is evident in a changing conceptualization of the goals of prevention and intervention that currently address competence and problems. Strategies for fostering resilience described in this article should be tested in future controlled psychotherapy trials to verify their efficacy on children's protective factors.


Estimation and prediction of blood volume in infants and children.
Blood volume was studied in 160 infants and children aged from one hour to 14 years. From these data linear and logarithmic regression equations relating blood volume to weight, height and surface area were calculated. Equations utilizing combined weight and logarithmic weight values were found to be the most accurate for predicting blood volume. Therefore, nomograms were constructed for reading blood volume directly from height and weight.


RNAi-mediated downregulation of oral cancer overexpressed 1 (ORAOV1) inhibits vascular endothelial cell proliferation, migration, invasion, and tube formation.
Oral squamous cell carcinoma (OSCC) is one of the top ten tumors threatening human health. Oral cancer overexpressed 1 (ORAOV1) identified within chromosomal region 11q13, one of the most frequently amplified regions in OSCC, has been suggested as a novel candidate oncogene in OSCC, regulating cell cycle, apoptosis, and angiogenesis. In this study, we investigated the role of ORAOV1 in OSCC-induced angiogenesis in vitro. EA.hy926 human endothelial cells were co-cultured with OSCC cells (HSC-3 and SCC-25) transfected with ORAOV1-specific shRNA to downregulate ORAOV1 expression, and analyzed for proliferation, migration, invasion, and tube formation by specific assays. EA.hy926 endothelial cells co-cultured with ORAOV1-deficient OSCC cells exhibited significantly lower proliferation, migration, and invasion, as well as the activity in tube formation compared to that in the control cells. Our results show, for the first time, that ORAOV1 expressed by OSCC cells promotes tube formation by endothelial cells, indicating its involvement in OSCC angiogenesis. Considering the importance of neovascularization in tumor development and metastasis, these findings suggest that targeting ORAOV1 may be a potential therapeutic strategy against OSCC.


Design of ambulatory center energizes patient-focused care.
Ambulatory care center that delivers comprehensive out-patient medical care is described on a 50 acre site and uses a large, spacious atrium as the focus of patient management, including the movement of patients to different services.


Investigations of host defence in patients with sickle cell disease.
Parameters of host defence were investigated in 30 patients with sickle cell disease (SCD). A newly devised perfusion system was used to study the kinetics in whole blood of leucocyte adherence, phagocytosis, killing and solubilization of a mixture of Staph. aureus and Str. pneumoniae, and secretion of lactoferrin. A skin window technique was used to examine the accumulation of leucocytes at inflammatory foci and their subsequent rate of movement through a filter. Serum concentrations of C3, C4, total haemolytic complement and immunoglobulins were also measured. The rate of neutrophil migration into filters was slightly reduced in patients with SCD. The proportion of monocytes that emigrated from the skin windows and their rate of migration were markedly diminished. The adhesion of neutrophils and their ability to kill staphylococci were also reduced, particularly in patients of the haemoglobin (Hb) SS and Hb S-beta-thalassaemia genotypes. Neutrophil function was mostly impaired in patients with the greatest frequency of bacterial infection. The rate of clearance of pneumococci was related to the concentration of type specific immunoglobulin G but not M. Serum concentrations of immunoglobulins and complement were normal. We were unable to define a defect of host defence of sufficient magnitude to explain the susceptibility of these patients to severe infection.


Drug-induced toxic epidermal necrolysis in children.
Nine pediatric patients with drug-induced toxic epidermal necrolysis were treated with a regimen of basic burn care and without the use of steroids or topical or systemic antimicrobial agents. Although septic complications continue to occur frequently, infections are better tolerated when potential iatrogenic sources of decreased host resistance, such as steroids, are eliminated. Neutropenia, gram-negative sepsis, and mortality were all greatly reduced with this regimen, while healing was neither impaired nor prolonged. Thus, steroids and sulfa-containing topical agents should be avoided in the treatment of this disorder.


Psychopathological description of hyperventilation-induced panic attacks: a comparison with spontaneous panic attacks.
Our aim was to describe the clinical features of hyperventilation-induced panic attacks (HPA) in panic disorder patients - DSM-IV - and to compare them with their spontaneous panic attacks and with spontaneous panic attacks in panic disorder (PD) patients not sensible to the hyperventilation challenge test. We reexamined 88 previously studied PD patients when they were submitted to a hyperventilation challenge test. They were induced to hyperventilate (30 breaths/min) for 4 min and anxiety scales were applied before and after the test. A total of 51.1% (n = 45) PD patients had a panic attack after hyperventilating - HPA (chi(2) = 13.11, d.f. = 1, p = 0.017). The clinical symptoms of the most severe panic attack were recorded by the HPA patient and by the PD patients not sensible to this test (non-HPA; n = 43, 48.9%) in a diary during a 1-week period and then compared. The HPA group had more respiratory symptoms (chi(2) = 15.26, d.f. = 1, p < 0.001), fulfilling the criteria for the respiratory PD subtype (75.6%), the disorder started later (Mann-Whitney, p < 0.001), had a higher familial prevalence of PD (chi(2) = 19.45, d.f. = 1, p = 0.036), and had more previous depressive episodes (chi(2) = 18.74, d.f. = 1, p < 0.001). The HPA group had similar symptomatology in spontaneous attacks and HPA. The HPA group may be regarded as a subgroup of the respiratory panic disorder subtype with diagnostic and therapeutic implications.


Induction of Th1-type immunity and tumor protection with a prostate-specific antigen DNA vaccine.
Prostate specific antigen (PSA) is a serum marker that is widely used in the detection and monitoring of prostate cancer. Though PSA is a self-antigen, T cell responses to PSA epitopes have been detected in healthy men and prostate cancer patients, suggesting it may be used as a target for active immunotherapy of prostate cancer. A PSA DNA vaccine (pPSA) was evaluated in mice and monkeys for its ability to induce antigen-specific immune responses. Mice immunized intradermally with pPSA demonstrated strong PSA-specific humoral and cellular immunity. The anti-PSA immune responses were skewed toward Th1, as shown by high IFNgamma and IL-2 production. The immune response was sufficient to protect mice from challenge with PSA-expressing tumor cells. Tumor protection was durable in the absence of additional vaccination, as demonstrated by protection of vaccinated mice from tumor rechallenge. Furthermore, pPSA vaccination induced PSA-specific antibody titers in male cynomolgus monkeys, which express a closely related PSA gene. These results demonstrate that vaccination with pPSA may be able to break tolerance and can induce an immune response that mediates tumor protection.


A Novel Technique of Endoscopic Papillectomy with Hybrid Endoscopic Submucosal Dissection for Ampullary Tumors: A Proof-of-Concept Study (with Video).
Endoscopic papillectomy (EP) carries a potential risk of procedure-related adverse events and incomplete resection. Since hybrid endoscopic submucosal dissection (ESD) had been established as an alternative option for relatively large and difficult gastrointestinal tumors, we evaluated a novel EP with hybrid ESD (hybrid ESD-EP) for curative safe margin in this proof-of-concept study. A total of eight cases who underwent hybrid ESD-EP between 2018 and 2020 were identified from our prospectively maintained database. Hybrid ESD-EP involved a (sub)circumferential incision with partial submucosal dissection, and subsequent snare resection of ampullary tumors, which was performed by two endoscopists with expertise in ESD or endoscopic retrograde cholangiopancreatography. Demographic data and clinicopathological outcomes were retrospectively evaluated. En bloc resection was achieved by hybrid ESD-EP in all eight cases, with the median procedure time of 112 (range: 65-170) minutes. The median diameters of the resected specimens and tumors were 18 and 12 mm, respectively. All lateral margins were clear, whereas vertical margin was uncertain in three (38%), resulting in the complete resection rate of 63%. Postoperative bleeding and pancreatitis developed in each one (13%). No tumor recurrence was observed even in those cases with uncertain vertical margin, after a median follow-up of 244 (range, 97-678) days. Hybrid ESD-EP seems to be feasible and promising in ensuring the lateral resection margin. However, further investigations, especially to secure the vertical margin and to shorten the procedure time, should be required.


Dermatologic laser surgery.
The laser is becoming an increasingly important surgical and medical instrument in the field of dermatology. Developments in the areas of laser technology and application have significantly broadened its clinical use over the past several years. These rapid changes and advances in the use of the laser for cutaneous processes may make it difficult for the physician to maintain a current base of knowledge in this area. Relevant laser systems and their clinical applications are presented and discussed, as are investigational laser systems, which may be of importance for the future. This will allow the dermatologic surgeon the opportunity to update concepts and evaluate the various laser systems and their cutaneous applications.


Effect of N-acetylcysteine combined with infliximab on toxic epidermal necrolysis. A proof-of-concept study.
The pathophysiology of toxic epidermal necrolysis (TEN) is thought to be related to a drug-induced oxidative stress combined with TNFα overexpression by keratinocytes. None of the current treatments for TEN including systemic corticosteroids, cyclosporine and intravenous administration of immunoglobulins has proven superior over supportive care only. A total of 10 TEN patients were enrolled to be treated at admission in burn units with the antioxidant N-acetylcysteine [NAC, 150mg/kg in a 20-h intravenous (IV) administration], or the combination of the same IV NAC perfusion with the anti-TNFα antibody infliximab (Remicade(®)), administered at a 5mg/kg dosage as a single 2-h IV administration. TEN was confirmed by a skin biopsy taken from a bullous lesion. At entry in the trial and 48h later, the illness auxiliary score (IAS) of clinical severity was determined and the extent in altered skin area (erythema and blisters) was assessed as a relative body area. Skin biopsies of both clinically uninvolved and erythematous areas were collected and immunohistochemistry was performed for assessing the density of inflammatory cells (CD8+ T cells, CD68+ macrophages) and keratinocytes enriched in intracellular calcium (Ca(++)) identified by the Mac387 anti-calprotectin antibody. No unexpected drug-induced adverse event was noticed. After 48h of both treatment modalities, improvements were not observed in the extent of skin involvement and in IAS. Immunohistopathology showed the absence of reduction in the amount of intraepidermal inflammatory cells. An increased intracellular Ca(++) load in clinically uninvolved keratinocytes and in erythematous epidermis was noticed. This latter finding suggested the progression in the way of the apoptotic process. On burn unit discharge, the survival in each modality of treatment was not improved compared to the expected outcomes determined from the IAS at admission. In this proof-to-concept attempt, NAC treatment or its combination with infliximab did not appear to reverse the evolving TEN process.


[Characteristics of bone marrow compensatory erythropoiesis in hereditary spherocytosis].
Objective: To reveal the compensatory features of bone marrow (BM) erythropoiesis in hereditary spherocytosis (HS) and to explore the effect of diferent hemoglobin levels on this compensation. Methods: Clinical and laboratory data of patients with HS were collected, and the peripheral blood absolute reticulocytes counts value was taken as the surrogate parameter to evaluate the ability of erythropoiesis compensation. BM erythropoiesis compensation in HS with diferent degrees of anemia were evaluated. Results: ①Three hundred and two patients were enrolled, including 115 with compensated hemolytic disease, 74 with mild anemia, 90 with moderate anemia, and 23 with severe anemia. ②Hemoglobin (HGB) was negatively correlated with serum erythropoietin in the decompensated hemolytic anemia group (EPO; rs=-0.585, P<0.001) . ③The median absolute reticulocyte count (ARC) of HS patients was 0.34 (0.27, 0.44) ×10(12)/L, up to 4.25 times that of normal people. The maximum ARC was 0.81×10(12)/L, about 10 times that of normal people. The median ARC of patients with compensated hemolytic disease was 0.29 (0.22, 0.38) ×10(12)/L, up to 3.63 times that of normal people. The median ARC of patients with hemolytic anemia was 0.38 (0.30, 0.46) ×10(12)/L, which was significantly higher than the patients with compensated hemolytic disease, up to 4.75 times that of normal people (z=4.999, P=0.003) . ④ ARC was negatively correlated with HGB in the compensated hemolytic disease group (rs=-0.177, P=0.002) and positively correlated with HGB in the decompensated hemolytic anemia group (rs=0.191, P=0.009) . There was no significant difference in the ARC among patients with mild, moderate, and severe anemia (χ(2)=4.588, P=0.101) . ⑤The median immature reticulocyte production index of the mild, moderate, and severe anemia groups was 13.1% (9.1%, 18.4%) , 17.0% (13.4%, 20.8%) , and 17.8% (14.6%, 21.8%) , respectively; the mild anemia group had lower index values than the moderate and severe anemia groups (P(adj) values were both<0.05) , but there was no significant difference between the latter groups (P(adj)=1.000) . The median immature reticulocyte count of patients in the mild, moderate, and severe groups was 5.09 (2.60, 7.74) ×10(10)/L, 6.24 (4.34, 8.83) ×10(10)/L, and 7.00 (3.07, 8.22) ×10(10)/L, respectively; there was no significant difference among the groups (χ(2)=3.081, P=0.214) . Conclusion: HGB can be maintained at a normal level through bone marrow erythropoiesis, while red blood cells are reduced in HS. However, once anemia develops, the bone marrow exerts its maximum erythropoiesis capacity and does not increase, regardless of anemia aggravation or serum EPO increase.


Salvage Surgery for Bronchopleural Fistula Due to Durvalumab and Chemoradiation.
Herein we describe a case of salvage lobectomy for a bronchopleural fistula secondary to durvalumab after chemoradiotherapy in initially unresectable non-small cell lung cancer. A 75-year-old man with adenocarcinoma, c-stage IIIB (T4N2M0) in the upper lobe of the right lung, received durvalumab after chemoradiotherapy. Fourteen months after durvalumab was discontinued, pneumonitis developed followed by peripheral bronchopleural fistula, and he unavoidably underwent salvage right upper lobectomy. The postoperative course was uneventful, and air leakage ceased. After chemoradiotherapy and durvalumab, refractory air leakage may develop that may require salvage surgery.


Separable roles of hippocampal granule cells in forgetting and pyramidal cells in remembering spatial information.
To investigate the roles individual hippocampal cell groups play in processing of spatial information for memory, we administered low-intensity electrical stimulation to the granule cells, CA3 and CA1 pyramidal cells of the dorsal hippocampus at selected times before and after acquisition of the solution to a radial maze win-stay task. Stimulation of any of the 3 cells populations yielded a variable duration anterograde disruption of memory performance, while stimulation of dentate gyrus granule cells alone produced a declarative memory-specific retrograde amnesia. The amnestic effect of granule cell stimulation was not associated with after discharges in the hippocampus and was prevented by systemic administration of the opiate antagonist naloxone. Our results support the view that this electrical stimulus does not disrupt, but rather, activates the normal function of the granule cell system, resulting in erasure of information held in declarative memory. In contrast, similar activation of the pyramidal cell system does not yield retrograde amnesia, suggesting a normal role for these cells in promoting memory for spatial information.


Synthesis of 5-(4-alkylsulfanyl-[1, 2, 5]thiadiazol-3-yl)-3-methyl-1, 2, 3, 4-tetrahydropyrimidine oxalate salts and their evaluation as muscarinic receptor agonists.
The synthesis and biological test of 5-(4-alkylsulfanyl-[1, 2, 5]thiadiazol-3-yl)-3-me-thyl-1, 2, 3, 4-tetrahydropyrimidine oxalate salts 7 as muscarinic receptor agonists are described. The key intermediate 4 was obtained by a modified Strecker reaction and cyclization, and the 3-methyl-1, 2, 3, 4-tetrahydropyrimidines were obtained by subsequent substitution, quarternization, and reduction. The final products 7 were obtained as oxalic acid salts. The prepared compounds were examined in vitro for their binding affinities to the cloned human muscarinic receptor by the [(3)H]-NMS binding assay.


The association of parity/live birth number with incident type 2 diabetes among women: over 15 years of follow-up in The Tehran Lipid and Glucose Study.
Childbearing may increase the future risk of developing type 2 diabetes mellitus (T2DM) in mothers. However, the issue is not clear completely and not investigated in the Middle East, a region with a high burden of T2DM. In the current study, we examined the association of parity/live birth number with incident T2DM among Iranian women. The study population included 2552 women aged 30-65 years recruited in 1999-2001 and were followed for incident T2DM by 3-year intervals. Multivariable Cox proportional hazard models were applied to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) of the parity/live birth number for incident T2DM. Parity number was defined as the number of live childbirth (number of live birth) plus the number of stillbirth (defined as birth of an infant that died after the 20th week of pregnancy in the uterus). During a median follow-up of 15.4 years, 557 incident T2DM cases have occurred. After adjustment for potential T2DM risk factors and reproductive factors, each additional parity caused a 9% higher risk for incident T2DM. Moreover, compared to women with one parity, those with 3 and ≥ 4 parity had HRs of 1.73 [95% CI: 1.06-2.83] and 2.23 [1.36-3.65], respectively. After further adjustment for body mass index (BMI) and waist circumference, although the HRs were attenuated prominently, parity ≥ 4 was associated with significantly higher risk (HR: 1.72 [1.05-2.83]); even after further adjustment for triglycerides (TG)/ high-density lipoprotein cholesterol (HDL-C), the risk remained marginally significant (HR: 1.64 [1.00-2.70; P value: 0.051]). For the number of live birth, the results were also similar. Moreover, in a sensitivity analysis, when we considered BMI change during follow-up as another covariate, generally, the effect sizes did not change; the trend of HRs across categories of parity number remained marginally significant (P value: 0.064). During a long-term follow-up, after adjustment for potential T2DM risk factors, reproductive factors, obesity indices, and TG/HDL-C (insulin resistance surrogate), we demonstrated that higher parity/live birth numbers could be associated with increased risk of T2DM development among Iranian women. Moreover, even after further adjustment for BMI change, the suggestive higher risk was still found.


Association of human microRNAs miR-22 and miR-491 polymorphisms with panic disorder with or without agoraphobia in a Korean population.
The possible involvement of microRNAs (miRNA) in psychiatric disorders has been recently recognized. Several miRNA polymorphisms have been found to be associated with panic disorder (PD) in European populations. However, the association of miRNA polymorphisms on PD has not been reported in Asian populations. We evaluated the effect of miR-22 and miR-491 polymorphisms on susceptibility to PD in a Korean population. Genotyping for four polymorphic variants of the primary miRNA (pri-miRNA) regions of miR-22 (rs8076112 and rs6502892) and miR-491 (rs4977831 and rs2039391) was performed using blood samples of 341 Korean patients with PD and 229 healthy control subjects. To evaluate PD phenotypes, the Panic Disorder Severity Scale (PDSS) and Anxiety Sensitivity Inventory-Revised (ASI-R) were administered. Three single-nucleotide polymorphisms (SNPs) were found to be associated with PD: rs8076112 miR-22 and rs4977831 and miR-491 rs2039391. The rs8076112C/rs6502892C haplotypes of miR-22 and rs4977831G/rs2039391G and rs4977831A/rs2039391A haplotypes of miR-491 were significantly overrepresented in patients with PD than in healthy control subjects. In combination analysis, miR-22 rs8076112AC/rs6502892CC and rs8076112CC/rs6502892CC and miR-491 rs4977831AG/rs2039391AA were more frequent in patients with PD. Among the phenotype assessments, ASI-R scores were significantly associated with miR-22 rs6502892 in the subgroup with the agoraphobic phenotype. The results should be considered preliminary due to the relatively small sample size and the selection of only four SNPs. This is the first report to show possible associations of miR-22 and miR-491 with genetic susceptibility to PD in a Korean population.


Students' perspectives on the outcomes of the joint international programme for nurse practitioners.
Nurse practitioners (NPs) play a vital role in the primary healthcare systems in several countries. However, the NP system has either only recently been established or has yet to be set up in many Asian countries. The Joint International Programme for Nurse Practitioners (JIPNP), developed by a university in South Korea and one in the United States, has been running since 1998. The purpose of this study was to evaluate this programme by assessing student satisfaction through an e-mail-based questionnaire survey. The results of the survey indicated that the students found that this programme helped refine their professional skills. The students were most satisfied with the practicum. Their responses to the open-ended question regarding their individual experiences during the course indicated that they were most satisfied with their improvement in the following areas: understanding of the role of the NP, the responsibility and limitations of NPs with regard to diagnosing and prescribing, familiarity with culture and language, and understanding of healthcare systems. They were also pleased with the teaching methods deployed. It is anticipated that the graduates who completed this programme will help develop the NP system in Korea by assuming leading positions in many NP fields in South Korea.


Postgraduate training in orthodontics in the UK.
This article briefly describes the postgraduate pathways in orthodontics in the United Kingdom. It is hoped that this will assist potential trainees who want to pursue postgraduate training to understand the different career pathways available to them. It may also add to the knowledge of some senior colleagues who can explain these career opportunities to their students, as not all potential trainees will have the opportunity to read this article. In the following pages we will discuss the different pathways leading to specialist qualification in orthodontics, in the United Kingdom.


A review: dietary and endogenously formed N-nitroso compounds and risk of childhood brain tumors.
Maternal dietary exposure to N-nitroso compounds (NOC) or to their precursors during pregnancy has been associated with risk of childhood brain tumors. Cured meat is one source of exposure to dietary NOC and their precursors. Most epidemiological studies that have examined the role of maternal consumption of cured meats during pregnancy have found a significant positive association between maternal intake of cured meat and the risk of childhood brain tumor (CBT). NOC consist of two main groups, N-nitrosamines and N-nitrosamides. The carcinogenicity profiles of NOC suggest that N-nitrosamides rather than N-nitrosamines are the compounds that may be associated with CBT and that they should be investigated more closely in epidemiological studies. We present a review of the chemical and carcinogenic properties of NOC in connection with the findings of case-control studies. This approach may be helpful in determining the essential information that must be collected in future epidemiological studies on CBT.


Distribution modeling of nanoparticles for brachytherapy of human eye tumor.
Due to their unique properties, gold nanoparticles (GNPs) have been proposed to be used for a wide range of applications, especially for photon radiation therapy. In addition to experimental works, there are worthwhile simulation-based studies focused on the investigation of the effect of parameters governing the dose enhancement due to the presence of GNPs in tissue. In a recently published study, we found that the distribution of GNPs in a single cell plays an important role in nucleus dose enhancement. The present work investigates the sensitivity of dose enhancement of a macroscopic phantom to the modeling of GNPs at the cellular level by using the MCNPX Monte Carlo code. A human eye phantom containing the realistic structures and materials was simulated, with a typical tumor located in its corner filled with three different patterns of distribution of GNPs around the nuclei of the cells. The primary photons emit from a COMS eye plaque brachytherapy containing thirteen 131Cs seeds in the vicinity of the tumor. The study was extended to estimate dose enhancement for various concentration, size, and density of the GNPs accumulated around the nuclei of the tumor. Moreover, the dose delivered to the healthy eye structures for different models has been investigated and discussed. The results show obvious differences between the dose enhancements in the tumor depending on the modeling of GNPs. The results emphasized that an appropriate small-scale model for the distribution of GNPs in the cell would be of high importance to estimate the degree of dose enhancement in a macroscopic phantom to provide a trustworthy prediction to move towards clinical application.


Effects of Bay k 8644, a dihydropyridine analog, on [3H]nitrendipine binding to canine cardiac sarcolemma and the relationship to a positive inotropic effect.
Equilibrium dissociation constants of Bay k 8644, a calcium agonist, and nitrendipine, a calcium antagonist, were determined in canine cardiac sarcolemma. The equilibrium dissociation constant for Bay k 8644 was compared to the concentration that produced a fifty percent increase, and the equilibrium dissociation constant for nitrendipine was compared to the concentration that produced a fifty percent decrease, in contractile force in canine heart trabecular muscle. Both saturation and inhibition binding data suggest that Bay k 8644 and nitrendipine bind to and compete for a high affinity dihydropyridine-binding site present in isolated cardiac sarcolemma preparations. The equilibrium dissociation constant (7-10 nM) and concentration that produced a fifty percent increase in contractile force in the canine trabecular muscle (30 +/- 8 nM) of Bay k 8644 were in a similar concentration range, but the equilibrium dissociation constant (0.29 +/- 0.025 nM) of nitrendipine binding was more than a thousand-fold lower than the concentration that produced a fifty percent decrease in contractile force in canine trabecular muscle (613 +/- 109 nM). These data suggest that binding of Bay k 8644 to high affinity binding sites is pharmacologically relevant, and is related to a positive inotropic effect.


Intravital microscopic analysis of vascular perfusion and macromolecule extravasation after photodynamic vascular targeting therapy.
Photodynamic therapy (PDT), involving the combination of a photosensitizer and light, is being evaluated as a vascular disrupting therapy and drug delivery enhancement modality based on its effects on vascular perfusion and barrier function. Since tumor vasculature is the common route for the delivery of both blood and therapeutic agents, it is important to compare the effects of PDT on blood perfusion and substance transport. Tumor blood cell velocity and the extravasation of high molecular weight dextran molecules were continuously monitored by intravital fluorescence microscopy for up to 60 min after PDT using three doses of verteporfin in the MatLyLu prostate tumor model. PDT induced tumor perfusion disruption via thrombus formation. PDT using a higher dose of verteporfin was more effective in inhibiting blood perfusion while a lower dose verteporfin-PDT was more potent in enhancing dextran extravasation. The increase in dextran extravasation induced by PDT was dependent upon dextran molecular weight. A lower molecular weight dextran obtained a higher tumor accumulation after PDT than a higher molecular weight dextran. PDT with verteporfin had different effects on tumor vascular perfusion versus the extravasation of macromolecules. Optimal PDT conditions should be adjusted based on the therapeutic application.


Transport and distribution of residual nitrogen in ion-adsorption rare earth tailings.
A large amount of nitrogen remains in ion-absorption rare earth tailings with in-situ leaching technology, and it continually ends up in groundwater sources. However, the distribution and transport of ammonium nitrogen (NH4+-N) and nitrate nitrogen (NO3--N) across tailings with both depth and hill slopes is still unknown. In this study, the amount of NH4+-N and nitrate nitrogen (NO3--N) was determined in tailings, and a soil column leaching experiment, served to assess the transport and distribution following mine closure. Firstly, a high concentration of NH4+-N in the leachate at the initial leaching stage was detected, up to 2000 mg L-1, and the concentration of NH4+-N clearly diminished as time passed. Meanwhile, the NH4+-N contents remained relatively high in soil. Secondly, both the content of NH4+-N and NO3--N varied greatly according to vertical distribution after leaching lasting several years. The amounts of NH4+-N and NO3--N in surface soil were much smaller than those in deep soil, with 3-4 orders of magnitude variation with depth. Thirdly, when disturbed by NH4+-N, the pH not only diminished but also changed irregularly as depth increased. Fourthly, although the amount of NO3--N was smaller than that of NH4+-N, both their distribution trend was similar with depth. In fact, NH4+-N and NO3--N were significantly correlated but this declined from the knap to the piedmont. Based on these results, it is suggested that mining activity could cause nitrogen to be dominated by NH4+-N and acidification in a tailing even if leaching occurs over several years. NO3--N derived from NH4+-N transports easily and it becomes the main nitrogen pollutant with the potential to be a long-lasting threat to the environment around a mine.


Simultaneous expression and transportation of insulin by supramolecular polysaccharide nanocluster.
Drug/gene transportation systems with stimuli-responsive release behaviors are becoming research hotspots in biochemical and biomedical fields. In this work, a glucose-responsive supramolecular nanocluster was successfully constructed by the intermolecular complexation of phenylboronic acid modified β-cyclodextrin with adamantane modified polyethylenimine, which could be used as a biocompatible carrier for insulin and pCMV3-C-GFPSpark-Ins DNA which could express insulin co-delivery. Benefiting from the response capability of phenylboronic acid moiety toward glucose, the encapsulated insulin could be specifically released and the corresponding targeted DNA could efficiently express insulin in HepG2 cell, accompanied by the high-level insulin release in vitro. Our results demonstrate that the simultaneous insulin drug delivery and insulin gene transfection in a controlled mode may have great potential in the clinical diabetes treatments.


Alterations of choline acetyltransferase, phosphoinositide hydrolysis, and cytoskeletal proteins in rat brain in response to colchicine administration.
Colchicine, a microtubule disrupting toxin, was administered intracerebroventricularly to rats, followed by measurements of (i) the activity of choline acetyltransferase, a biochemical marker of cholinergic neurons, (ii) cytoskeletal protein concentrations, including tau, MAP-2, spectrin, and tubulin, and (iii) the activity of the second messenger-generating system, receptor-coupled phosphoinositide hydrolysis. One week after colchicine treatment there was a 60% decrease in choline acetyltransferase activity in the hippocampus, which was followed by a gradual increase in only a 29% deficit after 12 weeks. In the striatum and cerebral cortex, choline acetyltransferase activity was slightly reduced (by 13% and 19%, respectively) 1 week after colchicine treatment followed by increases to control values. The concentrations of tau and tubulin in the hippocampus were unaltered by colchicine treatment, and MAP-2 and spectrin were only slightly reduced 4 weeks after colchicine. Hippocampal phosphoinositide hydrolysis induced by norepinephrine was elevated approximately 28% 1 and 2 weeks after colchicine treatment and that induced by ibotenate was increased by 53% 2 weeks after colchicine. These results demonstrate that colchicine causes a severe depletion of choline acetyltransferase 1 week after administration. There was not a significant reduction of the concentration of any of the cytoskeletal proteins after 1 week, possibly due to the cell-selectivity of the toxic effect of colchicine, but there was a delayed, and temporary, decline of MAP-2 and spectrin concentrations. Associated with the decreased choline acetyltransferase activity after 1 week was an enhanced phosphoinositide hydrolysis in response to norepinephrine, and after 2 weeks there were enhanced responses to norepinephrine and to ibotenate.(ABSTRACT TRUNCATED AT 250 WORDS)


Long-term results and tolerability of tandem peptide receptor radionuclide therapy with 90Y/177Lu-DOTATATE in neuroendocrine tumors with respect to the primary location: a 10-year study.
The peptide receptor radionuclide therapy (PRRT) with 90Y and 177Lu is a form of molecular targeted therapy for inoperable or disseminated neuroendocrine tumors (NET). The aim of the study was to evaluate clinical results and long-term side effects of tandem 90Y /177Lu-DOTATATE therapy in patients with NET. Additionally, we evaluated clinical results with reference to the primary site. 59 patients with disseminated NET were included in the study prospectively. 3-5 cycles of combined 1:1 90Y/177Lu-DOTATATE (total injected activity 11.1-16.65 GBq) with mixed amino acids for kidney protection were performed. During a median follow-up of 75.8 months, the PFS was 32.2 months, and the OS was 82 months; 25 patients died. Depending on primary tumor's site, the PFS and the OS for pancreatic NET vs. small bowel, NET vs. large bowel, NET were 30.4 vs. 29.5 vs. 40.3 and 78.9 vs. 58.1 vs. 82.5, respectively. The observed 5-year overall survival was 63%, and a 2-year risk of progression was 39.4%. The following imaging response was observed: CR in 2%, PR in 22%, SD in 65%, and PD in 6% patients. The disease control rate was 89%. The objective response rate was 24%. The PRRT was well tolerated by all patients. One patient (2%) revealed MDS, and one patient (2%) grade 3 nephrotoxicity. No other grade 3 and 4 hematological or renal toxicity was observed. These results indicated the tandem 90Y/177Lu-DOTATATE therapy for patients with disseminated/inoperable NET as highly effective and safe, considering long-term side effects. In the majority of patients, clinical improvement was observed.


Autosomal dominant epilepsy with febrile seizures plus with missense mutations of the (Na+)-channel alpha 1 subunit gene, SCN1A.
Evidence that febrile seizures have a strong genetic predisposition has been well documented. In families of probands with multiple febrile convulsions, an autosomal dominant inheritance with reduced penetrance is suspected. Four candidate loci for febrile seizures have been suggested to date; FEB1 on 8q13-q21, FEB2 on 19p, FEB3 on 2q23-q24, and FEB4 on 5q14-15. A missense mutation was identified in the voltage-gated sodium (Na(+))-channel beta 1 subunit gene, SCN1B at chromosome 19p13.1 in generalized epilepsy with the febrile seizures plus type 1 (GEFS+1) family. Several missense mutations of the (Na(+))-channel alpha 1 subunit (Nav1.1) gene, SCN1A were also identified in GEFS+2 families at chromosome 2q23-q24.3. The aim of this report is precisely to describe the phenotypes of Japanese patients with novel SCN1A mutations and to reevaluate the entity of GEFS+. Four family members over three generations and one isolated (phenotypically sporadic) case with SCN1A mutations were clinically investigated. The common seizure type in these patients was febrile and afebrile generalized tonic-clonic seizures (FS+). In addition to FS+, partial epilepsy phenotypes were suspected in all affected family members and electroencephalographically confirmed in three patients of two families. GEFS+ is genetically and clinically heterogeneous, and associated with generalized epilepsy and partial epilepsy as well. The spectrum of GEFS+ should be expanded to include partial epilepsies and better to be termed autosomal dominant epilepsy with febrile seizures plus (ADEFS+).


Cell wall distraction and biofilm inhibition of marine Streptomyces derived angucycline in methicillin resistant Staphylococcus aureus.
The emergence of life threatening antibiotic resistant pathogens and its associated mortality and morbidity necessitates many new antibiotics from diverse ecological habitats. Marine sponge associated microbes are promising to provide such antimicrobial compounds. In the present study, we report antibacterial and anti-biofilm potential of the angucycline antibiotic 8-O-metyltetrangomycin from Streptomyces sp. SBRK2 isolated from a marine sponge of Gulf of Mannar, Rameswaram, India. Our screening program to tackle methicillin-resistant Staphylococcus aureus (MRSA) drug resistance from marine sponge associated actinobacteria yielded the bioactive strain SBRK2. Based on 16S rRNA gene phylogenetic analysis the isolate was found to closely related with Streptomyces longispororuber NBRC 13488T. In vitro production by agar plate fermentation, solvent based extraction, TLC, HPLC purification and LC-MS based de-replication revealed the bioactive compound as 8-O-metyltetrangomycin. The antibacterial minimum inhibitory concentrations against MRSA was identified as 2 μg/mL. Sub-inhibitory concentration of the compound 8-O-metyltetrangomycin reduced the biofilm formation of S. aureus ATCC25923 and increased the cell surface hydrophobicity index. Scanning electron microscopic observation of the sub-inhibitory concentration exposure revealed a wrinkled membrane surface and slight cellular damage shows the cell wall distracting property of the compound. Zebrafish embryo based toxicity assays exhibited 100 μg/mL of compound as maximal non-lethal concentration which had demonstrated the positive relationship in safety index. The angucycline compound 8-O-metyltetrangomycin could be a potential candidate for the development of anti-biofilm agents against drug resistant pathogens.


Effect of phosphate on cadmium immobilized by microbial-induced carbonate precipitation: Mobilization or immobilization?
Microbial-induced carbonate precipitation (MICP) is a promising technology to immobilize/remediate heavy metals (HMs) like cadmium (Cd). However, the long-term stability of MICP-immobilized HMs is unclear, especially in farmland where chemical fertilization is necessary. Therefore, we performed MICP treatment on soils contaminated with various Cd compounds (CdCO3, CdS, and CdCl2) and added diammonium phosphate (DAP) to explore the impact of phosphate on the MICP-immobilized Cd. The results showed that MICP treatment was practical to immobilize the exchangeable Cd but to mobilize the carbonate and Fe/Mn oxide-bound Cd. After applying DAP, soil pH declined due to ammonium nitrification. At high P/Ca molar ratios (1/2 and 1), partial previously immobilized Cd was released due to the carbonate dissolution. Contrarily, exchangeable Cd transformed to less mobilizable Fe/Mn oxide-bound at low P/Ca molar ratios (1/4 and 1/8). Meanwhile, other treatments were also helpful in avoiding the release of immobilized Cd, such as applying non-ammonium phosphate and adding lime material after soil acidification. Our investigation suggested that the long-term stability of HMs in remediated sites should be carefully evaluated, especially in agricultural areas with phosphate and nitrogen fertilizer input.


Cryo-EM structure of the mitochondrial protein-import channel TOM complex at near-atomic resolution.
Nearly all mitochondrial proteins are encoded by the nuclear genome and imported into mitochondria after synthesis on cytosolic ribosomes. These precursor proteins are translocated into mitochondria by the TOM complex, a protein-conducting channel in the mitochondrial outer membrane. We have determined high-resolution cryo-EM structures of the core TOM complex from Saccharomyces cerevisiae in dimeric and tetrameric forms. Dimeric TOM consists of two copies each of five proteins arranged in two-fold symmetry: pore-forming β-barrel protein Tom40 and four auxiliary α-helical transmembrane proteins. The pore of each Tom40 has an overall negatively charged inner surface attributed to multiple functionally important acidic patches. The tetrameric complex is essentially a dimer of dimeric TOM, which may be capable of forming higher-order oligomers. Our study reveals the detailed molecular organization of the TOM complex and provides new insights about the mechanism of protein translocation into mitochondria.


Angle closure secondary to lens remnants in a patient with presumed aphakia: case report.
Eyes with a short axial length or anterior chamber depth often develop narrowed anterior chamber angles in association with an enlarging crystalline lens. We report a case of a patient who presented in angle closure, with a distant history of prior intervention for congenital cataracts and was presumed to be aphakic. A 78-year-old male presented with acute onset unilateral eye pain and blurred vision. He was found to have increased intraocular pressure, anteriorly bowed iris, and angle closure. Despite prior documentation of aphakia after treatment for congenital cataracts, detailed workup revealed residual crystalline lens material pushing the peripheral iris anteriorly. Further history confirmed that the patient underwent a procedure in the 1940's to remove lens material centrally but was not truly aphakic. The patient was treated with anterior chamber paracentesis and intraocular pressure lowering drops. His intraocular pressure remains controlled with medical therapy alone. Patients that appear to be aphakic centrally may still present with angle closure secondary to residual peripheral lens material. This case highlights the importance of keeping this etiology on the differential in a patient with presumed aphakia.


GazeAlyze: a MATLAB toolbox for the analysis of eye movement data.
This article presents GazeAlyze, a software package, written as a MATLAB (MathWorks Inc., Natick, MA) toolbox developed for the analysis of eye movement data. GazeAlyze was developed for the batch processing of multiple data files and was designed as a framework with extendable modules. GazeAlyze encompasses the main functions of the entire processing queue of eye movement data to static visual stimuli. This includes detecting and filtering artifacts, detecting events, generating regions of interest, generating spread sheets for further statistical analysis, and providing methods for the visualization of results, such as path plots and fixation heat maps. All functions can be controlled through graphical user interfaces. GazeAlyze includes functions for correcting eye movement data for the displacement of the head relative to the camera after calibration in fixed head mounts. The preprocessing and event detection methods in GazeAlyze are based on the software ILAB 3.6.8 Gitelman (Behav Res Methods Instrum Comput 34(4), 605-612, 2002). GazeAlyze is distributed free of charge under the terms of the GNU public license and allows code modifications to be made so that the program's performance can be adjusted according to a user's scientific requirements.


Exchange integral for a variety of tetranuclear ferredoxins.
The temperature dependence of EPR spectra of oxidized [4Fe-4S](-1,-2) ferredoxins (previously designated HiPIP) and a reduced [4Fe-4S](-2,-3) ferredoxin have been analyzed so as to determine the energy of a low-lying excited electronic state. The values obtained were: Center S-3 from beef heart, 44 cm-1; Center S-3 from mung bean, 53 cm-1; the [4Fe-4S](-1,-2) ferredoxin from Thermus thermophilus, 78 cm-1; Center N-2 of NADH ubiquinone reductase, 83 cm-1. Increasing axial distortion in the EPR spectra of the [4Fe-4S](-1,-2), ferrodoxins was associated with higher energy differences. Center N-2, a [Fe-4S](-2,-3) iron-sulfur cluster does not fit this relationship.


Influence of Body Mass Index on the Surgical Outcomes of Flexible Ureteroscopy for Pediatric Upper Urinary Tract Aalculi-A Single Surgeon Experience.
To examine the association between body mass index (BMI) and surgical outcomes of flexible ureteroscopy (FURS) for pediatric upper urinary tract calculi and to estimate the influence of BMI percentile on the learning curve of pediatric FURS. We reviewed our prospectively maintained database containing children who had kidney or ureteral stones from June 2014 to April 2019. We calculated BMI and plotted it on the Centers for Disease Control and Prevention growth chart for sex and age to estimate BMI percentile. Patient demographics, intraoperative data, stone characteristics, stone-free rate (SFR), and complication rate (CR) were analyzed. Learning curves stratified by BMI percentile groups were generated. The final analysis included 161 children, of whom 63 (39.1%) had upper body weight percentile (UBW), 64 (39.8%) had normal body weight percentile (NBW), and 34 (21.1%) had lower body weight percentile (LBW). The median stone burden of the 3 groups were 1.14 (IQR 0.50-3.41), 1.13 (IQR 0.70-3.14), and 0.95 (IQR 0.50-1.77), respectively (P = .17). The SFRs were 90.5% (57/63) in the UBW group, 81.2% (52/64) in the NBW group and 70.6% (24/34) in the LBW group (P = .04). The CRs were 15.9% (10/63), 21.9% (14/64), and 29.4% (10/34), respectively (P = .29). The learning curves showed that the SFR of FURS could be improved after about 100 cases. And decreasing BMI could steepen the learning curve of SFR. BMI is associated with the SFR of FURS. LBW children had the lowest SFR compared to UBW and NBW children. Lower BMI percentile makes the success of FURS more challenging.


mRNA vaccination with charge-altering releasable transporters elicits human T cell responses and cures established tumors in mice.
In vivo delivery of antigen-encoding mRNA is a promising approach to personalized cancer treatment. The therapeutic efficacy of mRNA vaccines is contingent on safe and efficient gene delivery, biological stability of the mRNA, and the immunological properties of the vaccine. Here we describe the development and evaluation of a versatile and highly efficient mRNA vaccine-delivery system that employs charge-altering releasable transporters (CARTs) to deliver antigen-coding mRNA to antigen-presenting cells (APCs). We demonstrate in human peripheral blood mononuclear cells that CART vaccines can activate a robust antigen-specific immune response against mRNA-encoded viral epitopes. In an established mouse model, we demonstrate that CARTs preferentially target professional APCs in secondary lymphoid organs upon i.v. injections and target local APCs upon s.c. injection. Finally, we show that CARTs coformulated with mRNA and a Toll-like receptor ligand simultaneously transfect and activate target cells to generate an immune response that can treat and cure mice with large, established tumors.


Effect of chloroquine on membrane permeability in yeast--release of cellular coproporphyrin.
1. The influx and efflux of labelled substances with and without chloroquine was studied in yeast cells. 2. The uptake of delta-aminolevulinic acid by Saccharomyces cerevisiae is characterized by a KT of 3-4 mM and Jmax of 1.0-1.2 mumol min-1 g dry weight-1. 3. A method for loading yeast with labelled coproporphyrin is suggested. 4. The uptake of sorbitol and coproporphyrin was slightly stimulated, while the uptake of 6-deoxyglucose was slightly, that of 2-aminoisobutyric acid and leucine strongly inhibited by chloroquine. 5. The efflux of coproporphyrin, 2-aminoisobutyric acid and sorbitol was stimulated while that of leucine was not influenced by chloroquine. 6. The result showed that chloroquine influenced directly but nonspecifically the membrane permeability, apparently mainly that of the vacuolar membrane.


Primary prevention in patients with hypertension: comments on the clinical implications of the MAPHY Study. Metoprolol Atherosclerosis Prevention in Hypertensives Study.
The MAPHY Study was conducted to investigate whether metoprolol, a relatively beta 1-selective beta-blocker, given as initial antihypertensive treatment, would lower cardiovascular complications of high blood pressure to a greater extent than do thiazide diuretics as initial treatment at similar blood pressure control. In a controlled, randomized, stratified primary prevention study (n = 3234; 16,180 patient-years) in white men (40 to 64 years old) with mild to moderate uncomplicated hypertension, a treatment schedule starting antihypertensive treatment with metoprolol was compared with treatment starting with a thiazide diuretic. The two treatment groups were well matched in blood pressure and other major clinical characteristics at randomization, as well as at the last follow-up during treatment. Total mortality was significantly lower in patients randomized to metoprolol. At the median follow-up time (4.2 years) 28 deaths occurred in the metoprolol group and 54 in the diuretic group--a difference of 48% in total mortality (95% confidence limit, ranging from 68% to 17%), mainly because of a highly significant reduction in coronary heart disease at this point in time. The benefit demonstrated with metoprolol seems to have important implications for clinical practice and public health policy, since hypertension and coronary heart disease are major health problems worldwide.


Oxidation reactivity of zinc-cysteine clusters in metallothionein.
Evaluating the reactivity of the metal-thiolate clusters in metallothionein (MT) is a key step in understanding the biological functions of this protein. The effects of the metal clustering and protein environment on the thiolate reactivity with hydrogen peroxide (H(2)O(2)) were investigated by performing quantum theory calculations with chemical accuracy at two levels of complexity. At the first level, the reactivity with H(2)O(2) of a model system ([(Zn)(3)(MeS)(9)](3-), MeS is methanethiolate) of the β domain cluster of MT was evaluated using density functional theory (DFT) with the mPW1PW91 functional. At the second level of complexity, the protein environment was included in the reactant system and the calculations were performed with the hybrid ONIOM method combining the DFT-mPW1PW91 and the semiempirical PM6 levels of theory. In these conditions, the energy barrier for the oxidation of the most reactive terminal thiolate was 21.5 kcal mol(-1). This is 3 kcal mol(-1) higher than that calculated for the terminal thiolate in the model system [(Zn)(3)(MeS)(9)](3-) and about 7 kcal mol(-1) higher than that obtained for the free thiolate. In spite of this rise of the energy barrier induced by the protein environment, the thiolate oxidation by H(2)O(2) is confirmed as a possible way for metal release from MT. On the other hand, the results suggest that the antioxidant role of MT in the living cell cannot be as important as that of glutathione (which bears a free thiol).


Repeated stress increases the density of angiotensin II binding sites in rat paraventricular nucleus and subfornical organ.
We have studied the properties of angiotensin II binding sites in the paraventricular nucleus, subfornical organ and anterior pituitary lobe of rats subjected to repeated immobilization stress. This treatment produced significant increase in the density of angiotensin II binding sites in these two nuclei without any significant alteration in binding affinity. Repeated stress did not alter angiotensin II binding properties in the anterior pituitary lobe. Our results suggest that brain angiotensin binding sites may have a role in regulation of the stress response.


Cognitive and behavior deficits in sickle cell mice are associated with profound neuropathologic changes in hippocampus and cerebellum.
Strokes are perhaps the most serious complications of sickle cell disease (SCD) and by the fifth decade occur in approximately 25% of patients. While most patients do not develop strokes, mounting evidence indicates that even without brain abnormalities on imaging studies, SCD patients can present profound neurocognitive dysfunction. We sought to evaluate the neurocognitive behavior profile of humanized SCD mice (Townes, BERK) and to identify hematologic and neuropathologic abnormalities associated with the behavioral alterations observed in these mice. Heterozygous and homozygous Townes mice displayed severe cognitive deficits shown by significant delays in spatial learning compared to controls. Homozygous Townes also had increased depression- and anxiety-like behaviors as well as reduced performance on voluntary wheel running compared to controls. Behavior deficits observed in Townes were also seen in BERKs. Interestingly, most deficits in homozygotes were observed in older mice and were associated with worsening anemia. Further, neuropathologic abnormalities including the presence of large bands of dark/pyknotic (shrunken) neurons in CA1 and CA3 fields of hippocampus and evidence of neuronal dropout in cerebellum were present in homozygotes but not control Townes. These observations suggest that cognitive and behavioral deficits in SCD mice mirror those described in SCD patients and that aging, anemia, and profound neuropathologic changes in hippocampus and cerebellum are possible biologic correlates of those deficits. These findings support using SCD mice for studies of cognitive deficits in SCD and point to vulnerable brain areas with susceptibility to neuronal injury in SCD and to mechanisms that potentially underlie those deficits.


Non-clinical pharmacology and toxicology studies of bevacizumab biosimilar LY01008.
LY01008 was a biosimilar of Avastin® developed by Shandong Boan Biotechnology. To support the clinical trial and marketing application of LY01008 as a biosimilar, a series of non-clinical pharmacodynamics (PD), pharmacokinetics (PK), and toxicological studies have been conducted. The PD study results showed that LY01008 had similar pharmacodynamic effects with Avastin in VEGF (vascular endothelial growth factor) binding activity, inhibitory effect on angiogenesis and vascular permeability, and anti-tumor activities in nude mouse models alone or combined with chemotherapeutic agents. PK study showed that LY01008 had similar PK parameters with Avastin at the same doses, and the relative bioavailability of LY01008 was 111.4%. The maximum tolerated dose of LY01008 in the single-dose toxicity study of cynomolgus monkeys was greater than 258 mg/kg. LY01008 had no effects on central nervous system, cardiovascular system and respiratory system in cynomolgus monkeys. LY01008 had no hemolytic effect in vitro and no local irritation in cynomolgus monkeys. The immunogenicity of LY01008 was no higher than that of Avastin in cynomolgus monkeys. In the one-month multiple-dose toxicity study in cynomolgus monkeys, the toxicokinetics profiles of LY01008 was similar with Avastin, the characteristics of the toxic reactions were the same and the extent was similar between LY01008 and Avastin, and no new toxic reactions were observed on LY01008. In conclusion, LY01008 had a good safety profile, and was biosimilar with Avastin in the comparative studies of pharmacodynamics, pharmacokinetics, toxicokinetics and toxicology, which supported the clinical trial and marketing application of LY01008 as a biosimilar of Avastin.


Signet ring cell carcinoma of bladder.
The ninth reported case of primary signet ring cell carcinoma of the bladder is described. This particular tumor, whose histology was examined repeatedly during its evolution, showed an interesting change in microscopic appearance after irradiation. The emerging features of signet ring cell carcinoma of the bladder are discussed.


Highly Stretchable and Conductive Self-Healing Hydrogels for Temperature and Strain Sensing and Chronic Wound Treatment.
Flexible bioelectronics for biomedical applications requires a stretchable, conductive, self-healable, and biocompatible material that can be obtained by cost-effective chemicals and strategies. Herein, we synthesized polypyrrole or Zn-functionalized chitosan molecules, which are cross-linked with poly(vinyl alcohol) to form a hydrogel through dynamic di-diol complexations, hydrogen bonding, and zinc-based coordination bonds. These multiple dynamic interactions endow the material with excellent stretchability and autonomous self-healing ability. The choice of Food and Drug Administration (FDA)-approved materials (poly(vinyl alcohol) and chitosan) as the matrix materials ensures the good biocompatibility of the hydrogel. The conductivity contributed by the polypyrrole allowed the hydrogel to sense strain and temperature, and the coordinated Zn significantly enhanced the antibacterial activity of the hydrogel. Moreover, using a diabetic rat model, we have proved that this hydrogel is capable of promoting the healing of the infected chronic wounds with electrical stimulation.


Structural aberrations of the X chromosome in man.
Among 209 patients with Shereshevsky-Turner syndrome, 69 women with structural aberrations of X chromosome were detected: 46,X,i(Xq) - 11; 45,X/46,X,i(Xq) - 24; 45,X/46,X,r(X) - 14; 45,X/46,X,f(X or Y) - 10; 45,X/46,X,del(Xq) - 4; 45,X/46,X,del(Xp) - 2; 45,X/46,X,idic(X) - 2; 46,X,idic(X) - 1; and 46,X,t(X,2) - 1. All the patients with structural abnormalities of X chromosome were short in stature, but in no group was it as low on the average as in 45,X cases. Somatic signs were noticed in all structural changes of X, but they were less frequent and less pronounced. In some patients with r(X) and i(Xq), spontaneous menstrual bleeding and breast development was found. The structurally abnormal X chromosome appears to be functionally inactive, the phenotype of patients with structural rearrangements being close to the phenotype of patients with X monosomy. At the same time, the abnormal X might have certain effects in early embryogenesis which mitigated the further development of the Shereshevsky-Turner syndrome.


[Genetic counseling in Germany: development of demand].
With the Act on Genetic Testing (GenDG), the German legislator has issued far-reaching regulations for human genetic services, including genetic counseling. This paper presents data on the use of human genetic counseling in the years before and after the entry into force of GenDG in order to provide an informed assessment of the possible effects of the law. Over a period of 13 years (2005 to 2017), the human genetic counseling services provided within the framework of the statutory health insurance and billable by EBM via the Kassenärztliche associations were recorded via a database query at the Central Institute of the National Association of Statutory Health Insurance Physicians (ZI-KBV) and via individual Kassenärztliche Vereinigungen Deutschlands. For the discussion of the observable development of using genetic counseling and possible future development, additional data on the referral behavior, the waiting times, processing time, and reasons for consultations were extracted from the GenBIn database. Demand for genetic counseling has steadily increased at an average rate of approximately 6% per year since 2009. This increase started well before the enactment of the GenDG and may be attributed to a multiplicity of factors. Change in demand for genetic counseling is characterized by increasing self-referrals and by increasing referrals by specialists other than obstetricians/gynecologists. Waiting times between 2011 and 2016/2017 have increased. While demand has been growing, the number of key service providers, the contracted medical specialists in human genetics, has remained almost constant. It is foreseeable that capacity limits will be reached if both trends continue.


Human milk feeding and direct breastfeeding improve outcomes for infants with single ventricle congenital heart disease: Propensity score matched analysis of the NPC-QIC registry.
Infants with single ventricle (SV) congenital heart disease (CHD) undergo three staged surgeries/interventions, with risk for morbidity and mortality. We estimated the effect of human milk (HM) and direct breastfeeding (BF) on outcomes including necrotizing enterocolitis (NEC), infection-related complications, length of stay (LOS), and mortality. We analyzed the National Pediatric Cardiology Quality Improvement Collaborative registry (2016-2021), examining HM/BF groups during stage 1 (S1P) and stage 2 (S2P) palliations. We calculated propensity scores for feeding exposures, then fitted Poisson and logistic regression models to compare outcomes between propensity-matched cohorts. Participants included 2491 infants (68 sites). Estimates for all outcomes were better in HM/BF groups. Infants fed exclusive HM before S1P had lower odds of preoperative NEC (OR=0.37, 95% CI=0.17-0.84, p=0.017) and shorter S1P LOS (RR=0.87, 0.78-0.98, p=0.027). During the S1P hospitalization, infants with high HM had lower odds of postoperative NEC (OR=0.28, 0.15-0.50, p<0.001) and sepsis (0.29, 0.13-0.65, p=0.003), and shorter S1P LOS (RR=0.75, 0.66-0.86, p<0.001). At S2P, infants with any HM (0.82, 0.69-0.97, p=0.018) and any BF (0.71, 0.57-0.89, p=0.003) experienced shorter LOS. Infants with SV CHD in high HM and BF groups experienced multiple significantly better outcomes. Given our findings of improved health, strategies to increase the rates of HM/BF in these patients should be implemented. Future research should replicate these findings with granular feeding data and in broader CHD populations, and should examine mechanisms (eg, HM components; microbiome) by which HM/BF benefits these infants. What is new?: This is the first large, multisite study examining the impact of human milk and breastfeeding on outcomes for infants with single ventricle congenital heart disease.All outcome estimates were better in high human milk and breastfeeding groups, with significantly lower odds of necrotizing enterocolitis, sepsis, and infection-related complications; and significantly shorter length of stay at both the neonatal stage 1 palliation and the subsequent stage 2 palliation.All estimates of all-cause mortality were substantially lower in human milk and breastfeeding groups, with clinically important estimates of 75%-100% lower odds of mortality in direct breastfeeding groups.What are the clinical implications?: There is a critical need for improved, condition-specific lactation support to address the low prevalence of human milk and breastfeeding for infants with single ventricle congenital heart disease.Increasing the dose and duration of human milk and direct breastfeeding has strong potential to substantially improve the health outcomes of these vulnerable infants.


Postinspiratory-ramp activity of diaphragm under inspiratory resistive load.
Inspiratory ramp activity, postramp activity (PRA) of diaphragm and respiratory flow were studied in anesthetized rabbits and conscious humans under inspiratory resistive load. Under load with chloralose-urethane anesthesia neural inspiration lasted 124 +/- 12 msec more than under control conditions and end of mechanical inspiration lagged behind that of neural inspiration by 114 +/- 4 msec. With pentobarbital-urethane anesthesia corresponding changes under load were 67 +/- 18 msec and 44 +/- 6 msec; after SO2 block of slowly adapting stretch receptors in bronchi corresponding changes under load were nil and 65 +/- 7 msec. In humans corresponding changes under load were about 192 and 127 msec. Both in rabbits and humans time course of PRA under load was essentially unchanged. Hence, under load a considerable part of PRA (or all of it under pentobarbital-urethane anesthesia without SO2 block) occurred during inspiration and that occurring during expiration was smaller than under control conditions. Consequently, under load braking of expiratory flow was smaller and peak expiratory flow occurred earlier than under control conditions.


[Laparoscopic appendectomy. Surgical technique description and literature review].
The laparoscopic appendectomy (LA) is considered a surgical process of controversy, although it offers better recovery and better aesthetic results, and allows visualization of the abdominal cavity. In this work, we presented a revision of the topic and the experience of our group in the Endoscopic Surgery Unit at the Dr. Manuel Gea González General Hospital and at the Hospital Médica Sur, both in Mexico City. We conducted a retrospective study, randomized, which included patients with LA from January 1, 1993 to December 31, 2002. All were operated on by the same group of surgeons. No exclusion criterion existed for the procedure. The surgical technique used in studied patients is described. We operated on 218 patients; 69% were women of 10 to 78 years of age with a 27 years-of-age average. Surgical time varied from 20 to 90 min with an average of 35 min. Hospitalization lasted 1 to 5 days, with an average of 2 days. We found peritonitis in 23%. In 202 patients, diagnosis of appendicitis was confirmed; only in 16 patients was diagnosis different. Surgical technique was the same during the entire time. We used the Gea knot with polypropylene to tie off the appendix and appendicular artery. In cases of peritonitis, irrigation was not used, but only aspiration, cleaning with gauzes, and placement of drainages. Mortality was not reported. LA offers advantages over AA whenever it is carried out by an experienced laparoscopic surgeon. Peritonitis is not a contraindication for LA, and it is not necessary to irrigate with solution for cleaning the peritoneal cavity.


A self-administered Timeline Followback to measure variations in underage drinkers' alcohol intake and binge drinking.
Underage drinkers typically have not developed regular patterns of drinking and so are likely to exhibit situational variation in alcohol intake, including binge drinking. Information about such variation is not well captured by quantity/frequency (QF) measures, which require that drinkers blend information over time to derive a representative estimate of "typical" drinking. The Timeline Followback (TLFB) method is designed to retrospectively capture situational variations in drinking during a specific period of time. We compared our newly-developed Self-administered TLFB (STLFB) measure to a QF measure for reporting alcohol intake. Our sample of 429 (men=204; women=225) underage (i.e., age 18-20 years) drinkers completed the two drinking measures and reported on alcohol problems. The STLFB and QF measures converged in assessing typical daily intake, but the STLFB provided more information about situational variations in alcohol use and better identification of regular versus intermittent binge drinkers. Regular binge drinkers reported more alcohol problems. The STLFB is an easy-to-administer measure of variations in alcohol intake, which can be useful for understanding drinking behavior.


Fabrication of Co(Ni)-P surface bonding states on core-shell Co(OH)2@P-NiCo-LDH towards electrocatalytic hydrogen evolution reaction.
The application of NiCo-LDHs in the hydrogen evolution reaction (HER) is rarely reported because it is usually served as an effective electrocatalyst for oxygen evolution reaction (OER) owing to the essential chemical and electric structure features. It still is a challenge to realize the effective HER over NiCo-LDHs unless modified with noble metals. In this work, the noble-metal-free core-shell Co(OH)2@P-NiCo-LDH hybrid with dodecahedral hierarchical structure is prepared by the sacrificial template method along with subsequent phosphating process, which is available for the electrocatalytic HER. The fabricated Co(Ni)δ+-Pδ- bonding states on the surface of outer shell NiCo-LDH not only provide the abundant active centers but also reduce interfacial charge transfer resistance to enhance the HER activity. Meanwhile, the unique core-shell dodecahedral structure effectively avoids the agglomeration and restacking of NiCo-LDH nanosheets to improve the electrochemical stability. This work exploits a novel noble-metal-free modification strategy to expand the application of NiCo-LDH in HER.


Brain region-specific regulation of urocortin 1 innervation and corticotropin-releasing factor receptor type 2 binding by ethanol exposure.
Ethanol administration and consumption selectively activates the urocortin 1 (Ucn1)-expressing neurons of the Edinger-Westphal nucleus. We investigated whether repeated ethanol exposure affects Ucn1 and Ucn1-responsive corticotropin-releasing factor type-2 receptors (CRF2). Male C57BL/6J and DBA/2J mice were exposed to 2 g/kg ethanol via intraperitoneal injection once per day for 14, seven, or zero days. Ucn1 immunoreactivity was measured in the lateral septum, dorsal raphe, and Edinger-Westphal nucleus. In a separate experiment, C57BL/6J mice were exposed to ethanol for seven, one, or zero days, and CRF2 receptor binding was measured in the lateral septum and dorsal raphe by receptor autoradiography. Ethanol exposure induced parallel changes in Ucn1 immunoreactive terminal fibers in the lateral septum and dorsal raphe of both strains. Seven ethanol exposures but not one ethanol exposure significantly increased CRF2 receptor binding in the dorsal raphe and slightly increased CRF2 receptor binding in the lateral septum. These results provide evidence that the Ucn1/CRF2 receptor system can be modified by ethanol exposure. They additionally suggest that this system may be involved in behavioral changes during alcoholism.


Effect of berberine on the antioxidant status, ultrastructural modifications and protein bound carbohydrates in azoxymethane-induced colon cancer in rats.
Chemically induced carcinogenesis models in the rat are widely used for studying the biology of cancer and for developing and evaluating cancer prevention strategies. The azoxymethane(AOM)-induced rat colon tumor is a valuable tool for studying the interaction between tumor development and exogenous factors. Malignant conditions are characterized by enhanced levels of lipid peroxidative products, protein bound carbohydrates indicative of membrane damage and an ineffective antioxidant scavenging system. In the present study, AOM-induced rat showed lipid peroxidative damage, alterations in the membrane glycoprotein component and antioxidant defense system, along with ultrastructural changes like disruption in goblet cell and its membrane, swollen mitochondria with cristae dispersed, elongated endoplasmic reticulum and other features of neoplastic invasion. Berberine significantly attenuated the increases in lipid peroxidation, protein bound carbohydrates and enhanced the antioxidative status. The present study suggests that berberine prevents the appearance of malignant morphology and ultrastructural changes of AOM induced cancer by producing apoptosis like changes. Thus berberine inhibits neoplastic transformation by the induction of antioxidant defence system and ability to induce apoptotic like changes--thus elucidating its anti cancer role.


Molecular interactions of plant viral satellites.
Plant viral satellites fall under the category of subviral agents. Their genomes are composed of small RNA or DNA molecules a few hundred nucleotides in length and contain an assortment of highly complex and overlapping functions. Each lacks the ability to either replicate or undergo encapsidation or both in the absence of a helper virus (HV). As the number of known satellites increases steadily, our knowledge regarding their sequence conservation strategies, means of replication and specific interactions with host and helper viruses is improving. This review demonstrates that the molecular interactions of these satellites are unique and highly complex, largely influenced by the highly specific host plants and helper viruses that they associate with. Circularized forms of single-stranded RNA are of particular interest, as they have recently been found to play a variety of novel cellular functions. Linear forms of satRNA are also of great significance as they may complement the helper virus genome in exacerbating symptoms, or in certain instances, actively compete against it, thus reducing symptom severity. This review serves to describe the current literature with respect to these molecular mechanisms in detail as well as to discuss recent insights into this emerging field in terms of evolution, classification and symptom development. The review concludes with a discussion of future steps in plant viral satellite research and development.


Adenovirus 2 Ip+ locus codes for a 19 kd tumor antigen that plays an essential role in cell transformation.
Adenovirus 2 large plaque (Ip) mutants produce large clear plaques on human KB cells. These mutants are shown to be defective in inducing transformation of the established rat embryo cell line 3Y1. The Ip mutation was localized within one of the two transforming early gene blocks, E1b (map position 4.5 to 11.2) which codes for two major T antigens of 53 kd and 19 kd by marker transfer. The mutational defects in mutants Ip3 and Ip5 were analyzed by DNA sequence analysis and by analysis of viral E1 proteins. These results reveal that Ip3 and Ip5 mutations map within the 19 kd tumor antigen coding region. Mutant Ip3 has a single base pair change at the N terminus of 19 kd polypeptide, resulting in the substitution of valine for alanine. Mutant Ip5 has two mutational changes, one of which results in the substitution of tyrosine for aspartic acid near the N-terminal region. The second mutation changes the termination codon into a leucine codon, increasing the size of the 19 kd tumor antigen. These results provide direct genetic evidence for an essential role of the 19 kd tumor antigen in cell transformation and indicate that the N-terminal region of the 19 kd tumor antigen is an essential function domain for the induction of cell transformation.


Surgical excision of type 2 choroidal neovascularization in age-related macular degeneration.
To evaluate the visual acuity outcome and the influence of various factors on visual outcome in patients undergoing surgical removal of type 2 choroidal neovascular neovascularization (CNV) caused by age-related macular degeneration (AMD). We studied the records of 92 patients (92 eyes) who were followed for at least 1 year after surgical excision of CNV associated with AMD. The final visual acuity was 0.4 or better in 21%, 0.1 to 0.3 in 66%, and worse than 0.1 in 13% of the patients. Final visual acuity was improved in 62%, stable in 29%, and worse in 9%. Stepwise regression identified CNV size as a significant factor influencing final visual acuity (R = 0.287, P = 0.0045). Surgical excision of CNV for AMD is indicated for patients with subfoveal active type 2 CNV with a visual acuity of 0.3 or worse. To achieve better postoperative visual acuity it is important to operate on AMD patients in the early stage of CNV.


The impact of proton pump inhibitors on the pharmacokinetics of voriconazole in vitro and in vivo.
Voriconazole (VRC) and proton pump inhibitors (PPIs) have similar metabolic pathways. The objectives of the study are to evaluate the impact of PPIs on the pharmacokinetics of VRC. Human liver microsomes model was applied to assess the inhibitory effects of PPIs on the metabolism of VRC in vitro. A retrospective study was also carried out to explore the relationship between the plasma VRC trough concentrations and PPIs uses. Patients were divided into six groups: control (n = 166), lansoprazole (LAN, n = 38), esomeprazole (ESO, n = 19), omeprazole (OME, n = 45), pantoprazole (PAN, n = 43), and ilaprazole (ILA, n = 38) groups. All five PPIs showed concentration-dependent inhibitory effects on the VRC metabolism in human liver microsomes, among which LAN, OME and ESO were three of the most potent inhibitors. Consistently, co-administered with LAN, OME and ESO significantly increased the plasma VRC trough levels (p < 0.05), whereas there was no significant association between VRC concentrations and PAN or ILA use. Interestingly, patients in the PPIs groups were more likely to reach the therapeutic VRC range of 1-5.5 μg/mL in steady state when compared with control patients (75-81% VS 69%). In conclusion, although all PPIs showed inhibitory effects on the VRC metabolism in vitro, only LAN, OME and ESO significantly increased VRC plasma concentrations. This study should be helpful for choice of the type of PPIs for patients administered with VRC.


Optimization of mRNA extraction from human nasal mucosa biopsies for gene expression profile analysis by qRT-PCR.
Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) is the gold-standard method for analyzing modifications in gene expression in cells and tissues. However, large quantities of high-quality RNA samples are needed for analyzing the expression of multiple genes from one human tissue sample. Here, we provide an optimized protocol for extracting large amounts of RNA from human nasal mucosal biopsies. The quality and quantity of samples were sufficient for qRT-PCR analyses of the expressions of various genes, in duplicate. In contrast to other protocols, we optimized RNA isolation to increase the amount from nasal biopsy samples for analyses of multiple genes. In most previous publications, expressions of only one or a few genes, including housekeeping genes, were analyzed because the amount of biological material was small. We were able to improve our protocol with respect to the yield and quality of RNA. This is likely to produce better results from molecular analyses of very small biopsy samples of human nasal mucosa.


Migrasomes, a new mode of intercellular communication.
Migrasomes are newly discovered extracellular vesicles (EVs) that are formed in migrating cells and mediate intercellular communication. However, their size, biological generation, cargo packaging, transport, and effects on recipient cells by migrasomes are different from those of other EVs. In addition to mediating organ morphogenesis during zebrafish gastrulation, discarding damaged mitochondria, and lateral transport of mRNA and proteins, growing evidence has demonstrated that migrasomes mediate a variety of pathological processes. In this review, we summarize the discovery, mechanisms of formation, isolation, identification, and mediation of cellular communication in migrasomes. We discuss migrasome-mediated disease processes, such as osteoclast differentiation, proliferative vitreoretinopathy, tumor cell metastasis by PD-L1 transport, immune cell chemotaxis to the site of infection by chemokines, angiogenesis promotion via angiogenic factors by immune cells, and leukemic cells chemotaxis to the site of mesenchymal stromal cells. Moreover, as new EVs, we propose the potential of migrasomes for disease diagnosis and treatment. Video Abstract.


Role of interleukin-6 in the anemia of chronic disease.
To review evidence supporting the involvement of interleukin-6 in the pathophysiology of anemia of chronic disease, to discuss the possible molecular mechanisms driving this condition in patients with end-stage renal disease and rheumatoid arthritis, and to review clinical manifestations in these patients. A literature search was performed using MEDLINE and the reference lists of relevant review articles. The following key words were used in the MEDLINE search: interleukin-6, "anemia of chronic disease" OR "anemia of inflammation," "pathophysiology," "end-stage renal disease," and "rheumatoid arthritis." The search was limited to English-language articles. Interleukin-6 is a multifunctional cytokine that regulates the hepatic acute-phase response, the immune response, inflammation, and hematopoiesis. Interleukin-6 appears to be the central mediator of anemia of chronic disease in a range of inflammatory diseases, including end-stage renal disease and rheumatoid arthritis, through increased generation of hepcidin and the resultant alterations in iron metabolism. Clinically, patients with anemia of chronic disease are more likely to experience increased disease severity and duration than patients who have chronic disease without anemia. The integral role of interleukin-6 in the pathogenesis of anemia of chronic disease suggests that it could be an important therapeutic target. Currently available treatments target interleukin-1, and tumor necrosis factor-alpha and its receptors, and have been only partially successful. Given the complexity of the cytokine pathways that are involved in the pathogenesis of inflammatory disease, further studies are required to test other molecular targets.


Ethnicity and the experience of work: job stress and satisfaction of minority ethnic teachers in the UK.
This paper presents the findings of a nationwide investigation into the mental well-being and job satisfaction of minority ethnic teachers in the UK. Data were collected via a questionnaire containing both open and closed questions. The sample, totalling 208 participants was derived from the National Union of Teachers (NUT) database of minority ethnic teachers and an advertisement in the NUT's Teacher magazine. Univariate analysis of the results revealed that this group of teachers, as compared with other groups were experiencing poorer mental health and lower job satisfaction. Multivariate analysis revealed four reliable factors regarding the 'sources of stress' these minority ethnic teachers perceived they were experiencing. They are the 'hierarchy and culture of the school', workload', 'cultural barriers', and the 'lack of status and promotion'. Some minority ethnic teachers reported that ethnic discrimination on a daily basis or at least several times per week was a contributory factor in their experience of stress. Many of the teachers believed they worked within an institutionally racist environment. Multiple regression analysis discovered that 'total stress', 'total self-esteem', 'working conditions job satisfaction' and 'total discrimination' were the major predictors of mental ill-health in the minority ethnic teachers. Job dissatisfaction was predicted by 'total discrimination', 'workload', 'total general health', 'resolution strategy', and the 'lack of status and promotion'.