Does the Lipid Bilayer Orchestrate Access and Binding of Ligands to Transmembrane Orthosteric/Allosteric Sites of G Protein-Coupled Receptors?
The ligand-binding sites of many G protein-coupled receptors (GPCRs) are situated around and deeply embedded within the central pocket formed by their seven transmembrane-spanning α-helical domains. Generally, these binding sites are assumed accessible to endogenous ligands from the aqueous phase. Recent advances in the structural biology of GPCRs, along with biophysical and computational studies, suggest that amphiphilic and lipophilic molecules may gain access to these receptors by first partitioning into the membrane and then reaching the binding site via lateral diffusion through the lipid bilayer. In addition, several crystal structures of class A and class B GPCRs bound to their ligands offer unprecedented details on the existence of lipid-facing allosteric binding sites outside the transmembrane helices that can only be reached via lipid pathways. The highly organized structure of the lipid bilayer may direct lipophilic or amphiphilic drugs to a specific depth within the bilayer, changing local concentration of the drug near the binding site and affecting its binding kinetics. Additionally, the constraints of the lipid bilayer, including its composition and biophysical properties, may play a critical role in "pre-organizing" ligand molecules in an optimal orientation and conformation to facilitate receptor binding. Despite its clear involvement in molecular recognition processes, the critical role of the membrane in binding ligands to lipid-exposed transmembrane binding sites remains poorly understood and warrants comprehensive investigation. Understanding the mechanistic basis of the structure-membrane interaction relationship of drugs will not only provide useful insights about receptor binding kinetics but will also enhance our ability to take advantage of the apparent membrane contributions when designing drugs that target transmembrane proteins with improved efficacy and safety. In this minireview, we summarize recent structural and computational studies on membrane contributions to binding processes, elucidating both lipid pathways of ligand access and binding mechanisms for several orthosteric and allosteric ligands of class A and class B GPCRs.


New insights into p53 function from structural studies.
Recent structural analysis of p53 has greatly enhanced our understanding of the biochemical activities of this protein by presenting us with a detailed picture of the chemical groups in the protein that are involved in protein stability, conformation and functional interactions. The current structures form the basis for the design of potential therapeutics which could, for example, revert a DNA-binding mutant back to a DNA-binding competent conformation. The structure of the tet domain forms the basis for designing an active therapeutic p53 with an oligomerization domain which would not cross react with a DNA-binding mutant p53. However, as useful as these structures have been in providing insight into the structure/function relationship for p53, a complete understanding of this protein awaits more detailed information on the full-length protein. In this respect, one of the most useful roles for future structural studies will be to help identify the nature of the conformational transition between latent and active p53, and how it can be modulated.


Anti-emetic doses of dexamethasone suppress cortisol response in laparoscopic cholecystectomy.
The efficacy of dexamethasone alone or in combination with a serotonergic antagonist to prevent nausea and vomiting in laparoscopic cholecystectomy is well established, but few data exist regarding its effects on perioperative cortisol and glucose levels. Fourteen non-diabetic subjects having elective laparoscopic choleycystectomy and standardised general anaesthesia were randomised to receive 8 mg of intravenous dexamethasone and tropisetron or tropisetron alone. Plasma cortisol and glucose were measured preinduction, at five and 24 hours postoperatively. There was no difference in plasma cortisol at five hours postoperatively in patients who received dexamethasone, but by 24 hours there was marked suppression compared to the control group (P < 0.005) to less than 5% of the preoperative value. There was a small but statistically significant elevation in blood glucose at 24 hours (P < 0.01) in the dexamethasone-treated group. In patients undergoing laparoscopic cholecystectomy, an anti-emetic dose of dexamethasone (8 mg) markedly suppresses plasma cortisol at 24 hours and causes a minor elevation in blood glucose.


PEs: an efficient solution to primary care maldistribution.
Specializing in often-neglected features of medical practice, physician extenders emphasize the family approach to health education and promote wellness and prevention. Continued federal support of physician extender programs is necessary.


Global distribution of Polaromonas phylotypes--evidence for a highly successful dispersal capacity.
Bacteria from the genus Polaromonas are dominant phylotypes in clone libraries and culture collections from polar and high-elevation environments. Although Polaromonas has been found on six continents, we do not know if the same phylotypes exist in all locations or if they exhibit genetic isolation by distance patterns. To examine their biogeographic distribution, we analyzed all available, long-read 16S rRNA gene sequences of Polaromonas phylotypes from glacial and periglacial environments across the globe. Using genetic isolation by geographic distance analyses, including Mantel tests and Mantel correlograms, we found that Polaromonas phylotypes are globally distributed showing weak isolation by distance patterns at global scales. More focused analyses using discrete, equally sampled distances classes, revealed that only two distance classes (out of 12 total) showed significant spatial structuring. Overall, our analyses show that most Polaromonas phylotypes are truly globally distributed, but that some, as yet unknown, environmental variable may be selecting for unique phylotypes at a minority of our global sites. Analyses of aerobiological and genomic data suggest that Polaromonas phylotypes are globally distributed as dormant cells through high-elevation air currents; Polaromonas phylotypes are common in air and snow samples from high altitudes, and a glacial-ice metagenome and the two sequenced Polaromonas genomes contain the gene hipA, suggesting that Polaromonas can form dormant cells.


Adaptive sweat gland response after spinal cord injury.
To compare the eccrine gland cholinergic sensitivity of upper (UE) and lower (LE) extremities in untrained able-bodied individuals (UAB), untrained individuals with spinal cord injury (U/SCI), and trained wheelchair athletes (E/SCI). Static group comparison. SCI population. A total of 25 men (mean age, 27.8+/-5.0 y; mean height, 175.8+/-9.1cm; mean weight, 76.2+/-7.7 kg) were categorized into 3 groups UAB (n=10), U/SCI (n=10), and E/SCI (n=5). Individuals with SCI had injuries ranging from C4-8. Peripheral sweat production was induced by using pilocarpine iontophoresis at surface landmarks relative to the flexor carpi radialis and medial gastrocnemius muscles. Peripheral sweat rate (SR), sweat gland density (SGD), and sweat per gland (S/G) were calculated for both UE and LE. Peripheral SR for the UAB in both UE (7.58+/-1.99 g x m(-2) x min(-1)) and LE (4.42+/-1.23 g x m(-2) x min(-1)) were significantly greater than those for U/SCI (1.08+/-1.01 g x m(-2) x min(-1), .24+/-.35 g x m(-2) x min(-1), respectively) and E/SCI (3.61+/-2.1 g x m(-2) x min(-1),.71+/-.81 g x m(-2). min(-1), respectively). Furthermore, the UE versus LE SR ratio was calculated at 1.71:1 for UAB subjects, whereas U/SCI and E/SCI subjects showed a ratio of 4.50:1 and 5.07:1, respectively. UE SGD measures in U/SCI (83.20+/-39.84 glands/cm(2)) persons were significantly less than either the UAB (120.20+/-21.42 glands/cm(2)) or the E/SCI (120.80+/-21.56 glands/cm(2)). These results may indicate that sweat glands below the lesion are less sensitive to cholinergic activation, regardless of central or exogenous stimulation. However, glands above the level of the lesion may exhibit increased productivity when individuals are exposed to physical training and physiologic stress.


A Polypeptide of Tumor-Associated Antigen L6 with Intrinsic Adjuvant Activity Enhances Antitumor Immunity.
Peptide vaccines are safe, and aim to elicit and expand tumor-specific immunity so as to eradicate tumors. However, achieving strong and long-lasting anti-tumor immunity with peptide vaccines for the antigen-specific treatment of cancer is challenging, in part because their efficacy depends on strong adjuvants or immunomodulators. We approached this problem by conjugating an epitope-based cancer vaccine with a lipidated sequence (an immunomodulator) to elicit a strong immune response. Lipidated and non-lipidated polyepitope proteins were generated that contained the universal T helper cell epitope (pan-DR), B cell epitopes, and the extended loop sequence of extracellular domain 2 of tumor-associated antigen L6 (TAL6). We show that the lipidated polyepitope cancer vaccine can activate bone marrow-derived dendritic cells, and trigger effective antigen-specific antibody and T helper cell responses, more effectively than the non-lipidated vaccine. Moreover, potent T cell immune responses were elicited in mice inoculated with the lipidated polyepitope cancer vaccine, providing protective antitumor immunity in mice bearing TAL6 tumors. Our study demonstrates that a lipidated polyepitope cancer vaccine could be employed to generate potent anti-tumor immune responses, including humoral and cellular immunity, which could be beneficial in the treatment of TAL6+ cancer.


The role of phenotypic plasticity in the establishment of range margins.
It has been argued that adaptive phenotypic plasticity may facilitate range expansions over spatially and temporally variable environments. However, plasticity may induce fitness costs. This may hinder the evolution of plasticity. Earlier modelling studies examined the role of plasticity during range expansions of populations with fixed genetic variance. However, genetic variance evolves in natural populations. This may critically alter model outcomes. We ask: how does the capacity for plasticity in populations with evolving genetic variance alter range margins that populations without the capacity for plasticity are expected to attain? We answered this question using computer simulations and analytical approximations. We found a critical plasticity cost above which the capacity for plasticity has no impact on the expected range of the population. Below the critical cost, by contrast, plasticity facilitates range expansion, extending the range in comparison to that expected for populations without plasticity. We further found that populations may evolve plasticity to buffer temporal environmental fluctuations, but only when the plasticity cost is below the critical cost. Thus, the cost of plasticity is a key factor involved in range expansions of populations with the potential to express plastic response in the adaptive trait. This article is part of the theme issue 'Species' ranges in the face of changing environments (part I)'.


[Could a health service worker with viral hepatitis be a danger to patients? An attempt to assess the problem in Poland].
The paper presents current problems regarding the risk for viral hepatitis transmission to patients by health service workers infected with this disease. In Poland, the scientific assessment of this problem has not as yet been undertaken. Moreover, there is a lack of preventive measures and legal regulations in this area. In our country, the stay in a hospital is accompanied by a high risk of viral hepatitis infection (hospital infections make 60% of all HBV cases). In view of the HBV and HCV preventive programs carried out in many countries, the Polish specialists should turned attention to the virus carriers among health service workers. The authors propose to establish a legal framework aimed at reducing the risk of patient' infection with viral hepatitis by a health service worker. The experience of other countries presented in this paper may prove to be a useful guide in taking appropriate steps.


The role of US examination in the management of acute abdomen.
Acute abdomen is a medical emergency, in which there is sudden and severe pain in abdomen of recent onset with accompanying signs and symptoms that focus on an abdominal involvement. It can represent a wide spectrum of conditions, ranging from a benign and self-limiting disease to a surgical emergency. Nevertheless, only one quarter of patients who have previously been classified with an acute abdomen actually receive surgical treatment, so the clinical dilemma is if the patients need surgical treatment or not and, furthermore, in which cases the surgical option needs to be urgently adopted. Due to this reason a thorough and logical approach to the diagnosis of abdominal pain is necessary. Some Authors assert that the location of pain is a useful starting point and will guide a further evaluation. However some causes are more frequent in the paediatric population (like appendicitis or adenomesenteritis) or are strictly related to the gender (i.e. gynaechologic causes). It is also important to consider special populations such as the elderly or oncologic patients, who may present with atypical symptoms of a disease. These considerations also reflect a different diagnostic approach. Today, surely the integrated imaging, and in particular the use of multidetector Computed Tomography (MDCT) has revolutionised the clinical approach to this condition, simplyfing the diagnosis but burdening the radiologists with the problems related to the clinical management. However although CT emerging as a modality of choice for evaluation of the acute abdomen, ultrasonography (US) remains the primary imaging technique in the majority of cases, especially in young and female patients, when the limitation of the radiation exposure should be mandatory, limiting the use of CT in cases of nondiagnostic US and in all cases where there is a discrepancy between the clinical symptoms and negative imaging at US.


Identification of members of several homeobox genes in a planarian using a ligation-mediated polymerase chain reaction technique.
I have used a novel single-sided specific polymerase chain reaction (PCR) strategy inspired by ligation-mediated PCR to clone fragments of divergent homeobox genes from a flatworm, the planarian Polycelis nigra. Eight homeobox-containing fragments were amplified, belonging to the Hox, msh, NK-1 and NK-2 classes. Together with the results obtained from several genomes of platyhelminths, my screening shows the presence of the same array of homeodomain developmental regulators in planarians, traditionally regarded as primitive metazoans in terms of body plan, as in coelomate organisms. However, the presence of a Ubx/abd-A homolog may indicate that platyhelminths are more closely related to protostomes than to deuterostomes and supports the idea that flatworms have inherited an elaborate HOX cluster (seven or eight genes) from their ancestor. Likely homologs of the fly genes tinman, bagpipe and S59 suggest that the mesoderm might be patterned by the same genes in all bilaterally symmetrical animals. Finally, a msh-like gene, a family known to be involved in inductive mechanisms in vertebrates, has been found. These results support the hypothesis that the tremendous diversity of metazoan body plans is specified by a largely conserved array of homeobox-containing developmental genes.


Recurrence of pigmented villonodular synovitis of the knee 17 years after the initial treatment. A case report.
Pigmented villonodular synovitis (PVNS) is a benign proliferative process involving the synovial membranes bursas or tendon sheaths. The rarity of the process, the absence of well-documented long follow-up examinations, as well as reports on different forms of the disease from multiple anatomic sites has lead to some confusion. This is a case report of a 44-year-old woman with recurrence of PVNS of her right knee, 17 years after the initial limited knee synovectomy. It was proven impossible to differentiate the disease from soft-tissue malignancy despite the use of all the available investigation methods, including nuclear magnetic resonance. Eventually the diagnosis was made by an open biopsy. Marginal excision of the soft-tissue mass from the popliteal fossa was performed initially, followed by a total knee synovectomy within the next two weeks. The patient remains recurrence free 32 months after the operation.


Role of the phosphatidylinositol 3-kinase/Akt and mTOR/P70S6-kinase pathways in the proliferation and apoptosis in multiple myeloma.
Multiple myeloma (MM) is a plasma cell malignancy preliminary localized in the bone marrow and characterized by its capacity to disseminate. IL-6 and IGF-1 have been shown to mediate proliferative and anti-apoptotic signals in plasmocytes. However, in primary plasma-cell leukemia (PCL) and in end-stage aggressive extramedullar disease, the cytokine requirement for both effects may be not mandatory. This suggests that constitutive activation of signaling pathways occurs. One of the signaling pathways whose deregulation may play an oncogenic role in MM is the phosphatidylinositol 3-kinase (PI 3-K) pathway. In human growth factor-independent MM cell lines OPM2 and RPMI8226, we show that the PI 3-K inhibitors LY294002 and Wortmannin strongly inhibited cell proliferation, whereas inhibition of the mammalian Target Of Rapamycin (mTOR)/P70-S6-kinase (P70(S6K)) pathway with rapamycin or of the Mitogen-Activated Protein Kinase (MAPK) pathway with PD98059 had minimal effect on proliferation. In both cell lines, constitutive activation of the PI 3-K/Akt/FKHRL-1, mTOR/P70(S6K) and MAPK pathways was detected. LY294002 inhibited phosphorylation of Akt, FKHRL-1 and P70(S6K) but had no effect on ERK1/2 phosphorylation, indicating that the PI 3-K and MAPK pathways are independent. IGF-1 but not IL-6 increased phosphorylation of Akt, FKHRL-1 and P70(S6K). Purified plasmocytes from four patients with MM and two patients with primary PCL were studied. In three of them including the two patients with PCL, constitutive phosphorylation of Akt, FKHRL-1 and P70(S6K) was present, inhibited by LY294002 and enhanced by IGF-1. In these patients with constitutive Akt activation, normal PTEN expression was detected. PI 3-K inhibition induced caspase-dependent apoptosis as confirmed by inhibition with the large spectrum caspase inhibitor Z-VAD-FMK and cleavage of pro-caspase-3. Both cell lines spontaneously expressed Skp2 and cyclin D1 proteins at high levels but no p27(Kip1) protein. In the presence of LY294002, cell-cycle arrest in G0/G1 was observed, p27(Kip1) protein expression was up-regulated whereas the expression of both Skp2 and cyclin D1 dramatically diminished. PI 3-K-dependent GSK-3alpha/beta constitutive phosphorylation was also detected in OPM2 cells that may contribute to high cyclin D1 expression. Overall, our results suggest that PI 3-K has a major role in the control of proliferation and apoptosis of growth factor-independent MM cell lines. Most of the biological effects of PI 3-K activation in these cell lines may be mediated by the opposite modulation of p27(Kip1) and Skp2 protein expression. Moreover, constitutive activation of this pathway is a frequent event in the biology of MM in vivo and may be more frequently observed in PCL.


Catalytic enantioselective synthesis of tetrasubstituted chromanones via palladium-catalyzed asymmetric conjugate arylation using chiral pyridine-dihydroisoquinoline ligands.
Highly enantioselective conjugate addition reactions of arylboronic acids to 2-substituted chromones catalyzed by palladium complexes with new chiral Pyridine-Dihydroisoquinoline (PyDHIQ) ligands have been developed. These reactions provide highly enantioselective access to chromanones containing tetrasubstituted stereocenters. Various arylboronic acids and 2-substituted chromones can be used in the catalytic reaction to afford the chiral tetrasubstituted chromanones in good yields and excellent enantioselectivities (25 examples, up to 98% yields, up to 99% ee).


Randomised controlled trial of cognitive-behavioural therapy in early schizophrenia: acute-phase outcomes.
Cognitive-behavioural therapy (CBT) improves persistent psychotic symptoms. To test the effectiveness of added CBT in accelerating remission from acute psychotic symptoms in early schizophrenia. A 5-week CBT programme plus routine care was compared with supportive counselling plus routine care and routine care alone in a multi-centre trial randomising 315 people with DSM-IV schizophrenia and related disorders in their first (83%) or second acute admission. Outcome assessments were blinded. Linear regression over 70 days showed predicted trends towards faster improvement in the CBT group. Uncorrected univariate comparisons showed significant benefits at 4 but not 6 weeks for CBT v. routine care alone on Positive and Negative Syndrome Scale total and positive sub-scale scores and delusion score and benefits v. supportive counselling for auditory hallucinations score. CBT shows transient advantages over routine care alone or supportive counselling in speeding remission from acute symptoms in early schizophrenia.


Development of a panel of three multiplex allele-specific qRT-PCR assays for quick differentiation of recombinant variants and Omicron subvariants of SARS-CoV-2.
Quick differentiation of the circulating variants and the emerging recombinant variants of SARS-CoV-2 is essential to monitor their transmission. However, the widely used gene sequencing method is time-consuming and costly when facing the viral recombinant variants, because partial or whole genome sequencing is required. Allele-specific real time RT-PCR (qRT-PCR) represents a quick and cost-effective method in SNP genotyping and has been successfully applied for SARS-CoV-2 variant screening. In the present study, we developed a panel of 3 multiplex allele-specific qRT-PCR assays targeting 12 key differential mutations for quick differentiation of SARS-CoV-2 recombinant variants (XD and XE) and Omicron subvariants (BA.1 and BA.2). Two parallel multiplex qRT-PCR reactions were designed to separately target the protype allele and the mutated allele of the four mutations in each allele-specific qRT-PCR assay. The variation of Cp values (ΔCp) between the two multiplex qRT-PCR reactions was applied for mutation determination. The developed multiplex allele-specific qRT-PCR assays exhibited outstanding analytical sensitivities (with limits of detection [LoDs] of 2.97-27.43 copies per reaction), wide linear detection ranges (107-100 copies per reaction), good amplification efficiencies (82% to 95%), good reproducibility (Coefficient of Variations (CVs) < 5% in both intra-assay and inter-assay tests) and clinical performances (99.5%-100% consistency with Sanger sequencing). The developed multiplex allele-specific qRT-PCR assays in this study provide an alternative tool for quick differentiation of SARS-CoV-2 recombinant variants (XD and XE) and Omicron subvariants (BA.1 and BA.2).


The bacteriology of diabetic foot ulcers, with a special reference to multidrug resistant strains.
A diabetic foot infection is one of the most feared complications of Diabetes mellitus. Many studies have reported on the bacteriology of Diabetic Foot Infections (DFIs) over the past 25 years, but the results have been varied and often contradictory. This study was carried out to determine the bacterial profiles of infected ulcers and the antibiotic resistance pattern of the isolates. Samples were collected from 50 patients with diabetic foot ulcers by using sterile swabs and they were processed. A total of 75 bacterial isolates were obtained from 50 patients with diabetic foot ulcers. The age group of these patients ranged from 35 to 80 years and the maximum number of patients was in the age group of 60 to 65 years. Gram negative bacilli were more prevalent (65.1%) than gram positive cocci (34.9%). The commonest isolate was Pseudomonas spp (16%), followed by Escherichia coli (14.6%) and Staphylococcus aureus (13.3%).The antibiotic sensivity profiles of the bacteria were also studied. 37.5% of the gram negative bacilli were ESBL producers and 31% were carbapenemase producers. This study showed a preponderance of gram negative bacilli among the isolates from the diabetic foot ulcers. Knowledge on the antibiotic sensitivity pattern of the isolates will be helpful in determining the drugs for the empirical treatment of diabetic ulcers.


Rhenium-catalyzed allylation of C-H bonds of benzoic and acrylic acids.
We have succeeded in the allylation of aromatic and olefinic C-H bonds of benzoic and acrylic acids using a rhenium catalyst, Re(2)(CO)(10). In this reaction, isomerization of the introduced allyl group to the 1-propenyl group did not occur.


[Comparison of Effects of Exhalation and Inhalation Breathing Exercises on Pulmonary Function and Complications in Elderly Patients with Upper-abdominal Surgery].
The purpose of this study was to identify the effects of exhalation breathing exercises using expirometer and that of inhalation breathing exercises using incentive spirometry on pulmonary function and complications in elderly patients with upper-abdominal surgery. The research design was a nonequivalent control group non-synchronized design. Participants were 63 patients who underwent upper-abdominal surgery under general anesthesia (32 in experiment group, 31 in control group). They were recruited at P university hospital from August 1 to November 30, 2015. Effects were evaluated by measuring pulmonary functions (Forced Vital Capacity [FVC], Forced Expiratory Volume in 1 second [FEV1]) and pulmonary complications. Data were analyzed using SPSS/WIN 18.0 program. There was no difference in FVC between the experimental group and the control group, but FEV1 in the experimental group increased significantly compared to the control group by time change (p=.001). Also, there were no pulmonary complications in the experimental group but there were 5 cases (16.1%)(p=.018) in the control group. Findings indicate that exhalation breathing exercises by elderly patients following upper-abdominal surgery is an effective nursing intervention in enhancing pulmonary function and preventing pulmonary complications.


Elevation in viral entry genes and innate immunity compromise underlying increased infectivity and severity of COVID-19 in cancer patients.
Multiple studies have reported a doubling in risk of Coronavirus Disease-2019 (COVID-19) among cancer patients. Here, we examine the potential biological rationale behind this recurrent epidemiological observation. By leveraging large-scale genome-wide transcriptional data of normal and malignant tissues from adults and children, we found evidence of increased expression of SARS-CoV-2 viral entry genes in the cancer state, particularly in respiratory, gastrointestinal, and genitourinary tract tissues, with decreased expression in pediatric vs. adult samples. Additionally, by interrogating the temporal effects of radiotherapy on human peripheral blood mononuclear and mucosal cells, we observed important treatment-related alterations in host innate immunity, specifically type I interferon responses. Overall, cancers enhance expression of critical viral entry genes, and innate viral defenses can be dysregulated transiently during radiation treatments. These factors may contribute to the observed increased susceptibility to SARS-CoV-2 entry and severity of COVID-19 in cancer patients.


The number of trait loci in late-onset Alzheimer disease.
Although it is clear that apoE plays an important role in the genetics of late-onset Alzheimer disease (AD), evidence exists that additional genes may play a role in AD, and estimates of the total contribution of apoE to the variance in onset of AD vary widely. Unfortunately, little information is available on the number and contribution of additional genes. We estimated the number of additional quantitative-trait loci and their contribution to the variance in age at onset of AD, as well as the contribution of apoE and sex, in an oligogenic segregation analysis of 75 families (742 individuals) ascertained for members with late-onset AD. We found evidence that four additional loci make a contribution to the variance in age at onset of late-onset AD that is similar to or greater in magnitude than that made by apoE, with one locus making a contribution several times greater than that of apoE. Additionally, we confirmed previous findings of a dose effect for the apoE varepsilon4 allele, a protective effect for the varepsilon2 allele, evidence for allelic interactions at the apoE locus, and a small protective effect for males. Furthermore, although we estimate that the apoE genotype can make a difference of </=17 years in age at onset of AD, our estimate of the contribution of apoE (7%-9%) to total variation in onset of AD is somewhat smaller than that which has previously been reported. Our results suggest that several genes that have not yet been localized may play a larger role than does apoE in late-onset AD.


The epidermolysis bullosa acquisita antigen (type VII collagen) is present in human colon and patients with crohn's disease have autoantibodies to type VII collagen.
Epidermolysis bullosa acquisita is an autoimmune blistering disease of the skin characterized by IgG autoantibodies against type VII collagen. Systemic diseases are often associated with epidermolysis bullosa acquisita, Crohn's disease being the most frequent. This study sought to determine if type VII collagen, the epidermolysis bullosa acquisita autoantigen, was present in normal human colon by western blotting and immunofluorescence. The 290 kDa type VII collagen alpha chain was demonstrated by western blotting in four normal intraoperative colon specimens. Antibodies to type VII collagen labeled the junction between the intestinal epithelium and the lamina propria. We also used an enzyme-linked immunosorbent assay to test sera from patients with Crohn's disease (n = 19), ulcerative colitis (n = 31), celiac disease (n = 17), rheumatoid arthritis (n = 15), and normal controls (n = 16). It was found that 13 of 19 patients with Crohn's disease and four of 31 patients with ulcerative colitis demonstrated reactivity to type VII collagen. Sera from control subjects, patients with celiac disease or rheumatoid arthritis were negative. The sera from Crohn's disease patients also reacted with type VII collagen by immunoblot analysis. It was concluded that patients with inflammatory bowel disease may have IgG autoantibodies to type VII collagen, which exists in both the skin and the gut.


Fusarium Mycotoxins, Their Metabolites (Free, Emerging, and Masked), Food Safety Concerns, and Health Impacts.
The genus Fusarium produces a number of mycotoxins of diverse chemical structures. Fusariotoxins are secondary metabolites produced by toxigenic fungi of the genus Fusarium. The important and commonly encountered fusariotoxins are trichothecenes, fumonisins, and zearalenone. Fusarium mycotoxins pose varying toxicities to humans and/or animals after consumption of contaminated grain. They can cause acute or chronic illness and, in some cases, death. For instance, a range of Fusarium mycotoxins can alter different intestinal defense mechanisms, such as the epithelial integrity, cell proliferation, mucus layer, immunoglobulins, and cytokine production. Of recent concern is the occurrence of emerging and masked Fusarium mycotoxins in agricultural commodities, which may contribute to toxic health effects, although the metabolic fate of masked mycotoxins still remains a matter of scientific discussion. These mycotoxins have attracted attention worldwide because of their impact on human and animal health, animal productivity, and the associated economic losses. In this paper, we review Fusarium mycotoxins and their metabolites with the aim of summarizing the baseline information on the types, occurrence, and health impacts of these mycotoxins in order to encourage much-needed research on integrated management of this unavoidable food contaminant as concerns for food safety continues to grow worldwide.


Radioimmunoassay screening of dried blood spot materials for benzoylecgonine.
Residual samples of blood spots, which are routinely collected on almost all newborns in the United States, can be used to determine seroprevalence information on newborns and maternal exposures to various substances, including drugs of abuse. By modifying a commercial radioimmunoassay (RIA) kit for urinary samples, one can use blood spotted on filter paper as a matrix to quantitate the cocaine metabolite benzoylecgonine (BE). BE is stable for long periods of time in blood spots and we were able to quantitatively extract it with aqueous buffer. There were no matrix effects of the blood spot eluate on the RIA, and excess lipid in the blood did not alter measurement of BE. By using standards made up of BE in negative blood spot eluate and calibrators of blood that were spiked with BE and then spotted on filter paper to determine extraction efficiency, low levels of BE in blood could be measured. The limit of detection was 5 ng/mL, and the limit of quantitation was 10 ng/mL. Levels of BE in blood collected at autopsy in eluates of blood spots were measured, and they established excellent correlation (r2 = 0.93) with gas chromatography/mass spectrometry measurements. To test this technology, residual blood spots on 545 infants from three states were analyzed for BE.


Subchronic and developmental toxicity studies of vaporized diisopropyl ether in rats.
Two inhalation studies were performed with a vaporized sample of commercial-grade diisopropyl ether (DIPE). In the subchronic study, Sprague-Dawley rats (14/sex) were exposed to 0 (both untreated and sham-exposed controls), 480, 3300, or 7100 ppm DIPE for 6 h/d, 5 d/wk, for approximately 90 d. DIPE itself accounted for 91-95% of the vapors, with the remainder being a mixture of 27-29 compounds. Exposure to DIPE did not adversely affect clinical signs, body weight, serum chemistry, hematology, or the number of sperm or spermatids. Exposure of males to 7100 ppm resulted in hypertrophy of liver cells associated with increased liver weight and in increased kidney weight with an increased incidence of hyaline droplets in proximal tubules of the kidney. Females had increased weight of both liver and kidney, although kidney increased only in relation to sham-exposed controls and no morphological changes were observed in either organ. At 3300 ppm, weights of liver and kidney were again increased in males; the liver weights were increased in females only compared to sham-exposed controls and not untreated controls. No abnormalities were observed morphologically. No changes were observed with 480 ppm. In the developmental toxicity study, pregnant Sprague-Dawley rats (22/group) were exposed to 0 (both untreated and sham-exposed controls), 430, 3095, or 6745 ppm for 6 h/d on gestation d 6-15. Animals were sacrificed on gestation d 20. With 6745 ppm, dams had a slight reduction in body weight gain and a significant decrease in food consumption. A concentration-related increase in the incidence of rudimentary 14th ribs was observed, but its significance was uncertain. There was no apparent toxicity, either maternal or fetal, at the lowest exposure concentration. Both studies indicated a low order of toxicity for DIPE.


Seasonal variations of symptoms and occurrence of human bladder carcinomas.
A longitudinal study of transitional cell carcinomas of the urinary bladder has been performed in a region of Western Norway. Hematuria and infection were the first symptoms of neoplasia of the urinary bladder, showing marked and reproducible seasonal variations. Most of the tumours showed the first symptom at the end of the year. They also had a higher histological grade (WHO) than those occurring at other times of the year. This indicates that environmental factors dependent on the season may be of importance for the manifestation of malignant growth of the urinary bladder.


Knowledge Gaps: A Challenge for Agent-Based Automatic Task Completion.
The study of human cognition and the study of artificial intelligence (AI) have a symbiotic relationship, with advancements in one field often informing or creating new work in the other. Human cognition has many capabilities modern AI systems cannot compete with. One such capability is the detection, identification, and resolution of knowledge gaps (KGs). Using these capabilities as inspiration, we examine how to incorporate detection, identification, and resolution of KGs in artificial agents. We present a paradigm that enables research on the understanding of KGs for visual-linguistic communication. We leverage and enhance and existing KG taxonomy to identify possible KGs that can occur for visual question answer (VQA) tasks and use these findings to develop a classifier to identify questions that could be engineered to contain specific KG types for other VQA datasets. Additionally, we examine the performance of different VQA models through the lens of KGs.


Submammary granular parakeratosis: an acquired punctate hyperkeratosis of exogenic origin.
Granular parakeratosis is a histologic phenomenon that produces a characteristic clinical picture with multiple brownish and hyperkeratotic papules. In all 6 published cases of localized axillary parakeratosis, excessive use of different topical preparations (cream-type products, deodorants that include roll-on and stick types, antiperspirants, shampoos, bath soaps) was reported by the authors. The exact pathogenic causal relationships have not yet been resolved. In the case reported below, we demonstrate for the first time that the macro- and micromorphological entities can also occur in the submammary region.


An overview on biological markers in reproductive and developmental toxicology: concepts, definitions and use in risk assessment.
Reproduction and development are complex couple-dependent processes. Risk assessment for these health outcomes requires the use of biomarkers to link exposures to disease. Biological markers of susceptibility, external dose, internal dose, biologically effective dose, early or late biological responses, altered reproductive or developmental function, and reproductive or developmental disease are introduced. Using these biomarkers it is possible to define a biologically based risk assessment methodology for reproductive and developmental toxicity. Risk assessment for reproductive toxicity requires definition of male and female fecundity, couple-specific factors, spontaneous abortion rate, and other factors. Using using sperm count as a biomarker for male fecundity, an example of a reproductive risk assessment using biomarkers is performed.


Coordinate synthesis of stromelysin, interleukin-1, and oncogene proteins in experimental osteoarthritis. An immunohistochemical study.
Metalloproteases appear to play an important role in the pathophysiology of osteoarthritis (OA) and their expression is believed to be regulated by cytokines such as interleukin-1 (IL-1). Nuclear oncogene products are suggested as mediators through which IL-1 induces metalloprotease gene expression. Little data are available on the in vivo involvement of these agents in the pathophysiology of OA. This study examined by immunohistochemistry, using specific antibodies, the distribution of stromelysin, IL-1 alpha, IL-1 beta, and oncogene products (c-FOS, c-JUN, and c-MYC) in synovium and cartilage from normal and experimental canine models of OA. In the OA synovium, stromelysin and IL-1 were localized in the cytoplasm of superficial synovial lining cells, infiltrating mononuclear cells, and endothelial and smooth muscle cells of the blood vessels, whereas oncoproteins were detected predominantly in the synovial lining cells. Normal synovial membranes demonstrated low levels of specific staining in synovial lining cells with occasional staining of blood vessel cells for IL-1 alpha, IL-1 beta, and stromelysin. In OA cartilage, chondrocytes at the superficial and middle layers as well as in fibrillated areas were found to be involved in the synthesis of stromelysin, IL-1, and oncoproteins. Diffuse staining of stromelysin and IL-1 beta in OA cartilage matrix was also identified. In normal cartilage, only a few chondrocytes at the superficial layer showed a low level of antigens. These results demonstrate the in vivo concomitant cellular and/or matrical presence of stromelysin, IL-1, and oncogene proteins in tissues from experimentally induced OA with the most intense staining at the sites of cartilage erosion and synovial proliferation. These findings suggest that they may be involved in the pathophysiology of OA, and that the regulatory mechanisms involved in the expression of these proteins may be associated.


Cardiac risk stratification for postmyocardial infarction dental patients.
Traditional dental management guidelines of myocardial infarction survivors mandate a 6-month waiting period before elective treatment can be considered. Technological advances in cardiac disease diagnosis, management, and revascularization treatment may make this older mandatory 6-month waiting period obsolete. The purposes of this literature review are to provide an overview of the historical development of cardiac risk stratification and discuss current developments and guidelines in cardiac risk assessment. We hope that this review and update will stimulate the development of updated dental guidelines for treating the cardiac patient.


Renal Na+-K+-ATPase in Okamoto and Dahl hypertensive rats.
Plasma renin activity (PRA, ng AI/ml/hr), plasma aldosterone (PA, ng%) and renal Na+-K+-ATPase (micron m PO 4/mg protein/hr) were measured in tow groups of eight spontaneously hypertensive rats (SHR), two groups of eight Dahl salt hypertensive rats (SS), and their controls (16 normal Wistar and 16 salt-resistant rats). Measurements were made in one group after 2 weeks on a normal (0.48% sodium) and in the other group after 2 weeks on a low (0.01% sodium) sodium diet. After a normal sodium diet, PRA and PA were lower in both groups of hypertensive rats than in control normotensive animals. Renal NA+-K+-ATPase was lower in SS than in controls: in SHR it was not different from control. On a sodium-free diet, SHR exhibited a rise in renal Na+-K+-ATPase but PRA and PA remained low. In contrast, under similar conditions PRA, PA, and renal Na+-K-ATPase increased in SS rats, although to a lesser extent than in SR. These results suggest that under basal conditions and after low salt diet, renal Na+-K+-ATPase activity in SHR behaves as it does in normal rats. However, the changes are independent of PA in SHR. The reduction in PRA and PA in SS suggest volume expansion hypertension. IN SHR, volume expansion is not present, and renal Na+-K+-ATPase is not altered. Enzyme activity is lower in SS than in SHR and control. This suggests that some factor that results from volume expansion may be responsible for inhibition of renal Na+-K+-ATPase.


Reverse transcription-polymerase chain reaction detection of prostate-specific antigen, prostate-specific membrane antigen, and prostate stem cell antigen in one milliliter of peripheral blood: value for the staging of prostate cancer.
There have been several studies on the presence of circulating tumor cells in the peripheral blood of patients with malignant tumors including prostate cancer (PCa) using reverse transcription-PCR (RT-PCR). One of the aims of these studies was to obtain high sensitivity that would enable early-stage diagnosis. However, they varied in their detection rates, and the methods were rather complicated. We have improved the RT-PCR assay combining three prostate-associated molecules, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), and prostate stem cell antigen (PSCA) to reveal patients with poor prognosis. Peripheral blood samples were obtained from 129 patients including 58 cases of PCa and 71 cases of nonmalignant disorders. Total RNA was extracted from 1 ml of whole blood using a commercially available kit. The sensitivity of PSA-, PSMA-, and PSCA-nested RT-PCR was verified with positive signals of a single LNCaP cell in 1 ml of female blood sample. PSA-, PSMA-, and PSCA-mRNA were detected in 7 (12.1%), 12 (20.7%), and 8 (13.8%) PCa, and in 1, 2, and 0 samples in nonmalignant disorders, respectively. Among 58 PCa patients, each PCR indicated the prognostic value in the hierarchy of PSCA>PSA>PSMA RT-PCR, and extraprostatic cases with positive PSCA PCR indicated lower disease-progression-free survival than those with negative PSCA PCR. The present findings suggest that PSCA PCR would be most promising for the molecular staging of PCa. The present RT-PCR is a highly cost-effective and rapid procedure, enabling the molecular staging of PCa with RT-PCR as a diagnostic routine.


Effect of gibberellin a(3) on the endoplasmic reticulum and on the formation of glyoxysomes in the endosperm of germinating castor bean.
Seedlings of castor bean (Ricinus communis cv. Hale) were exposed to a range of concentrations of gibberellin A(3) (GA(3)). Treatments for 20 hours with GA(3) concentrations of 0.5 muM or higher resulted in increased levels of NADH-cytochrome c reductase, phosphorylcholine glyceride transferase, and malate synthase in endoplasmic reticulum (ER) isolated from endosperm on linear sucrose gradients. GA(3) treatment also resulted in increased RNA associated with ER. Malate synthase and catalase in crude homogenates were enhanced by 1 to 100 muM GA(3) concentrations. Isocitrate lyase, citrate synthase, malate synthase, catalase, and glycolate oxidase in isolated glyoxysomes were enhanced by 60, 20, 18, 40, and 28%, respectively, over controls. Treatment with abscisic acid led to decreased levels of glyoxysomal enzymes and reduced glyoxysomal protein. The effect of GA(3) and abscisic acid on the specific activities of glyoxysomes of different densities suggests that GA(3) influences enzyme levels and glyoxysome assembly.


Experimental charge density in an oxidized trinuclear iron complex using 15 K synchrotron and 100 K conventional single-crystal X-ray diffraction.
The experimental electron density distribution in a crystal consisting of the simplest conceivable trinuclear carboxylate-bridged iron-mu3-oxo dianion with two alpha-picolinium cations has been determined using both synchrotron (15 K) and conventional (100 K) X-ray diffraction data. The constituent trinuclear oxo-centered molecule consists of six mu2-bridging formate groups between the iron pairs, while the axial ligand for all iron atoms is another formate group. The compound {[FeO(HCOO)5(HCOO)3]2-.H2O.2(alpha-CH3NC5H5)}, (1) crystallizes in the monoclinic space group P2(1)/m with charge assisted hydrogen bonds linking the alpha-picolinium cations to the trinuclear groups. The chemical bonding in the weakly asymmetric Fe3O-core of 1 has been examined through the use of the quantum theory of atoms in molecules, and in combination with experimental d-orbital populations, a significant electron sharing is observed between the Fe atoms and the central oxygen. The central oxygen exhibits clear sp2 hybridization, and the iron atoms have valence shell charge concentrations in all metal-ligand bond directions. The relative bond strengths are evaluated based upon the charge density distribution and found to be in accordance with the geometrical results. Integrated group charges follow expectations from formal chemical valences.


Right ventricular adaptation to increased afterload after orthotopic cardiac transplantation in the setting of recipient chronic pulmonary hypertension.
Right ventricular (RV) failure remains an important risk factor for early morbidity and mortality after orthotopic cardiac transplantation and is most commonly related to preexistent chronic pulmonary hypertension (CPH) in the recipient, which occurs secondary to long-standing congestive heart failure. This study was designed to assess the compensatory mechanisms of the acutely transplanted RV in the setting of recipient CPH using a canine model of bicaval cardiac transplantation (TX) and monocrotaline pyrrole (MCTP)-induced CPH. Twenty adult mongrel dogs were used for 10 successfully completed TX experiments. Recipients received an injection of 3 mg/kg MCTP 4 months before TX. RV function was assessed with load-insensitive means (preload recruitable stroke work), and Fourier analysis was used to calculate RV hydraulic power and transpulmonary efficiency. At the time of TX, significant increases in the mean pulmonary artery pressure, mean right ventricular pressure, and pulmonary vascular resistance were observed in recipients compared with donors and were further significantly increased after cardiopulmonary bypass. Significant increases in RV preload recruitable stroke work and RV hydraulic power were observed after TX compared with before TX and occurred in association with significant decreases in transpulmonary efficiency. Significant increases in pulmonary hemodynamic indexes occurred after MCTP injection and were further significantly increased after cardiopulmonary bypass. In the setting of recipient CPH, RV performance adapts acutely after bicaval TX with significant increases in power and contractility. However, a significant decrease in transpulmonary efficiency was also observed, which may improve over time as the RV adapts to the increased afterload.


A connectionist study on the role of pitch in infant-directed speech.
Infant-directed speech (IDS) is believed to facilitate language learning. However, the benefit may be either due to clearer acoustic correlates to linguistic structures, or simply increased attention from infants induced by IDS exaggerated prosody. This study investigated the pure effect of IDS pitch on lexical tone learning, with attentional/affective factors removed by using artificial neural networks. Following training with the pitch of Mandarin tones in IDS versus adult-directed speech, the networks yielded equal tonal categorization for both registers. IDS pitch produced no additional linguistic support. IDS pitch appears to strictly play the non-linguistic role of attention/affect, which may indirectly benefit learning.


The histidine kinase-related domain of Arabidopsis phytochrome a controls the spectral sensitivity and the subcellular distribution of the photoreceptor.
Phytochrome A (phyA) is the primary photoreceptor for sensing extremely low amounts of light and for mediating various far-red light-induced responses in higher plants. Translocation from the cytosol to the nucleus is an essential step in phyA signal transduction. EID1 (for EMPFINDLICHER IM DUNKELROTEN LICHT1) is an F-box protein that functions as a negative regulator in far-red light signaling downstream of the phyA in Arabidopsis (Arabidopsis thaliana). To identify factors involved in EID1-dependent light signal transduction, pools of ethylmethylsulfonate-treated eid1-3 seeds were screened for seedlings that suppress the hypersensitive phenotype of the mutant. The phenotype of the suppressor mutant presented here is caused by a missense mutation in the PHYA gene that leads to an amino acid transition in its histidine kinase-related domain. The novel phyA-402 allele alters the spectral sensitivity and the persistence of far-red light-induced high-irradiance responses. The strong eid1-3 suppressor phenotype of phyA-402 contrasts with the moderate phenotype observed when phyA-402 is introgressed into the wild-type background, which indicates that the mutation mainly alters functions in an EID1-dependent signaling cascade. The mutation specifically inhibits nuclear accumulation of the photoreceptor molecule upon red light irradiation, even though it still interacts with FHY1 (for far-red long hypocotyl 1) and FHL (for FHY1-like protein), two factors that are essential for nuclear accumulation of phyA. Degradation of the mutated phyA is unaltered even under light conditions that inhibit its nuclear accumulation, indicating that phyA degradation may occur mostly in the cytoplasm.


The role of the low density lipoprotein receptor pathway in the delivery of lipophilic photosensitizers in the photodynamic therapy of tumours.
Lipoproteins are now recognized as major blood carriers of many hydrophobic porphyrins and related chromophores which are being investigated as possible photosensitizers in the photodynamic therapy of tumours. In vitro and in vivo studies have demonstrated the role of the low density lipoprotein (LDL) receptor pathway in the delivery of photosensitizers to tumour cells and its importance in porphyrin accumulation by tumours. Lysosomes, which are involved in the cellular processing of LDL, are important intracellular targets in the LDL-porphyrin-induced phototoxicity. The use of the LDL receptor pathway as a tool for enhancing the selectivity of photosensitizer delivery to tumour cells appears to be a promising field of research in the photodynamic therapy of tumours.


[A kind of improved operative correction of severe inverted nipples].
To introduce a kind of improved operative method for correction of severe inverted nipples. Sixteen patients (aged 28-38 years) with 31 inborn inverted nipples were operated with the improved method from January 2002 to March 2005. The nipple was incised horizontally through the middle, the fibrotic band was released, and the tissue below the nipple was contracted. Then the defective space with mammary gland flap. Finally, a Z-shaped suture was employed at each base of nipple. The shapes of these nipples were satisfactory. No infection and necrosis of the nipples occurred. The follow-up showed that the results were perfect in the sensory and elective functions of nipples. All the corrected nipples did not have recurrence except one nipple. The scars in the local site were limited and inconspicuous. Severe inverted nipples can be corrected effectively by the combined improved incisions and Z-shaped sutures. This method is suitable for the female patients with no requirement for feeding.


Microbial transformation of aliphatic aldehydes by Bacillus megaterium to 2,3-dialkylacroleins.
The biotransformation of a series of aliphatic aldehydes (C(8)-C(12)) by Bacillus megaterium isolated from strawberry leaf surfaces was investigated. Products were isolated by liquid/liquid extraction and analyzed by gas chromatography (GC) combined with mass spectrometry (MS). In addition to aliphatic alcohols and the remaining aldehydes, major transformation products included the corresponding acids as well as 2,3-dialkylacroleins, dehydrated aldol addition products, which were detected for the first time as biotransformation products. To verify the structures, 2,3-dialkylacroleins were chemically synthesized from the appropriate aldehydes by base-catalyzed aldol condensation reactions and characterized by (1)H and (13)C NMR spectroscopy. Time-course studies showed that the maximum yield of the acrolein derivatives was obtained after 6 days of incubation.


Peripheral giant cell granuloma of maxilla.
It is often documented that chronic irritation could be an etiology of oral cancer; yet out of negligence little heed is paid to any sort of discomfort until it grows to a sizable mass where it is difficult to go unnoticed. Intraorally, the source of irritation could vary from a jagged tooth edge to a chronic cheek bite. Furthermore, the removable prosthesis and orthodontic appliances can lead to alteration of the oral mucosal tissue. This case report represents one such sequelae of an ill-fitting dental prosthesis. Surprisingly and unfortunately, the broken denture was still in use by the patient until the damage caused by it could no further be ignored, it is then that the patient sought medical help. A histopathological diagnosis of reparative giant cell granuloma was made, and the patient was treated by successful excision of the same.


Detecting item bias with the Rasch model.
The purpose of this article is to introduce the concept of item bias, highlighting the differences between the definition of the term as it is used within Rasch measurement and the definition of the term as it is used in the true-score model, non-model based approaches, or multi-item parameter latent trait models. The discussion continues with a description of alternative methods of assessing item bias within the Rasch measurement framework and discusses the power of these methods to detect the presence of item bias. The discussion concludes with several examples drawn from a number of different mathematics tests. This includes a comparison of the Rasch separate calibration t-test and the Mantel-Haenszel approaches.


Recording from an Identified Neuron Efficiently Reveals Hazard for Brain Function in Risk Assessment.
Modern societies use a continuously growing number of chemicals. Because these are released into the environment and are taken up by humans, rigorous (but practicable) risk assessment must precede the approval of new substances for commerce. A number of tests is applicable, but it has been very difficult to efficiently assay the effect of chemicals on communication and information processing in vivo in the adult vertebrate brain. Here, we suggest a straightforward way to rapidly and accurately detect effects of chemical exposure on action potential generation, synaptic transmission, central information processing, and even processing in sensory systems in vivo by recording from a single neuron. The approach is possible in an identified neuron in the hindbrain of fish that integrates various sources of information and whose properties are ideal for rapid analysis of the various effects chemicals can have on the nervous system. The analysis uses fish but, as we discuss here, key neuronal functions are conserved and differences can only be due to differences in metabolism or passage into the brain, factors that can easily be determined. Speed and efficiency of the method, therefore, make it suitable to provide information in risk assessment, as we illustrate here with the effects of bisphenols on adult brain function.


Divergent effects of 4-1BB antibodies on antitumor immunity and on tumor-reactive T-cell generation.
4-1BB is an inducible receptor-like protein expressed rapidly by both CD4 and CD8 T-cells after activation. 4-1BB cross-linking, either by binding to 4-1BBL or by antibody ligation, delivers a costimulatory signal to enhance T-cell activation and proliferation. Previous studies have demonstrated that the administration of 4-1BB monoclonal antibodies (mAbs) induces antitumor immune responses. In the current study using several murine tumors, we examined the systemic effects of 4-1BB mAb on the growth of s.c., intracranial (i.c.), and pulmonary metastases. In addition, the effects of 4-1BB mAb on the generation of antitumor effector T cells were examined. Treatment of 3-day i.c. MCA 205 sarcoma and GL261 glioma with the antibody resulted in prolongation of survival and cure of disease in some mice, whereas only minimal therapeutic effects were observed in established s.c. and pulmonary tumors. No antitumor effects against the poorly immunogenic B16/D5 melanoma were observed. Interestingly, successful treatment of i.c. tumors induced concomitant regression of s.c. tumors. Experiments using severe combined immunodeficient mice and mice depleted of either CD4 or CD8 T cells demonstrated T-cell dependence of the antitumor effects. For generation of effector T cells in the tumor-draining lymph nodes (LNs), administration of 4-1BB mAb had adverse effects, despite the apparent hypertrophy of the LNs. During in vitro activation of tumor-draining LN T cells with anti-CD3 and interleukin 2, the 4-1BB mAb augmented proliferation, resulting in an increase in CD8 T cells. However, they were less therapeutic than not treated LN cells. In adoptive immunotherapy, the coadministration of 4-1BB mAb enhanced the therapeutic efficacy. These results thus demonstrate the limits and potential advantages of 4-1BB antibody interactions with antitumor T cells in vivo and in vitro and suggest that therapeutic interactions of the antibody may be used in a variety of immunotherapeutic approaches.


Pixel and Feature Transfer Fusion for Unsupervised Cross-Dataset Person Reidentification.
Recently, unsupervised cross-dataset person reidentification (Re-ID) has attracted more and more attention, which aims to transfer knowledge of a labeled source domain to an unlabeled target domain. There are two common frameworks: one is pixel-alignment of transferring low-level knowledge, and the other is feature-alignment of transferring high-level knowledge. In this article, we propose a novel recurrent autoencoder (RAE) framework to unify these two kinds of methods and inherit their merits. Specifically, the proposed RAE includes three modules, i.e., a feature-transfer (FT) module, a pixel-transfer (PT) module, and a fusion module. The FT module utilizes an encoder to map source and target images to a shared feature space. In the space, not only features are identity-discriminative but also the gap between source and target features is reduced. The PT module takes a decoder to reconstruct original images with its features. Here, we hope that the images reconstructed from target features are in the source style. Thus, the low-level knowledge can be propagated to the target domain. After transferring both high- and low-level knowledge with the two proposed modules above, we design another bilinear pooling layer to fuse both kinds of knowledge. Extensive experiments on Market-1501, DukeMTMC-ReID, and MSMT17 datasets show that our method significantly outperforms either pixel-alignment or feature-alignment Re-ID methods and achieves new state-of-the-art results.


Evaluation of procedures for the fixation and processing of human tissue for immunohistochemical analysis of human monoclonal antibodies.
Preliminary investigations suggested the importance of an evaluation of different tissue preparation methods frequently used for immunohistochemical analysis of human or murine monoclonal antibodies on human tissue. Colon adenocarcinomas and adjacent morphologically normal colon epithelia were analyzed with an indirect immunoperoxidase technique. Duplicate tissue specimens were (1) snap frozen and fixed in acetone, (2) formalin fixed and paraffin embedded, with or (3) without ensuing treatment with pronase, or (4) alcohol fixed and paraffin embedded. Three different human monoclonal anti-colon cancer IgM antibodies, COU-1, D4213, and F10279, were used in the present study. Endogenous immunoglobulin and the secretory-component-mediated IgG binding were blocked on frozen sections with Fab' anti-IgM and anti-SC antibody. Bound monoclonal antibody was detected with horseradish peroxidase-anti-IgM. COU-1 was found to stain frozen sections of all 25 cancer and adjacent normal colon epithelia. In contrast, on formalin-fixed, paraffin-embedded tissue, only 80% (20/25) of the colon cancer and 44% (11/25) of the adjacent normal colon epithelia were positive. After treatment of the formalin-fixed sections with pronase, all cancers and normal adjacent epithelia were stained, but the cancer cells were more intensely stained than the normal colon epithelial cells. On alcohol-fixed tissues, intense staining was found in all the colon carcinomas analyzed, whereas no staining was found of the adjacent normal colon epithelia, except for a few cells in some of the sections investigated.(ABSTRACT TRUNCATED AT 250 WORDS)


Gaining adequate interdental space with modified elastic separating rings: rationale and technique.
Restoring a tooth with an inadequate contact point and root proximity is a challenge to the practitioner. Ignoring such situations or making compromises in the treatment plan may hinder a successful treatment outcome. Treatment options include strategic extractions, sectional orthodontics, and minor orthodontic movements. The purpose of this article is to discuss the clinical problems and difficulties arising from this situation and to present a modified treatment modality through two case reports. Elastic separating rings, which open an interdental space for placing orthodontic appliances, can be modified to serve as a preprosthetic means for solving mesiodistal crowding of teeth in daily practice. The classic method is modified by the use of elastic rings in sequentially increased thickness, so that the space gained with one ring is followed and increased with a thicker one. An orthodontic background and special instruments are not necessary.


A revised model of the inner-hair cell and auditory-nerve complex.
A revised computational model of the inner-hair cell (IHC) and auditory-nerve (AN) complex is presented and evaluated. Building on previous models, the algorithm is intended as a component for use in more comprehensive models of the auditory periphery. It combines smaller components that aim to be faithful to physiology in so far as is practicable and known. Transduction between cochlear mechanical motion and IHC receptor potential (RP) is simulated using a modification of an existing biophysical IHC model. Changes in RP control the opening of calcium ion channels near the synapse, and local calcium levels determine the probability of the release of neurotransmitter. AN adaptation results from transmitter depletion. The exact timing of AN action potentials is determined by the quantal and stochastic release of neurotransmitter into the cleft. The model reproduces a wide range of animal RP and AN observations. When the input to the model is taken from a suitably nonlinear simulation of the motion of the cochlear partition, the new algorithm is able to simulate the rate-intensity functions of low-, medium-, and high-spontaneous rate AN fibers in response to stimulation both at best frequency and at other frequencies. The variation in fiber type arises in large part from the manipulation of a single parameter in the model: maximum calcium conductance. The model also reproduces quantitatively phase-locking characteristics, relative refractory effects, mean-to-variance ratio, and first- and second-order discharge history effects.


Lipoatrophic panniculitis: case report and review of the literature.
Lipoatrophic panniculitis (LP) is a rare disease of childhood characterized by eruption of tender erythematous nodules and plaques followed by circumferential bands of lipoatrophy often seen on the arms and legs. This condition has also been known as lipophagic panniculitis of childhood, annular atrophy of the ankles, and partial lipodystrophy. A previously healthy 8-year-old boy was evaluated for tender, raised plaques on the ankles, which progressed to circumferential atrophy of the distal lower extremities. Biopsy specimen analysis revealed a dense mixed infiltrate extending into the subcutaneous tissue as well as lipophages within the fatty lobules. A diagnosis of LP was made, and the patient began treatment with prednisone and hydroxychloroquine. Methotrexate was added later to the regimen as a steroid-sparing agent, and the dose was increased over the course of 3 months, by which time the cutaneous disease progression was nearly halted. However, the patient continued to have lower leg pain with bone changes demonstrated on magnetic resonance imaging. We report this case and review of the literature to call attention to the clinical features of LP and its association with skeletal changes. Our patient's response to combination therapy is of interest and contributes to the limited literature about management of this disease.


The high-affinity nAChR partial agonists varenicline and sazetidine-A exhibit reinforcing properties in rats.
Varenicline (Chantix®, Champix®) is a nicotinic acetylcholine receptor (nAChR) partial agonist clinically approved for smoking cessation, yet its potential abuse liability properties have not been fully characterized. The nAChR ligand sazetidine-A has been reported as a selective full or partial agonist at α4β2* nAChR subtypes in in vitro studies. In the present studies, varenicline, sazetidine-A and nicotine exhibited inverted U-shaped dose-response functions under fixed-ratio (peak responding at 30, 60 and 10-30 μg/kg/inf, respectively) or progressive-ratio (peak responding at 30-60, 30-100 and 30 μg/kg/inf, respectively) schedules in rats trained to self-administer nicotine. Varenicline (ED(50) 0.2 mg/kg) and sazetidine-A (ED(50) 0.44 mg/kg) fully substituted for nicotine (ED(50) 0.09 mg/kg) in rats trained to discriminate nicotine (0.4 mg/kg, i.p.) from saline. The reinforcing and discriminative stimulus (DS) properties of sazetidine-A, varenicline and nicotine were attenuated by acute pretreatment with the non-selective neuronal non-competitive nAChR antagonist mecamylamine or the α4* nAChR-selective antagonist dihydro-β-erythroidine, but not by the α7 nAChR subtype antagonist methyllycaconitine. Drug-naïve rats acquired stable self-administration of varenicline (30 μg/kg/inf), and sazetidine-A (60 μg/kg/inf), at doses that supported peak responding under a fixed-ratio 3 schedule in nicotine-trained rats. Nonetheless, self-administration and re-acquisition of varenicline and sazetidine-A were less robust than nicotine. Thus, partial activation of α4β2* nAChRs by varenicline or sazetidine-A is sufficient to mimic the DS and reinforcing properties of nicotine in nicotine-experienced rats, although the reinforcing properties of partial agonists are diminished in nicotine-naïve rats. Future studies should assess nicotine withdrawal measures in animals chronically exposed to varenicline or sazetidine-A.


Chemical composition, bactericidal kinetics, mechanism of action, and anti-inflammatory activity of Isodon melissoides (Benth.) H. Hara essential oil.
Present study was aimed to investigate the antibacterial activity, bactericidal mechanism of action, killing kinetics and anti-inflammatory activity of Isodon melissoides (Benth.) H. Hara essential oil. The gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC-MS) analysis revealed the presence of carvacrol (45.4%), p-cymene (11.6%) and thymol (11.3%) as major constituents of the oil. The oil displayed broad spectrum significant antibacterial activity (MIC: 0.13-8.33 ppm; MBC: 0.13->33.34 ppm) against test strains. The oil exhibited a time and dose-dependent bactericidal effect. The oil disrupted the cell membrane by changing the cell membrane permeability. The essential oil significantly decreased the overproduction of proinflammatory cytokines in LPS-induced inflammation in HaCaT cells without any cytotoxic effect. I. melissoides essential oil can be a promising alternative antimicrobial agent for the control of methicillin resistant staphylococci and other pathogenic bacteria tested, and also useful for the topical anti-inflammatory properties.


Retroperitoneal hematoma as a serious complication of endovascular aneurysmal coiling.
Retroperitoneal hematoma (RH) due to radiologic intervention for an intracranial lesion is relatively rare, difficult to diagnose, and can be life-threatening. We report a case of RH that developed in a patient on anticoagulant therapy following endovascular coiling of a ruptured anterior communicating artery (AcoA) aneurysm. An 82-year-old man presented with a 12-day history of headache. Computed tomography (CT) on admission demonstrated slight subarachnoid hemorrhage, and left carotid angiography revealed an AcoA aneurysm. The next day, the aneurysm was occluded with coils via the femoral approach under general anesthesia. The patient received a bolus of 5,000 units of heparin immediately following the procedure, and an infusion rate of 10,000 units/day was initiated. The patient gradually became hypotensive 25 hours after coiling. Abdominal CT showed a huge, high-density soft-tissue mass filling the right side of the retroperitoneum space. The patient eventually died of multiple organ failure five days after coiling. RH after interventional radiology for neurological disease is relatively rare and can be difficult to diagnose if consciousness is disturbed. This case demonstrates the importance of performing routine physical examinations, sequentially measuring the hematocrit and closely monitoring systemic blood pressures following interventional radiologic procedures in patients with abnormal mental status.


["Chinese hat" flap].
A design of a new type of advancement flap is described and called "chinese hat" flap. It uses the properties of the traditional advancement flap combined with rotation at the two distal edges, that overlap the height of the defect like two claws. This allows direct suture for larger defects with a prominent dog ear that become level with time. Two cases illustrate the procedure.


Clinicopathology and immunohistochemistry of genital warts.
Genital warts are a fairly common sexually transmitted disease and often affect young, illiterate subjects of poor socioeconomic status in India. The mean incubation period was 2.4 months, and the duration varied from 10 days to 4 years. Hyperplastic sessile, verruca vulgaris-like, and flat variants of warts were seen in that sequence, with distinct clinical features. The epidermal and the dermal changes were largely identical; however, the flat type in the epidermal changes were inconspicuous. Koilocytosis was a pathognomonic feature in 28, whereas human papilloma virus (HPV) antigens were detected through peroxidase-antiperoxidase technique in 18 patients.


Efficacy and Safety of TACE Combined With Sorafenib Plus Immune Checkpoint Inhibitors for the Treatment of Intermediate and Advanced TACE-Refractory Hepatocellular Carcinoma: A Retrospective Study.
Purpose: The study aims to retrospectively investigate the efficacy and safety of sorafenib combined with transarterial chemoembolization (TACE) (TACE+Sor) vs. TACE combined with sorafenib plus immune checkpoint inhibitors (TACE+Sor+ICIs) in treating intermediate and advanced TACE-refractory hepatocellular carcinoma (HCC). Materials and Methods: This study was approved by the ethics committee of Lisui Hospital, Zhejiang University, China. From January 2016 to June 2020, 51 eligible patients with intermediate or advanced TACE-refractory HCC received TACE+Sor (n = 29) or TACE+Sor+ICIs (n = 22). The differences in tumor response, adverse events (AEs), progression-free survival (PFS), and overall survival (OS) were compared between the two groups. Factors affecting PFS and OS were determined by Cox regression. Results: The disease control rate was higher in the TACE+Sor+ICIs group than in the TACE+Sor group (81.82 vs. 55.17%, P = 0.046). Compared with the TACE+Sor group, PFS and OS were prolonged in the TACE+Sor+ICIs group (median PFS: 16.26 vs. 7.30 months, P < 0.001; median OS: 23.3 vs. 13.8 months, P = 0.012). Multivariate analysis showed that BCLC stage, alpha-fetoprotein and treatment were independent factors of PFS; BCLC, Child-Pugh class, ablation after disease progression and treatment were independent predictive factors of OS. Four patients in the TACE+Sor+ICIs group and three patients in the TACE+Sor group suffered from dose reduction or interruption (18.18 vs. 10.34%, P = 0.421). The incidence of ICI-related AEs in the TACE+Sor+ICIs group was well-controlled. Conclusion: The therapeutic schedule of TACE+Sor+ICIs demonstrated efficacy and safety in intermediate and advanced TACE-refractory HCC.


Regionalized sequence of myocardial cell growth and proliferation characterizes early chamber formation.
Increase in cell size and proliferation of myocytes are key processes in cardiac morphogenesis, yet their regionalization during development of the heart has been described only anecdotally. We have made quantitative reconstructions of embryonic chicken hearts ranging in stage from the fusion of the heart-forming fields to early formation of the chambers. These reconstructions reveal that the early heart tube is recruited from a pool of rapidly proliferating cardiac precursor cells. The proliferation of these small precursor cells ceases as they differentiate into overt cardiomyocytes, producing a slowly proliferating straight heart tube composed of cells increasing in size. The largest cells were found at the ventral side of the heart tube, which corresponds to the site of the forming ventricle, as well as the site where proliferation is reinitiated. The significance of these observations is 2-fold. First, they support a model of early cardiac morphogenesis in 2 stages. Second, they demonstrate that regional increase in size of myocytes contributes significantly to chamber formation.


Role of magnetic resonance imaging in the staging of gastrointestinal neoplasms.
A concise review is presented of the current applications and techniques of magnetic resonance imaging (MRI) in the field of diagnostic imaging of oncologic disease of the gastrointestinal tract, with a summary of the diagnostic possibilities of MRI in the various types of tumor pathology of the gastrointestinal tract. We conclude with a review of the specific situations in which MRI presents distinct advantages over other diagnostic imaging methods (such as computed tomography and ultrasound).


Tuning donut profile for spatial resolution in stimulated emission depletion microscopy.
In stimulated emission depletion (STED)-based or up-conversion depletion-based super-resolution optical microscopy, the donut-shaped depletion beam profile is of critical importance to its resolution. In this study, we investigate the transformation of the donut-shaped depletion beam focused by a high numerical aperture (NA) microscope objective, and model STED point spread function (PSF) as a function of donut beam profile. We show experimentally that the intensity profile of the dark kernel of the donut can be approximated as a parabolic function, whose slope is determined by the donut beam size before the objective back aperture, or the effective NA. Based on this, we derive the mathematical expression for continuous wave (CW) STED PSF as a function of focal plane donut and excitation beam profiles, as well as dye properties. We find that the effective NA and the residual intensity at the center are critical factors for STED imaging quality and the resolution. The effective NA is critical for STED resolution in that it not only determines the donut shape but also the area the depletion laser power is dispersed. An improperly expanded depletion beam will have negligible improvement in resolution. The polarization of the depletion beam also plays an important role as it affects the residual intensity in the center of the donut. Finally, we construct a CW STED microscope operating at 488 nm excitation and 592 nm depletion with a resolution of 70 nm. Our study provides detailed insight to the property of donut beam, and parameters that are important for the optimal performance of STED microscopes. This paper will provide a useful guide for the construction and future development of STED microscopes.


A new injectable calcium phosphate biomaterial for immediate bone filling of extraction sockets: a preliminary study in dogs.
Many different bone substitutes, such as autografts, allografts or synthetic biomaterials have been proposed to restore alveolar bone loss and support efficient placement of dental implants. This experimental study evaluated the osteoconductive properties of an injectable bone substitute (IBS) composed of a polymeric carrier and a calcium phosphate mineral phase, used to fill mandibular and maxillary canine extraction sockets. The polymer was a cellulose derivative (methyl-hydroxy-propyl-cellulose, MHPC), and the mineral phase consisted of granules of biphasic calcium phosphate (BCP) ceramics 200 to 500 microm in diameter. Mandibular and maxillary premolars extracted from 3 dogs (a total of 60 extraction sites) were immediately treated with the IBS or left unfilled as control sites. Animals were sacrificed 3 months after implantation and all extraction sockets were prepared for histological evaluation. Qualitative histological studies showed that the IBS was able to support the extensive apposition of well-mineralized newly formed lamellar bone over the entire socket surface and appeared to prevent alveolar ridge bone loss in treated extraction sites. Quantitative evaluation showed that the amount of newly formed bone was significantly higher in mandibular than maxillary extraction sockets for both treated and control sites. An injectable bone substitute composed of a polymeric carrier and calcium phosphate was effective in enhancing the bone fill of extraction sockets. This approach may prove promising for periodontal lesions. The material expressed osteoconductive capacities, and the biological properties of the mineral phase were conserved.


PSE: a tool for browsing a large amount of MEDLINE/PubMed abstracts with gene names and common words as the keywords.
MEDLINE/PubMed (hereinafter called PubMed) is one of the most important literature databases for the biological and medical sciences, but it is impossible to read all related records due to the sheer size of the repository. We usually have to repeatedly enter keywords in a trial-and-error manner to extract useful records. Software which can reduce such a laborious task is therefore required. We developed a web-based software, the PubMed Sentence Extractor (PSE), which parses large number of PubMed abstracts, extracts and displays the co-occurrence sentences of gene names and other keywords, and some information from EntrezGene records. The result links to whole abstracts and other resources such as the Online Mendelian Inheritance in Men and Reference Sequence. While PSE executes at the sentence-level when evaluating the existence of keywords, the popular PubMed operates at the record-level. Therefore, the relationship between the two keywords, a gene name and a common word, is more accurately captured by PSE than PubMed. In addition, PSE shows the list of keywords and considers the synonyms and variations on gene names. Through these functions, PSE would reduce the task of searching through records for gene information. We developed PSE in order to extract useful records efficiently from PubMed. This system has four advantages over a simple PubMed search; the reduction in the amount of collected literatures, the showing of keyword lists, the consideration for synonyms and variations on gene names, and the links to external databases. We believe PSE is helpful in collecting necessary literatures efficiently in order to find research targets. PSE is freely available under the GPL licence as additional files to this manuscript.


[A control study of steroid withdrawal protection strategy after kidney transplantation in children].
Objective: To study the influence of steroid withdrawal protection strategy on height growth in pediatric patients after kidney transplantation. Methods: The prospective cohort study enrolled 40 stage 5 chronic kidney disease children receiving kidney transplantation from July 2017 to September 2022 at Guangzhou Women and Children's Medical Center. Based on the primary preoperative disease, patients with immune abnormality-associated glomerular diseases or unknown causes were assigned to the steroid maintenance group, in which patients received steroid tapering within 3 months after surgery to a maintenance dose of 2.5 to 5.0 mg/d. While patients with hereditary kidney disease or congenital urinary malformations were assigned to the steroid withdrawal group, in which patients had steroids tapered off within 3 months. The characteristics of height catch-up growth and clinical data were compared between the 2 groups at baseline, 6, 12, 18 and 24 months after kidney transplantation. T-test, repeated measurement of variance analysis, Mann-Whitney U test, and Fisher exact test were used for the comparison between the 2 groups. Results: Among the 40 children, 17 were males, 23 were females, 25 were in the steroid withdraw group ((7.8±2.8) years old when receiving kidney transplantation) and 15 cases were in the steroid maintenance group ((7.6±3.5) years old when receiving kidney transplantation). The study population was followed up for (26±12) months. The total dose per unit body weight of steroids in the steroid withdrawal group was lower than that in the steroid maintenance group ((0.13±0.06) vs. (0.36±0.19) mg/(kg·d), t=5.83, P<0.001). The height catch-up rate (ΔHtSDS) in the first year after kidney transplantation in the steroid withdraw and steroid maintenance groups was 1.0 (0.7, 1.4) and 0.4 (0.1, 1.0), respectively; in the second year, the ΔHtSDS in the steroid withdraw group was significantly higher than that in the steroid maintenance group (1.1 (0.2, 1.7) vs. 0.3 (0, 0.8), U=28.00, P=0.039). The HtSDS in the steroid withdrawal group at the five follow-up time points was -2.5±0.8, -2.0±0.8, -1.5±0.8, -1.3±0.9 and -0.5±0.3, respectively, while in the steroid maintenance was -2.4±1.3, -2.2±1.1, -2.0±1.0, -1.8±1.0 and -1.6±1.0, respectively. There were statistically significant differences in HtSDS at different follow-up time points in both 2 groups (F=19.81, P<0.01), but no statistical differences in overall impact between the 2 groups (F=1.13, P=0.204). The steroid treatment was interaction with the increase of follow-up time (F=3.62, P=0.009). At the 24th month after transplantation, the HtSDS in the steroid withdrawal group was significantly higher than that in the steroid maintenance group (P=0.047). Six patients in the steroid withdrawal group experienced antibody-mediated immune rejection (AMR), while 3 did in the steroid maintenance group. Moreover, there was no significant difference in AMR between the two groups (χ2=0.06, P=0.814). Conclusion: The steroid withdrawal protection strategy favors the height catch-up growth in pediatric patients after kidney transplantation and does not increase the risk of postoperative antibody-mediated immune rejection.


Articulatory rate and perceptual constancy in phonetic perception.
The perception of syllable-initial stop consonants as voiced (/b/, /d/, /g/) or voiceless (/p/, /t/, /k/) was shown to depend on the prevailing rate of articulation. Reducing the articulatory rate of a precursor phrase causes a greater proportion of test consonants to be identified as voiced. Subsequent experiments demonstrated that this effect depends almost entirely on variation in the duration of the syllable immediately preceding the test syllable; this, the duration of the intervening silent stop closure, and the duration of the test syllable itself all influenced the identification of the stop as voiced or voiceless. Variation in the tempo of a nonspeech melody produced no effect on the perception of embedded test syllables. Those manipulations which produce the major part of the influence of rate do so not by changing the context in which the stop is perceived, but rather by changing temporal concomitants of the constriction, occlusion, and release phases of the articulation of the stop itself. For this reason, an explanation for such effects based on extrinsic timing in perception is found to be wanting. Timing should, in the main, be regarded as intrinsic to the acoustical specifications of phonetic events, a view that is compatible with recent reformulations of the problem of timing control in speech production.


Cerebrovascular autoregulation is profoundly impaired in mice overexpressing amyloid precursor protein.
The amyloid-beta (A beta) peptide, which is derived from the amyloid precursor protein (APP), is involved in the pathogenesis of Alzheimer's dementia and impairs endothelium-dependent vasodilation in cerebral vessels. We investigated whether cerebrovascular autoregulation, i.e., the ability of the cerebral circulation to maintain flow in the face of changes in mean arterial pressure (MAP), is impaired in transgenic mice that overexpress APP and A beta. Neocortical cerebral blood flow (CBF) was monitored by laser-Doppler flowmetry in anesthetized APP(+) and APP(-) mice. MAP was elevated by intravenous infusion of phenylephrine and reduced by controlled exsanguination. In APP(-) mice, autoregulation was preserved. However, in APP(+) mice, autoregulation was markedly disrupted. The magnitude of the disruption was linearly related to brain A beta concentration. The failure of autoregulation was paralleled by impairment of the CBF response to endothelium-dependent vasodilators. Thus A beta disrupts a critical homeostatic mechanism of the cerebral circulation and renders CBF highly dependent on MAP. The resulting alterations in cerebral perfusion may play a role in the brain dysfunction and periventricular white-matter changes associated with Alzheimer's dementia.


Sex differences in depressive symptoms and tolerability after treatment with selective serotonin reuptake inhibitor antidepressants: Secondary analyses of the GENPOD trial.
Differences in serotonergic neurotransmission could lead to sex differences in depressive symptoms and tolerability after treatment with selective serotonin reuptake inhibitors (SSRIs). We investigated whether women have greater reductions in depressive symptoms than men after treatment with an SSRI (citalopram) compared with a noradrenaline reuptake inhibitor (reboxetine) control, and after antidepressant treatment irrespective of class. We also investigated tolerability and the influence of menopausal status. Secondary analyses of the GENPOD (GENetic and clinical Predictors Of treatment response in Depression) trial. Six hundred and one people with depression were recruited from UK primary care and randomized to citalopram or reboxetine. Beck Depression Inventory (BDI-II) score at 6 weeks was the primary outcome. Secondary outcomes included BDI-II score at 12 weeks, and physical symptoms and treatment discontinuation. We calculated main effects and interaction terms using linear and logistic regression models. There was no evidence that women experienced greater reductions in depressive symptoms than men when treated with citalopram compared with reboxetine. We also found no evidence of sex differences at six or 12 weeks (irrespective of antidepressant class): men scored -0.31 (95% confidence interval (CI) -2.23 to 1.62) BDI-II points lower than women at six weeks and -0.44 (95% CI -2.62 to 1.74) points lower at 12 weeks. There was no evidence of sex differences in physical symptoms or treatment discontinuation and no evidence for an influence of menopausal status. Citalopram was not more effective in women compared with men and there was no difference in tolerability. Women and men had similar prognosis after SSRI treatment and similar prognosis regardless of antidepressant class. Findings were unaltered by menopausal status.


[The differentiation of the antigens making up the circulating immune complexes].
A simple method for the detection and analysis of circulating immune complexes (CIC) in specimens of biological fluids is proposed. The method was approved in the examination of patients with chronic infections caused by mycoplasmas and Streptococcus pyogenes L-forms. The method made it possible to diagnose infectious diseases accompanied by the formation of immune complexes and to study the dynamics of the processes of the accumulation and elimination of CIC in the course of the disease. Thus, the detection rate of specific antigens (Ag) incorporated into CIC in patients with mycoplasmal pneumonia exceeded 90 %. In children aged up to 1 year this rate decreased to 40 %. The diagnostic value of the determination of specific Ag incorporated into CIC was shown in streptococcal infections caused by S.pyogenes L-forms, viz. in frequently relapsing erysipelas, as well as in subacute rheumatism and in infectious allergic myocarditis.


Meanings of balance for people with long-term illnesses.
The aim of this study was to elucidate meanings of balance in everyday life for people with long-term illnesses living at home. People living with long-term illnesses are in need of help and support to manage their daily lives. Twelve adults with extensive needs for help and support were interviewed. A phenomenological hermeneutic interpretation was used to analyse the interview texts. The findings show that balance in everyday life for people with long-term illnesses means striving for independence through care and support in their surroundings. It was also important to be able to choose how their lives would be and to be with others who listened and understood them for who they are. By building a relationship with patients at an early stage of their illnesses, nursing staff have an opportunity to understand what people who are living at home with long-term illnesses need in order to achieve balance in their everyday lives.


The complete chloroplast genome of Rhododendron platypodum (Ericaceae): an endemic and endangered species from China.
Rhododendron platypodum Diels (Ericaceae) is a Chinese endemic and endangered species with high ornamental value. Here the complete chloroplast (cp) genome of R. platypodum was assembled and characterized. The cp genome is in a total length of 201,047 bp with the typical quadripartite structure of angiosperms, containing two inverted repeats (IRs) of 44,650 bp separated by a large single-copy (LSC) region of 109,134 bp and a small single-copy (SSC) region of 2613 bp. The whole cp genome of R. platypodum contains 143 genes, including 93 protein-coding genes, 42 transfer RNA genes and 8 ribosomal RNA genes. Phylogenetic analysis based on the coding sequences of cp genome within the Ericaceae family suggests that R. platypodum is closely related to R. delavayi.


Catecholamine responsive adenylate cyclase in human myocardial preparations. Properties and optimalization of assay conditions.
In the present study we have investigated the conditions for optimal adenylate cyclase (AC) activity in preparations of human myocardial biopsies, with emphasis on both basal enzyme activity and isoproterenol response. Different preparation procedures (homogenates, membrane particles) of the same biopsy showed no difference in relative response to isoproterenol, although absolute activities, using protein concentration for normalization, showed some variance. The AC-receptor complexes of the preparations were also stable when stored on ice for 3 h, and both basal and stimulated AC activities were constant at a wide range of protein concentrations (2.9-31.9 micrograms/tube), and throughout 92 min incubation. The effects of varying Mg2+, guanyl nucleotides (GTP, GMP-P(NH)P), and ATP concentrations on myocardial AC activities were also investigated under both basal conditions as well as after isoproterenol stimulation. The apparent Km for the substrate (Mg X ATP) binding to the AC was approximately 0.1 mmol/l. Isoproterenol stimulated the AC activity by increasing Vmax (41 to 142 pmol/mg protein X min) without any change in the apparent Km. Maximal relative activation by isoproterenol was achieved at pH 6.5-7.0. The concentration of isoproterenol causing half maximal AC stimulation was approximately 0.1 micrograms/ml (2 X 10(-4) mmol/l). Half maximal inhibition of isoproterenol (4 micrograms/ml) stimulated AC activity was obtained by 0.025 micrograms/ml propranolol (8 X 10(-5) mmol/l). The sensitivity and precision of this assay should make it possible to measure AC activity as well as isoproterenol response in very small quantities of myocardial tissue. This could provide a method for studying receptor functions of sick hearts by endomyocardial biopsies.


Factor XIII deficiency: new nonsense and deletion mutations in the human factor XIIIA gene.
We identified five disease-causing mutations in six factor XIII deficient patients from four unrelated families: two novel nonsense mutations (nucleotide 979C-->T corresponding to Arg326Stop; and nucleotide 2075G-->A corresponding to Trp691 Stop), one novel deletion of a single nucleotide (nucleotide 708G or 709G), one previously reported missense mutation (nucleotide 888C-->G corresponding to Ser295Arg), and a previously reported splice site mutation (nucleotide 319G-->T at the last position of exon 3). The phenotypic consequences of these mutations are discussed.


CD36-Binding Amphiphilic Nanoparticles for Attenuation of Alpha Synuclein-Induced Microglial Activation.
Neuroinflammation is one of the hallmarks contributing to Parkinson's Disease (PD) pathology, where microglial activation occurs as one of the earliest events, triggered by extracellular alpha synuclein (aSYN) binding to the CD36 receptor. Here, CD36-binding nanoparticles (NPs) containing synthetic tartaric acid-based amphiphilic polymers (AMs) were rationally designed to inhibit this aSYN-CD36 binding. In silico docking revealed that four AMs with varying alkyl side chain lengths presented differential levels of CD36 binding affinity and that an optimal alkyl chain length would promote the strongest inhibitory activity towards aSYN-CD36 interactions. In vitro competitive binding assays indicated that the inhibitory activity of AM-based NPs plateaued at intermediate side chain lengths of 12- and 18-carbons, supporting the in silico docking predictions. These 12- and 18-carbon length AM NPs also had significantly stronger effects on reducing aSYN internalization and inhibiting the production of the proinflammatory molecules TNF-α and nitric oxide from aSYN-challenged microglia. All four NPs modulated the gene expression of aSYN-challenged microglia, downregulating the expression of the proinflammatory genes TNF, IL-6, and IL-1β, and upregulating the expression of the anti-inflammatory genes TGF-β and Arg1. Overall, this work represents a novel polymeric nanotechnology platform that can be used to modulate aSYN-induced microglial activation in PD.