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Objectives To evaluate the ability of post-chemo-radiotherapy (CRT) diffusion-weighted-MRI apparent diffusion coefficient (ADCmean) and 18F-FDG PET maximum standardized uptake value (SUVmax) to predict disease-free survival (DFS) in head and neck squamous cell carcinoma (HNSCC), and to determine whether this ability is influenced by human papillomavirus oropharyngeal cancer (HPV-OPC) status. Methods This prospective cohort observational study included 65 participants (53 male, mean ± SD age 59.9 ± 7.9 years, 46 HPV-OPC) with stage III or IV HNSCC. Primary tumour and nodal ADCmean (pre-treatment, 6- and 12-weeks post-CRT) and SUVmax (12-weeks post-CRT) were measured. Variables were compared with 2-year DFS (independent t-test/Mann–Whitney test) and overall DFS (Cox regression), before and after accounting for HPV-OPC status. Variables were also compared between HPV-OPC and other HNSCC subgroups after stratifying for DFS. Results Absolute post-CRT ADCmean values predicted 2-year DFS and overall DFS for all participants (p = 0.03/0.03, 6-week node; p = 0.02/0.03 12-week primary tumour) but not in the HPV-OPC subgroup. In participants with DFS, percentage interval changes in primary tumour ADCmean at 6- and 12-weeks were higher in HPV-OPC than other HNSCC (p = 0.01, 6 weeks; p = 0.005, 12 weeks). The 12-week post-CRT SUVmax did not predict DFS. Conclusion Absolute post-CRT ADCmean values predicted DFS in HNSCC but not in the HPV-OPC subgroup. Amongst participants with DFS, post-CRT percentage interval changes in primary tumour ADCmean were significantly higher in HPV-OPC than in other HNSCC. Knowledge of HPV-OPC status is crucial to the clinical utilisation of post-CRT DWI-MRI for the prediction of outcomes.

Background Pancreatic ductal adenocarcinoma (PDAC) does not respond to immune checkpoint inhibitors (ICI) therapy as single agent treatments including anti-PD-1 antibody. One of the mechanisms for the resistance of PDAC to ICI is now attributed to the immunosuppressive microenvironment (TME) in PDAC. Myeloid cells are thought to be the predominant immunosuppressive cells in the TME. Human interleukin-8 (IL-8) is a pro-inflammatory chemokine in the CXC family and has the capability of recruiting myeloid cells into the TME to promote tumor progression and immune escape. Therefore, several anti-IL-8 blockade antibodies were developed including HuMax-IL8 and B108-IL8, which both are fully human IgG1 kappa monoclonal antibodies. We therefore tested whether anti-IL-8 antibodies can potentiate anti-tumor activity of anti-PD-1 antibody in a humanized model of PDAC. Materials and Methods We reconstituted the immune system of the NGS mice with ex vivo activated human T cells and a combination of CD14+ and CD16+ myeloid cells after the mice were orthotopically implanted with human PDAC cells. 10x single nuclei RNA-Seq data processing was further performed to analyze differentially expressed genes among certain cell clusters. Results Our results showed that anti-PD-1 antibody alone had a minimal anti-tumor activity when mice was reconstituted with ex vivo activated T cells. Interestingly, the infusion of the combination of CD14+ and CD16+ myeloid cells together with anti-PD-1 antibody resulted in a modest anti-tumor activity. Adding either HuMAX-IL8 or B108-IL8 led to a significantly enhanced anti-tumor activity. Both CD14+ and CD16+ myeloid cells appeared to be needed for the full anti-tumor activity of IL-8 blockade because mice infused with only CD14+ myeloid cells did not respond to IL-8 blockade and mice infused with only CD16+ myeloid cells responded partially to IL-8 blockade. This result suggested that the target of IL-8 is mainly present in CD16+ myeloid cells and is likely to be granulocytes. Tumor infiltrating immune cells were isolated and demonstrated that IL-8 blockade increases CD45+CD11b+CD15+CD14- myeloid cells, which is known to comprise neutrophils and granulocytic myeloid derived suppressive cells (G-MDSC), in the tumors. Reconstitution of the mice with myeloid cells led to a decrease of CD8+ T cells in the tumors; however, IL-8 blockade brought the CD8+ T cell number back to the baseline. Consistent with an effect of IL-8 blockade on the increase of CD15+CD14- myeloid cells, single nuclear RNA sequencing analysis of the tumor tissues showed that the innate immune response and cytokine response pathways in the myeloid cell cluster were activated by IL-8 blockade. Conclusions This result suggested that IL-8 blockade did not simply inhibit myeloid cells as previously anticipated, but potentiated myeloid cells for the innate immune response and concomitant production of type I cytokines. Such immune responses may subsequently activate the effector T cells as the single nuclear RNA sequencing analysis demonstrated enhanced activation signals in the T cell cluster from the tumors treated by anti-IL-8 antibodies. Taken together, this study supports further testing of anti-IL-8 antibodies including B108-IL8 and HuMax-IL8 in combination with anti-PD-1 antibodies for PDAC treatment. Disclosure Information P. Li: None. N. Rozich: None. J. Wang: None. J. Gai: None. J. Wang: None. Y. Xu: None. B. Herbst: None. R. Yu: A. Employment (full or part-time); Significant; NovaRock. S. Muth: None. N. Niu: None. K. Li: None. V. Fune: None. A. Osipov: None. C. Wolfgang: None. M. Lei: A. Employment (full or part-time); Significant; NovaRock. T. Liang: None. L. Zheng: B. Research Grant (principal investigator, collaborator or consultant and pending grants as well as grants already received); Significant; Bristol-Meyer Squibb, Merck, iTeos, Amgen, NovaRock, Inxmed, Halozyme. E. Ownership Interest (stock, stock options, patent or other intellectual property); Significant; Alphamab, Mingruzhiyao. F. Consultant/Advisory Board; Significant; Biosion, Alphamab, NovaRock, Akrevia/Xilio, Ambrx, Novagenesis, Datarevive, Snow Lake Capitals, Mingruzhiyao. Other; Significant; Aduro Biotech.

Cell migration plays an important role in physiological and pathological processes where the fibrillar morphology of extracellular matrice (ECM) could regulate the migration dynamics. To mimic the morphological characteristics of fibrillar matrix structures, low-voltage continuous electrospinning was adapted to construct straight, wavy, looped and gridded fibre patterns made of polystyrene (of fibre diameter ca. 3 μm). With microfibres deposited onto non-passivated surfaces, cells were permitted to explore their different shapes in response to the directly-adhered fibre, as well as to the neighbouring patterns. For all the patterns studied, analysing cellular migration dynamics of MDA-MB-231 (a highly migratory breast cancer cell line) demonstrated a switch in behaviour when the pattern features approach the upper limit of the cell minor axis. Our findings suggest that, although cells dynamically adjust their shapes in response to different fibrillar environments during migration, their ability to divert from an existing fibre track is limited by the size along the cell minor axis. We therefore conclude that the upper limit of cell minor axis might act as a guide for the design of microfibre patterns for different purposes of cell migration.

BACKGROUND: In December 2020, Moderna released the mRNA-1273 vaccine. The most common side effects are headache, muscle pain, redness, swelling, and tenderness at the injection site. In addition, there have been dermatological adverse events, such as hypersensitivity reactions. Although rare, various bullous eruptions have been described following vaccination. Bullous pemphigoid has been reported to occur most often after receipt of influenza and the diphtheria-tetanus-pertussis vaccine. To the best of our knowledge, there have been no reports of bullous drug eruptions resulting from mRNA vaccines. CASE SUMMARY: A 66-years-old obese Guyanese male presented with a bullous rash following receipt of a commercial COVID-19 mRNA vaccine. He received the first dose uneventfully. However, within 24 h of receiving the second dose, he developed fever, myalgias, and malaise accompanied by a painful blistering rash of his torso, arms, and legs. His fever and myalgias improved after 24 h, but his painful rash did not, and five days after the initial symptoms, he presented to the hospital. There were many violaceous, poorly demarcated patches on his trunk, arms, and thighs on examination, many of which had large flaccid bullae within, and a few areas on his buttocks, posterior shoulder, and scrotum were eroded. The exam was also significant for lower extremity muscle tenderness, stiffness with preserved strength. A skin biopsy showed epidermal necrosis and sparse perivascular dermatitis concerning Stevens-Johnson syndrome or erythema multiforme. However, in the absence of mucous membrane involvement or targetoid lesions, the diagnosis of an extensive bullous fixed drug eruption was made. CONCLUSION: This case illustrates that the bullae eruption occurred as a result of receiving the Moderna vaccination.

Background This study investigated the psychometric properties of a Chinese version of the short form of Metacognitions Questionnaire (MCQ-30), which is an effective tool to assess metacognitive beliefs that have been linked to mental disturbance/disorder. We also tested whether metacognition factors would correlate with Internet gaming disorder (IGD) tendency. Methods Six hundred and eighty Chinese university students (Mean age=19.72, SD=1.38; 31.9% male) voluntarily completed an anonymous online questionnaire. Using standard translation procedures, the MCQ-30 items were translated from English into Chinese language and then back-translated. Depression, anxiety, and IGD tendency were assessed by validated scales. Results Confirmatory factor analysis (CFA) results supported the five-factor structure of the MCQ-30, with satisfactory model fit and good reliability. Criterion-related validity was also supported by significant positive correlations between the five factors and depression/anxiety symptoms. All of the five metacognition factors were also significantly correlated with IGD tendency, with positive beliefs about worry and cognitive confidence being the most salient factor after controlling for gender and age. Limitations This study did not examine the test–retest reliability of this scale, and the generalizability of the findings to other age groups remains unclear. Conclusions This study showed satisfactory psychometric properties of a Chinese version of the MCQ-30 and is the first to demonstrate a positive association between specific metacognition factors and IGD tendency among Chinese people. The latter finding supports an application of metacognitive therapy to the treatment of behavioral addictions, including IGD.

To investigate whether ERK/MNK/eIF4E contributes chemoresistance in ovarian cancer. The phosphorylated levels of Erk, Mnk, and eIF4E were systematically analyzed in ovarian cancer patients before and after chemotherapy, and ovarian cancer cells exposed to short- and long-term chemo-agent treatment. The roles of Erk/Mnk/eIF4E were investigated using pharmacological and genetic approaches. Increased phosphorylation levels of ERK, Mnk1, and eIF4E were observed in ovarian cancer cell exposed to chemotherapeutic agents, and paclitaxel-resistant SK-OV-3-r cells, and is a common response of ovarian cancer patients undergoing chemotherapy. MEK inhibitor U0126 inhibits basal and chemodrug-induced phosphorylation of ERK as well as Mnk1 and eIF4E, suggesting that Mnk1/eIF4E are the downstream signaling of ERK pathway and chemotherapy agents activate ERK/MNK/eIF4E in a MEK-dependent manner. eIF4E overexpression promotes ovarian cancer cell growth without affecting migration. In addition, ovarian cancer cells with eIF4E overexpression are more resistant to chemotherapeutic agents in aspect of growth inhibition and apoptosis induction compared to control cells. In contrast, eIF4E depletion augments chemotherapeutic agents’ effect in ovarian cancer cells. These demonstrate that eIF4E play roles in growth and chemoresistance in ovarian cancer. MEK inhibitor U0126 also significantly enhances chemotherapeutic agents’ inhibitory effects. Our work shows that ERK/Mnk/eIF4E activation is critically involved in ovarian cancer chemoresistance and inhibiting ERK/Mnk/eIF4E broadly sensitizes ovarian cancer response to chemotherapy.

Quantitative measurement of small forces and small displacement using holographic optical tweezers (HOTs) is finding increasing applications due to the features of non-contact and high accuracy manipulation. Although hologram optimization algorithms have been widely reported, the holographic optical trapping performance relying on the algorithms has not been studied systematically. In this paper, we investigated the force measuring the performance of various types of HOTs generated with six different hologram algorithms (GSW, GAA, GS, SR, S, and RM). To do this, we built up a HOT instrument and compared the light fields’ intensity distribution, trap stiffness, efficiency, and calculation time of multi-point trap arrays generated by six hologram algorithms with this setup. Our work will provide a better understanding of the performance of different hologram algorithms in HOTs.

Ketosis is a prevalent periparturient metabolic disorder and we hypothesize that lipopolysaccharide (LPS) infiltration may play a key role in its etiology. Study objectives were to characterize biomarkers of inflammation during the transition period in healthy and clinically diagnosed ketotic cows. Cows were retrospectively categorized into one of two groups: healthy and clinically diagnosed ketotic. Two data sets were utilized; the first dataset (Study A) was obtained as a subset of cows (n=16) enrolled in a larger experiment conducted at the Iowa State University Dairy utilizing Holstein cows (8 healthy; 8 ketotic), and the second dataset (Study B; 22 healthy; 22 ketotic) was obtained from a commercial farm. For both experiments, blood samples were collected prior to and following calving. Ketotic cows in both studies had reduced milk production compared to healthy cows (P0.01). Post-calving, ketotic cows had increased serum amyloid A (4.2 and 1.8 fold in studies A and B, respectively; P=0.03 and P=0.04), haptoglobin (6 fold and ~4 fold; P=0.04 and P=0.03), and lipopolysaccharide binding protein (66 and 45%; P0.01 and P=0.02) compared with their healthy counterparts. Antepartum circulating LPS in ketotic cows was increased (2.3 fold; P=0.01) compared to healthy cows in Study B. In summary, increased biomarkers of inflammation appear to be closely associated with ketosis in transition dairy cows.

To examine how tobacco use and depression/anxiety disorders are related to disturbed sleep in college students.85,138 undergraduate respondents (66.3% female, 74.5% white, non-Hispanic, ages 18-25) from the Spring 2011 American College Health Association-National College Health Assessment II database.Multivariate analyses of tobacco use (none, intermediate, daily) and mental health (diagnosed and/or symptomatic depression or anxiety) were used to predict sleep disturbance.Daily tobacco use was associated with more sleep problems than binge drinking, illegal drug use, obesity, gender, and working20 hours/week. Students with depression or anxiety reported more sleep disturbances than individuals without either disorder, and tobacco use in this population was associated with the most sleep problems.Tobacco use and depression/anxiety disorders are both independently associated with more sleep problems in college students. Students with depression and/or anxiety are more likely to be daily tobacco users, which likely exacerbates their sleep problems.

Summary The aim of this study was to investigate the association between the single-nucleotide polymorphisms (SNPs) of the interleukin 22 (IL-22) gene and systemic lupus erythematosus (SLE) in a Chinese population. Three IL-22 SNPs (rs2227485, rs2227513 and rs2227491) were genotyped using SNaPshot SNP genotyping assays and identified by sequencing in 314 SLE patients and 411 healthy controls. The IL-22 level of serum was assessed by enzyme-linked immunosorbent assay (ELISA) kits. Data were analysed by spss version 17.0 software. We found that rs2227513 was associated with an increased risk of SLE [AG versus AA: adjusted odds ratio (aOR) = 2·24, 95% confidence interval (CI) = 1·22–4·12, P = 0·010; G versus· A: adjusted OR = 2·18, 95% CI = 1·20-3·97, P = 0·011]. Further analysis in patients with SLE showed that the AG genotype and G allele were associated with an increased risk of renal disorder in SLE (G versus A: aOR = 3·09, 95% CI = 1·30–7·33, P = 0·011; AG versus· AA: aOR = 3·25, 95% CI = 1·35–7·85, P = 0·009). In addition, the concentration of IL-22 was significantly lower in the rs2227513 AG genotype compared with AA genotype (P = 0·028). These results suggest that rs2227513 polymorphism might contribute to SLE susceptibility, probably by decreasing the expression of IL-22.

This is the official guideline endorsed by the specialty associations involved in the care of head and neck cancer patients in the UK. Although much commoner in the eastern hemisphere, with an age-standardised incidence rate of 0.39 per 100 000 population, cancers of the nasopharynx form one of the rarer subsites in the head and neck.1 This paper provides recommendations on the work up and management of nasopharyngeal cancer based on the existing evidence base for this condition.Recommendations• Patients with nasopharyngeal carcinoma (NPC) should be assessed with rigid and fibre-optic nasendoscopy. (R)• Nasopharyngeal biopsies should be preferably carried out endoscopically. (R)• Multislice computed tomographic (CT) scan of head, neck and chest should be carried out in all patients and magnetic resonance imaging (MRI) where appropriate to optimise staging. (R)• Radiotherapy (RT) is the mainstay for the radical treatment for NPC. (R)• Concurrent chemoradiotherapy offers significant improvement in overall survival in stage III and IV diseases. (R)• Surgery should only be used to obtain tissue for diagnosis and to deal with otitis media with effusion. (R)• Radiation therapy is the treatment of choice for stage I and II disease. (R)• Intensity modulated radiation therapy techniques should be employed. (R)• Concurrent chemotherapy with radiation therapy is the treatment of choice for stage III and IV disease. (R)• Patients with NPC should be followed-up and assessed with rigid and/or fibre-optic nasendoscopy. (G)• Positron emission tomography–computed tomography (PET–CT), CT or MRI scan should be carried out at three months from completion of treatment to assess response. (R)• Multislice CT scan of head, neck and chest should be carried out in all patients and MRI scan whenever possible and specially in advanced cases with suspected recurrence. (R)• Surgery in form of nasopharyngectomy should be considered as a first line treatment of residual or recurrent disease at the primary site. (R)• Neck dissection remains the treatment of choice for residual or metastatic neck disease whenever possible. (R)• Re-irradiation should be considered as a second line of treatment in recurrent disease. (R)

Background Stenting of airway stenosis is a common procedure in specialized centers. The aim of this study was to summarize our clinical experience in ventilation strategy and anesthesia management of patients undergoing urgent tracheal stenting. Methods Clinical data of 22 patients with severe tracheal stenosis who underwent urgent endoscopic placement of a tracheal stent during a 2-year period were retrospectively reviewed. The efficacy and safety of different ventilation strategies and veno-arterial extracorporeal membrane oxygenation (ECMO), individualized based on the cause and location of tracheal narrowing, were evaluated. Results Sufficient ventilation was successfully established in all patients; ECMO was used in five patients with stenosis in the mid-trachea who were unable to tolerate conventional intubation; a laryngeal mask airway (LMA) was used in five patients with post-intubation tracheal stenosis; a cuffed tracheal tube was used in eight patients with lower tracheal stenosis; and low-frequency jet ventilation in rigid bronchoscopy was used in four patients with mid- or lower tracheal stenosis. Tracheal stents were successfully placed and there were significant improvements in dyspnea. There were significant increases in the partial pressure of carbon dioxide in patients ventilated with the LMA and cuffed tracheal tube. There was no hypoxia during the operative period. Conclusion Establishment of effective airway ventilation in patients with severe tracheal stenosis should be based on the cause, location, and severity of tracheal narrowing. Veno- arterial ECMO may be considered in patients with severe stenosis, if they are judged unable to tolerate conventional ventilation or jet ventilation.

Loss of function of mutated solute carrier family 12 member 3 (SLC12A3) gene is the most frequent etiology for Gitelman syndrome (GS), which is mainly manifested by hypokalemia, hypomagnesemia and hypocalciuria. We report the genetic characteristics of one suspicious Chinese GS pedigree by gene sequencing. Complete sequencing analysis of the SLC12A3 gene revealed that both the proband and his elder sister had a novel homozygous SLC12A3 mutation: c.2099T>C and p.Leu700Pro. Moreover, the SLC12A3 genes of his mother and daughter encoded the same mutated heterozygote. It was noted that in this pedigree, only the proband complained about recurrent episodes of bilateral lower limb weakness over 8 years, while his elder sister, mother and daughter did not present symptoms. The inconsistent clinical features of this pedigree implied that besides diverse phenotypes possibly originated from the same genotype, gender difference may also dominate the variant GS phenotypes. Further genetic and proteomic research are needed to investigate the precise mechanisms of GS, including the study of specific ethnicities.

Chromosomal aberrations are a hallmark of human cancers, with complex cytogenetic rearrangements leading to genetic changes permissive for cancer initiation and progression. Protection of Telomere 1 (POT1) is an essential component of the shelterin complex and functions to maintain chromosome stability by repressing the activation of aberrant DNA damage and repair responses at telomeres. Sporadic and familial mutations in the oligosaccharide-oligonucleotide (OB) folds of POT1 have been identified in many human cancers, but the mechanism underlying how hPOT1 mutations initiate tumorigenesis has remained unclear. Here we show that the human POT1's OB-folds are essential for the protection of newly replicated telomeres. Oncogenic mutations in hPOT1 OB-fold fail to bind to single-stranded telomeric DNA, eliciting a DNA damage response at telomeres that promote inappropriate chromosome fusions via the mutagenic alternative non-homologous end joining (A-NHEJ) pathway. hPOT1 mutations also result in telomere elongation and the formation of transplantable hematopoietic malignancies. Strikingly, conditional deletion of both mPot1a and p53 in mouse mammary epithelium resulted in development of highly invasive breast carcinomas and the formation of whole chromosomes containing massive arrays of telomeric fusions indicative of multiple breakage-fusion-bridge cycles. Our results reveal that hPOT1 OB-folds are required to protect and prevent newly replicated telomeres from engaging in A-NHEJ mediated fusions that would otherwise promote genome instability to fuel tumorigenesis.

This paper proposed a relative value method for measuring the indicators of cardiac reserve and investigated the application on monitoring and evaluating cardiac function for pregnant women. A heart sound sensor is placed at the precordial region to detect phonocardiogram. In order to access the cardiac reserve mobilization level during pregnancy, the cardiac reserve indicators of 1,683 normal pregnant women, 96 abnormal cases with different obstetric complications and 624 non-pregnant women were measured, analyzed and compared. The result shows that the differences between the indicators of pregnant and non-pregnant women were significant (p < 0.05). The ratio of diastolic to systolic duration (D/S) was obviously declined with the increase of gestational weeks and the occurrence of obstetric complication. This very encouraging result indicates that the D/S can be used as an indicator for evaluating the cardiac safety of parturition, which provides a reference for cardiac safety assessment of pregnant women.

OBJETIVO: O objetivo deste estudo foi avaliar o impacto do Programa de Incentivo ao Combate às Carências Nutricionais no município de Assis, estado de São Paulo. MÉTODOS: Foram acompanhadas 132 crianças menores de 24 meses, que permaneceram por um ano no Programa de Incentivo ao Combate às Carências Nutricionais. O estado nutricional foi avaliado pelo índice Peso para Idade expresso em escore-Z da mediana da população de referência do Nacional Center Health Statistics no início, aos 2, 6, 9 e 12 meses de acompanhamento. RESULTADOS: Entre as crianças estudadas, 70% foram classificadas como desnutridas ou em risco nutricional no momento do ingresso no Programa Incentivo ao Combate às Carências Nutricionais. Em relação ao incremento de peso, comparando-se o início e após 12 meses, 64% das crianças ganharam peso. Analisando-se a variação média do escore-Z de Peso para Idade nos intervalos de tempo estudados, segundo as faixas etárias e o estado nutricional na entrada ao programa, observou-se que as crianças menores de um ano de idade foram as que apresentaram um significativo incremento de peso. CONCLUSÃO: Pode-se concluir que as crianças com déficits nutricionais e menores de um ano de idade foram as que mais se beneficiaram do programa de suplementação alimentar Incentivo ao Combate às Carências Nutricionais.

We previously reported that polo-like kinase 1 (Plk1) depletion by lentivirus-based RNA interference led to mitotic arrest and apoptosis in cancer cells, whereas normal diploid cell lines, hTERT-RPE1 and MCF10A, survived a similar level of depletion. To study homogeneous cell lines, we generated several Plk1-depleted hTERT-RPE1 and MCF10A clones that were derived from single cells depleted of Plk1. We found that in the long term, Plk1 depletion slowed proliferation of hTERT-RPE1 cells, apparently due to attenuated progression through S phase. These cells had altered morphology and were elongated compared with control. In contrast, MCF10A clones with mild levels of depletion showed no obvious phenotype. They appeared to have normal proliferation rates with no cell-cycle arrest. However, one MCF10A clone, which was severely depleted of Plk1, although viable, showed sporadic G2/M arrest and apoptosis. This MCF10A clone and all the hTERT-RPE1 clones displayed evidence of DNA-damage checkpoint activation. These data further support the interpretation that cancer cell lines have a much greater requirement for Plk1 than normal nontransformed diploid cells.

Background The pathogenesis of endometriosis remains poorly defined. The aberrant angiogenesis that occurs in eutopic endometrium may play a role in the lesion formation and survival. The difference in gene expression profile between human endometrial endothelial cells (HEECs) from eutopic endometria of patients with and without endometriosis would be a factor that affects the occurrence of endometriosis. Methods To explore the difference, we performed in vitro culture and identified the endothelial origin, as well as observed growth features, of HEECs from the two different sources. We also identified their differences in gene expression profiles by combined suppression subtractive hybridization (SSH) with Genechip, and confirmed the results by quantitative reverse transcription-polymerase chain reaction. Results The HEECs derived from endometriosis patients exhibited a potent survival ability in vitro compared with those from non-endometriosis patients. In the HEECs from EM patients, an altered secretion pattern of extracellular matrix (ECM) components and up-regulation of GREM1 were found. These findings may be used to interpret the remarkable change of phenotype in HEECs from endometriosis patients. The synergistic action of these differentially expressed genes is to promote cell proliferation and concomitantly to inhibit apoptosis. Among the up-regulated ECM genes, TSP2 was the only one which exhibits the capacity to suppress angiogenesis; it may therefore function as an antagonist to the aberrant angiogenesis and may confine its extent and severity. Conclusion It may be postulated that differential regulation of some of these genes in eutopic HEECs plays a facilitating role during the peritoneal vascularization of ectopic endometrial lesions by enhancing angiogenic activity via a paracrine effect.

The displacement adsorption enthalpies (ΔH) of the refolding of lysozyme (Lys) denatured by 1.8 mol L–1 guanidine hydrochloride (GuHCl) on a moderately hydrophobic surface at 298 K, pH 7.0 and various (NH4)2SO4 concentrations were determined by using a Micro DSC-III calorimeter. The study shows that the effect of salt concentrations on the three fractions of the enthalpy is that with increasing (NH4)2SO4 concentrations, the molecular conformation enthalpy of the adsorbed Lys has probably no distinct change at 1.8 mol L–1 GuHCl; the adsorption affinity enthalpy (exothermic) becomes more negative; and the dehydration enthalpy (endothermic) decreases. At lower salt concentrations, the dehydration, especially squeezing water molecules led by molecular conformation, which leads to an entropy-driving process, predominates over the adsorption affinity (also including the orderly orientation of molecular conformation), while at higher salt concentrations, the latter is prior to the former for contribution to ΔH and induces an enthalpy-driving process. Also, the optimal NH4)2SO4 concentration favoring refolding and renaturing of Lys denatured by 1.8 mol L–1 GuHCl was found.

The Web Services standard is becoming the lingua franca for loosely coupled distributed applications. As the number of nodes and the complexity of these applications grow over the coming years, it will become more challenging for developers to understand, debug, and optimize them. In this paper, we describe Web Services Navigator, a visualization tool that fosters better understanding of service-oriented architecture (SOA) applications. We draw on our experience with real SOA applications to show how this tool has been applied to practical problems ranging from business logic misunderstandings to performance bottlenecks to syntax and semantic errors. Web Services Navigator helps to solve these problems by visualizing how applications really execute, enabling business owners, application designers, project managers, programmers, and operations staff to understand how their applications actually behave. We sketch the architecture of Web Services Navigator, outline how it reconstructs application execution from event logs, and describe how users interactively explore their applications using its five linked views.

Retinoids are vitamin A derivatives that exert major effects on growth control, epithelial differentiation and embryonic development and, accordingly, are highly teratogenic. In HL60 myelo-monocytic or F9 embryonal carcinoma cell lines, retinoic acid (RA) turns a transformed cell line into terminally differentiated non-proliferating cells. It was only in the mid 1980s that the cloning of the retinoic acid receptors gave a molecular basis to the pleiotropic action of these compound. Retinoic acid receptors (RARs) and retinoid X receptors (RXRs) are nuclear receptors, like those for steroids or thyroid hormones. They are hormone-activated transcription factors that bind to the promoters of their target genes as RAR/RXR heterodimers and repress or activate their transcription depending on the presence of the hormone. Both retinoic acid and its three RAR (α, β, γ) receptors have close links to cancer. Retinoids have significant antitumour activities in several clinical settings, in particular in cervical, skin, head and neck cancers, neuroblastoma and oral leukoplakia, where high doses have yielded significant improvements. A better understanding of the function targeted in the cancer cell and the receptor involved may lead to more specific drugs. Alterations of RARβ expression were reported in a variety of tumors and have suggested that it may act as a tumour suppressor. The clearest illustration of how alterations in RAR structure are linked to cancer is acute promyelocytic leukemia (APL), where a chromosome translocation fuses the PML gene to that of RARα, creating a PML/RARα fusion protein. Strikingly, in contrast, the previous malignancies that are RA-responsive, APL cells undergo terminal differentiation ex vivo or in vivo, similar to HL60 or F9 cells. APL therefore became the paradigm for differentiation therapy. Remarkably, APL is also exquisitely sensitive to arsenic trioxide and both RA and arsenic directly target the PML/RARα fusion protein, identifying the first example of oncogene-targeted therapies. Animal models have helped model the effects of these drugs and provide invaluable preclinical systems for one of the best understood malignancies to date.

Cushing's syndrome occurring during pregnancy is frequently due to an adrenal neoplasm. Adrenal gland tumors occasionally respond to luteinizing hormone (LH) or human chorionic gonadotropin (hCG). We report a case of Cushing's syndrome during and following pregnancy due to an adrenal carcinoma which expressed the LH/hCG receptor. The presence of these receptors may have led to the growth and function of the tumor during pregnancy.

The presence of luteinizing hormone receptors in human pineal glands from five females and three males, ranging in age from 61-89 yr, was examined by in situ hybridization and immunocytochemistry. The results demonstrated the presence of these receptors at the mRNA and protein levels in all the pineal glands examined. Pineal gland luteinizing hormone receptors could potentially be involved in the regulation of melatonin synthesis.

Magnetocardiography (MCG) is a highly reliable non-invasive technique that measures the magnetic field generated by the electrical activity of the heart. Previous studies on MCG have shown high sensitivity in detecting myocardial ischemia, especially in cases of inverse problem solution usage, e.g.

M2A antigen is an oncofetal antigen associated with germ cell neoplasia, present in testis on fetal gonocytes and re-expressed on carcinoma in situ (CIS) and germ cell tumours. We developed a panel of monoclonal antibodies (mAb), M2A (IgG2a), D1-26 (IgG2b) and D2-40 (IgG1), to this antigen in order to characterize its structure and study its distribution among germ cell tumours. M2A antigen was purified by sequential lectin and antibody affinity chromatography and characterized as a monomeric Mr 40 000 surface sialoglycoprotein, extensively glycosylated with O-linked carbohydrate structures, but devoid of N-linked sugars. Terminal sialic acid residues were required for reactivity with mAb M2A and D1-26, but not D2-40. Sections of 69 testicular germ cell tumours, fixed in formalin and embedded in paraffin, were stained with mAb D2-40 to examine the distribution of M2A antigen. Uniform membrane staining was observed in seminomas, and focal staining in 69% of embryonal carcinomas, 29% of teratomas and 25% of yolk sac tumours. CIS in the vicinity of all germ cell tumours also displayed uniform membrane staining. The characterization of M2A antigen, and the development of mAb which react with it in conventionally preserved archival specimens, provide important initiatives to study the origin and progression of germ cell neoplasia. © 1999 Cancer Research Campaign

The heart responds to an increase in sino-atrial node wall stress with an augmentation in rate of contraction. It has been suggested that swelling-activated ion channels may play a key role in that response. This paper investigates directly the effects of cell swelling on spontaneous activity of rabbit isolated sino-atrial node pacemaker cells. The main finding is that sino-atrial node cells, studied in current clamp mode using amphotericin-permeabilized patches, decrease their spontaneous pacemaker rate by 24.2 +/- 7.8% (P < 0.01, n = 9) during 75% hyposmotic swelling. This response is opposite to the predicted impact of volume-activation of sarcolemmal ion conductances. Computer modelling (OXSOFT Heart v4.8) suggests that swelling-induced dilution of the cytosol, reduction in intracellular potassium concentration, and decrease in the delayed rectifier potassium current, IK, are leading mechanisms in the response. This is supported by voltage-clamp data that show a swelling-induced positive shift in the reversal potential of IK by between 5 and 10 mV (n = 7) and a reduction in amplitude of its rapidly activating component, IKr, (n = 6). Thus, spontaneously active sino-atrial node cells reduce pacemaking rate during swelling. This response cannot be explained by the known volume-activated sarcolemmal ion conductances, but appears to be dictated by other mechanisms including dilution of the cytosol and reduction in IK. The results re-enforce the view that cardiac responses to cell volume changes may be quite different from those to longitudinal stretch.

Background Nutrition intake during growth strongly influences ovarian follicle development and egg laying in chicken hens, yet the underlying endocrine regulatory mechanism is still poorly understood. The relevant research progress is hindered by difficulties in detection of leptin gene and its expression in the chicken. However, a functional leptin receptor (LEPR) is present in the chicken which has been implicated to play a regulatory role in ovarian follicle development and egg laying. The present study targeted LEPR by immunizing against its extracellular domain (ECD), and examined the resultant ovarian follicle development and egg-laying rate in chicken hens. Methods Hens that have been immunized four times with chicken LEPR ECD were assessed for their egg laying rate and feed intake, numbers of ovarian follicles, gene expression profiles, serum lipid parameters, as well as STAT3 signaling pathway. Results Administrations of cLEPR ECD antigen resulted in marked reductions in laying rate that over time eventually recovered to the levels exhibited by the Control hens. Together with the decrease in egg laying rate, cLEPR-immunized hens also exhibited significant reductions in feed intake, plasma concentrations of glucose, triglyceride, high-density lipoprotein, and low-density lipoprotein. Parallelled by reductions in feed intake, mRNA gene expression levels of AgRP, orexin, and NPY were down regulated, but of POMC, MC4R and lepR up-regulated in Immunized hen hypothalamus. cLEPR-immunization also promoted expressions of apoptotic genes such as caspase3 in theca and fas in granulosa layer, but severely depressed IGF-I expression in both theca and granulosa layers. Conclusions Immunization against cLEPR ECD in egg-laying hens generated antibodies that mimic leptin bioactivity by enhancing leptin receptor transduction. This up-regulated apoptotic gene expression in ovarian follicles, negatively regulated the expression of genes that promote follicular development and hormone secretion, leading to follicle atresia and interruption of egg laying. The inhibition of progesterone secretion due to failure of follicle development also lowered feed intake. These results also demonstrate that immunization against cLEPR ECD may be utilized as a tool for studying bio-functions of cLEPR.

S100 protein is a 20 kDA calcium-binding protein that accumulates during CNS maturation in mammals. The human gene coding for the beta subunit of S100 protein (S100 beta) is located on chromosome 21, in a subtelomeric position in 21q22.3. In order to investigate the effect of trisomy 21 on S100 beta gene expression, we performed Southern, Northern and Western blot analysis on DNA, RNA and protein, respectively, extracted from the cerebellum of control and Down syndrome (DS) subjects aged 1-18 months. Southern blot analysis revealed a novel EcoRI polymorphism in the S100 beta gene in two of 15 DNA samples examined, and a 1.5 gene dosage for S100 beta in DS. Northern and Western blot analysis showed an approximately 10-fold increase in S100 beta mRNA and protein levels between 1 and 18 months. No differences in the rates of accumulation of S100 beta mRNA and protein were observed between DS and normal subjects. These results demonstrate an increase in S100 beta mRNA and protein levels during infancy indicative of postnatal astrocytic maturation and show that there is no gross deregulation in the expression of the S100 beta gene in DS as a consequence of trisomy 21.

Kornyei JL, Li X, Lei ZM, Rao ChV. Restoration of human chorionic gondadotropin response in human myometrial smooth muscle cells by treatment with follicle-stimulating hormone (FSH): evidence for the presence of FSH receptors in human myometrium. Eur J Endocrinol 1996;134:225–31. ISSN 0804–4643 Human myometrial smooth muscle cells contain receptors for human chorionic gonadotropin (hCG)/ luteinizing hormone (LH). Exogenous hCG and LH can cause a modest hyperplasia in myometrial smooth muscle cells in culture. This response is lost after about the third subculture of the cells. The present study investigated whether the loss of hCG response could be restored by co-culturing with human follicle stimulating hormone (FSH). The results showed that co-culturing with FSH can indeed restore a modest mitogenic response of hCG. However, FSH alone was not mitogenic. The FSH restoration of hCG response can be blocked by antibodies to FSH or hCG but not by non-specific rabbit IgG. The FSH treatment resulted in an increase of steady state levels of hCG/LH receptor mRNA and protein in myometrial smooth muscle cells. Since the FSH actions could be receptor mediated, we investigated the presence of FSH receptor mRNA transcripts and protein in freshly dispersed myometrial smooth muscle cells. Northern blotting demonstrated that myometrial smooth muscle cells, just as rat ovary, a classical target of FSH action, contain multiple FSH receptor mRNA transcripts. Western immunoblotting demonstrated that myometrial smooth muscle cells also contain a 60 kDa FSH receptor protein just as rat ovary and human granulosa cells used as positive control tissues. The immunocytochemistry also demonstrated that myometrial smooth muscle cells, as rat ovary and human granulosa cells, contain FSH receptor immunostaining. In summary, it is novel that FSH could restore the mitogenic response of hCG in human myometrial smooth muscle cells and these cells contain FSH receptors. These findings may have functional implications for direct regulation of human myometrium not only by hCG/LH but also by FSH. ChV Rao, Department of Obstetrics and Gynecology, 438 MDR Building, University of Louisville, School of Medicine, Louisville, Kentucky 40292, USA

It has been suggested that parenchymal viscoelasticity accounts for a large proportion of resistive pressure losses induced by smooth muscle agonists. We used morphometric methods to evaluate the relative contributions of parenchyma and airways to mechanical changes during methacholine- (MCh) induced bronchoconstriction. We measured tracheal pressure and tidal volume in 17 open-chest mechanically ventilated Sprague-Dawley rats (frequency = 1.5 Hz, tidal volume = 2 ml, positive end-expiratory pressure = 5 cmH2O) after saline (n = 7, C), 16 mg/ml MCh (n = 5, L), and 256 mg/ml MCh (n = 5, H) aerosols. We calculated pulmonary resistance (RL) and elastance (EL) by fitting the equation PL = VRL + VEL + K, where PL is transpulmonary pressure, V is flow, and V is volume. V was obtained numerically from V. Lungs were rapidly frozen in liquid N2 30 s after aerosolization and processed with freeze substitution. Using midsagittal slices of left lung, we measured airway narrowing as the ratio of lumen area (aBM) subtended by the basement membrane perimeter (LBM) to predicted maximal lumen area (ABM = LBM2/4 pi). Air space size was estimated as mean linear intercept (Lm) and parenchymal distortion as standard deviation of linear intercepts (SDi). Lm was increased in L and H animals, reflecting hyperinflation; aBM/ABM in 0.17- to 0.43-mm-diam airways decreased progressively from C to L to H animals, reflecting bronchoconstriction. By use of stepwise multiple linear regression, the contribution of Lm to variance in mechanics was found to consistently exceed or equal that of aBM/ABM. SDi contributed little once Lm was taken into account.(ABSTRACT TRUNCATED AT 250 WORDS)

The purpose of this study was to determine the relative contributions of airway wall edema and smooth muscle contraction to the early response (ER) of allergic bronchoconstriction. Brown Norway rats, 6 to 7 wk old, were sensitized with ovalbumin (OA). Anesthetized rats were challenged with either OA or saline 2 wk later. Pulmonary resistance (RL) was measured every minute until either it increased to 150% of the baseline, defined as a significant ER, or until 15 min elapsed. Eight OA-challenged test rats with a significant ER and eight saline-challenged control rats were used for morphometric studies. The lungs were quick-frozen with liquid nitrogen, processed with freeze substitution, and sagittal sections (5 microns) were stained with hematoxylin and eosin. The airway lumen subtended by the epithelial basement membrane (LuB) and cross sectional airway wall area (AW) of all airways were measured by camera lucida and digitization. The LuB and AW of each airway was standardized for size by dividing by the ideal airway lumen (LuBideal), which was calculated from the length of basement membrane, assuming a perfect circle in the unconstricted state. The cumulative frequency distribution of the LuB/LuBideal for the airways from test rats was shifted to the left compared with the control rats (p less than 0.01), indicating airway narrowing after challenge. Airway narrowing increased as a function of airway size. Cumulative frequency distributions of AW/LuBideal showed that there was a significant increase in the wall thickness of only the small airways of test animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Change in nuclear volume and chromatin conformation are generally considered to reflect altered gene expression in eukaryotic cells. The present studies were undertaken to investigate whether these nuclear parameters of luteal cells can be altered by hormone treatment in vitro or change during the estrous cycle. The nuclear volume of small luteal cells was significantly lower than that of large luteal cells during the cycle and pregnancy. The nuclear volumes of small and large luteal cells from pregnancy did not change during incubation without any hormone or with 10 nM prostaglandin (PG)F2α. However, incubation with 1 nM human chorionic gonadotropin (hCG) or 10 nM PGE1 resulted in a significant increase of nuclear volume of small luteal cells by 4 h and that of large luteal cells by 6 h. Small cells were more responsive to hCG than large luteal cells. The nuclear volumes of small and large luteal cells also significantly increased from early to mid luteal phase with no further change in late luteal phase. hCG and PGE1, as well as PGF2α, treatment resulted in a change of chromatin conformation of small and large luteal cells. Dibutyryl cyclic AMP (10 mM) mimicked the hormones by increasing nuclear volumes and changing the chromatin conformation of small and large luteal cells. Chromatin conformation of small and large luteal cells also changed from early to mid luteal phase and mid to late luteal phase. In conclusion, in vitro, hCG and PGs can regulate nuclear volume and/or chromatin conformation of small as well as large bovine luteal cells. In vivo, these nuclear changes occur during the periods of luteal growth, development and regression in the estrous cycle.

Rice is the most important staple food in the world. The rapid development of transgenic rice and its future commercialization have raised concerns regarding transgene flow and its potential environmental risk. It is known that rice is a self-pollinated crop; the outcrossing rate between common cultivars is generally less than 1%. In order to improve the detection sensitivity of rice transgene flow, a male sterile (ms) line BoA with a high outcrossing rate was used as a pollen detector in this study. A concentric circle design was adopted, in which the transgenic rice B2 containing bar gene as a pollen donor was planted in the center circle and the recipient BoA was planted in eight compass sectors. The frequency of transgene flow in compass sectors was analyzed by continuous sampling to generate cumulative data. The results of two years with sound reproducibility demonstrated that the rice gene flow was closely associated with the wind direction. According to the mean frequency of transgene flow, the eight sectors can be divided into two groups: a higher frequency group downstream of the prevailing wind (DPW) with a mean frequency ranging from 6.47 to 26.24%, and a lower frequency group lateral to or upstream of the prevailing wind (UPW) with a mean frequency of 0.39 to 3.03%. On the basis of the cumulative data, 90–96% of the cumulative gene flow events occurred in the four DPW sectors, while it was 4–10% in the four UPW sectors. By using these systematic data, simulation models and isograms of transgene flow in the eight compass sectors were calculated and drawn, respectively.

Schizophrenia is a common psychiatric disorder with a complex genetic etiology. To understand the genetic basis of this syndrome in Portuguese Island populations, we performed a genome-wide scan of 29 families with schizophrenia, which identified a single region on 5q31-5q35 with strong linkage (NPL=3.09, P=0.0012 at D5S820). Empirical simulations set a genome-wide threshold of NPL=3.10 for significant linkage. Additional support for this locus in schizophrenia comes from higher-density mapping and mapping of 11 additional families. The combined set of 40 families had a peak NPL=3.28 (P=0.00066) at markers D5S2112-D5S820. These data and previous linkage findings from other investigators provide strong and consistent evidence for this genomic region as a susceptibility locus for schizophrenia. Exploratory analyses of a novel phenotype, psychosis, in families with schizophrenia and bipolar disorder detected evidence for linkage to the same markers as found in schizophrenia (peak NPL=3.03, P=0.0012 at D5S820), suggesting that this locus may be responsible for the psychotic symptoms observed in both diseases. Molecular Psychiatry (2004) 9, 213-218. doi:10.1038/sj.mp.4001418 Published online 30 December 2003

Lei, M., H. Ghezzo, M. F. Chen, and D. H. Eidelman.Airway smooth muscle orientation in intraparenchymal airways. J. Appl. Physiol. 82(1): 70–77, 1997.—Airway smooth muscle (ASM) shortening is the central event leading to bronchoconstriction. The degree to which airway narrowing occurs as a consequence of shortening is a function of both the mechanical properties of the airway wall as well as the orientation of the muscle fibers. Although the latter is theoretically important, it has not been systematically measured to date. The purpose of this study was to determine the angle of orientation of ASM (θ) in normal lungs by using a morphometric approach. We analyzed the airway tree of the left lower lobes of four cats and one human. All material was fixed with 10% buffered Formalin at a pressure of 25 cmH2O for 48 h. The fixed material was dissected along the airway tree to permit isolation of generations 4–18 in the cats and generations 5–22 in the human specimen. Each airway generation was individually embedded in paraffin. Five-micrometer-thick serial sections were cut parallel to the airway long axis and stained with hematoxylin-phloxine-saffron. Each block yielded three to five sections containing ASM. To determine θ, we measured the orientation of ASM nuclei relative to the transverse axis of the airway by using a digitizing tablet and a light microscope (×250) equipped with a drawing tube attachment. Inspection of the sections revealed extensive ASM crisscrossing without a homogeneous orientation. The θ was clustered between −20° and 20° in all airway generations and did not vary much between generations in any of the cats or in the human specimen. When θ was expressed without regard to sign, the mean values were 13.2° in the cats and 13.1° in the human. This magnitude of obliquity is not likely to result in physiologically important changes in airway length during bronchoconstriction.

To investigate the possibility that altered airway-parenchymal interaction may account for bronchial hyperresponsiveness induced by cigarette smoke exposure, we tested the effect of administration of cigarette smoke (SM), elastase (EL), and both SM and EL on airway responsiveness in 41 Long-Evans male rats. Twelve were exposed to 30 puffs of SM for 15 weeks; 8 received a single intratracheal injection of EL (250 IU/kg); 9 received both EL and SM exposure (SE); 12 control rats were exposed to room air (CO). After 15 weeks, animals were anesthetized and mechanically ventilated (Vt = 2.5 ml, f = 80/min). Methacholine (MCh) dose-response curves (DRCs) were constructed by calculating pulmonary resistance (RL) after ultrasonic nebulization of saline followed by doubling concentrations of MCh (0.0625–256 mg/ml). Exposure to cigarette smoking, with or without elastase, led to a significant reduction in body weight and increased total lung capacity (TLC) compared to exposure to CO. However, there was no significant change in static compliance in the experimental groups, despite increased lung volume. The concentration resulting in a doubling of RL (EC2OORL) was significantly lower in rats treated with SM (n = 7) than CO (n = 8) (3.3 vs. 56.1 mg/ml, geometric mean, p < 0.01). The concentration at which a maximal RL was achieved was lower in SM than CO, EL, and SE (p < 0.05). To assess the possible influence of airway-parenchymal interaction on responsiveness, we measured RL both at functional residual capacity (FRC) and at a volume above FRC equivalent to 1 tidal volume. RL changed similarily in all groups. Despite similar effects on mechanics of both cigarette smoke exposure and elastase administration, only cigarette smoke-exposed animals exhibited evidence of hyperresponsiveness. In this model cigarette smoke-induced hyperresponsiveness is unrelated to changes in either lung elasticity or airway-parenchymal interaction.