CNN-Bi-LSTM: A Complex Environment-Oriented Cattle Behavior Classification Network Based on the Fusion of CNN and Bi-LSTM.

Cattle behavior classification technology holds a crucial position within the realm of smart cattle farming. Addressing the requisites of cattle behavior classification in the agricultural sector, this paper presents a novel cattle behavior classification network tailored for intricate environments. This network amalgamates the capabilities of CNN and Bi-LSTM. Initially, a data collection method is devised within an authentic farm setting, followed by the delineation of eight fundamental cattle behaviors. The foundational step involves utilizing VGG16 as the cornerstone of the CNN network, thereby extracting spatial feature vectors from each video data sequence. Subsequently, these features are channeled into a Bi-LSTM classification model, adept at unearthing semantic insights from temporal data in both directions. This process ensures precise recognition and categorization of cattle behaviors. To validate the model's efficacy, ablation experiments, generalization effect assessments, and comparative analyses under consistent experimental conditions are performed. These investigations, involving module replacements within the classification model and comprehensive analysis of ablation experiments, affirm the model's effectiveness. The self-constructed dataset about cattle is subjected to evaluation using cross-entropy loss, assessing the model's generalization efficacy across diverse subjects and viewing perspectives. Classification performance accuracy is quantified through the application of a confusion matrix. Furthermore, a set of comparison experiments is conducted, involving three pertinent deep learning models: MASK-RCNN, CNN-LSTM, and EfficientNet-LSTM. The outcomes of these experiments unequivocally substantiate the superiority of the proposed model. Empirical results underscore the CNN-Bi-LSTM model's commendable performance metrics: achieving 94.3% accuracy, 94.2% precision, and 93.4% recall while navigating challenges such as varying light conditions, occlusions, and environmental influences. The objective of this study is to employ a fusion of CNN and Bi-LSTM to autonomously extract features from multimodal data, thereby addressing the challenge of classifying cattle behaviors within intricate scenes. By surpassing the constraints imposed by conventional methodologies and the analysis of single-sensor data, this approach seeks to enhance the precision and generalizability of cattle behavior classification. The consequential practical, economic, and societal implications for the agricultural sector are of considerable significance.

Emulsion-templated porous polymers: drying condition-dependent properties.

Macroporous materials templated using high internal phase emulsions (HIPEs) are promising for various applications. To date, new strategies to create emulsion-templated porous materials and to tune their properties (especially wetting properties) are still highly required. Here, we report the fabrication of macroporous polymers from oil-in-water HIPEs, bereft of conventional monomers and crosslinking monomers, by simultaneous ring-opening polymerization and interface-catalyzed condensation, without heating or removal of oxygen. The resulting macroporous polymers showed drying condition-dependent wetting properties (e.g., hydrophilicity-oleophilicity from freezing drying, hydrophilicity-oleophobicity from vacuum drying, and amphiphobicity from heat drying), densities (from 0.019 to 0.350 g cc-1), and compressive properties. Hydrophilic-oleophilic and amphiphobic porous polymers turned hydrophilic-oleophobic simply by heating and protonation, respectively. The hydrophilic-oleophobic porous polymers could remove a small amount of water from oil-water mixtures (including surfactant-stabilized water-in-oil emulsions) by selective absorption and could remove water-soluble dyes from oil-water mixtures. Moreover, the transition in wetting properties enabled the removal of water and dyes in a controlled manner. The feature that combines simply preparation, tunable wetting properties and densities, robust compression, high absorption capacity (rate) and controllable absorption makes the porous polymers to be excellent candidates for the removal of water and water-soluble dyes from oil-water mixtures.

MiR-548a-3p regulates inflammatory response via TLR4/NF-κB signaling pathway in rheumatoid arthritis.

Currently published studies have implicated that microRNAs (miRNAs) including exosomes-encapsulated miRNAs play a critical role in rheumatoid arthritis (RA). Previously, we have found that exosomes-encapsulated miR-548a-3p was significantly decreased in serum samples from RA patients by miRNAs microarray analysis. However, little is known of the role of miR-548a-3p in the development and progression of RA. In this study, we aim to investigate the underlying molecular mechanisms of miR-548a-3p in RA, which will provide new insight into understanding the pathogenesis of RA and identifying novel therapeutics targets for this disease. As validated by quantitative real-time polymerase chain reaction (qRT-PCR), the expression of miR-548a-3p in serum exosomes and peripheral blood mononuclear cells (PBMCs) of RA patients (n = 76) was obviously down-regulated compared with healthy controls (n = 20). Serum exosomal miR-548a-3p was negatively associated with levels of CRP, RF, and ESR in serum of patients with RA. MiR-548a-3p could inhibit the proliferation and activation of pTHP-1 cells by regulating the TLR4/NF-κB signaling pathway. Accordingly, exosomes-delivered miR-548a-3p may be a critical factor predicting the disease activity of RA. MiR-548a-3p/TLR4/NF-κB axis can serve as promising targets for RA diagnosis and treatment.

Key genes and functional coexpression modules involved in the pathogenesis of systemic lupus erythematosus.

We performed a systematic review of genome-wide gene expression datasets to identify key genes and functional modules involved in the pathogenesis of systemic lupus erythematosus (SLE) at a systems level. Genome-wide gene expression datasets involving SLE patients were searched in Gene Expression Omnibus and ArrayExpress databases. Robust rank aggregation (RRA) analysis was used to integrate those public datasets and identify key genes associated with SLE. The weighted gene coexpression network analysis (WGCNA) was adapted to identify functional modules involved in SLE pathogenesis, and the gene ontology enrichment analysis was utilized to explore their functions. The aberrant expressions of several randomly selected key genes were further validated in SLE patients through quantitative real-time polymerase chain reaction. Fifteen genome-wide gene expression datasets were finally included, which involved a total of 1,778 SLE patients and 408 healthy controls. A large number of significantly upregulated or downregulated genes were identified through RRA analysis, and some of those genes were novel SLE gene signatures and their molecular roles in etiology of SLE remained vague. WGCNA further successfully identified six main functional modules involved in the pathogenesis of SLE. The most important functional module involved in SLE included 182 genes and mainly enriched in biological processes, including defense response to virus, interferon signaling pathway, and cytokine-mediated signaling pathway. This study identifies a number of key genes and functional coexpression modules involved in SLE, which provides deepening insights into the molecular mechanism of SLE at a systems level and also provides some promising therapeutic targets.

Baseline traits of patients presenting at a low vision clinic in Shanghai, China.

BACKGROUND: Low vision, along with cataract, trachoma, onchocerciasis, childhood blindness and refractive error, is one of the priorities in the global initiative, VISION 2020-The Right to Sight. The purpose of this study was to characterize the traits of patients presenting at a low vision clinic in China. METHODS: A retrospective study was conducted of the records of 299 patients who visited the Low Vision Clinic of Eye and ENT Hospital Affiliated to Fudan University from January 2009 to May 2014. Reviewed parameters included age, gender, education, occupation, cause of visual impairment and types of low vision aids (LVAs) dispensed. RESULTS: Of all the patients (193 male; aged from 3 to 96 years, with a mean of 29.74 ± 25.23 years), 43.48% experienced moderate visual impairment, 25.42% had severe visual impairment and 21.07% were blind. The four major causes of visual impairment were congenital cataract (14.38%), degenerative myopia (13.71%), juvenile macular degeneration (9.36%) and retinitis pigmentosa (9.36%). The most common causes of visual impairment were congenital cataract (22.67%) in 0-19-year-olds, retinitis pigmentosa (20.62%) in 20-59-year-olds, and age-related macular degeneration (36.54%) in the 60+ group. With the help of LVAs, a significant improvement of distance and/or near vision or visual field was observed in 243 patients, of whom 185 accepted LVAs and 58 patients refused due to high price, inconvenience, young age (≤ 6 y), clumsy appearance and ignorance. The most commonly dispensed LVAs were stand magnifiers (21.57%) followed by spectacle-type LVAs (19.21%). CONCLUSIONS: The majority of the patients in our low vision clinic were young, the main causes of visual impairment were congenital and hereditary diseases. Stand magnifiers were the most commonly dispensed LVAs. High price was the major reason for refusing LVAs.

Meta-analysis of methylprednisolone pulse therapy for Graves' ophthalmopathy.

A meta-analysis was performed to evaluate the efficacy of methylprednisolone pulse therapy for Graves' ophthalmopathy. Eight studies involving 376 patients were included. A higher effective rate was found for patients treated with intravenous glucocorticoids (IVGC) over oral glucocorticoids (OGC) (risk ratio [RR] = 1.48; 95% confidence interval [CI] = 1.18-1.86). The combined IVGC and orbital radiotherapy (OR) was markedly more effective than OGC+OR (RR = 1.40; 95% CI = 1.11-1.77). IVGC resulted in an obvious reduction of clinical activity score (CAS) compared with OGC, with a weighted mean difference (WMD) of 0.86 (95% CI = 0.53-1.18). The WMD for the reduction of the CAS between IVGC+OR and OGC+OR was 0.66 (95% CI = 0.30-1.02). IVGC is an effective treatment and cause fewer adverse events. Limiting the total cumulative dose of methylprednisolone, careful patient selection and monitoring the condition of patients during treatment are necessary.

Experimental data simulating lithium battery charging and discharging tests under different external constraint pressure conditions.

In this paper, the GSP655060Fe soft pack lithium-ion battery with a capacity of 1600 mAh is utilized, employing lithium iron phosphate as the positive electrode and graphite as the negative electrode. In order to comprehensively evaluate the performance of lithium batteries under the conditions of multi-application scenarios, the operating conditions of the battery were simulated under various external confinement pressures of 300 N, 400 N, 500 N, and 600 N, respectively, and the ambient temperatures of 10 ℃, 25 ℃, and 40 ℃, respectively, were controlled to thoroughly test the battery. One charge/discharge test was conducted on six batteries of the same model at multiplicities of 0.5 C, 1 C, 1.5 C, and 2 C, respectively. To ensure the accuracy and reliability of the experimental data, a Battery comprehensive tester Neware BTS-5V12A was utilized, which possesses high-precision voltage and current measurement capabilities with an error rate of only 0.05 %. This data plays an important role in battery research and development, new energy vehicles, electronic products, and other fields.

Interfacial engineering of CoP/CoS2 heterostructure for efficiently electrocatalytic pH-universal hydrogen production.

The design and development of high-performance, low-cost catalysts with long-term durability are crucial for hydrogen generation from water electrolysis. Interfacial engineering is an appealing strategy to boost the catalytic performance of electrode materials toward hydrogen evolution reaction (HER). Herein, we report a simple phosphidation followed by sulfidation treatment to construct heterogeneous cobalt phosphide-cobalt sulfide nanowire arrays on carbon cloth (CoP/CoS2/CC). When evaluated as catalysts toward the HER, the resultant CoP/CoS2/CC exhibits efficient pH-universal hydrogen production due to the heterostructure, synergistic contribution of CoP and CoS2, and conductive substrate. To attain a current density of 10 mA cm-2, overpotentials of only 111.2, 58.1, and 182.9 mV for CoP/CoS2/CC are required under alkaline, acidic, and neutral conditions, respectively. In particular, the as-prepared CoP/CoS2/CC shows markedly improved HER electroactivity in 1.0 M KOH, even outperforming commercial Pt-C/CC at a current density of >50 mA cm-2. In addition, the self-assembled CoP/CoS2||NiFe layered double hydroxide electrolyzer demonstrates efficient catalytic performance and long-time stability, excelling the benchmark Pt-C||IrO2. These findings indicate an effective pathway for the fabrication of high-performance heterogeneous electrocatalysts for hydrogen production in the future.

Prognosis of colon cancer patients based on enhancer RNAs-related genes.

PURPOSE: Colon cancer (CC) is a cancer of the large intestine with high prevalence and poor prognosis. enhancer RNAs. Therefore, valuable tools or biomarkers for predicting patient status, directing clinical practice, and reducing overtreatment are needed. Enhancer RNAs (eRNAs), a class of noncoding RNAs transcribed from enhancers, have been shown to function as regulators of oncogene or tumor suppressor gene expression. The aim of our study was to explore the potential roles of eRNAs and their target enhancer-related genes (ERGs) in the prognosis of CC. METHODS: Selected CC cases (stage I-III) from The Cancer Genome Atlas database were used as a training set, and cases from the Gene Expression Omnibus were used as the validation set. ERGs associated with prognosis were screened through three steps: potential, candidate, and prognosis ERGs. Multivariate Cox proportional hazards analysis was used to identify independent prognostic factors, and a nomogram was created. Calibration curves were drawn by comparing predicted and observed survival probability. For validation, the calibration curves and ROC analysis were also applied to two external validation sets. The biological significance and clinical application of the genes obtained were investigated. RESULTS: Based on the multiple tiers of strict screening, 11 prognostic ERGs were obtained, which were combined to obtain a prognosis signature. A compound nomogram integrating age, TNM classification, and the prognostic signature was constructed. The model was reliable in distinguishing the risk of patients with stage I-III CC, with AUCs of 0.78 and 0.70 at 5 and 7 years, respectively. There was good reproducibility in calibration curves. The prognostic model also yielded good prediction capability in the validation sets. CONCLUSION: In this study, the usefulness and specificity of the ERGs in prognosis were described, which should be considered a key feature in the clinical guidance of CC patients with early stage. We concluded that the major implications of the eRNAs and ERGs should be valued, which would be an emerging hallmark in the prognosis of cancer.

Identification of molecular subtyping system and four-gene prognostic signature with immune-related genes for uveal melanoma.

Immunotherapy is the most promising treatment for uveal melanoma patients with metastasis. Tumor microenvironment plays an essential role in tumor progression and greatly affects the efficacy of immunotherapy. This research constructed an immune-related subtyping system and discovered immune prognostic genes to further understand the immune mechanism in uveal melanoma. Immune-related genes were determined from literature. Gene expression profiles of uveal melanoma were clustered using consensus clustering based on immune-related genes. Subtypes were further divided by applying immune landscape, and weighted correlation network analysis was performed to construct immune gene modules. Univariate Cox regression analysis was conducted to generate a prognostic model. Enriched immune cells were determined after gene set enrichment analysis. Three major immune subtypes (IS1, IS2, and IS3) were identified, and IS2 could be further divided into IS2A and IS2B. The subtypes were closely associated with uveal melanoma prognosis. IS3 group had the most favorable prognosis and was sensitive to PD-1 inhibitor. Immune genes in IS1 group showed an overall higher expression than IS3 group. Six immune gene modules were identified, and the enrichment score of immune genes varied within immune subtypes. Four immune prognostic genes (IL32, IRF1, SNX20, and VAV1) were found to be closely related to survival. This novel immune subtyping system and immune landscape provide a new understanding of immunotherapy in uveal melanoma. The four prognostic genes can predict prognosis of uveal melanoma patients and contribute to new development of targeted drugs.

Integration of Bulk RNA Sequencing and Single-Cell RNA Sequencing to Reveal Uveal Melanoma Tumor Heterogeneity and Cells Related to Survival.

Molecular classification based on transcriptional characteristics is often used to study tumor heterogeneity. Human cancer has different cell populations with distinct transcription in tumors, and their heterogeneity is the focus of tumor therapy. Our purpose was to explore the tumor heterogeneity of uveal melanoma (UM) through RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq). Based on the consensus clustering assays of the prognosis-related immune gene set, the immune subtype (IS) of UM and its corresponding immune characteristics were comprehensively analyzed. The heterogeneous cell groups and corresponding marker genes of UM were identified from GSE138433 using scRNA-seq analysis. Pseudotime trajectory analysis and SCENIC analysis were conducted to explore the trajectory of cell differentiation and the regulatory network of single-cell transcription factors (TFs). Based on 37 immune gene sets, UM was divided into two different immune subtypes (IS1 and IS2). The two kinds of ISs have different characteristics in prognosis, immune-related molecules, immune score, and immune cell infiltration. According to 11,988 cells of scRNA-seq data from six UM samples, 11 cell clusters and 10 cell types were identified. The subsets of C1, C4, C5, C8, and C9 were related to the prognosis of UM, and different TF-target gene regulatory networks were involved. These five cell subsets differentiated into 3 different states. Our results provided valuable information about the heterogeneity of UM tumors and the expression patterns of TFs in different cell types.

PCDE-Sync: A Time Synchronization Mechanism Based on Partial Clustering and the Doppler Effect for Underwater Acoustic Networks.

Time synchronization is the basis of coordination and cooperation in underwater acoustic networks. However, because of the propagation delay, node mobility, and Doppler shift, it is impossible to balance the accuracy and energy consumption simply in water. As a promising technology, partial clustering has high convergence and makes breakthroughs in time synchronization. This paper proposes PCDE-Sync, a novel synchronization mechanism with partial clustering and the Doppler effect. Firstly, a clustering method built on the artificial fish swarm algorithm is presented. It models the cluster construction according to fish's preying, swarming, and following behaviors. Secondly, we design a synchronization mechanism to conduct clock correction and compensation by the Doppler effect. Finally, we compare the performance of PCDE-Sync with the most advanced protocols, namely MU-Sync, MM-Sync, and DE-Sync, in terms of the cumulative error after synchronization, the mean square error under different clock skew and that under distinctive node mobility, and energy consumption. The experimental results show that PCDE-Sync makes a trade-off between accuracy and complexity, which does well in solving synchronization issues.

Native MAG-1 antibody almost destroys human breast cancer xenografts.

A native form of mouse monoclonal IgG1 antibody called MAG-1, which recognizes an epitope on provasopressin, has been found to shrink and produce extensive necrosis of human breast tumor xenografts in nu/nu mice. We examined the ability of (90)Yttrium-labeled and native MAG-1 to affect the growth in nu/nu mice of cancer xenografts that were estrogen-responsive (from MCF-7 cells) and triple-negative (from MDA-MB231 cells). The growth rates of treated cells were compared to those receiving saline vehicle and those receiving (90)Yttrium-labeled and native forms of the ubiquitous antibody, MOPC21. Short-term treatments (4 doses over 6 days) not only with (90)Yttrium-MAG-1 but also native MAG-1 produced large reductions in size of rapidly growing tumors of both types, while both (90)Yttrium- MOPC21 and native MOPC21 had no effect. Native and (90)Yttrium-MAG-1 effects were similar, and arrested tumors recommenced growing soon after treatments stopped. Increasing native MAG-1 treatment to single dosing for 16 consecutive days shrank tumors of both types with no regrowth apparent over a 20-day post-treatment period of observation. Pathological examination of such tumors revealed they had undergone very extensive (>66%) necrosis.

Immune classification and identification of prognostic genes for uveal melanoma based on six immune cell signatures.

Cutaneous melanoma could be treated by immunotherapy, which only has limited efficacy on uveal melanoma (UM). UM immunotyping for predicting immunotherapeutic responses and guiding immunotherapy should be better understood. This study identified molecular subtypes and key genetic markers associated with immunotherapy through immunosignature analysis. We screened a 6-immune cell signature simultaneously correlated with UM prognosis. Three immune subtypes (IS) were determined based on the 6-immune cell signature. Overall survival (OS) of IS3 was the longest. Significant differences of linear discriminant analysis (LDA) score were detected among the three IS types. IS3 with the highest LDA score showed a low immunosuppression. IS1 with the lowest LDA score was more immunosuppressive. LDA score was significantly negatively correlated with most immune checkpoint-related genes, and could reflect UM patients' response to anti-PD1 immunotherapy. Weighted correlation network analysis (WGCNA) identified that salmon, purple, yellow modules were related to IS and screened 6 prognostic genes. Patients with high-expressed NME1 and TMEM255A developed poor prognosis, while those with high-expressed BEX5 and ROPN1 had better prognosis. There was no notable difference in OS between patients with high-expressed LRRN1 and ST13 and those with low-expressed LRRN1 and ST13. NME1, TMEM255A, Bex5 and ROPN1 showed potential prognostic significance in UM.

Breast cancer expresses functional NMDA receptors.

We demonstrate here that functional NMDAR1 and NMDAR2 receptors are expressed by Mcf-7 and SKBR3 breast cancer cell lines, and possibly by most or all high-grade breast tumors, and that these receptors are important for the growth of human breast cancer xenografts in mice. RT-PCR demonstrated mRNA for both NMDAR1 and NMDAR2 receptors are expressed in both Mcf-7 and SKBR3 cell lines, and these messages likely have sequences identical to those reported for human mRNAs. Proteins of the expected respective sizes 120 and 170 kD are generated from these mRNAs by the tumor cells. Cell growth was found to be significantly (P < 0.0001) impaired down to 10% of normal growth by the irreversible NMDAR1 antagonists MK-801 and memantine with IC 50s ranging from 600 to >800 microM and from 200 to 300 microM for the two lines. Paradoxically, memantine with a lower binding affinity had the greater influence of the two inhibitors on cell viability. Immunohistochemical examination of high-grade invasive ductal and lobular breast cancer with our polyclonal antibodies against a peptide (-Met-Ser-Ile-Tyr-Ser-Asp-Lys-Ser-Ile-His-) in the extracellular domain of the NMDAR1 receptor gave specific positive staining for the receptor in all 10 cases examined. Positive staining was chiefly concentrated at the membranes of these tumor tissues. No staining with these antibodies was found for normal breast and kidney tissues. When Mcf-7 cells were grown as tumor xenografts in nu/nu mice, the growth of these tumors was completely arrested by daily treatments with MK-801 over 5 days. All of these data point to active NMDAR receptors being expressed by most breast cancers, and having an important influence on their survival.

Demographic and clinical characteristics of a paediatric low vision population in a low vision clinic in China.

PURPOSE: The aim was to describe the characteristics of the paediatric population attending the low vision clinic of the Eye and ENT Hospital, located in Shanghai, China. METHODS: The clinical records of all the children attending the low vision clinic of Eye and ENT Hospital affiliated to Fudan University between January 1, 2009 and May 31, 2014 were retrospectively reviewed. The main data analysed were age, gender, education, visual demand, diagnosis, visual acuity and prescription of low vision aids. RESULTS: Of the 162 patients, 104 (64.20 per cent) were male. The age range of the study population was three to 20 years, with a mean of 10.73 ± 5.08 years. There were 43.21 per cent with moderate visual impairment, 26.54 per cent had severe visual impairment and 19.75 per cent were blind. The leading causes of visual impairment were congenital cataract (21.61 per cent), optic atrophy (14.20 per cent), macular dystrophy (11.73 per cent), nystagmus (9.88 per cent) and congenital glaucoma (9.26 per cent). The most frequently prescribed low vision devices for distant and near vision were binocular telescopes (23.57 per cent) and stand magnifiers (22.93 per cent), respectively. Young age (up to six years, 37.93 per cent), high cost (24.14 per cent), cosmetic reasons (17.24 per cent) and inconvenience (13.79 per cent) were the main reasons that children or parents refused to accept useful low vision aids. CONCLUSIONS: Congenital and hereditary diseases constituted the major causes of low vision in the study population. Strategies that make good-quality rehabilitation services available, affordable and accessible, especially in developing countries, will have the greatest impact on visual impairment. In China, both urban and rural, the coverage of low vision services should be strengthened.

Impact of long-term soft contact lens wear on epithelial flap production and postoperative recovery in laser-assisted subepithelial keratomileusis.

PURPOSE: To investigate the impact and postoperative clinical recovery of long-term soft contact lens wear on the epithelial flap made during laser-assisted subepithelial keratomileusis (LASEK). METHODS: In a prospective study, 371 patients (589 eyes) who underwent LASEK were divided into four groups (G1, G2, G3, G4) according to their length of soft contact lens wear. After the contact lens (CL) was removed 1 week after surgery, various symptoms - uncorrected visual acuity (UCVA), oedema of the corneal epithelium, spherical equivalent (SE) and haze degree - were recorded on day 1, and at 1 and 3 months postoperatively. RESULTS: There were no significant differences in corneal flap production among the first three groups that wore CLs, but various symptoms and UCVA were all different from the fourth group that did not wear CLs. There were statistically significant differences in oedema of corneal epithelium among the first three groups, and the degree of oedema was positively correlated with the CL wearing time. There were no significant differences in postoperative SE and haze in all four groups. CONCLUSIONS: Long-term soft CL wear can affect production of the epithelial flap and postoperative recovery, including various symptoms, oedema of the central corneal epithelium and visual acuity. In contrast, there was no effect of long-term CL wear on postoperative mean refractive spherical equivalent (MRSE) and haze.

NMDA receptors are expressed by small-cell lung cancer and are potential targets for effective treatment.

We previously showed that functional N-methyl-D-aspartate (NMDA) receptors are expressed by human neuroblastoma cells. In this study we demonstrate functional NMDAR1 and NMDAR2 receptors are expressed by small-cell lung cancer (SCLC) classical cell lines NCI H146, NCI H345, and DMS 53, by variant cell line NCI H82, and by most SCLC tumors, and that these receptors are important for the growth of human SCLC tumor xenografts in mice. Reverse transcription-polymerase chain reaction demonstrated mRNA for both receptors, with sequences identical to those for human mRNAs, are expressed in all four cell lines, and these generated proteins of the expected sizes 120 and 170 kDa. Cell viability tests showed cell growth was significantly (P < 0.0001) impaired by NMDAR1 antagonists MK-801 and memantine. Ifenprodil and Ro25-6981, NMDAR2B antagonists at the polyamine site, also significantly (P < 0.001) inhibited the growth/survival of these cells. Alternatively, the glycine-binding antagonist, L701, 324, increased viability to 140% and 120% in NCI H345 and NCI H82 cells after 48 hours of incubation. Immunohistochemistry of SCLC tumors with our polyclonal antibodies gave specific positive staining for the NMDAR1 receptor in 8 of 10 tissues examined. Small amounts of these same antibodies significantly reduced the growth of NCI-H345 cells up to 25% (P < 0.001). When NCI H345 cells were grown as tumor xenografts in mice, the growth of these tumors was reduced by 60% (P < 0.001) by treatments with MK-801 over five days. All of these data point to active NMDAR receptors possibly having an important influence on SCLC growth and survival.

Role of bradykinin B1 and B2 receptors in normal blood pressure regulation.

With inhibition or absence of the bradykinin B2 receptor (B2R), B1R is upregulated and assumes some of the hemodynamic properties of B2R, indicating that both participate in the maintenance of normal vasoregulation or to development of hypertension. Herein we further evaluate the role of bradykinin in normal blood pressure (BP) regulation and its relationship with other vasoactive factors by selectively blocking its receptors. Six groups of Wistar rats were treated for 3 wk: one control group with vehicle alone, one with concurrent administration of B1R antagonist R-954 (70 microg x kg(-1) x day(-1)) and B2R antagonist HOE-140 (500 microg x kg(-1) x day(-1)), one with R-954 alone, one with HOE 140 alone, one with concurrent administration of both R-954 and HOE-140 plus the angiotensin antagonist losartan (5 mg x kg(-1) x day(-1)), and one with only losartan. BP was measured continuously by radiotelemetry. Only combined administration of B1R and B2R antagonists produced a significant BP increase from a baseline of 107-119 mmHg at end point, which could be partly prevented by losartan and was not associated with change in catecholamines, suggesting no involvement of the sympathoadrenal system. The impact of blockade of bradykinin on other vasoregulating systems was assessed by evaluating gene expression of different vasoactive factors. There was upregulation of the eNOS, AT1 receptor, PGE2 receptor, and tissue kallikrein genes in cardiac and renal tissues, more pronounced when both bradykinin receptors were blocked; significant downregulation of AT2 receptor gene in renal tissues only; and no consistent changes in B1R and B2R genes in either tissue. The results indicate that both B1R and B2R contribute to the maintenance of normal BP, but one can compensate for inhibition of the other, and the chronic inhibition of both leads to significant upregulation in the genes of related vasoactive systems.