The UVSSA protein is part of a genome integrity homeostasis network with links to transcription-coupled DNA repair and ATM signaling.

SignificanceTranscription-coupled repair (TCR) involves four core proteins: CSA, CSB, USP7, and UVSSA. CSA and CSB are mutated in the severe human neurocutaneous disease Cockayne syndrome. In contrast UVSSA is a mild photosensitive disease in which a mutated protein sequence prevents recruitment of USP7 protease to deubiquitinate and stabilize CSB. We deleted the UVSSA protein using CRISPR-Cas9 in an aneuploid cell line, HEK293, and determined the functional consequences. The knockout cell line was sensitive to transcription-blocking lesions but not sensitive to oxidative agents or PARP inhibitors, unlike CSB. Knockout of UVSSA also activated ATM, like CSB, in transcription-arrested cells. The phenotype of UVSSA, especially its rarity, suggests that many TCR-deficient patients and tumors fail to be recognized clinically.

Patterns and factors associated with dental service utilization among insured people: a data mining approach.

BACKGROUND: Insurance databases contain valuable information related to the use of dental services. This data is instrumental in decision-making processes, enhancing risk assessment, and predicting outcomes. The objective of this study was to identify patterns and factors influencing the utilization of dental services among complementary insured individuals, employing a data mining methodology. METHODS: A secondary data analysis was conducted using a dental insurance dataset from Iran in 2022. The Cross-Industry Standard Process for Data Mining (CRISP-DM) was employed as a data mining approach for knowledge extraction from the database. The utilization of dental services was the outcome of interest, and independent variables were chosen based on the available information in the insurance dataset. Dental services were categorized into nine groups: diagnostic, preventive, periodontal, restorative, endodontic, prosthetic, implant, extraction/surgical, and orthodontic procedures. The independent variables included age, gender, family size, insurance history, franchise, insurance limit, and policyholder. A multinomial logistic regression model was utilized to investigate the factors associated with dental care utilization. All analyses were conducted using RapidMiner Version 2020. RESULTS: The analysis encompassed a total of 654,418 records, corresponding to 118,268 insured individuals. Predominantly, restorative treatments were the most utilized services, accounting for approximately 38% of all services, followed by diagnostic (18.35%) and endodontic (13.3%) care. Individuals aged between 36 and 60 years had the highest rate of utilization for any dental services. Additionally, families comprising three to four members, individuals with a one-year insurance history, people contracted with a 20% franchise, individuals with a high insurance limit, and insured individuals with a small policyholder, exhibited the highest rate of service usage compared to their counterparts. The regression model revealed that all independent variables were significantly associated with the use of dental services. However, the patterns of association varied among different service categories. CONCLUSIONS: Restorative treatments emerged as the most frequently used dental services among insured individuals, followed by diagnostic and endodontic procedures. The pattern of service utilization was influenced by the characteristics of the insured individuals and attributes related to their insurance.

Semen Protein CRISP3 Promotes Reproductive Performance of Boars through Immunomodulation.

Semen proteins play an important role in male reproductive performance and sperm fertilization ability and can be used as potential biomarkers to evaluate male fertility. The role of cysteine-rich secretory protein 3 (CRISP3) in male reproduction remains unknown. This study aimed to investigate the role of CRISP3 in the reproductive performance of boars. Our results showed that the CRISP3 protein content was significantly and positively correlated with boar fertility, sow delivery rate, and litter size. CRISP3 is highly expressed in the bulbourethral gland of adult boars and is enriched in the seminal plasma. It is localized in the post-acrosomal region of the sperm head and migrates to the anterior end of the tail after capacitation. The CRISP3 recombinant protein did not affect sperm motility and cleavage rate, but it significantly downregulated the mRNA expression of inflammatory factors IL-α, IL-1ß, and IL-6 and the protein expression of IL-α and IL-6 in lipopolysaccharide (LPS)-induced RAW264.7 cells, indicating that CRISP3 has an immunomodulatory function. In conclusion, our study suggests that semen CRISP3 protein levels positively correlate with reproductive performance, which may be achieved by regulating immune responses in the female reproductive tract.

Prostate secretory protein 94 inhibits sterol binding and export by the mammalian CAP protein CRISP2 in a calcium-sensitive manner.

Members of the CAP protein superfamily are present in all kingdoms of life and have been implicated in many different processes, including pathogen defense, immune evasion, sperm maturation, and cancer progression. Most CAP proteins are secreted glycoproteins and share a unique conserved αßα sandwich fold. The precise mode of action of this class of proteins, however, has remained elusive. Saccharomyces cerevisiae has three CAP family members, termed pathogen related in yeast (Pry). We have previously shown that Pry1 and Pry2 export sterols in vivo and that they bind sterols in vitro. This sterol binding and export function of yeast Pry proteins is conserved in the mammalian CRISP proteins and other CAP superfamily members. CRISP3 is an abundant protein of the human seminal plasma and interacts with prostate secretory protein of 94 amino acids (PSP94), another major protein component in the seminal plasma. Here we examine whether the interaction between CRISP proteins and PSP94 affects the sterol binding function of CAP family members. We show that coexpression of PSP94 with CAP proteins in yeast abolished their sterol export function and the interaction between PSP94 and CAP proteins inhibits sterol binding in vitro. In addition, mutations that affect the formation of the PSP94-CRISP2 heteromeric complex restore sterol binding. Of interest, we found the interaction of PSP94 with CRISP2 is sensitive to high calcium concentrations. The observation that PSP94 modulates the sterol binding function of CRISP2 in a calcium-dependent manner has potential implications for the role of PSP94 and CRISP2 in prostate physiology and progression of prostate cancer.

A cysteine-rich secretory protein involves in phytohormone melatonin mediated plant resistance to CGMMV.

BACKGROUND: Melatonin is considered to be a polyfunctional master regulator in animals and higher plants. Exogenous melatonin inhibits plant infection by multiple diseases; however, the role of melatonin in Cucumber green mottle mosaic virus (CGMMV) infection remains unknown. RESULTS: In this study, we demonstrated that exogenous melatonin treatment can effectively control CGMMV infection. The greatest control effect was achieved by 3 days of root irrigation at a melatonin concentration of 50 µM. Exogenous melatonin showed preventive and therapeutic effects against CGMMV infection at early stage in tobacco and cucumber. We utilized RNA sequencing technology to compare the expression profiles of mock-inoculated, CGMMV-infected, and melatonin+CGMMV-infected tobacco leaves. Defense-related gene CRISP1 was specifically upregulated in response to melatonin, but not to salicylic acid (SA). Silencing CRISP1 enhanced the preventive effects of melatonin on CGMMV infection, but had no effect on CGMMV infection. We also found exogenous melatonin has preventive effects against another Tobamovirus, Pepper mild mottle virus (PMMoV) infection. CONCLUSIONS: Together, these results indicate that exogenous melatonin controls two Tobamovirus infections and inhibition of CRISP1 enhanced melatonin control effects against CGMMV infection, which may lead to the development of a novel melatonin treatment for Tobamovirus control.

[Delivery of epididymis-specific mRNAs from mouse epididymosomes into N2a and TM4 cells].

OBJECTIVE: To investigate whether mouse epididymis-specific mRNAs Adam7 and Crisp1 can be delivered into N2a and TM4 cells, and to provide an experimental basis for exploring the function of epididymal mRNAs. METHODS: Using RT-PCR, we detected the presence of epididymis-specific genes (Adam7, Crisp1, Defb22, Wfdc2, and Wfdc9) in the testis, epididymis, epididymosome and sperm of adult male BALB/c mice as well as in the human testis, seminal vesicles and sperm. We isolated epididymosomes of BALB/c mice by low-speed centrifugation, filtration and ultracentrifugation, fluorescently labeled them by PKH26, co-incubated them for 1 hour with the N2a and TM4 cells after 24 hours of starvation culture, and observed whether they were fused with the N2a and TM4 cells and ingested using the epididymosomes without PKH26 labeling, PKH26 dye without epididymosomes, and non- epididymosome or -PKH26 dye as controls. Then we detected the epididymis-specific genes in the N2a and TM4 cells after 1-hour co-incubation by RT-PCR. RESULTS: Adam7 and Crisp1 were present in the mouse epididymis, epididymosomes and sperm, and in the human seminal vesicles and sperm as well, but not in the testes of either the mice or men. PKH26 and Hoechst33258 fluorescence double-labeling showed that the mouse epididymosomes were fused with the N2a and TM4 cells and ingested; RT-PCR revealed the mRNAs of Adam7 and Crisp1 in the N2a and TM4 cells after 1-hour co-incubation; and Western blot exhibited the CRISP1 protein in the N2a and TM4 cells incubated with epididymosomes. CONCLUSION: Epididymosomes can deliver epididymis-specific mRNAs Adam7 and Crisp1 into N2a and TM4 cells, where Crisp1 may be translated into proteins, though their function and significance need to be further studied.

Acrylamide mitigation and 2,4-decadienal elimination in potato-crisps using L-proline accompanied by modified processing conditions.

Presence of toxic compounds such as acrylamide and 2,4-decadienal in fried products are dependent on frying temperature and time and the frying oil. Combination treatments such as aqueous pre-treatments of potato slices prior to frying; addition of L-proline to pre-treated samples; moisture reduction of samples pre-frying, replacement of refined soybean oil by deodorized-virgin-coconut oil (DVCO) as frying medium; and modification of frying time-temperature regime, were implemented to mitigate acrylamide and 2,4-decadienal in fried potato-crips, concomitantly enhancing their organoleptic quality. Based on similarity values of % acrylamide mitigation, experimental conditions were classified into four main clusters and the optimized conditions of the combination treatments obtained by central composite rotatable design were: blanching at 70 °C for 20 min; addition of 2% L-proline to pre-treated potato slices; and deep-frying in DVCO under modified frying conditions (140 °C, 5 min) successfully alleviated acrylamide (~ 99%) (confirmed by HR-MS and quantified by RP-HPLC) and 2,4-decadienal (quantified by RP-HPLC) in the fried potato-crisps, improving them sensorically. High Pearson's correlation co-efficient (r = 0.9955) was obtained between sensory scores and texture profile analyses data of the fried crisps. This mitigation strategy can be successfully extrapolated to industrial-scale frying for enhanced safety and sensory appeal of fried products. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05328-6.

The fate of porcine sperm CRISP2 from the perinuclear theca before and after in vitro fertilization†.

In a previous study, we reported that porcine sperm cysteine-rich secretory protein 2 (CRISP2) is localized in the post-acrosomal sheath-perinuclear theca (PT) as reduction-sensitive oligomers. In the current study, the decondensation and removal of CRISP2 was investigated during in vitro sperm capacitation, after both the induction of the acrosome reaction and in vitro fertilization. Confocal immunofluorescent imaging revealed that additional CRISP2 fluorescence appeared on the apical ridge and on the equatorial segment (EqS) of the sperm head following capacitation, likely due to cholesterol removal. After an ionophore A23187-induced acrosome reaction, CRISP2 immunofluorescence disappeared from the apical ridge and the EqS area partly not only owing to the removal of the acrosomal shroud vesicles, but to its presence in a subdomain of EqS. The fate of sperm head CRISP2 was further examined post-fertilization. In vitro matured porcine oocytes were co-incubated with boar sperm cells for 6-8 h and the zygotes were processed for CRISP2 immunofluorescent staining. Notably, decondensation of CRISP2, and thus of the sperm PT, occurred while the sperm nucleus was still fully condensed. CRISP2 was no longer detectable in fertilized oocytes in which sperm nuclear decondensation and paternal pronucleus formation were apparent. This rapid dispersal of CRISP2 in the PT is likely regulated by redox reactions for which its cysteine-rich domain is sensitive. Reduction of disulfide bridges within CRISP2 oligomers may be instrumental for PT dispersal and elimination.

CRISP method with flipped classroom approach in ECG teaching of arrhythmia for trainee nurses: a randomized controlled study.

BACKGROUND: This study aimed to explored the effects of the Cardiac Rhythm Identification for Simple People (CRISP) method with flipped classroom approach for arrhythmia interpretation in electrocardiogram (ECG) by trainee nurses. METHODS: A total of 120 trainee nurses were enrolled and randomly divided into the experimental group and the control group using lecture-based learning method. We observed the effects of the two methods in ECG interpretation training and investigated the students' attitudes toward the teaching practices. RESULTS: After training, the ECG test scores in the experimental group were significantly higher than that of the control group. Six months later, the ECG test scores of the experimental group was still higher. Self-learning enthusiasm, understanding of teaching content, satisfaction of teaching mode, satisfaction of teaching effectiveness, and interest in learning ECG were significantly higher in the experimental group. CONCLUSION: CRISP method with flipped classroom approach is a new and effective mode worth trying in ECG teaching for trainee nurses.

Data-Driven Low-Frequency Oscillation Event Detection Strategy for Railway Electrification Networks.

Low-frequency oscillations (LFO) occur in railway electrification systems due to the incorporation of new trains with switching converters. As a result, the increased harmonic content can cause catenary stability problems under certain conditions. Most of the research published on this topic to date is focused on modelling the event and analysing it using frequency spectrums. However, in recent years, due to the new technologies linked to Big Data (BD) and data mining (DM), a new opportunity to study and detect LFO events by means of machine-learning (ML) methods has emerged. Trains continuously collect data from the most important catenary variables, which offers new resources for analysing this type of event. Therefore, this article presents the design and implementation of a data-driven LFO event detection strategy for AC railway network scenarios. Compared to previous investigations, a new approach to analyse and detect LFO events, based on field data and ML, is presented. To obtain the most appropriate detection approach for the context of this application, on the one hand, this investigation includes a comparison of machine-learning algorithms (support vector machine, logistic regression, random forest, k-nearest neighbours, naïve Bayes) which have been trained with real field data. On the other hand, an analysis of key parameters and features to optimize event detection is also included. Thus, the most significant result of this work is the high metric values of the solution, reaching values above 97% in accuracy and 93% in F-1 score with the random forest algorithm. In addition, the applicability and training of data-driven methods with real field data are demonstrated. This automatic detection strategy can help with speeding up and improving LFO detection tasks that used to be performed manually. Finally, it is worth mentioning that this research has been structured based on the CRISP-DM methodology, established as the de facto approach for industrial DM projects.

A Novel Low-Cost Bio-Sorbent Prepared from Crisp Persimmon Peel by Low-Temperature Pyrolysis for Adsorption of Organic Dyes.

In order to properly reuse food waste and remove various contaminants from wastewater, the development of green, sustainable and clean technologies has demonstrated potential in the efficient inhibition of secondary pollution to the environment. In this study, an economical and green method was used to prepare biochar from crisp persimmon peel (CPP) using flash-vacuum pyrolysis at different temperatures (200-700 °C; referred to as CPP200-CPP700). CPP200 has high polarity, low aromaticity and high oxygen-containing functional groups that exhibit superior MB adsorption capabilities. CPP200 that was prepared at a relatively low temperature of 200 °C exhibited a high adsorption capacity of 59.72 mg/g toward methylene blue (MB), which was relatively higher than that for alizarin yellow R (4.05 mg/g) and neutral red (39.08 mg/g), indicating that CPP200 possesses a higher adsorption selectivity for cationic dyes. Kinetics investigation revealed that the kinetic data of CPP200 for the adsorption of MB was better fitted by a linear pseudo-second-order model. Isothermal studies indicated that the linear Langmuir model was more suitable for describing the adsorption process. The adsorption thermodynamics illustrated that the adsorption of MB onto CPP200 was spontaneous and endothermic. EDS and IR analyses of CPP200 for both pre- and post-adsorption of MB showed that electrostatic interactions between oxygen-containing groups on biochar and target MB dominated the adsorption procedure, in addition to hydrogen bonding interactions. Reusability tests confirmed the excellent regeneration characteristics of CPP200, indicating that CPP200 may be used as a green, sustainable, highly efficient and recyclable adsorbent for the selective removal of cationic organic dyes.

Characterization of different oligomeric forms of CRISP2 in the perinuclear theca versus the fibrous tail structures of boar spermatozoa†.

Mammalian sperm carry a variety of highly condensed insoluble protein structures such as the perinuclear theca, the fibrous sheath and the outer dense fibers, which are essential to sperm function. We studied the role of cysteine rich secretory protein 2 (CRISP2); a known inducer of non-pathological protein amyloids, in pig sperm with a variety of techniques. CRISP2, which is synthesized during spermatogenesis, was localized by confocal immunofluorescent imaging in the tail and in the post-acrosomal region of the sperm head. High-resolution localization by immunogold labeling electron microscopy of ultrathin cryosections revealed that CRISP2 was present in the perinuclear theca and neck region of the sperm head, as well as in the outer dense fibers and the fibrous sheath of the sperm tail. Interestingly, we found that under native, non-reducing conditions CRISP2 formed oligomers both in the tail and the head but with different molecular weights and different biochemical properties. The tail oligomers were insensitive to reducing conditions but nearly complete dissociated into monomers under 8 M urea treatment, while the head 250 kDa CRISP2 positive oligomer completely dissociated into CRISP2 monomers under reducing conditions. The head specific dissociation of CRISP2 oligomer is likely a result of the reduction of various sulfhydryl groups in the cysteine rich domain of this protein. The sperm head CRISP2 shared typical solubilization characteristics with other perinuclear theca proteins as was shown with sequential detergent and salt treatments. Thus, CRISP2 is likely to participate in the formation of functional protein complexes in both the sperm tail and sperm head, but with differing oligomeric organization and biochemical properties. Future studies will be devoted to the understand the role of CRISP2 in sperm protein complexes formation and how this contributes to the fertilization processes.

A Decentralized Fuzzy Rule-Based Approach for Computing Topological Relations between Spatial Dynamic Continuous Phenomena with Vague Boundaries Using Sensor Data.

Sensor networks (SN) are increasingly used for the observation and monitoring of spatiotemporal phenomena and their dynamics such as pollution, noise and forest fires. In multisensory systems, a sensor node may be equipped with different sensing units to observe and detect several spatiotemporal phenomena at the same time. Simultaneous detection of different phenomena can be used to infer their spatial interactions over space and time. For this purpose, decentralized spatial computing approaches have shown their potential for effective reasoning on spatial phenomena within a sensor network. However, in most cases, spatial extents of continuous dynamic phenomena are uncertain, and their relations and interactions cannot be inferred by the existing approaches at the sensor node level. To address this limitation, in this paper, we propose and develop a decentralized fuzzy rule-based spatial reasoning approach to depict the spatial relations that hold between two evolving spatial phenomena with fuzzy boundaries. The proposed method benefits from a more adapted fuzzy-crisp representation of dynamic phenomena observed by SN where each vague phenomenon is composed of five distinguished zones including the kernel, conjecture and exterior zone and their boundaries. For each detected phenomenon, a sensor node will report one of these zones based on its location. Aggregation of the information reported from the sensor nodes allows reasoning on spatial relations between the observed phenomena and their evolution. Such spatial information provides users with more valuable near real-time information on the state of different phenomena that can be used for informed decision-making.

Data Mining for Cardiovascular Disease Prediction.

Cardiovascular diseases (CVDs) aredisorders of the heart and blood vessels and are a major cause of disability and premature death worldwide. Individuals at higher risk of developing CVD must be noticed at an early stage to prevent premature deaths. Advances in the field of computational intelligence, together with the vast amount of data produced daily in clinical settings, have made it possible to create recognition systems capable of identifying hidden patterns and useful information. This paper focuses on the application of Data Mining Techniques (DMTs) to clinical data collected during the medical examination in an attempt to predict whether or not an individual has a CVD. To this end, the CRossIndustry Standard Process for Data Mining (CRISP-DM) methodology was followed, in which five classifiers were applied, namely DT, Optimized DT, RI, RF, and DL. The models were mainly developed using the RapidMiner software with the assist of the WEKA tool and were analyzed based on accuracy, precision, sensitivity, and specificity. The results obtained were considered promising on the basis of the research for effective means of diagnosing CVD, with the best model being Optimized DT, which achieved the highest values for all the evaluation metrics, 73.54%, 75.82%, 68.89%, 78.16% and 0.788 for accuracy, precision, sensitivity, specificity, and AUC, respectively.

Evolutionary analysis of genes coding for Cysteine-RIch Secretory Proteins (CRISPs) in mammals.

BACKGROUND: Cysteine-RIch Secretory Proteins (CRISP) are expressed in the reproductive tract of mammalian males and are involved in fertilization and related processes. Due to their important role in sperm performance and sperm-egg interaction, these genes are likely to be exposed to strong selective pressures, including postcopulatory sexual selection and/or male-female coevolution. We here perform a comparative evolutionary analysis of Crisp genes in mammals. Currently, the nomenclature of CRISP genes is confusing, as a consequence of discrepancies between assignments of orthologs, particularly due to numbering of CRISP genes. This may generate problems when performing comparative evolutionary analyses of mammalian clades and species. To avoid such problems, we first carried out a study of possible orthologous relationships and putative origins of the known CRISP gene sequences. Furthermore, and with the aim to facilitate analyses, we here propose a different nomenclature for CRISP genes (EVAC1-4, "EVolutionarily-analyzed CRISP") to be used in an evolutionary context. RESULTS: We found differing selective pressures among Crisp genes. CRISP1/4 (EVAC1) and CRISP2 (EVAC2) orthologs are found across eutherian mammals and seem to be conserved in general, but show signs of positive selection in primate CRISP1/4 (EVAC1). Rodent Crisp1 (Evac3a) seems to evolve under a comparatively more relaxed constraint with positive selection on codon sites. Finally, murine Crisp3 (Evac4), which appears to be specific to the genus Mus, shows signs of possible positive selection. We further provide evidence for sexual selection on the sequence of one of these genes (Crisp1/4) that, unlike others, is thought to be exclusively expressed in male reproductive tissues. CONCLUSIONS: We found differing selective pressures among CRISP genes and sexual selection as a contributing factor in CRISP1/4 gene sequence evolution. Our evolutionary analysis of this unique set of genes contributes to a better understanding of Crisp function in particular and the influence of sexual selection on reproductive mechanisms in general.

Expression and purification of recombinant mouse CRISP4 using a baculovirus system.

Cysteine-rich secretory protein 4 (CRISP4) is a member of the CAP superfamily protein, is highly expressed in the male reproductive tract and is required for optimal mammalian fertility. CRISPs are characterized by the presence of 16 conserved cysteine residues which forms 8 disulphide bond spread across the N-terminal CAP domain, a hinge region and a C-terminal ion channel regulatory (ICR) domain. Previous attempts to purify recombinant CRISPs as a group have resulted in misfolded and/or insoluble recombinant proteins, protein aggregates or unusable low protein yield. Thus, defining the functions of CRISPs have been impeded. In this study, we report a three-step purification protocol for expression and purification of mouse CRISP4 protein in High Five™ cells using a baculovirus expression system. Recombinant mouse CRISP4 was recognized by western blotting and structurally characterized using Circular Dichroism (CD). Using the protocol described herein, we generated high yields of soluble and correctly folded recombinant mouse CRISP4.

Let-7 microRNA is a critical regulator in controlling the growth and function of silk gland in the silkworm.

The silk gland is characterized by high protein synthesis. However, the molecular mechanisms controlling silk gland growth and silk protein synthesis remain undetermined. Here we demonstrated that CRISPR/Cas9-based knockdown of let-7 or the whole cluster promoted endoreduplication and enlargement of the silk gland, accompanied by changing silk yield, whereas transgenic overexpression of let-7 led to atrophy and degeneration of the silk gland. Mechanistically, let-7 controls cell growth in the silk gland through coordinating nutrient metabolism processes and energy signalling pathways. Transgenic overexpression of pyruvate carboxylase, a novel target of let-7, resulted in enlargement of the silk glands, which is consistent with the abnormal phenotype of the let-7 knockdown. Overall, our data reveal a previously unknown miRNA-mediated regulation of silk gland growth and physiology and shed light on involvement of let-7 as a critical stabilizer and booster in carbohydrate metabolism, which may have important implications for understanding of the molecular mechanism and physiological function of specialized organs in other species.

Purification and Characterization of a New CRISP-Related Protein from Scapharca broughtonii and Its Immunomodulatory Activity.

More and more attention has been paid to bioactive compounds isolated from marine organisms or microorganisms in recent years. At the present study, a new protein coded as HPCG2, was purified from Scapharca broughtonii by stepwise chromatography methods. The molecular weight of HPCG2 was determined to be 30.71 kDa by MALDI-TOF-MS. The complete amino acid sequence of HPCG2 was obtained by tandem mass spectrometry combined with transcriptome database analysis, and its secondary structure was analyzed using circular dichroism. HPCG2 comprised 251 amino acids and contained 28.4% α-helix, 26% ß-sheet, 18.6% ß-turn, and 29.9% random coil. HPCG2 was predicted to be a cysteine-rich secretory protein-related (CRISP-related) protein by domain prediction. Moreover, HPCG2 was proved to possess the immunomodulatory effect on the murine immune cells. MTT assay showed that HPCG2 promoted the proliferation of splenic lymphocytes and the cytotoxicity of NK cells against YAC-1 cells. Flow cytometry test revealed that HPCG2 enhanced the phagocytic function of macrophages and polarized them into M1 type in RAW264.7 cells. In particular, Western blot analysis indicated that the immunomodulatory mechanism of HPCG2 was associated with the regulation on TLR4/JNK/ERK and STAT3 signaling pathways in RAW 264.7 cells. These results suggested that HPCG2 might be developed as a potential immunomodulatory agent or new functional product from marine organisms.

Complex Reporting of the COVID-19 Epidemic in the Czech Republic: Use of an Interactive Web-Based App in Practice.

BACKGROUND: The beginning of the coronavirus disease (COVID-19) epidemic dates back to December 31, 2019, when the first cases were reported in the People's Republic of China. In the Czech Republic, the first three cases of infection with the novel coronavirus were confirmed on March 1, 2020. The joint effort of state authorities and researchers gave rise to a unique team, which combines methodical knowledge of real-world processes with the know-how needed for effective processing, analysis, and online visualization of data. OBJECTIVE: Due to an urgent need for a tool that presents important reports based on valid data sources, a team of government experts and researchers focused on the design and development of a web app intended to provide a regularly updated overview of COVID-19 epidemiology in the Czech Republic to the general population. METHODS: The cross-industry standard process for data mining model was chosen for the complex solution of analytical processing and visualization of data that provides validated information on the COVID-19 epidemic across the Czech Republic. Great emphasis was put on the understanding and a correct implementation of all six steps (business understanding, data understanding, data preparation, modelling, evaluation, and deployment) needed in the process, including the infrastructure of a nationwide information system; the methodological setting of communication channels between all involved stakeholders; and data collection, processing, analysis, validation, and visualization. RESULTS: The web-based overview of the current spread of COVID-19 in the Czech Republic has been developed as an online platform providing a set of outputs in the form of tables, graphs, and maps intended for the general public. On March 12, 2020, the first version of the web portal, containing fourteen overviews divided into five topical sections, was released. The web portal's primary objective is to publish a well-arranged visualization and clear explanation of basic information consisting of the overall numbers of performed tests, confirmed cases of COVID-19, COVID-19-related deaths, the daily and cumulative overviews of people with a positive COVID-19 case, performed tests, location and country of infection of people with a positive COVID-19 case, hospitalizations of patients with COVID-19, and distribution of personal protective equipment. CONCLUSIONS: The online interactive overview of the current spread of COVID-19 in the Czech Republic was launched on March 11, 2020, and has immediately become the primary communication channel employed by the health care sector to present the current situation regarding the COVID-19 epidemic. This complex reporting of the COVID-19 epidemic in the Czech Republic also shows an effective way to interconnect knowledge held by various specialists, such as regional and national methodology experts (who report positive cases of the disease on a daily basis), with knowledge held by developers of central registries, analysts, developers of web apps, and leaders in the health care sector.

Low expression of CRISP3 predicts a favorable prognosis in patients with mammary carcinoma.

The discovery of cysteine-rich secretory protein 3 (CRISP3) has been made in human neutrophils for the first time. Cloning of the complementary DNA (cDNA) for CRISP3 was performed from a cDNA library of human bone marrow. In patients with mammary carcinoma, we found that lower expression of CRISP3 was connected to a significantly improved DFS (disease-free survival) and OS (overall survival). Furthermore, the CRISP3 protein level was significantly associated with negative ANXA1 protein level. In addition, the heterogeneous expression of CRISP3 had been exhibited in diverse mammary carcinoma cells. A significant higher mRNA and the protein level of CRISP3 were seen in T-47D as well as SK-BR-3 cells compared with those in other types of mammary carcinoma cells. Knockdown of CRISP3 in T-47D or SK-BR-3 cells resulted in the weakened migration or invasion abilities. Furthermore, CRISP3 knockdown significantly inhibited the ERK1/2 MAPK signaling pathway in T-47D or SK-BR-3 cells. Research results indicated that the lowering in the expression of CRISP3 is likely to serve as an unprecedented approach for the treatment of mammary carcinoma.