Exploring the JAK/STAT Signaling Pathway in Hepatocellular Carcinoma: Unraveling Signaling Complexity and Therapeutic Implications.

Hepatocellular Carcinoma (HCC) continues to pose a substantial global health challenge due to its high incidence and limited therapeutic options. In recent years, the Janus Kinase (JAK) and Signal Transducer and Activator of Transcription (STAT) pathway has emerged as a critical signaling cascade in HCC pathogenesis. The review commences with an overview of the JAK/STAT pathway, delving into the dynamic interplay between the JAK/STAT pathway and its numerous upstream activators, such as cytokines and growth factors enriched in pathogenic livers afflicted with chronic inflammation and cirrhosis. This paper also elucidates how the persistent activation of JAK/STAT signaling leads to diverse oncogenic processes during hepatocarcinogenesis, including uncontrolled cell proliferation, evasion of apoptosis, and immune escape. In the context of therapeutic implications, this review summarizes recent advancements in targeting the JAK/STAT pathway for HCC treatment. Preclinical and clinical studies investigating inhibitors and modulators of JAK/STAT signaling are discussed, highlighting their potential in suppressing the deadly disease. The insights presented herein underscore the necessity for continued research into targeting the JAK/STAT signaling pathway as a promising avenue for HCC therapy.

Adaptive horizontal transfer of a bacterial gene to an invasive insect pest of coffee.

Horizontal gene transfer (HGT) involves the nonsexual transmission of genetic material across species boundaries. Although often detected in prokaryotes, examples of HGT involving animals are relatively rare, and any evolutionary advantage conferred to the recipient is typically obscure. We identified a gene (HhMAN1) from the coffee berry borer beetle, Hypothenemus hampei, a devastating pest of coffee, which shows clear evidence of HGT from bacteria. HhMAN1 encodes a mannanase, representing a class of glycosyl hydrolases that has not previously been reported in insects. Recombinant HhMAN1 protein hydrolyzes coffee berry galactomannan, the major storage polysaccharide in this species and the presumed food of H. hampei. HhMAN1 was found to be widespread in a broad biogeographic survey of H. hampei accessions, indicating that the HGT event occurred before radiation of the insect from West Africa to Asia and South America. However, the gene was not detected in the closely related species H. obscurus (the tropical nut borer or "false berry borer"), which does not colonize coffee beans. Thus, HGT of HhMAN1 from bacteria represents a likely adaptation to a specific ecological niche and may have been promoted by intensive agricultural practices.

Mining secreted proteins that function in pepper fruit development and ripening using a yeast secretion trap (YST).

Plant cells secrete diverse sets of constitutively- and conditionally-expressed proteins under various environmental and developmental states. Secreted protein populations, or secretomes have multiple functions, including defense responses, signaling, metabolic processes, and developmental regulation. To identify genes encoding secreted proteins that function in fruit development and ripening, a yeast secretion trap (YST) screen was employed using pepper (Capsicum annuum) fruit cDNAs. The YST screen revealed 80 pepper fruit-related genes (CaPFRs) encoding secreted proteins including cell wall proteins, several of which have not been previously described. Transient GFP-fusion assay and an in planta secretion trap were used to validate the secretion of proteins encoded by selected YST clones. In addition, RNA gel blot analyses provided further insights into their expression and regulation during fruit development and ripening. Integrating our data, we conclude that the YST provides a valuable functional genomics tool for the identification of substantial numbers of novel secreted plant proteins that are associated with biological processes, including fruit development and ripening.

A secreted effector protein (SNE1) from Phytophthora infestans is a broadly acting suppressor of programmed cell death.

Evasion or active suppression of host defenses are critical strategies employed by biotrophic phytopathogens and hemibiotrophs whose infection mechanism includes sequential biotrophic and necrotrophic stages. Although defense suppression by secreted effector proteins has been well studied in bacteria, equivalent systems in fungi and oomycetes are poorly understood. We report the characterization of SNE1 (suppressor of necrosis 1), a gene encoding a secreted protein from the hemibiotrophic oomycete Phytophthora infestans that is specifically expressed at the transcriptional level during biotrophic growth within the host plant tomato (Solanum lycopersicum). Using transient expression assays, we show that SNE1 suppresses the action of secreted cell death-inducing effectors from Phytophthora that are expressed during the necrotrophic growth phase, as well as programmed cell death mediated by a range of Avr-R protein interactions. We also report that SNE1 contains predicted NLS motifs and translocates to the plant nucleus in transient expression studies. A conceptual model is presented in which the sequential coordinated secretion of antagonistic effectors by P. infestans first suppresses, but then induces, host cell death, thereby providing a highly regulated means to control the transition from biotrophy to necrotrophy.

Effect of the Lapping Platen Groove Density on the Characteristics of Microabrasive-Based Lapping.

Microabrasive-based lapping is widely used in the manufacturing of single-crystal substrates such as sapphire, SiC, and GaN. Although many studies have been conducted to improve the lapping process characteristics, most of them focused on process conditions or consumables. In this study, the effect of the lapping platen groove density on the lapping characteristics was studied using a sapphire substrate. Groove density was defined as the ratio of groove width to groove pitch, and the displacement of the lapping head was measured to calculate the oil film thickness. It was confirmed that, for groove densities below 0.30, hydroplaning occurs when the oil film thickness increases. When the oil film thickness is larger than the abrasive particle size, the material removal rate is low because the abrasive does not participate in the lapping process. When the oil film was developed, the experimental results showed a high surface roughness and poor flatness of the substrate, as only large abrasive particles participated in the lapping process. Therefore, to improve the lapping characteristics, it is important to reduce the groove density by reducing the groove pitch, which prevents the development of the oil film.

Comparison of daily physical activity parameters using objective methods between overweight and normal-weight children.

PURPOSE: The purpose of the present study was to determine if there were any differences in various aspects of physical activity such as energy expenditure, intensity, and type of activity between normal-weight and overweight boys. METHODS: Children aged 9-12 years were recruited from 2 elementary schools located in different urban districts in Republic of Korea. The present study included 45 Korean boys, of which 19 were normal-weight and 26 were overweight. Daily physical activity was estimated over the same 1-week study period under free-living conditions using the doubly labeled water (DLW) method and a tri-axial accelerometer. Resting metabolic rate (RMR) was measured using the Douglas bag method and open-circuit indirect calorimetry. We calculated the physical activity level (PAL) as the total energy expenditure (TEE)/RMR. RESULTS: PAL was not significantly different between the groups. In the accelerometer data, time spent in locomotive moderate-to-vigorous physical activity (MVPA) was significantly lower in overweight boys than in normal-weight subjects, whereas other variables including non-locomotive activity did not differ between groups. In addition, among all participants, time spent in total locomotive activity was significantly associated with PAL. Time spent in locomotive MVPA was significantly associated with PAL. CONCLUSION: Overweight boys may be less physically active based on locomotive MVPA, which was positively related with PAL. Our findings suggest that the contribution of locomotive MVPA to the increase in PAL was relatively significant.

Erratum: Validation of Dietary Reference Intakes for predicting energy requirements in elementary school-age children.

[This corrects the article on p. 336 in vol. 12, PMID: 30090171.].

Validation of Dietary Reference Intakes for predicting energy requirements in elementary school-age children.

BACKGROUND/OBJECTIVES: Dietary Reference Intakes (DRI) for energy are derived from total energy expenditure (TEE) measured using the doubly labelled water (DLW) method. The objective of this study was to assess the validity of DRI for predicting the energy requirements of elementary school-age children. SUBJECTS/METHODS: The present study involved 25 elementary school-age children aged between 9 and 11 years. TEE was assessed by the DLW method, and the results were compared with the TEE predicted by the DRI equations in order to evaluate accuracy. RESULTS: The subjects' TEE measured by the DLW method was 1,925.2 ± 380.9 kcal/day in boys and 1,930.0 ± 279.4 kcal/day in girls, whereas resting energy expenditure was 1,220.2 ± 176.9 kcal/day in boys and 1,245.9 ± 171.3 kcal/day for girls. The physical activity level was 1.58 ± 0.20 in boys and 1.55 ± 0.13 in girls. The mean bias between the predicted and measured TEE was 12.6% in boys and -1.6% in girls, and the percentage of accurate predictions was 28.6% and 63.6%, respectively. In boys, the equation resulted in underprediction of TEE among the subjects having low TEE values, whereas there was overprediction among subjects having high TEE values as shown by the Bland-Altman plot. On the contrary, this proportional bias was not observed in girls. CONCLUSIONS: The findings of this study suggest that the DRI equation for energy could result in the overestimation of energy requirements in elementary school-age boys. In the case of girls, the equations could be accurate at the group level. However, the DRI appears to be invalid for individual girls, as more than one third of girls had their TEE inaccurately predicted. We recommend more studies for confirmation of these results.

A functional screen to characterize the secretomes of eukaryotic pathogens and their hosts in planta.

Complex suites of proteins that are secreted by plants and phytopathogens into the plant apoplast play crucial roles in surveillance, assault, defense, and counter-defense. High-throughput genome-scale strategies are being developed to better understand the nature of these "secretomes" and the identity of pathogen-derived effector proteins that subvert plant defenses and promote pathogenicity. Although combined bioinformatic and experimental approaches recently have provided comprehensive coverage of secreted proteins from bacterial phytopathogens, far less is known about the secretomes and batteries of effectors of eukaryotic phytopathogens; notably fungi and oomycetes. The yeast secretion trap (YST) represents a potentially valuable technique to simultaneously target pathogen and host secretomes in infected plant material. A YST screen, using a new vector system, was applied to study the interaction between tomato (Solanum lycopersicum) and the oomycete Phytophthora infestans, revealing sets of genes encoding secreted proteins from both pathogen and host. Most of those from the oomycete had no identifiable function and were detectable in planta only during pathogenesis, underlining the value of YST as a tool to identify new candidate effectors and pathogenicity factors. In addition, the majority of the P. infestans proteins had homologs in the genomes of the related oomycetes R. sojae and P. ramorum.

Association between daily step counts and physical activity level among Korean elementary schoolchildren.

PURPOSE: The purpose of the current study was to investigate steps per day (steps/d) and physical activity level (PAL) in Korean elementary school children having normal weight (normal-weight). We also clarified whether a gender difference exited between steps/d and PAL. METHODS: Children aged 9 to 12 y were recruited from two elementary schools located in different urban districts in Korea. The present study included 33 Korean children, of which 18 were normal-weight boys and 15 were normal-weight girls. During the same 1 week study period under free-living conditions the total energy expenditure (TEE) and step counts were estimated using the doubly labeled water (DLW) method and an accelerometer, respectively. We calculated PAL as the TEE/ resting metabolic rate. RESULTS: The range of PAL was 1.25 - 1.93 with a mean value of 1.57. None of the variables of energy expenditure was significantly different by sex. However, steps/d were significantly higher in boys than in girls. When adjusting regression analysis by gender, steps/ d were positively associated with PAL among all subjects (r = 0.56, P < 0.01). Furthermore, steps/d were positively associated with PAL in boys (r = 0.68, P < 0.01), but not in girls (r = 0.27, P = 0.34). CONCLUSION: Our results suggest that locomotive activity may be the main contributor to the individual PAL differences for elementary school boys, while non-locomotive activity may be the main contributor for elementary school girls.

Digging deeper into the plant cell wall proteome.

The proteome of the plant cell wall/apoplast is less well characterized than those of other subcellular compartments. This largely reflects the many technical challenges involved in extracting and identifying extracellular proteins, many of which resist isolation and identification, and in capturing a population that is both comprehensive and relatively uncontaminated with intracellular proteins. However, a range of disruptive techniques, involving tissue homogenization and subsequent sequential extraction and non-disruptive approaches has been developed. These approaches have been complemented more recently by other genome-scale screens, such as secretion traps that reveal the genes encoding proteins with N-terminal signal peptides that are targeted to the secretory pathway, many of which are subsequently localized in the wall. While the size and complexity of the wall proteome is still unresolved, the combination of experimental tools and computational prediction is rapidly expanding the catalog of known wall-localized proteins, suggesting the unexpected extracellular localization of other polypeptides and providing the basis for further exploration of plant wall structure and function.

Individuality of handwriting.

Motivated by several rulings in United States courts concerning expert testimony in general, and handwriting testimony in particular, we undertook a study to objectively validate the hypothesis that handwriting is individual. Handwriting samples of 1,500 individuals, representative of the U.S. population with respect to gender, age, ethnic groups, etc., were obtained. Analyzing differences in handwriting was done by using computer algorithms for extracting features from scanned images of handwriting. Attributes characteristic of the handwriting were obtained, e.g., line separation, slant, character shapes, etc. These attributes, which are a subset of attributes used by forensic document examiners (FDEs), were used to quantitatively establish individuality by using machine learning approaches. Using global attributes of handwriting and very few characters in the writing, the ability to determine the writer with a high degree of confidence was established. The work is a step towards providing scientific support for admitting handwriting evidence in court. The mathematical approach and the resulting software also have the promise of aiding the FDE.

A yeast secretion trap assay for identification of secreted proteins from eukaryotic phytopathogens and their plant hosts.

Secreted proteins from plants and phytopathogens play important roles in their interactions and contribute to elaborate mechanisms of attack, defense, and counter-defense, as well as surveillance and signaling. There is therefore considerable interest in developing techniques to characterize "secretomes." Here, we describe the use of the yeast secretion trap (YST) functional screen to isolate and identify secreted proteins that are accumulated and detected in the extracellular matrix of eukaryotes. This method involves fusing cDNAs generated or derived from plants, pathogens, or infected tissue to a yeast (Saccharomyces cerevisiae) invertase (suc2) reporter gene lacking its signal peptide, transforming the resulting fusion library into an invertase-deficient yeast strain, and plating the transformants on a sucrose selection medium. A yeast transformant containing a cDNA that encodes a secreted protein can rescue the mutant and the plasmid DNA can then be sequenced to identify the secreted protein. The YST screen can be a very powerful tool in the study of dynamics of plant host-pathogen interactions.

Accuracy of five implant impression technique: effect of splinting materials and methods.

PURPOSE: The aim of this study was to evaluate the effect of dimensional stability of splinting material on the accuracy of master casts. MATERIALS AND METHODS: A stainless steel metal model with 6 implants embedded was used as a master model. Implant level impressions were made after square impression copings were splinted using 5 different techniques as follows. (1) Splinted with autopolymerizing resin and sectioned, reconnected to compensate polymerization shrinkage before the impression procedure. (2) Splinted with autopolymerizing resin just before impression procedure. (3) Primary impression made with impression plaster and secondary impression were made over with polyether impression material. (4) Splinted with impression plaster. (5) Splinted with VPS bite registration material. From master model, 5 impressions and 5 experimental casts, total 25 casts were made for each of 5 splinting methods. The distortion values of each splinting methods were measured using coordinate measuring machine, capable of recordings in the x-, y-, z-axes. A one-way analysis of variance (ANOVA) at a confidence level of 95% was used to evaluate the data and Tukey's studentized range test was used to determine significant differences between the groups. RESULTS: Group 1 showed best accuracy followed by Group 3 & 4. Group 2 and 5 showed relatively larger distortion value than other groups. No significant difference was found between group 3, 4, 5 in x-axis, group 2, 3, 4 in y-axis and group 1, 3, 4, 5 in z-axis (P<.0001). CONCLUSION: Both Splinting impression copings with autopolymerizing resin following compensation of polymerization shrinkage and splinting method with impression plaster can enhance the accuracy of master cast and impression plaster can be used simple and effective splinting material for implant impression procedure.

Characterization of the plant cell wall proteome using high-throughput screens.

Plant cell wall proteins play essential roles in many important biological processes, and yet there is still not a comprehensive catalogue of the cell wall proteome, or "secretome". Here, we describe three procedures, including a yeast secretion trap (YST), Agrobacterium-mediated transient expression using a necrosis-inducing protein (NIP) and protein localization assay using a fluorescent protein, to identify and confirm the localization of cell wall proteins. The approaches are orthogonal and collectively provide a powerful suite of approaches to complement more commonly used strategies to isolate plant cell wall-associated proteins.

Mediation of the transition from biotrophy to necrotrophy in hemibiotrophic plant pathogens by secreted effector proteins.

Hemibiotrophs, such as Phytophthora infestans, exhibit distinct phases of their life cycle: an early asymptomatic biotrophic phase and a late necrotrophic stage that is characterized by tissue degradation and disease symptoms. To date, little is known of the molecular mechanisms that promote each distinct phase, nor those that mediate the transition between the two. We hypothesized that these phytopathogens might secrete distinct classes of effector proteins that first suppress plant defense responses and associated programmed cell death (PCD), and later induce large scale necrosis. To this end, we have identified proteins that are secreted by P. infestans early or late in the infection cycle. Recently we described the characterization of SNE1, which is specifically expressed during early biotrophic growth in the host plant tomato (Solanum lycopersicum). We found that SNE1 suppresses the action of necrosis-inducing effectors (Nep1-like proteins), including PiNPP1.1 and PsojNIP, which are secreted by Phytophthora during necrotrophic growth, as well as PCD mediated by a broad spectrum of Avr-R protein interactions. This suggests that SNE1 and PiNPP1.1 act antagonistically, thereby providing a highly regulated means to control the transition from biotrophy to necrotrophy.

Identification of eukaryotic secreted and cell surface proteins using the yeast secretion trap screen.

Secreted and cell surface proteins play essential roles in numerous essential biological processes in eukaryotic organisms, but are often more difficult to isolate and identify than proteins that are localized in intracellular compartments. However, several high-throughput 'gene-trap' techniques have been developed to characterize these 'secretomes', including the yeast secretion trap (YST) screen. This method involves fusing cDNA libraries from the tissue or cell type of interest to a yeast (Saccharomyces cerevisiae) invertase reporter gene, transforming the resulting fusion library into an invertase-deficient yeast strain and plating the transformants on a medium containing sucrose as the sole carbon source. A yeast cell with a transgene encoding a secreted or cell surface protein can synthesize a secreted invertase fusion protein that can rescue the mutant, and the plasmid DNA can then be sequenced to identify the gene that encodes it. We describe a recently improved version of this screen, which allows the identification of genes encoding secreted proteins in 1-2 months.

A coupled yeast signal sequence trap and transient plant expression strategy to identify genes encoding secreted proteins from peach pistils.

Many developmental processes and induced plant responses have been identified that are directly or indirectly influenced by wall-localized, or apoplastic, molecular interactions and signalling pathways. The yeast-based signal sequence trap (YSST) is a potentially valuable experimental tool to characterize the proteome of the wall and apoplast, or 'secretome', although few studies have been performed with plants and to date this strategy has not been coupled with a subsequent analysis to confirm extracellular localization of candidate proteins in planta. This current report describes the use of the YSST, together with transient expression of a selection of identified proteins as fusions with the reporter GFP, focusing on the complex extracellular interactions between peach (Prunus persica) pollen and pistil tissues. The coupled YSST and GFP localization assay was also used to confirm the extracellular localization of a recently identified pistil-specific basic RNase protein (PA1), as has been observed with S-RNases that are involved in self-incompatibility. This pilot YSST screen of pollinated and unpollinated pistil cDNAs revealed a diverse set of predicted cell wall-localized or plasma membrane-bound proteins, several of which have not previously been described. Transient GFP-fusion assays and RNA gel blot analyses were used to confirm their subcellular localization and to provide further insights into their expression or regulation, respectively. These results demonstrated that the YSST strategy represents an effective means either to confirm the extracellular localization of a specific candidate secreted protein, as demonstrated here with PA1, or to conduct a screen for new extracellular proteins.