Câncer de pele não melanoma: revisão integrativa / Non-melanoma skin cancer: an integrative review

Introdução: O câncer de pele não melanoma compeende grupo de neoplasias com alta incidência na população mundial. É dividido em carcinoma basocelular e de células escamosas. Por ser de grande prevalência, entender o processo de oncogênese e a relação com íons, proteínas e receptores celulares no câncer de pele não melanoma pode contribuir para que novas terapêuticas sejam avaliadas. Objetivo: Entender o processo da oncogênese dos tumores de pele não melanomas e sua relação com a imunolocalização do IP3R. Métodos: Revisão integrativa da literatura com síntese de evidências. A base de dados foi o PubMed; a estratégia de busca: "carcinoma espinocelular, AND/ OR carcinoma basocelular, AND/OR IP3R, AND/OR imunoistoquímica". Foram considerados para revisão os trabalhos publicados entre 2018 e 2023. Foram incluídos 40 trabalhos, integralmente lidos e resumidos. Resultados: Câncer de pele não melanoma são os tumores malignos mais comuns em todo o mundo, sendo 75-80% o carcinoma basocelular, e até 25% o de células escamosas. As interações moleculares de forma geral, envolvem grande participação de moléculas supressoras tumorais, assim como de procto-oncogenes. Além disso, canais iônicos voltagem dependente controlam o fluxo citosólico de íons, dentre eles o cálcio. O IP3R (receptor do fosfatidil inositol-3) permite a saída de cálcio do retículo endoplasmático para que seja utilizado pela célula para atividades fisiológias como proliferação, angiogênese, motilidade e capacidade de invasão. Conclusão: O IP3R, pelas características de expressão imunoistoquímica, parece estar relacionado também, à fisiopatologia do câncer de pele não melanoma.

Introduction: Non-melanoma skin cancer comprises a group of neoplasms with a high incidence in the world population. It is divided into basal cell carcinoma and squamous cell carcinoma. As it is highly prevalent, understanding the process of oncogenesis and the relationship with ions, proteins and cellular receptors in nonmelanoma skin cancer can contribute to the evaluation of new therapies. Objective: To understand the oncogenesis process of non-melanoma skin tumors and its relationship with the immunolocalization of IP3R. Methods: Integrative literature review with evidence synthesis. The database was PubMed; the search strategy: "squamous cell carcinoma, AND/OR basal cell carcinoma, AND/OR IP3R, AND/OR immunohistochemistry". Works published between 2018 and 2023 were considered for review; 40 works were included, fully read and summarized. Results: Non-melanoma skin cancer is the most common malignant tumor worldwide, with 75-80% being basal cell carcinoma, and up to 25% being cell carcinoma. Molecular interactions in general involve a large participation of tumor suppressor molecules, as well as procto-oncogenes. Furthermore, voltage-dependent ion channels control the cytosolic flow of ions, including calcium. The IP3R (phosphatidyl inositol-3 receptor) allows the exit of calcium from the endoplasmic reticulum so that it can be used by the cell for physiological activities such as proliferation, angiogenesis, motility and invasion capacity. Conclusion: The IP3R, due to its immunohistochemical expression characteristics, appears may also be related to the pathophysiology of nonmelanoma skin cancer.

IN VITRO GERMINATION AND VEGETATIVE PROPAGATION THROUGH BUD DEVELOPMENT OF SACHA INCHI (Plukenetia volubilis) / Germinación y propagación vegetativa in vitro a través del desarrollo de yemas de Sacha Inchi (Plukenetia volubilis)

ABSTRACT This study describes the in vitro seed germination and micropropagation of Plukenetia volubilis (sacha inchi), an oilseed crop rich in omega-3 fatty acids, with health benefits and several industrial applications. Seed germination was evaluated in different culture media (MS and 1/2 MS), seed coat presence/absence and culture temperature (18 °C and 28 °C). Micropropagation was performed using axillary bud development (ABD) on nodal segments from in vitro seedlings. KIN, BAP and 2-ip were evaluated for ABD, and the effect of modified MS in 453 mg L-1 CaCl2 and 351.62 mg L-1 MgSO4 on ABD and shoot survival was assessed to improve the process. Finally, six treatments were evaluated to optimize ABD and shoot leaf formation. Seed germination of 91.6 % was achieved in MS at 28 °C when the seed coat was removed. ABD was obtained in 45 % and 40 % with 0.4 mg L-1 KIN and 0.6 mg L-1 2-ip, respectively, with the least CAL. The modification in 453 mg L-1 CaCl2 then allowed 76 % ABD and 82 % explant survival. ABD response was optimized to 95 % and 2.45 leaves with MS medium + CaCl2 modification + 10 % coconut water + 0.4 mg L-1 KIN. The same results were obtained by replacing the latter with 0.6 mg L-1 2-ip. Rooting was achieved in MS without PGR, and acclimatization was successful. The results indicate that plant production via germination and vegetative propagation is effective for commercial purposes.

RESUMEN Este estudio describe la germinación in vitro y micropropagación de Plukenetia volubilis, un cultivo oleaginoso rico en omega-3 benéfico para la salud y con múltiples aplicaciones industriales. Se evaluó en la germinación diferentes medios de cultivo (MS y 1/2 MS), presencia-ausencia de testa y temperatura de cultivo (18 ° C y 28 ° C). La micropropagación se realizó vía yemas axilares (ABD) de plántulas in vitro. Se evaluó el efecto de KIN, BAP y 2-ip sobre ABD, seguidamente, para mejorar el proceso se evaluó el efecto de MS modificado en 453 mg L-1 CaCl2 y 351.62 mg L-1 MgSO4 sobre ABD y supervivencia del brote. Finalmente, se evaluaron seis tratamientos para optimizar ABD y la formación de hojas. Se logró una germinación 91,6 % en MS a 28 °C cuando se retiró la testa. Se obtuvo 45 % y 40 % de ABD con 0,4 mg L-1 KIN y 0,6 mg L-1 2-ip respectivamente, ambos con la menor CAL. Posteriormente, la modificación de CaCl2 permitió 76 % ABD y 82 % de supervivencia. Se optimizó ABD al 95 % con 2,45 hojas por brote con el medio: MS + modificación de CaCl2 + 10 % de agua de coco + 0.4 mg L-1 KIN, los mismos resultados se obtuvieron cambiando este último con 0,6 mg L-1 2-ip, se logró enraizamiento en MS sin PGR, y la aclimatización fue exitosa. Los resultados indican que la producción de plantas vía germinación y propagación vegetativa es efectiva con fines comerciales.

Peroxidase from foxtail millet bran exerts anti-colorectal cancer activity via targeting cell-surface GRP78 to inactivate STAT3 pathway

Molecular targeted therapy has become an emerging promising strategy in cancer treatment, and screening the agents targeting at cancer cell specific targets is very desirable for cancer treatment. Our previous study firstly found that a secretory peroxidase of class III derived from foxtail millet bran (FMBP) exhibited excellent targeting anti-colorectal cancer (CRC) activity in vivo and in vitro, whereas its underlying target remains unclear. The highlight of present study focuses on the finding that cell surface glucose-regulated protein 78 (csGRP78) abnormally located on CRC is positively correlated with the anti-CRC effects of FMBP, indicating it serves as a potential target of FMBP against CRC. Further, we demonstrated that the combination of FMBP with the nucleotide binding domain (NBD) of csGRP78 interfered with the downstream activation of signal transducer and activator of transcription 3 (STAT3) in CRC cells, thus promoting the intracellular accumulation of reactive oxygen species (ROS) and cell grown inhibition. These phenomena were further confirmed in nude mice tumor model. Collectively, our study highlights csGRP78 acts as an underlying target of FMBP against CRC, uncovering the clinical potential of FMBP as a targeted agent for CRC in the future.

Regulation of calcium oscillations by IP3R and RyR in neonatal rat cardiomyocytes / 西安交通大学学报(医学版)

【Objective】 To explore the roles of 1,4,5-trisphosphate receptor (IP3Rs) and ryanodine receptors (RyRs), the two main Ca2+ release channels in sarcoplasmic reticulum (SR), in the regulation of intracellular Ca2+ oscillations in neonatal rat cardiomyocytes (NRCMs). 【Methods】 We isolated and cultured NRCMs for different days, then loaded them with Ca2+ indicator fura-2 and performed real-time fluorescent imaging. To distinguish the effects of IP3Rs and RyRs, NRCMs were pre-treated with phenylephrine (PE, IP3Rs agonist), caffeine (RyRs agonist), 2-APB (IP3Rs antagonist), and tetracaine (RyRs antagonists), respectively. 【Results】 The cultured monolayer NRCMs showed spontaneous synchronized Ca2+ oscillations. PE activation or 2-APB blockade of IP3Rs increased or reduced the frequency of Ca2+ oscillations in NRCMs, accordingly, with no significant effect on the amplitude of Ca2+ oscillations. Activation of RyRs with caffeine increased the frequency of Ca2+ oscillations, but unsynchronized the intercellular rhythm of calcium release and beating pace, while blocking RyRs with tetracaine completely abolished the Ca2+ oscillations and beats in NRCMs. In addition, the effect of PE stimulation on Ca2+ oscillation frequency gradually decreased along with cultured days. 【Conclusion】 IP3Rs regulate the rhythm of calcium oscillations, whereas RyRs are the main channel for bulky store Ca2+ release.

Intervention of total saponins of Panax japonicas on non-alcoholic steatohepatitis in mice by regulating miR-199/SIRTl/MFN2 / 中国药理学通报

Aim To discuss the effect of miR-199/ SIRT1/MFN2 signaling pathway on the progression of NASH and its related mechanisms.Methods 45 BALB/e mice were randomly divided into normal group, high fat diet(HFD) group, total saponins of panax japonicas ( TSPJ ) low-dose group ( 15 mg • kg-1) and TSPJ high-dose group (45 mg • kg"1 ).Normal group was given normal diet, while HFD group, TSPJ low-dose and high-dose groups were given high-fat diet.The mice were intragastrioally given 15 and 45 mg 'kg"1 TSPJ (dissolved in saline) daily in TSPJ low-dose and high-dose groups, while those in other groups were intragastric ally given the same a- mount of saline daily.After seven months, they were sacrificed for serum collection and hepatic tissue col¬lection.Results HE staining showed that liver lipido¬sis and inflammation were obvious in HFD group.while liver lipidosis anrl inflammation were alleviated in TSPJ group.MFN2 and SIRT1 levels significantly de¬creased.TNF-a, 1L-1 p , SREBP, ChREBP levels sig¬nificantly increased in HFD group.After treated with TSPJ, SIRT1 and MFN2 levels were significantly up- regulated , while TNF-a, IL-ip, ChREBP and SREBP levels were significantly down-regulated.The Immuno¬fluorescence results showed that the fluorescence inten¬sity of MFN2 and SIR 11 increased in TSPJ low-dose and high-dose groups.At mRNA level, miR-199 had a negative regulatory relationship with SIRT1.Conclu¬sions TSPJ can alleviate NASH induced by high fat diet through miR-199/SIRTl/MFN2 signaling path¬way.

Theoretical Study of the Function of the IP3 Receptor/BK Channel Complex in a Single Neuron / Estudio Teórico de la Función del Complejo Receptor-IP3/Canales BK en Neurona Única

ABSTRACT Large conductance calcium-activated potassium (BK) channels carry out many functions in the central nervous system. These channels open in response to increased cytosolic calcium ([Ca2+]cyt) concentration. The influx of calcium ions to the cytosol can occur through voltage-gated calcium channels (VGCCs) on the plasma membrane and/ or through IP3 receptors (IP3-Rs) and ryanodine receptors (RyRs) on the endoplasmic reticulum membrane. The BK channel/IP3-R/RyR interaction has been widely reported in smooth muscle but scarcely investigated in relation to neurons. The aim of this study was to theoretically explore the function of the BK/IP3-R complex by means of a computational model of a neuron that replicates the interaction between the release of Ca2+ from the endoplasmic reticulum (through IP3-Rs) and the opening of the BK channels. The mathematical models are based on the Hodgkin-Huxley formalism and the Goldbeter model. These models were implemented on Visual Basic® and differential equations were solved numerically. Distinct conditions were contemplated for BK conductance and the efflux of endoplasmic Ca2+ to the cytosol. An abrupt rise in [Ca2+]cyt (≥ 5 μM) and short duration (spark) was found to activate BK channels and either pause or stop the action potential train.

RESUMEN Los canales de potasio activados por calcio de gran conductancia (canales BK) cumplen múltiples funciones en el sistema nervioso central. Estos canales se abren en respuesta al incremento de la concentración de calcio citosólico ([Ca2+]cyt). La entrada de Ca2+ puede ocurrir a través de canales de calcio dependientes de voltaje (VGCCs) localizados en la membrana plasmática y por eflujo de Ca2+ del retículo endoplásmico (ER) causado por 1,4,5-Trifosfato (IP3) o rianodina (RyR). La interacción BK/IP3/RyR ha sido ampliamente estudiada en músculo liso, pero escasamente en neuronas. El objetivo de este estudio fue explorar teóricamente la función del complejo BK/IP3-R mediante un modelo computacional de una neurona que replica la interacción entre la liberación de Ca2+ del retículo endoplásmico (a través de IP3-Rs) y la apertura de los canales BK. Los modelos matemáticos se basan en el formalismo de Hodgkin-Huxley y el modelo de Goldbeter. Estos modelos fueron implementados en Visual Basic® y las ecuaciones diferenciales fueron resueltas por métodos numéricos. Se contemplaron distintas condiciones para la conductancia del canal BK y la salida de Ca2+ endoplásmico al citosol. Los resultados muestran que un incremento abrupto de [Ca2+] cyt (≥ 5 μM) y de corta duración (spark) activa los canales BK y producen una pausa o detiene el tren de potenciales de acción.

The ATP Level in the mPFC Mediates the Antidepressant Effect of Calorie Restriction / 神经科学通报·英文版

Food deprivation can rescue obesity and overweight-induced mood disorders, and promote mood performance in normal subjects. Animal studies and clinical research have revealed the antidepressant-like effect of calorie restriction, but little is known about the mechanism of calorie restriction-induced mood modification. Previous studies have found that astrocytes modulate depressive-like behaviors. Inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is the predominant isoform in mediating astrocyte Ca

International communication system of acupuncture-moxibustion based on OSI model / 中国针灸

Borrowing the open system interconnection (OSI) model of internet protocol and the 5W theory in communication studies, the links in the international communication of acupuncture-moxibustion can be classified into seven layers: physical layer, data link layer, network layer, transport layer, session layer, presentation layer and application layer, therefore it is built an OSI model of international communication system of acupuncture-moxibustion. It is pointed out that present international communication system of acupuncture-moxibustion is similar to the user datagram protocol (UDP) in internet technology. Evidence-based medicine (EBM) plays a key role to modernize acupuncture- moxibustion theory based upon its clinical effects. According to phenomenon-taking by classified analogy, it is found that the PICO model of EBM agrees pretty well with the "three-way handshakes" mechanism of the internet transmission control protocol (TCP), which is promising to construct an international discourse of acupuncture-moxibustion compatible with western medicine. Thus it will benefit to explore the scientific connotation of acupuncture-moxibustion theory and significantly improve the international prestige of acupuncture-moxibustion.

Genetic study on a consanguineous Chinese family with premature ovarian insufficiency caused by a missense mutation of PSMC3IP gene / 中国医师杂志

Objective:To explore the genetic etiology for a premature ovarian insufficiency (POI) patient from a consanguineous Chinese family, and to provide basis for genetic counseling and fertility counseling.Methods:Whole-exome sequencing was performed using DNA extracted from the blood sample of POI patient. Suspected pathogenic mutation was analyzed by bioinformatics methods and verified by Sanger sequencing. The pathogenicity of the variation was assessed according to the ACMG genetic variation classification criteria and guidelines.Results:A homozygous variation, c. 32G>T (p.G11V), of PSMC3IP was identified in the patient. Bioinformatics analysis revealed that the variation was conserved in different animal species, and this variation was classified as possible pathogenic variation according to the ACMG genetic variation classification criteria and guidelines.Conclusions:The homozygous missense variation of PSMC3IP is the cause of the POI patient in this family. We are reporting for the first time the missense variation in PSMC3IP gene caused POI, which enriched the mutation spectrum of PSMC3IP and provided the basis for genetic counseling and fertility guidance of this family.

Pharmacologically targeting molecular motor promotes mitochondrial fission for anti-cancer

Mitochondrial shape rapidly changes by dynamic balance of fusion and fission to adjust to constantly changing energy demands of cancer cells. Mitochondrial dynamics balance is exactly regulated by molecular motor consisted of myosin and actin cytoskeleton proteins. Thus, targeting myosin-actin molecular motor is considered as a promising strategy for anti-cancer. In this study, we performed a proof-of-concept study with a natural-derived small-molecule J13 to test the feasibility of anti-cancer therapeutics

The ATP Level in the mPFC Mediates the Antidepressant Effect of Calorie Restriction / 神经科学通报·英文版

Food deprivation can rescue obesity and overweight-induced mood disorders, and promote mood performance in normal subjects. Animal studies and clinical research have revealed the antidepressant-like effect of calorie restriction, but little is known about the mechanism of calorie restriction-induced mood modification. Previous studies have found that astrocytes modulate depressive-like behaviors. Inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is the predominant isoform in mediating astrocyte Ca

Effects of Huangpu Tongqiao Capsules on EGFR-PLCγ signal pathway of hippocampus in rats with Alzheimer's disease / 中国中药杂志

Huangpu Tongqiao Capsules(HPTQC), with the functions of invigorating Qi and kidney, eliminating phlegm and removing blood stasis, have the effect of treating Alzheimer's disease(AD), but its mechanism needs further exploration. To explore the relationship between the therapeutic mechanism of HPTQC on Alzheimer's disease and EGFR-PLCγ signal pathway, 40 healthy male SD rats were selected and divided into 4 groups randomly: sham operation group(sham), model group(model), HPTQC group(HPTQC), and nimodipine group(NMP). AD rat model was established by intraperitoneal injection of D-galactose combined with an intracerebral injection of amyloid-β peptide(25-35). After 28 days of administration, Morris water maze test and HE staining showed that the learning and memory ability of AD rats were significantly decreased(P<0.01), and hippocampal neurons were obviously da-maged. However, HPTQC could improve the learning and memory ability of AD rats(P<0.05) and reduce the damage of hippocampal neurons. Immunofluorescence test results showed that the expression levels of EGFR and p-Tau in hippocampal CA1 region of AD rats were significantly increased(P<0.01), and HPTQC could reduce the expression of EGFR and p-Tau in hippocampus of AD rats(P<0.01). Western blot results showed that the protein expression levels of EGFR, PLCγ, IP3 R and p-Tau in hippocampus of AD rats were significantly increased(P<0.01), and HPTQC could reduce the protein expression of EGFR, PLCγ, IP3 R and p-Tau in AD rats(P<0.05). RT-PCR results showed that the mRNA levels of EGFR, PLCγ, IP3 R and Tau in hippocampus of AD rats were significantly increased(P<0.01), and HPTQC could reduce the mRNA levels of EGFR, PLCγ, IP3 R and Tau in AD rats(P<0.05). The results indicate that HPTQC can improve the learning and memory ability of AD rats, and its mechanism of action may be related to regulating EGFR-PLCγ signal pathway.

Concentrations of cytokinin and the use of agricultural residues in the in vitro propagation media of highbush blueberry / Concentrações de citocinina e uso de resíduos agrícolas no meio de multiplicação in vitro de mirtileiro

ABSTRACT: The objective of the research was to evaluate cytokinin concentrations and agricultural residues as physical structuring agents of the culture medium in the in vitro propagation of highbush blueberry (Vaccinium corymbosum 'Duke'). To that end, three different concentrations of 2-isopentenyladenine (2iP) (0, 5, and 10 mg L-1)in woody plant medium (WPM) and five structuring agents (in natura rice husks, carbonized rice husks, coconut fiber, S-10 Beifort®, and agar)were tested. After 60days of culture, the following parameters were evaluated: survival (%),sprout and shoot lengths (cm),propagation rate, number of sprouts, and shoot dry mass (g). The substrates used as structuring agents were analyzed in terms of pH and electrical conductivity. The in natura rice husk, S-10 Beifort®, and carbonized rice husk did not differ from the agar in terms of the sprout length. The shoots were longer (1.64 cm) in cultures within natura rice husk than in other treatments. In the absence of 2iP, S-10 Beifort® resulted in the highest propagation rate (2.97). Concerning the number of sprouts, S-10 Beifort®, in natura rice husks, and carbonized rice husks did not differ from agar. However, when the regulator was added, the agar performed better in comparison to the other residues. Thus, in natura rice husks, carbonized rice husks, and S-10 Beifort®are potential physical structuring agents of the culture medium that can be used without 2iP.

RESUMO: O objetivo deste trabalho foi avaliar concentrações de citocinina e resíduos agrícolas como agentes físicos de estruturação do meio na multiplicação in vitro de mirtileiro (Vaccinium corymbosum) 'Duke'. Para isso, foram testadas três concentrações de 2-isopenteniladenina (2iP) (0; 5 e 10 mg L-1) no meio WPM (Wood Plant Media); e cinco agentes estruturantes (casca de arroz in natura, casca de arroz carbonizada, fibra de coco, S-10 Beifort® e ágar). Após 60 dias foram avaliados: sobrevivência (%); comprimento de brotações e de parte aérea (cm); taxa de multiplicação; número de brotações e massa seca de parte aérea (g). Analisou-se os substratos utilizados como agentes estruturantes quanto ao pH e condutividade elétrica. A casca de arroz in natura, o S-10 Beifort®, e a casca de arroz carbonizada não diferiram do ágar quanto ao comprimento de brotações. Na casca de arroz in natura obteve-se o maior comprimento de parte aérea (1,64 cm), diferindo dos demais tratamentos. Na ausência de 2iP o S-10 Beifort® apresentou a maior taxa de multiplicação (2,97). Para o número de brotações o S-10 Beifort®, a casca de arroz in natura e carbonizada não diferiram do ágar. Entretanto, quando acrescido de regulador, o ágar apresenta um desempenho maior em relação aos resíduos. Sendo assim, a casca de arroz in natura, a casca de arroz carbonizada, e o S-10 Beifort® apresentam-se como potencias estruturadores físicos do meio de cultura, podendo ser utilizados sem o uso de 2iP.

Interaction between wheat translationally controlled tumor protein TCTP and SNF1-related protein kinase SnRK1 / 生物工程学报

Translationally controlled tumor proteins (TCTP) and SNF1- related protein kinase (SnRK1) are conserved and widely present in eukaryotic cells. TCTP regulates cell division, plant growth and development, and mediates plant resistance against pathogen infection. SnRK1 participates in a range of physiological processes including sugar metabolism and resistance to abiotic and biotic stresses. Previous work in our laboratory demonstrated that wheat TCTP can respond to Puccinia triticina infection and induce host defense responses. In order to further investigate the mechanism of TaTCTP in wheat resistance to Puccinia triticina infection, we used TAP (tandem affinity purification) and mass spectrometry to screen the potential interactants of TaTCTP. A SNF1- related protein kinase (SnRK1) was identified as a potential interacting protein of TaTCTP. The results of yeast two-hybrid assay showed that TCTP could interact with SnRK1 in yeast, and the yeast carrying TCTP and SnRK1 could grow on SD/-Leu/-Trp/-His/-Ade (SD/-LWHA) medium. The fluorescence signal of the interaction between TCTP and SnRK1 was found to be distributed in the cytoplasm in the Bi-fluorescense complementation experiment. Co-IP experiments further showed that TCTP and SnRK1 could interact in plant cells. This study lays an important foundation for further studying the mechanism of TaTCTP in the interaction between wheat and Puccinia triticina, and it play a great influence on further improving the molecular mechanism of wheat resistant to Puccinia triticina.

Expression of GTF2IP23 in breast cancer and it mediated regulation of GTF2I / 中华肿瘤杂志

Objective@#To investigate the expression level of antisense transcript of pseudogene, general transcription factor Ⅱi psedugen23 (GTF2IP23), in breast cancer and its effect on the host gene general transcription factor Ⅱi (GTF2I).@*Methods@#The expressions of GTF2IP23 and GTF2I were detected in 40 cases of invasive breast cancer tumors and their counterparts by using quantitative real-time polymerase chain reaction (qRT-PCR). The effects of GTF2IP23 on the expression of GTF2I gene and cell proliferation and migration were analyzed by overexpression of GTF2IP23 in breast cancer cells.@*Results@#The expression of GTF2IP23 mRNA in breast cancer tissues was significantly higher than that in adjacent tissues (P<0.001), while the expression of GTF2I mRNA was significantly lower than that in adjacent tissues (P=0.007). The expression of GTF2IP23 was negatively correlated with GTF2I (r=-0.335, P=0.025). The expression of GTF2IP23 in breast cancer cells was significantly higher than in normal breast cells (P<0.01), while GTF2I expression in breast cancer cells was significantly lower than that in normal breast cells (P<0.01). Overexpression of GTF2IP23 in ZR-75-30 cells significantly reduced the expression of GTF2I (P=0.034) and enhanced cell proliferation (P=0.017) and migration (P=0.026) capacity.@*Conclusions@#GTF2IP23 is distinctly upregulated in breast cancer, it inhibits the expression of real gene GTF2I and promotes the proliferation of breast cancer cells.

Aurone derivatives as Vps34 inhibitors that modulate autophagy

We report in this study the identification of a natural product-like antagonist () of Vps34 as a potent autophagy modulator structure-based virtual screening. Aurone derivative strongly inhibited Vps34 activity in cell-free and cell-based assays. Significantly, prevents autophagy in human cells induced either by starvation or by an mTOR inhibitor. modeling and kinetic data revealed that could function as an ATP-competitive inhibitor of Vps34. Moreover, it suppressed autophagy and without inducing heart or liver damage in mice. could be utilized as a new motif for more selective and efficacious antagonists of Vps34 for the potential treatment of autophagy-related human diseases.

Protein-protein interactions of the nicotinamide/nicotinate mononucleotide adenylyltransferase of Leishmania braziliensis

BACKGROUND Nicotinamide adenine dinucleotide (NAD) plays a central role in energy metabolism and integrates cellular metabolism with signalling and gene expression. NAD biosynthesis depends on the enzyme nicotinamide/nicotinate mononucleotide adenylyltransferase (NMNAT; EC: 2.7.7.1/18), in which converge the de novo and salvage pathways. OBJECTIVE The purpose of this study was to analyse the protein-protein interactions (PPI) of NMNAT of Leishmania braziliensis (LbNMNAT) in promastigotes. METHODS Transgenic lines of L. braziliensis promastigotes were established by transfection with the pSP72αneoαLbNMNAT-GFP vector. Soluble protein extracts were prepared, co-immunoprecipitation assays were performed, and the co-immunoprecipitates were analysed by mass spectrometry. Furthermore, bioinformatics tools such as network analysis were applied to generate a PPI network. FINDINGS Proteins involved in protein folding, redox homeostasis, and translation were found to interact with the LbNMNAT protein. The PPI network indicated enzymes of the nicotinate and nicotinamide metabolic routes, as well as RNA-binding proteins, the latter being the point of convergence between our experimental and computational results. MAIN CONCLUSION We constructed a model of PPI of LbNMNAT and showed its association with proteins involved in various functions such as protein folding, redox homeostasis, translation, and NAD synthesis.

High-throughput screening of Methylobacterium extorquens for high production of pyrroloquinoline quinone / 生物工程学报

Pyrroloquinoline quinone (PQQ) is a bacterial dehydrogenase coenzyme. PQQ can promote body growth and regulate the function of free radical level of the body. It could be applied in food, medicine and other fields. Due to the extremely high cost of chemical synthesis, the production of PQQ by microbial fermentation attracted more and more attention. At present, the production titer of PQQ by fermentation method is too low to achieve industrial application. Due to the lack of a thorough understanding of the PQQ biosynthesis and its regulation mechanisms, and the lack of necessary genetic engineering modification methods for wild type strains, metabolic engineering of microorganisms to enhance PQQ production still lacks essential requirements. In this study, a PQQ-producing bacterium, Methylobacterium extorquens I-F2, was employed as a model strain. By integration of Atmospheric and room temperature plasma (ARTP) mutagenesis, flow cytometry sorting and high-throughput screening strategies, optimization of sample preparation and flow sorting process, a high-titer PQQ mutant strain was obtained. The titer of PQQ was increased by 98.02% compared with that of M. extorqunens I-F2. The process described here showed that the combination of the flow cytometry with high-throughput screening method can be used to obtain high-titer mutants more simply and rapidly, compared with genetic engineering and traditional screening methods.

2- Isopentenyladenine in the induction of direct somatic embryogenesis capacity of Coffea arabica L / 2- Isopenteniladenina na capacidade de indução da embriogênese somática direta de Coffea arabica L

ABSTRACT: This study evaluated the effect of different concentrations of 2-isopentenyladenine (2-iP) on the direct somatic embryogenesis capacity of the Mundo Novo cultivar of Coffea arabica. Leaf explants were cultivated with half the MS salt concentration and the addition of sucrose (20gL-1) and 2-iP (0; 2.5; 5; 7.5 and 10µM). The 2-iP doses of 7.5 and 10µM produced the greatest responses with respect to the percentage of explants with embryogenic structures and the size of the embryogenic structures. However, the greatest production of somatic embryos occurred on the explants treated with 10µM of 2-iP, followed by 7.5µM, whereas their production was absent or reduced with 0 and 5µM, respectively.

RESUMO: Este estudo avaliou o efeito de diferentes concentrações de 2-isopentalinadenina (2-iP) na capacidade de embriogênese somática direta da cultivar Mundo Novo de Coffea arabica. Explantes foliares foram cultivados em meio com metade da concentração de sais de MS e adição de sacarose (20gL-1) e de 2-iP (0; 2,5; 5; 7,5 e 10µM). Os tratamentos com 7,5 e 10µM de 2-iP induziram respostas mais elevadas de porcentagem de explantes com estruturas embriogênicas e de tamanho de estrutura embriogênica. Porém, os explantes tratados 10µM de 2-iP tiveram maior produção de embriões somáticos, seguido de 7,5µM, enquanto nos tratamentos controle e com 5µM, as respostas foram respectivamente de ausência e de baixa formação destes.

Screening and verifying the proteins interacting with phosphorylation cluster of DNA-PKcs by yeast two-hybrid assay / 中华放射医学与防护杂志

Objective To screen and verify the proteins interacting with phosphorylation cluster of DNA dependent protein kinase catalytic subunit((DNA-PKcs) by yeast two-hybrid assay.Methods To know the proteins interacting with DNA-PKcs phosphorylation cluster,yeast two-hybrid assay was applied to screen the cDNA library of human hepatic tissue with a previously constructed plasmid pGBKT7-DPC.The positive clones were further identified by PCR,rotary validation and sequence analysis.Then the eukaryotic expression vectors of the bait protein and screened positive clone proteins were constructed and transfected into human embryonic kidney 293T cells to detect whether the proteins could been expressed correctly.At last,the bait protein and screened positive clone proteins were co-transfected into 293T cells and protein interaction was detected with Co-Immunoprecipitation (Co-IP) assay.Results After two rounds of screening using the yeast two-hybrid assay,12 candidate clones were obtained.Then 7 clones with different insert fragments were identified by PCR,and 3 positive proteins interacted with DNA-PKcs phosphorylation cluster were further verified by rotary validation.Sequencing analysis demonstrated that these 3 proteins were MBNL1,SIK2 and YY1AP1,respectively.Accordingly,the eukaryotic expression vectors of bait protein and 3 positive clone proteins were constructed successfully and expressed correctly in 293T ceils.Finally,the Co-IP assay confirmed that these 3 positive clone proteins could interact with DNA-PKcs phosphorylation cluster.Conclusions Proteins interacting with DNA-PKcs phosphorylation cluster are successfully screened and identified.