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Muscle growth stands as a pivotal economic trait within pig production, governed by a complex interplay of multiple genes, each playing a role in its quantitative manifestation. Understanding the intricate regulatory mechanisms of porcine muscle development is crucial for enhancing both pork yield and quality. This study used the GSE99749 dataset downloaded from the GEO database, conducting a detailed analysis of the RNA-seq results from the longissimus dorsi muscle (LD) of Tibetan pigs (TP), Wujin pigs (WJ) and large white pigs (LW) at 60 days of gestation, representing diverse body sizes and growth rates. Comparative analyses between TPvsWJ and TPvsLW, along with differential gene expression (DEG) analysis, functional enrichment analysis, and protein–protein interaction (PPI) network analysis, revealed 1048 and 1157 significantly differentially expressed genes (p < 0.001) in TPvsWJ and TPvsLW, respectively. With stricter screening criteria, 37 DEGs were found to overlap between the 2 groups. PPI analysis identified MYL5, MYL4, and ACTC1 as the three core genes. This article focuses on exploring the MYL4 gene. Molecular-level experimental validation, through overexpression and interference of the MYL4 gene combined with EDU staining experiments, demonstrated that overexpression of MYL4 significantly promoted the proliferation of porcine skeletal muscle satellite cells (PSMSC), while interference with MYL4 inhibited their proliferation. Furthermore, by examining the effects of overexpressing and interfering with the MYL4 gene on the muscle hypertrophy marker Fst gene and the muscle degradation marker FOXO3 gene, the pivotal role of the MYL4 gene in promoting muscle growth and preventing muscle degradation was further confirmed. These findings offer a new perspective on the molecular mechanisms behind porcine muscle growth and development, furnishing valuable data and insights for muscle biology research.

With the rapid development of 3D reconstruction, especially the emergence of algorithms such as NeRF and 3DGS, 3D reconstruction has become a popular research topic in recent years. 3D reconstruction technology provides crucial support for training extensive computer vision models and advancing the development of general artificial intelligence. With the development of deep learning and GPU technology, the demand for high-precision and high-efficiency 3D reconstruction information is increasing, especially in the fields of unmanned systems, human-computer interaction, virtual reality, and medicine. The rapid development of 3D reconstruction is becoming inevitable. This survey categorizes the various methods and technologies used in 3D reconstruction. It explores and classifies them based on three aspects: traditional static, dynamic, and machine learning. Furthermore, it compares and discusses these methods. At the end of the survey, which includes a detailed analysis of the trends and challenges in 3D reconstruction development, we aim to provide a comprehensive introduction for individuals who are currently engaged in or planning to conduct research on 3D reconstruction. Our goal is to help them gain a comprehensive understanding of the relevant knowledge related to 3D reconstruction.

Productively infected cells are generally thought to arise from HIV infection of activated CD4+ T cells, and these infected activated cells are thought to be a recurring source of latently infected cells when a portion of the population transitions to a resting state. We discovered and report here that productively and latently infected cells can instead originate from direct infection of resting CD4+ T cell populations in lymphoid tissues in Fiebig I, the earliest stage of detectable HIV infection. We found that direct infection of resting CD4+ T cells was correlated with the availability of susceptible target cells in lymphoid tissues largely restricted to resting CD4+ T cells in which expression of pTEFb enabled productive infection, and we documented persistence of HIV-producing resting T cells during antiretroviral therapy (ART). Thus, we provide evidence of a mechanism by which direct infection of resting T cells in lymphoid tissues to generate productively and latently infected cells creates a mechanism by which the productively infected cells can replenish both populations and maintain two sources of virus from which HIV infection can rebound, even if ART is instituted at the earliest stage of detectable infection.

In recent years, the concept of the negative resistor (NR) has been proposed and applied in the magnetic coupled wireless power transfer (MC-WPT) system to improve the stability of power transfer. However, it remains a fundamental challenge to find the characteristic differences and applications of the four basic NR-based MC-WPT topologies. In this paper, the circuit models of the four basic NR-based MC-WPT topologies: series-series (SS), series-parallel (SP), parallel-parallel (PP), and parallel-series (PS) are established, and their characteristics are analyzed and compared by using circuit theory, which enables engineers to determine and select topologies to suit different applications and requirements. Theoretical analysis shows that the compensation methods of the transmitter and receiver of the NR-based MC-WPT system would affect the operating frequency and output characteristics of the system. Moreover, by comparative analysis, the conditions and applicable ranges for a stable output of different topologies were provided. Finally, the experimental prototypes are set up by using the power electronic inverter and operational amplifier, and the voltage gain ratios of the four basic NR-based WPT topologies under the variations of transfer distance and load are observed. The experimental results validate the correctness of the theoretical analysis.

Interleukin-10 (IL-10) is an immunosuppressive cytokine that signals through STAT3 to regulate T follicular helper (Tfh) cell differentiation and germinal center formation. In SIV-infected macaques, levels of IL-10 in plasma and lymph nodes (LNs) were induced by infection and not normalized with antiretroviral therapy (ART). During chronic infection, plasma IL-10 and transcriptomic signatures of IL-10 signaling were correlated with the cell-associated SIV-DNA content within LN CD4+ memory subsets, including Tfh cells, and predicted the frequency of CD4+ Tfh cells and their cell-associated SIV-DNA content during ART, respectively. In ART-treated rhesus macaques, cells harboring SIV-DNA by DNAscope were preferentially found in the LN B cell follicle in proximity to IL-10. Finally, we demonstrated that the in vivo neutralization of soluble IL-10 in ART-treated, SIV-infected macaques reduced B cell follicle maintenance and, by extension, LN memory CD4+ T cells, including Tfh cells and those expressing PD-1 and CTLA-4. Thus, these data support a role for IL-10 in maintaining a pool of target cells in lymphoid tissue that serve as a niche for viral persistence. Targeting IL-10 signaling to impair CD4+ T cell survival and improve antiviral immune responses may represent a novel approach to limit viral persistence in ART-suppressed people living with HIV.

Latency reversal strategies for HIV cure using inhibitor of apoptosis protein (IAP) antagonists (IAPi) induce unprecedented levels of latent reservoir expression without immunotoxicity during suppressive antiretroviral therapy (ART). However, full targeting of the reservoir may require combinatorial approaches. A Jurkat latency model screen for IAPi combination partners demonstrated synergistic latency reversal with bromodomain (BD) and extraterminal domain protein inhibitors (BETi). Mechanistic investigations using CRISPR-CAS9 and single-cell RNA-Seq informed comprehensive ex vivo evaluations of IAPi plus pan-BET, bD-selective BET, or selective BET isoform targeting in CD4+ T cells from ART-suppressed donors. IAPi+BETi treatment resulted in striking induction of cell-associated HIV gag RNA, but lesser induction of fully elongated and tat-rev RNA compared with T cell activation–positive controls. IAPi+BETi resulted in HIV protein induction in bulk cultures of CD4+ T cells using an ultrasensitive p24 assay, but did not result in enhanced viral outgrowth frequency using a standard quantitative viral outgrowth assay. This study defines HIV transcriptional elongation and splicing as important barriers to latent HIV protein expression following latency reversal, delineates the roles of BET proteins and their BDs in HIV latency, and provides a rationale for exploration of IAPi+BETi in animal models of HIV latency.

Greater than 90% of HIV-1 proviruses are thought to be defective and incapable of viral replication. While replication competent proviruses are of primary concern with respect to disease progression or transmission, studies have shown that even defective proviruses are not silent and can produce viral proteins, which may contribute to inflammation and immune responses. Viral protein expression also has implications for immune-based HIV-1 clearance strategies, which rely on antigen recognition. Thus, sensitive assays aimed at quantifying both replication-competent proviruses and defective, yet translationally competent proviruses are needed to understand the contribution of viral protein to HIV-1 pathogenesis and determine the effectiveness of HIV-1 cure interventions. Previously, we reported a modified HIV-1 gag p24 digital enzyme-linked immunosorbent assay with single molecule array (Simoa) detection of cell-associated viral protein. Here we report a novel p24 protein enrichment method coupled with the digital immunoassay to further extend the sensitivity and specificity of viral protein detection. Immunocapture of HIV gag p24 followed by elution in a Simoa-compatible format resulted in higher protein recovery and lower background from various biological matrices and sample volumes. Quantification of as little as 1 fg of p24 protein from cell lysates from cells isolated from peripheral blood or tissues from ART-suppressed HIV participants, as well as simian–human immunodeficiency virus–infected non-human primates (NHPs), with high recovery and reproducibility is demonstrated here. The application of these enhanced methods to patient-derived samples has potential to further the study of the persistent HIV state and examine in vitro response to therapies, as well as ex vivo study of translationally competent cells from a variety of donors.

The human–machine dialogue system allows people and the machine to communicate basically. This technology is also attracting more and more attention. It is conducive to the realisation of resource sharing, inheritance, and protection of Dongba culture worldwide to analyse the Naxi speech recognition for the protection and inheritance needs of Naxi Dongba culture. The model was established for Naxi speaker recognition, and the method is studied with text-independent speaker speech voice recognition. The practical analysis method was used to compare the application effect of Gaussian mixture model (GMM) and hidden Markov model (HMM) in the processing of speaker recognition in Naxi speaker's pronunciation. It is very useful to provide a theoretical basis for the establishment of a Naxi language-based human–computer interaction system to find out the speaker's voice characteristics, personality differences, and different regions. This is a theoretically necessary analysis of ethnic minority languages and the protection of ethnic and folk cultures. It can also provide practical significance for carrying out ethnic cultural differences analysis.

It is necessary to test the reliability of the operation of the machine itself for meeting the high-speed, ultra-precision, flexible, and other modern manufacturing and processing needs. In practical applications, the obtained reliability detection information presents a variety of uncertainties such as randomness, imprecision, incompleteness and so on, due to the complexity of the machining environment, the limitations of the sensor itself and the imperfections of the information acquisition technology. The uncertainty reasoning method is proposed to test reliability and accuracy precision. Based on the multi-sensor information characteristic, the rule of the credibility sets is established to achieve the operational reliability analysis. It can realise the uncertainty reasoning for the modal and reliability rules. Examples of analysis show that during the high-end CNC machine tools operational reliability analysis, it has a practical effect to study the uncertainty reasoning method of the modal and reliability propositional rule and the analysis basis can be provided for the precision degradation and fault diagnosis.

Zinc-coated steel sheets, a kind of high-quality sheet, is widely used in all sorts of manufacturing industry. The fault on its surface is one of the factors which exert bad effects on its quality. Due to its complex texture, the experts first put forward to the defect segmentation methods of Discrete Fourier Spectrum Residuals and Gabor Filtering Image Fusion. Then, they analysed the grey form of the faults and the deficiencies of the two methods and made some improvements on them using Morphological black hat operation and eventually achieved the goal of effective defect segmentation. According to some results of experiments, the improved method is of high accuracy in the defect segmentation and to some extent can serve as the reference method to examine the faults of the zinc-coated sheets.

In order to improve the operation reliability of high-grade computer numerical control (CNC) machine tools and reduce the maintenance time of failures, the remote state monitoring and intelligent maintenance system for CNC machine tools was proposed. Through collecting the basic information of equipment, maintenance information, and the information of the vibration, temperature, main current and power of the machine tool based on soft-bus technology, the system analysed the real-time state data, got the reliability parameters, extracted the fault features and established the maintenance knowledge base. Combining the dynamic monitoring data and static data, the system evaluated the key parts of machine tools. By the fault knowledge base, the system analysed the reason for failure and recommended the maintenance method. With the function modules, the system can improve the intelligent maintenance level of CNC machine tools and improve the using efficiency of machine tools.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein that regulates hepatic low-density lipoprotein receptor (LDLR) levels in humans. PCSK9 has also been shown to regulate the levels of additional membrane-bound proteins in vitro, including the very low-density lipoprotein receptor (VLDLR), apolipoprotein E receptor 2 (ApoER2) and the β-site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1), which are all highly expressed in the CNS and have been implicated in Alzheimer's disease. To better understand the role of PCSK9 in regulating these additional target proteins in vivo, their steady-state levels were measured in the brain of wild-type, PCSK9-deficient, and human PCSK9 overexpressing transgenic mice. We found that while PCSK9 directly bound to recombinant LDLR, VLDLR, and apoER2 protein in vitro, changes in PCSK9 expression did not alter the level of these receptors in the mouse brain. In addition, we found no evidence that PCSK9 regulates BACE1 levels or APP processing in the mouse brain. In conclusion, our results suggest that while PCSK9 plays an important role in regulating circulating LDL cholesterol levels by reducing the number of hepatic LDLRs, it does not appear to modulate the levels of LDLR and other membrane-bound proteins in the adult mouse brain.

Image segmentation is a significant step in image analysis and computer vision. Many entropy-based approaches have been presented on this topic. Among them, Tsallis entropy isone of the best-performing methods. In this paper, the surface defect images of galvanised steel sheets were studied. Atwo-dimensional asymmetric Tsallis cross-entropy image segmentation algorithm based on chaotic bee colony algorithm optimisation was used to investigate the segmentation of surface defects under complex texture backgrounds. On the basis of Tsallis entropy threshold segmentation, a more concise expression form was used to define the asymmetric Tsallis cross-entropy in order to reduce the calculation complexity of the algorithm. The chaotic algorithm was combined with the artificial bee colony (ABC) algorithm to construct the chaotic bee colony algorithm, so that the optimal threshold of Tsallis entropy could be quickly identified. The experimental results showed that compared with the maximum Shannon entropy algorithm, the calculation time of this algorithm decreased by about 58% and the threshold value increased by about (26%, 54%). Compared with the two-dimensional Tsallis cross-entropy algorithm, the calculation time of this algorithm decreased by about 35% and about 20% was improved in the g-axis direction only. Compared with the two-dimensional asymmetric Tsallis cross-entropy algorithm, the calculation time of this algorithm decreased by about 30% and the threshold values of the two algorithms were almost the same. The algorithm proposed in this paper can rapidly and effectively segment defect targets, making it a more suitable method for detecting surface defects in factories with a rapid production pace. [ABSTRACT FROM AUTHOR]

Background: The latent reservoir in resting CD4+ T cells presents a major barrier to HIV cure. Latency-reversing agents are therefore being developed with the ultimate goal of disrupting the latent state, resulting in induction of HIV expression and clearance of infected cells. Histone deacetylase inhibitors (HDACi) have received a significant amount of attention for their potential as latency-reversing agents. Results: Here, we have investigated the in vitro and systemic in vivo effect of panobinostat, a clinically relevant HDACi, on HIV latency. We showed that panobinostat induces histone acetylation in human PBMCs. Further, we showed that panobinostat induced HIV RNA expression and allowed the outgrowth of replication-competent virus ex vivo from resting CD4+ T cells of HIV-infected patients on suppressive antiretroviral therapy (ART). Next, we demonstrated that panobinostat induced systemic histone acetylation in vivo in the tissues of BLT humanized mice. Finally, in HIV-infected, ART-suppressed BLT mice, we evaluated the effect of panobinostat on systemic cell-associated HIV RNA and DNA levels and the total frequency of latently infected resting CD4+ T cells. Our data indicate that panobinostat treatment resulted in systemic increases in cellular levels of histone acetylation, a key biomarker for in vivo activity. However, panobinostat did not affect the levels of cell-associated HIV RNA, HIV DNA, or latently infected resting CD4+ T cells. Conclusion: We have demonstrated robust levels of systemic histone acetylation after panobinostat treatment of BLT humanized mice; and we did not observe a detectable change in the levels of cell-associated HIV RNA, HIV DNA, or latently infected resting CD4+ T cells in HIV-infected, ART-suppressed BLT mice. These results are consistent with the modest effects noted in vitro and suggest that combination therapies may be necessary to reverse latency and enable clearance. Animal models will contribute to the progress towards an HIV cure. [ABSTRACT FROM AUTHOR]

β-site amyloid precursor protein-cleaving enzyme 1 (BACE1) plays an important role in the development of Alzheimer's disease (AD), freeing the amyloid-β (Aβ) N-terminus from the amyloid-β protein precursor (AβPP), the first step in Aβ formation. Increased BACE1 activity in AD brain or cerebrospinal fluid (CSF) has been reported. Other studies, however, found either no change or a decrease with AD diagnosis in either BACE1 activity or sAβPPβ, the N-terminal secreted product of BACE1 (sBACE1) activity on AβPP. Here, sBACE1 enzymatic activity and secreted AβPPβ (sAβPPβ) were measured in Alzheimer's Disease Neuroimaging Initiative-1 (ADNI-1) baseline CSF samples and no statistically significant changes were found in either measure comparing healthy control, mild cognitively impaired, or AD individual samples. While CSF sBACE1 activity and sAβPPβ demonstrated a moderate yet significant degree of correlation with each other, there was no correlation of either analyte to CSF Aβ peptide ending at residue 42. Surprisingly, a stronger correlation was demonstrated between CSF sBACE1 activity and tau, which was comparable to that between CSF Aβ₄₂ and tau. Unlike for these latter two analytes, receiver-operator characteristic curves demonstrate that neither CSF sBACE1 activity nor sAβPPβ concentrations can be used to differentiate between healthy elderly and AD individuals. [ABSTRACT FROM AUTHOR]

Concerning the problem of large rotating machinery like wind turbine which runs in low speed and non-stationary state, this research mainly focuses on separating fault trend feature from non-fault feature and the method of state deterioration evolution based on bi-spectrum. Firstly, the experimental signal such as low-speed startup vibration signal of rotor test rig in the normal state and a plurality of unbalanced state have been collected. Bi-spectrum method is applied to extract fault feature which submerged in complex background noise. On the basis of bi-spectrum analysis, the fault feature evolutionary matrix is defined to represent the state of equipment deterioration. The eigenvalues of fault feature evolutionary matrix are computed and fitted to a normal distribution curve, from which the mean value and variance are taken as fault feature parameters. Fault feature parameters are verified effectively by experiments. Finally, depending on fault feature parameters, graphical representation of state deterioration evolution is established. It is beneficial to provide guidelines for equipment deterioration trend. This method is applied to analyze the real vibration signal of wind turbine with the type of WD646/600 KW, and actual equipment condition verified the effectiveness of the proposed method. [ABSTRACT FROM PUBLISHER]

BACE, a β-secretase, is an attractive potential disease-modifying therapeutic strategy for Alzheimer's disease (AD) as it results directly in the decrease of amyloid precursor protein (APP) processing through the β-secretase pathway and a lowering of CNS amyloid-β (Aβ) levels. The interaction of the β-secretase and α-secretase pathway-mediated processing of APP in the rhesus monkey (nonhuman primate; NHP) CNS is not understood. We hypothesized that CNS inhibition of BACE would result in decreased newly generated Aβ and soluble APPβ (sAPPβ), with increased newly generated sAPPα. A stable isotope labeling kinetics experiment in NHPs was performed with a 13C6-leucine infusion protocol to evaluate effects of BACE inhibition on CNS APP processing by measuring the kinetics of sAPPα, sAPPβ, and Aβ in CSF. EachNHPreceived a low, medium, or high dose of MBI-5 (BACE inhibitor) or vehicle in a four-way crossover design. CSF sAPPα, sAPPβ, and Aβ were measured by ELISA and newly incorporated label following immunoprecipitation and liquid chromatography-mass spectrometry. Concentrations, kinetics, and amount of newly generated APP fragments were calculated. sAPPβ and sAPPα kinetics were similar, but both significantly slower than Aβ. BACE inhibition resulted in decreased labeled sAPPβ and Aβ in CSF, without observable changes in labeled CSF sAPPα. ELISA concentrations of sAPPβ and Aβ both decreased and sAPPα increased. sAPPα increased by ELISA, with no difference by labeled sAPPα kinetics indicating increases in product may be due to APP shunting from the β-secretase to the α-secretase pathway. These results provide a quantitative understanding of pharmacodynamic effects of BACE inhibition on NHP CNS, which can inform about target development. [ABSTRACT FROM AUTHOR]

Ahallmark of Alzheimer's disease (AD) brain is the amyloidβ (Aβ) plaque, which is comprised of Aβ peptides. Multiple lines of evidence suggest that Aβ oligomers are more toxic than other peptide forms. We sought to develop a robust assay to quantify oligomers from CSF. Antibody 19.3 was compared in one-site and competitive ELISAs for oligomer binding specificity. A two-site ELISA for oligomers was developed using 19.3 coupled to a sensitive, bead-based fluorescent platform able to detect single photons of emitted light. The two-site ELISA wasβ2500>selective for Aβ oligomers over Aβ monomers with a limit of detection~0.09 pg/ml in human CSF. The lower limit of reliable quantification of the assay was 0.18 pg/ml and the antibody pairs recognized Aβ multimers comprised of either synthetic standards, or endogenous oligomers isolated from confirmed human AD and healthy control brain. Using the assay, a significant 3- to 5-fold increase in Aβ oligomers in human AD CSF compared with comparably aged controls was demonstrated. The increase was seen in three separatehumancohorts, totaling 63ADand 54 controls. CSF oligomers ranged between 0.1 and 10 pg/ml.Aβ oligomer levels did not strongly associate with age or gender, but had an inverse correlation withMMSEscore. The C statistic for theAβ oligomer ROC curve was 0.86, with 80% sensitivity and 88% specificity to detect AD, suggesting reasonable discriminatory power for theADstate and the potential for utility as a diagnostic marker. [ABSTRACT FROM AUTHOR]

The article focuses on a study which examined the vagus and phrenic nerves from adult cadavers. Results of the study showed that there was a slight difference in the track of the right and left vagus nerve in the chest. Information on the positional point of the vagus and phrenic nerves on the clavicle is provided. The study observed that both nerves are suitable for end-to-end anastomosis. It also found similar quantity of nerve fibers in the two nerves.

The accumulation of amyloid β(Aβ) in Alzheimer's disease is caused by an imbalance of production and clearance, which leads to increased soluble Aβspecies and extracellular plaque formation in the brain. Multiple Aβ-lowering therapies are currently in development: an important goal is to characterize the molecular mechanisms of action and effects on physiological processing of Aβ, as well as other amyloid precursor protein (APP) metabolites, in models which approximate human Aβphysiology. To this end, we report the translation of the human in vivo stable-isotope-labeling kinetics (SILK) method to a rhesus monkey cisterna magna ported (CMP) nonhuman primate model, and use the model to test the mechanisms of action of a γ-secretase inhibitor (GSI). A major concern of inhibiting the enzymes which produce Aβ(β- and γ-secretase) is that precursors of Aβ may accumulate and cause a rapid increase in Aβ production when enzyme inhibition discontinues. In this study, the GSI MK-0752 was administered to conscious CMP rhesus monkeys in conjunction with in vivo stable-isotope-labeling, and dose-dependently reduced newly generated CNS Aβ. In contrast to systemic Aβ metabolism, CNS Aβ production was not increased after the GSI was cleared. These results indicate that most of the CNS APP was metabolized to products other than Aβ, including C-terminal truncated forms of Aβ: 1-14, 1-15 and 1-16; this demonstrates an alternative degradation pathway for CNS amyloid precursor protein during γ-secretase inhibition. [ABSTRACT FROM AUTHOR]

Background: We used a sensitive and specific site amyloid precursor protein (APP)-cleaving enzyme 1 (BACE1) assay to determine the relationship between BACE1 activity in cerebrospinal fluid (CSF) and markers of APP metabolism and axonal degeneration in early and late stages of Alzheimer disease (AD). Objective: To assess CSF BACE1 activity in AD. Design: Case-control and longitudinal follow-up study. Setting: Specialized memory clinic. Patients: Eighty-seven subjects with AD, 33 cognitively normal control subjects, and 113 subjects with mild cognitive impairment (MCI), who were followed up for 3 to 6 years. Main Outcome Measures: Cerebrospinal fluid BACE1 activity in relation to diagnosis and CSF levels of secreted APP and amyloid β protein (Aβ) isoforms and the axonal degeneration marker total tau. Results: Subjects with AD had higher CSF BACE1 activity (median, 30 pM [range, 11-96 pM]) than controls (median, 23pM[range, 8-43 pM]) (P=.02). Subjects with MCI who progressed to AD during the follow-up period had higher baseline BACE1 activity (median, 35 pM [range, 18-71 pM]) than subjects with MCI who remained stable (median, 29 pM [range, 14-83 pM]) (P<.001) and subjects with MCI who developed other forms of dementia (median, 20 pM [range, 10-56 pM]) (P<.001). BACE1 activity correlated positively with CSF levels of secreted APP isoforms and Aβ40 in the AD and control groups and in all MCI subgroups (P<.05) except the MCI subgroup that developed AD. Strong positive correlations were found between CSF BACE1 activity and total tau levels in all MCI subgroups (r≥0.57, P≤.009). Conclusion: Elevated BACE1 activity may contribute to the amyloidogenic process in sporadic AD and is associated with the intensity of axonal degeneration. [ABSTRACT FROM AUTHOR]

Productively infected cells are generally thought to arise from HIV infection of activated CD4+ T cells, and these infected activated cells are thought to be a recurring source of latently infected cells when a portion of the population transitions to a resting state. We discovered and report here that productively and latently infected cells can instead originate from direct infection of resting CD4+ T cell populations in lymphoid tissues in Fiebig I, the earliest stage of detectable HIV infection. We found that direct infection of resting CD4+ T cells was correlated with the availability of susceptible target cells in lymphoid tissues largely restricted to resting CD4+ T cells in which expression of pTEFb enabled productive infection, and we documented persistence of HIV-producing resting T cells during antiretroviral therapy (ART). Thus, we provide evidence of a mechanism by which direct infection of resting T cells in lymphoid tissues to generate productively and latently infected cells creates a mechanism by which the productively infected cells can replenish both populations and maintain two sources of virus from which HIV infection can rebound, even if ART is instituted at the earliest stage of detectable infection. [ABSTRACT FROM AUTHOR]

Anti-retroviral therapy (ART) generally suppresses HIV replication to undetectable levels in peripheral blood, but immune activation associated with increased morbidity and mortality is sustained during ART, and infection rebounds when treatment is interrupted. To identify drivers of immune activation and potential sources of viral rebound, we modified RNAscope in situ hybridization to visualize HIV-producing cells as a standard against which to compare the following assays of potential sources of immune activation and virus rebound following treatment interruption: (i) envelope detection by induced transcription-based sequencing (EDITS) assay; (ii) HIVFlow; (iii) Flow-FISH assays that can scan tissues and cell suspensions to detect rare cells expressing env mRNA, gag mRNA/Gag protein and p24; and (iv) an ultrasensitive immunoassay that detects p24 in cell/tissue lysates at subfemtomolar levels. We show that the sensitivities of these assays are sufficient to detect one rare HIVproducing/env mRNA+/p24+ cell in one million uninfected cells. These high-throughput technologies provide contemporary tools to detect and characterize rare cells producing virus and viral antigens as potential sources of immune activation and viral rebound. [ABSTRACT FROM AUTHOR]

Interleukin-10 (IL-10) is an immunosuppressive cytokine that signals through STAT3 to regulate T follicular helper (Tfh) cell differentiation and germinal center formation. In SIV-infected macaques, levels of IL-10 in plasma and lymph nodes (LNs) were induced by infection and not normalized with antiretroviral therapy (ART). During chronic infection, plasma IL-10 and transcriptomic signatures of IL-10 signaling were correlated with the cell-associated SIV-DNA content within LN CD4+ memory subsets, including Tfh cells, and predicted the frequency of CD4+ Tfh cells and their cell-associated SIV-DNA content during ART, respectively. In ART-treated rhesus macaques, cells harboring SIV-DNA by DNAscope were preferentially found in the LN B cell follicle in proximity to IL-10. Finally, we demonstrated that the in vivo neutralization of soluble IL-10 in ART-treated, SIV-infected macaques reduced B cell follicle maintenance and, by extension, LN memory CD4+ T cells, including Tfh cells and those expressing PD-1 and CTLA-4. Thus, these data support a role for IL-10 in maintaining a pool of target cells in lymphoid tissue that serve as a niche for viral persistence. Targeting IL-10 signaling to impair CD4+ T cell survival and improve antiviral immune responses may represent a novel approach to limit viral persistence in ART-suppressed people living with HIV. [ABSTRACT FROM AUTHOR]

Latency reversal strategies for HIV cure using inhibitor of apoptosis protein (IAP) antagonists (IAPi) induce unprecedented levels of latent reservoir expression without immunotoxicity during suppressive antiretroviral therapy (ART). However, full targeting of the reservoir may require combinatorial approaches. A Jurkat latency model screen for IAPi combination partners demonstrated synergistic latency reversal with bromodomain (BD) and extraterminal domain protein inhibitors (BETi). Mechanistic investigations using CRISPR-CAS9 and single-cell RNA-Seq informed comprehensive ex vivo evaluations of IAPi plus pan-BET, bD-selective BET, or selective BET isoform targeting in CD4+ T cells from ART-suppressed donors. IAPi+BETi treatment resulted in striking induction of cell-associated HIV gag RNA, but lesser induction of fully elongated and tat-rev RNA compared with T cell activation-positive controls. IAPi+BETi resulted in HIV protein induction in bulk cultures of CD4+ T cells using an ultrasensitive p24 assay, but did not result in enhanced viral outgrowth frequency using a standard quantitative viral outgrowth assay. This study defines HIV transcriptional elongation and splicing as important barriers to latent HIV protein expression following latency reversal, delineates the roles of BET proteins and their BDs in HIV latency, and provides a rationale for exploration of IAPi+BETi in animal models of HIV latency. [ABSTRACT FROM AUTHOR]

Histone deacetylase inhibitors (HDACi) are the most widely studied HIV latency reversing agents (LRAs). The HDACi suberoylanilidehydroxamic acid (vorinostat, VOR) has been employed in several clinical HIV latency reversal studies as well as in vitro models of HIV latency and shown to effectively induce HIV RNA and protein expression. Despite these findings, response to HDACi can vary, particularly with intermittent dosing, and information is lacking on the relationship between host transcriptional response and HIV latency reversal. Herein, we report on global gene expression responses to VOR and examined the longevity of the transcriptional response in various cellular models. We find many genes are modulated at 6 h post-VOR in HCT116, Jurkat and primary resting CD4 T cells yet return to baseline levels after an 18 h VOR-free period. With repeat exposure to VOR in resting CD4 T cells, we find similar and consistent transcriptional changes 6 h following each serial treatment. In addition, serial exposure in HIV infected suppressed donor CD4 T cells showed consistent transcriptional changes after each exposure to VOR. We identified five host genes that were strongly and consistently modulated following HDAC inhibition: 3 up-regulated (H1F0, IRGM, WIPI49) and 2 down-regulated (PHF15 and PRDM10). These genes demonstrated consistent modulation in PBMC samples from HIV+ participants who received either single or multiple doses of 400 mg VOR. Interestingly, host transcriptional response did not predict induction of cell-associated HIV RNA, suggesting other cellular factors play key roles in HIV latency reversal in vivo despite robust HDACi pharmacological activity. IMPORTANCE Histone deacetylase inhibitors are widely studied HIV latency reversing agents (LRAs). VOR, an HDACi, induces histone acetylation, chromatin remodeling, and modulates host and HIV gene expression. However, the relationship between these events are poorly defined, and clinical studies suggest diminished HIV reactivation in resting CD4 T cells with daily exposures to VOR. Our study provides evidence that VOR induces a consistent level of host cell gene transcription, following intermittent exposure. In addition, a gene signature was identified in response to VOR exposure that was conserved across single and serial exposures both in vitro and in vivo, indicating that VOR can consistently and reproducibly modulate transcriptional host responses. However, as responses to HIV response to HDACis decline over time, other factors modulate viral reactivation in vivo despite robust HDAC activity. The identified host gene VOR biomarkers can be used for monitoring the pharmacodynamic activity of HDAC inhibitors. [ABSTRACT FROM AUTHOR]

Clearance of HIV-infected germinal center (GC) CD4+ follicular helper T cells (Tfh) after combination antiretroviral therapy (ART) is essential to an HIV cure. Blocking B cell lymphoma 6 (BCL6, the master transcription factor for Tfh cells) represses HIV infection of tonsillar CD4+ Tfh ex vivo, reduces GC formation and limits immune activation in vivo. We assessed the anti-HIV activity of a novel BCL6 inhibitor, FX1, in Tfh/non-Tfh CD4+ T cells, and its impact on T cell activation and SAMHD1 phosphorylation (Thr592). FX1 repressed HIV-1 infection of peripheral CD4+ T cells, tonsillar Tfh/non-Tfh CD4+ T cells (p<0.05), total elongated-and multi-spliced HIV-1 RNA production during the first round of viral life cycle (p<0.01). Using purified circulating CD4+ T cells from uninfected donors, we demonstrate that FX1 treatment resulted in down-regulation pSAMHD1 expression (p<0.05) and T cell activation (HLA-DR, CD25, Ki67, p<0.05) ex vivo corresponding with inhibition of HIV-1 and HIV-2 replication. Ex vivo HIV-1 reactivation using purified peripheral CD4+ T cells from HIV-infected ART-suppressed donors was also blocked by FX1 treatment (p<0.01). Our results indicate that BCL6 function contributes to Tfh/non-Tfh CD4+ T cell activation and cellular susceptibility to HIV infection. BCL6 inhibition represents a novel therapeutic strategy to potentiate HIV suppression in Tfh/non-Tfh CD4+ T cells without reactivation of latent virus. [ABSTRACT FROM AUTHOR]

Extracellular deposits of aggregated amyloid-β (Aβ) peptides are a hallmark of Alzheimer disease; thus, inhibition of Aβ production and/or aggregation is an appealing strategy to thwart the onset and progression of this disease. The release of Aβ requires processing of the amyloid precursor protein (APP) by both β- and γ-secretase. Using an assay that incorporates full-length recombinant APP as a substrate for β-secretase (BACE), we have identified a series of compounds that inhibit APP processing, but do not affect the cleavage of peptide substrates by BACE1. These molecules also inhibit the processing of APP and Afl by BACE2 and selectively inhibit the production of Aβ42 species by γ-secretase in assays using CTF99. The compounds bind directly to APP, likely within the Aβ domain, and therefore, unlike previously described inhibitors of the secretase enzymes, their mechanism of action is mediated through APP. These studies demonstrate that APP binding agents can affect its processing through multiple pathways, providing proof of concept for novel strategies aimed at selectively modulating Aβ production. [ABSTRACT FROM AUTHOR]