Multiple Markers Research Articles

Nano-magnetic aptamer sensor incorporating AND logic recognition-launched hybridization chain reaction for organ origin identification of circulating tumor cells

Circulating tumor cells (CTCs) are effective bio-markers for early diagnosis and disease monitoring of cancer, but there is lack of simple and effective strategy to achieve the detection of multiple markers on CTCs while enriching, especially the identification of organ origin, limits their broad clinical applications. Here we present a nano-magnetic aptamer sensor platform to capture broad spectrum of epithelial CTCs and simultaneously identify their multiple organ origins by AND logic recognition-launched hybridization chain reaction. The platform exhibited excellent capture and enrichment capabilities for epithelial CTCs, and showed the ability to precisely identify their organ origin markers (GPC3 for liver and TTF1 for lung), even able to trace metastatic cancer to related organs. The developed nano-magnetic aptamer sensor platform provides a feasible strategy for multi-marker accurate detection of CTCs and a promising auxiliary tool for the precise treatment of cancer.

Genomic variation across Chinook salmon populations reveals effects of a duplication on migration alleles and supports fine scale structure.

The distribution of ecotypic variation in natural populations is influenced by neutral and adaptive evolutionary forces that are challenging to disentangle. This study provides a high-resolution portrait of genomic variation in Chinook salmon (Oncorhynchus tshawytscha) with emphasis on a region of major effect for ecotypic variation in migration timing. With a filtered data set of ~13million single nucleotide polymorphisms (SNPs) from low-coverage whole genome resequencing of 53 populations (3566 barcoded individuals), we contrasted patterns of genomic structure within and among major lineages and examined the extent of a selective sweep at a major effect region underlying migration timing (GREB1L/ROCK1). Neutral variation provided support for fine-scale structure of populations, while allele frequency variation in GREB1L/ROCK1 was highly correlated with mean return timing for early and late migrating populations within each of the lineages (r2 = .58-.95; p< .001). However, the extent of selection within the genomic region controlling migration timing was much narrower in one lineage (interior stream-type) compared to the other two major lineages, which corresponded to the breadth of phenotypic variation in migration timing observed among lineages. Evidence of a duplicated block within GREB1L/ROCK1 may be responsible for reduced recombination in this portion of the genome and contributes to phenotypic variation within and across lineages. Lastly, SNP positions across GREB1L/ROCK1 were assessed for their utility in discriminating migration timing among lineages, and we recommend multiple markers nearest the duplication to provide highest accuracy in conservation applications such as those that aim to protect early migrating Chinook salmon. These results highlight the need to investigate variation throughout the genome and the effects of structural variants on ecologically relevant phenotypic variation in natural species.

Recognizing Minor Leukemic Populations with Monocytic Features in Mixed-Phenotype Acute Leukemia by Flow Cell Sorting Followed by Cytogenetic and Molecular Studies: Report of Five Exemplary Cases

Mixed-phenotype acute leukemia (MPAL), a rare and heterogeneous category of acute leukemia, is characterized by cross-lineage antigen expression. Leukemic blasts in MPAL can be represented either by one population with multiple markers of different lineages or by several single-lineage populations. In some cases, a major blast population may coexist with a smaller population that has minor immunophenotypic abnormalities and may be missed even by an experienced pathologist. To avoid misdiagnosis, we suggest sorting doubtful populations and leukemic blasts and searching for similar genetic aberrations. Using this approach, we examined questionable monocytic populations in five patients with dominant leukemic populations of B-lymphoblastic origin. Cell populations were isolated either for fluorescence in situ hybridization or for clonality assessment by multiplex PCR or next-generation sequencing. In all cases, monocytic cells shared the same gene rearrangements with dominant leukemic populations, unequivocally confirming the same leukemic origin. This approach is able to identify implicit cases of MPAL and therefore leads to the necessary clinical management for patients.

The importance of targeting multiple risk markers in patients with type 2 diabetes: A post-hoc study from the CANVAS programme.

To investigate the extent to which improvements in multiple cardiovascular risk markers are associated with a lower risk of cardiovascular and kidney outcomes in patients with type 2 diabetes and high cardiovascular risk participating in the CANVAS programme. Clinically relevant improvements in cardiovascular risk factors were defined as a reduction in glycated haemoglobin ≥1.0%, systolic blood pressure ≥10 mmHg, body weight ≥3kg, urinary-albumin-creatinine ratio ≥30%, uric acid ≥0.5mg/dl, and an increase in haemoglobin of ≥1.0g/dl from baseline to week 26. Participants were categorized according to the number of improvements in cardiovascular risk markers: zero, one, two, three, or four or more risk marker improvements. The Cox proportional hazard regression adjusted for treatment assignment, demographic variables and laboratory measurements was performed to determine the association between the number of risk marker improvements and risk of a composite cardiovascular, heart failure or kidney outcomes. We included 9487 (93.5%) participants with available data at baseline and week 26. After week 26, 566 composite cardiovascular, 370 heart failure/cardiovascular death and 153 composite kidney outcomes occurred. The multivariable adjusted hazard ratios associated with four or more improvements in risk markers versus no risk marker improvement were 0.67 (95% CI 0.48, 0.92), 0.58 (95% CI 0.39, 0.87) and 0.49 (95% CI 0.25, 0.96) for the three outcomes respectively. We observed a trend of decreased hazard ratios across subgroups of increasing number of risk marker improvements (p for trend=.008, .02 and .047, respectively). In patients with type 2 diabetes, improvements in multiple risk markers were associated with a reduced risk of cardiovascular and kidney outcomes as compared with no risk marker improvement.

Comprehensive Analysis Based on the Cancer Immunotherapy and Immune Activation of Gastric Cancer Patients.

When it comes to aggressiveness and prognosis, immune cells play an important role in the microenvironment of gastric cancer (GC). Currently, there is no well-established evidence that immune status typing is reliable as a prognostic tool for gastric cancer. This study aimed to develop a genetic signature based on immune status typing for the stratification of gastric cancer risk. TCGA data were used for gene expression and clinical characteristics analysis. A ssGSEA algorithm was applied to type the gastric cancer cohorts. A multivariate and univariate Cox regression and a lasso regression were conducted to determine which genes are associated with gastric cancer prognosis. Finally, we were able to produce a 6-gene prognostic prediction model using immune-related genes. Further analysis revealed that the prognostic prediction model is closely related to the prognosis of patients with GC. Nomograms incorporating genetic signatures and risk factors produced better calibration results. The relationship between the risk score and gastric cancer T stage was also significantly correlated with multiple immune markers related to specific immune cell subsets. According to these results, patients' outcomes and tumor immune cell infiltration correlate with risk scores. In addition, immune cellular-based genetic signatures can contribute to improved risk stratification for gastric cancer. Clinical decisions regarding immunotherapy and followup can be guided by these features.

Immortalization of mouse primary astrocytes

In cell culture studies, immortalized primary cells have become a useful tool to investigate the molecular and cellular functions of different types of cells. Several immortalization agents, such as human telomerase reverse transcriptase (hTERT) and Simian Virus 40 (SV40) T antigens, are commonly used for primary cell immortalization. Astrocytes, as the most abundant glial cell type in the central nervous system, are promising therapeutical targets for many neuronal disorders, such as Alzheimer’s disease and Parkinson’s disease. Immortalized primary astrocytes can provide useful information for astrocytes biology, astrocytes-neuron interactions, glial interactions and astrocytes-associated neuronal diseases. In this study, we successfully purified primary astrocytes with immuno-panning method and examined the astrocyte functions after immortalization through both hTERT and SV40 Large-T antigens. As expected, both immortalized astrocytes presented unlimited lifespan and highly expressed multiple astrocyte-specific markers. However, SV40 Large-T antigen, but not hTERT, immortalized astrocytes displayed fast ATP-induced calcium wave in culture. Hence, SV40 Large-T antigen could be a better choice for primary astrocyte immortalization, which closely mimics the cell biology of primary astrocytes in culture. In summary, the purification and immortalization of primary astrocytes presented in this study can be used for studying astrocyte biology under physiological and pathological conditions.

Abstract P1-04-16: A nomogram based on clinicopathologic factors and pretreatment systemic inflammation response index to predict pathological complete response of neoadjuvant systemic therapy in HER2-positive breast cancer: A dual-center study

Abstract Purpose: Multiple pretreatment systemic inflammatory markers (SIMs) have been reported as predictors of pathological complete response (pCR) after neoadjuvant systemic therapy (NST) in patients with breast cancer (BC). However, the most significant SIM remains to be conclusively identified, and variations among different molecular subtypes remain unknown. The objective of the study was to identify the most significant SIM in patients with human epidermal growth factor receptor 2 (HER2) positive BC, and to construct a pCR-predictive nomogram combining it with other clinicopathological factors. Methods: We retrospectively reviewed the findings for 240 patients with stage I–III HER2-positive BC who underwent NST and subsequent surgery at Kaohsiung and Taichung Veterans General Hospital from 2011 to 2021. Clinicopathological factors were analyzed by stepwise logistic regression with forward selection. The data were used to construct a nomogram plot for determining the probability of pCR. Results: Among the pretreatment SIMs, only SIRI was significantly related to pCR, with an optimal cutoff value of 1.27 × 10^9/L. Stepwise logistic analyses indicated that clinical N stage, HER2 immunohistochemistry (IHC) score, hormone receptor status, targeted therapy regimen, and SIRI were independent predictors of pCR, with an area under the curve (AUC) of 0.722. The Hosmer–Lemeshow test and calibration curve revealed that the predictive ability was a good fit to actual observations. A nomogram was constructed based on the logistic model. Conclusions: Pretreatment SIRI &amp;lt; 1.27 is predictive of pCR in HER2-positive BC. Our nomogram could efficiently predict pCR and facilitate clinical decision-making before neoadjuvant treatment. Univariable and multivariable analyses for predictive factors of pathological complete response of neoadjuvant systemic therapy in HER2-positive breast cancer Citation Format: Hong Yu Wu, Yen-Dun Tzeng, Jie Ru Yang, Chih Chiang Hung. A nomogram based on clinicopathologic factors and pretreatment systemic inflammation response index to predict pathological complete response of neoadjuvant systemic therapy in HER2-positive breast cancer: A dual-center study [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P1-04-16.

Evaluation of DNA metabarcoding using Oxford Nanopore sequencing for authentication of mixed seafood products

Seafood is the world's most highly traded food commodity. Due to the non-stop demand of the ever-increasing human population, the supply of certain species is not guaranteed, and mislabelling practices arise. Fillets and heavily processed products that cannot be easily authenticated are particularly vulnerable to fraudulent practices. In order to face this challenge, as a cost-effective, rapid, and easy handling tool, the MinION sequencer from Oxford Nanopore Technologies (ONT) coupled with DNA (meta)barcoding represents a suitable option to assess seafood fraud. In this study, we evaluated the feasibility of MinION sequencing to authenticate mixed seafood products by (1) evaluating MinION generated barcodes from single species that have already been Sanger sequenced and (2) performing MinION sequencing of mixed samples containing commercial seafood species in different amounts to evaluate whether all species can be detected, even when they are low abundant. Using the mitochondrial DNA gene markers Cytb and COI, 86% and 76% of the single species were correctly identified, respectively, of which 26% and 29% also showed non-target species. For the mixed samples, not all species were detected regardless of the composition of the mixtures or the marker gene used. Nevertheless, the MinION sequencing platform can be considered an additional tool to complement currently available approaches to authenticate mixed seafood samples providing that adequate measures are taken to reduce false positives and false negatives. • ONT-based metabarcoding detects 76% (COI) and 86% (Cytb) of tested seafood species. • The detection of non-target species impact interpretation of DNA metabarcoding data. • Multiple marker genes increase the detection of fish species with DNA metabarcoding.

Diagnosis of malignant pleural effusion with combinations of multiple tumor markers: A comparison study of five machine learning models.

To evaluate the diagnostic value of combinations of tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 125, CA153, and CA19-9 in identifying malignant pleural effusion (MPE) from non-malignant pleural effusion (non-MPE) using machine learning, and compare the performance of popular machine learning methods. A total of 319 samples were collected from patients with pleural effusion in Beijing and Wuhan, China, from January 2018 to June 2020. Five machine learning methods including Logistic regression, extreme gradient boosting (XGBoost), Bayesian additive regression tree, random forest, and support vector machine were applied to evaluate the diagnostic performance. Sensitivity, specificity, Youden's index, and the area under the receiver operating characteristic curve (AUC) were used to evaluate the performance of different diagnostic models. For diagnostic models with a single tumor marker, the model using CEA, constructed by XGBoost, performed best (AUC = 0.895, sensitivity = 0.80), and the model with CA153, also by XGBoost, showed the largest specificity 0.98. Among all combinations of tumor markers, the combination of CEA and CA153 achieved the best performance (AUC = 0.921, sensitivity = 0.85) in identifying MPE under the diagnostic model constructed by XGBoost. Diagnostic models for MPE with a combination of multiple tumor markers outperformed the models with a single tumor marker, particularly in sensitivity. Using machine learning methods, especially XGBoost, could comprehensively improve the diagnostic accuracy of MPE.

Seed Protein Genetics Linked with Nitrogen and Phosphorus Translocation Efficiency in Soybean

Soybean (Glycine max (L.) Merr.) is an important nutritional crop with high seed protein content. Production of high protein concentrations relies on sufficient nutrient supplies, especially of nitrogen (N) and phosphorus (P). Although the genetic basis for seed quality traits has been well studied, little information exists on any genetic connections between seed quality and nutrient supplies in soybean. Here, a recombinant inbred line (RIL) population of 179 progeny was generated using HC6 and JD17 as parents contrasting in seed quality and N and P translocation efficiencies. Seed protein and N and P translocation efficiencies were higher in HC6 than in JD17. Meanwhile, positive correlations were observed between seed protein content and translocation efficiency of N and P in RILs, implying that high N and P translocation efficiencies might facilitate seed protein accumulation. A genetic map was constructed using 5250 SNP markers covering a genetic distance of 3154.83 cM. A total of 6 loci for quality and 13 loci for N and P translocation efficiency were detected. Among them, two fragments on chromosome 6 and chromosome 20 contained multiple significant markers for both quality and N and P translocation efficiencies, with the respective observed LOD values ranging from 2.98 to 5.61, and 3.01 to 11.91, while the respective PVE values ranged from 8.2% to 13.9%, and 8.3% to 28.0%. Interestingly, one significant locus on chromosome 20 appears to be the product of a transposable element (TE) InDel in Glyma.20G085100, with progeny lacking the TE also exhibiting higher N and P translocation efficiencies, along with higher seed protein contents. Taken together, these results provide genetic evidence that increasing N and P translocation efficiencies may lead to increasing protein contents in soybean seeds. Furthermore, a TE InDel may be used as a genetic marker for breeding elite soybean cultivars with high protein content and N and P translocation efficiencies.

CSF Aβ42 and Aβ42/Aβ40 Ratio in Alzheimer's Disease and Frontotemporal Dementias.

Alzheimer's disease dementia (ADD) may manifest with atypical phenotypes, resembling behavioral variant frontotemporal dementia (bvFTD) and corticobasal syndrome (CBS), phenotypes which typically have an underlying frontotemporal lobar degeneration with tau proteinopathy (FTLD-tau), such as Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), or FTLD with TDP-43 proteinopathy (FTLD-TDP). CSF biomarkers total and phosphorylated tau (τT and τP-181), and amyloid beta with 42 and 40 amino acids (Aβ42 and Aβ40) are biomarkers of AD pathology. The primary aim of this study was to compare the diagnostic accuracy of Aβ42 to Aβ42/Aβ40 ratio in: (a) differentiating ADD vs. frontotemporal dementias; (b) patients with AD pathology vs. non-AD pathologies; (c) compare biomarker ratios and composite markers to single CSF biomarkers in the differentiation of AD from FTD; Methods: In total, 263 subjects were included (ADD: n = 98; bvFTD: n = 49; PSP: n = 50; CBD: n = 45; controls: n = 21). CSF biomarkers were measured by commercially available ELISAs (EUROIMMUN). Multiple biomarker ratios (Aβ42/Aβ40; τT/τP-181; τT/Aβ42; τP-181/Aβ42) and composite markers (t-tau: τT/(Aβ42/Aβ40); p-tau: τP-181/(Aβ42/Aβ40) were calculated. ROC curve analysis was performed to compare AUCs of Aβ42 and Aβ42/Aβ40 ratio and relevant composite markers between ADD and FTD, as defined clinically. BIOMARKAPD/ABSI criteria (abnormal τT, τP-181 Aβ42, and Aβ42/Aβ40 ratio) were used to re-classify all patients into AD pathology vs. non-AD pathologies, and ROC curve analysis was repeated to compare Aβ42 and Aβ42/Aβ40; Results: Aβ42 did not differ from Aβ42/Aβ40 ratio in the differentiation of ADD from FTD (AUCs 0.752 and 0.788 respectively; p = 0.212). The τT/Aβ42 ratio provided maximal discrimination between ADD and FTD (AUC:0.893; sensitivity 88.8%, specificity 80%). BIOMARKAPD/ABSI criteria classified 60 patients as having AD pathology and 211 as non-AD. A total of 22 had discrepant results and were excluded. Aβ42/Aβ40 ratio was superior to Aβ42 in the differentiation of AD pathology from non-AD pathology (AUCs: 0.939 and 0.831, respectively; p < 0.001). In general, biomarker ratios and composite markers were superior to single CSF biomarkers in both analyses. Aβ42/Aβ40 ratio is superior to Aβ42 in identifying AD pathology, irrespective of the clinical phenotype. CSF biomarker ratios and composite markers provide higher diagnostic accuracy compared to single CSF biomarkers.

Estimating phylogenies from genomes: A beginners review of commonly used genomic data in vertebrate phylogenomics.

Despite the increasing feasibility of sequencing whole genomes from diverse taxa, a persistent problem in phylogenomics is the selection of appropriate genetic markers or loci for a given taxonomic group or research question. In this review, we aim to streamline the decision-making process when selecting specific markers to use in phylogenomic studies by introducing commonly used types of genomic markers, their evolutionary characteristics, and their associated uses in phylogenomics. Specifically, we review the utilities of ultraconserved elements (including flanking regions), anchored hybrid enrichment loci, conserved nonexonic elements, untranslated regions, introns, exons, mitochondrial DNA, single nucleotide polymorphisms, and anonymous regions (nonspecific regions that are evenly or randomly distributed across the genome). These various genomic elements and regions differ in their substitution rates, likelihood of neutrality or of being strongly linked to loci under selection, and mode of inheritance, each of which are important considerations in phylogenomic reconstruction. These features may give each type of marker important advantages and disadvantages depending on the biological question, number of taxa sampled, evolutionary timescale, cost effectiveness, and analytical methods used. We provide a concise outline as a resource to efficiently consider key aspects of each type of genetic marker. There are many factors to consider when designing phylogenomic studies, and this review may serve as a primer when weighing options between multiple potential phylogenomic markers.

Differentiation of Adipose-Derived Stem Cells into Smooth Muscle Cells in an Internal Anal Sphincter-Targeting Anal Incontinence Rat Model.

Studies on development of an anal incontinence (AI) model targeting smooth muscle cells (SMCs) of the internal anal sphincter (IAS) have not been reported. The differentiation of implanted human adipose-derived stem cells (hADScs) into SMCs in an IAS-targeting AI model has also not been demonstrated. We aimed to develop an IAS-targeting AI animal model and to determine the differentiation of hADScs into SMCs in an established model. The IAS-targeting AI model was developed by inducing cryoinjury at the inner side of the muscular layer via posterior intersphincteric dissection in Sprague-Dawley rats. Dil-stained hADScs were implanted at the IAS injury site. Multiple markers for SMCs were used to confirm molecular changes before and after cell implantation. Analyses were performed using H&E, immunofluorescence, Masson's trichrome staining, and quantitative RT-PCR. Impaired smooth muscle layers accompanying other intact layers were identified in the cryoinjury group. Specific SMC markers, including SM22α, calponin, caldesmon, SMMHC, smoothelin, and SDF-1 were significantly decreased in the cryoinjured group compared with levels in the control group. However, CoL1A1 was increased significantly in the cryoinjured group. In the hADSc-treated group, higher levels of SMMHC, smoothelin, SM22α, and α-SMA were observed at two weeks after implantation than at one week after implantation. Cell tracking revealed that Dil-stained cells were located at the site of augmented SMCs. This study first demonstrated that implanted hADSc restored impaired SMCs at the injury site, showing stem cell fate corresponding to the established IAS-specific AI model.

Morphology and morphometry of interneuron subpopulations of the marmoset monkey (Callithrix jacchus) striatum

The mammalian striatum has long been considered a homogeneous entity. However, neuroanatomical and histochemical studies reveal that the striatum is much more heterogeneous than previously suspected. The caudate (Cd) and putamen (Pu) are composed of two chemical compartments: the matrix and the striosomes. Striatal interneurons have been classified into a variety of morphological and neurochemical subtypes. In this study, we compared the distribution of multiple neurochemical markers in the striatum of marmosets and described the morphology of different types of striatum interneurons. The immunoreactivities of choline-acetyl transferase (ChAT), neuropeptide Y (NPY), nitric oxide synthase (NOS), calretinin (CR), parvalbumin (PV) were analyzed along the entire rostrocaudal extent of the marmoset striatum. Calbindin immunohistochemistry is useful in identifying medium spiny neurons (MSNs), with efficient soma staining. Based on the size of the CB-positive cells, considered medium-sized, as expected, cholinergic cells are larger in area and diameter than the other subpopulations investigated, followed by NOS, NPY, PV and CR. In adjacent CB and PV-stained sections, the matrix and striosomes were clearly distinguished. The matrix is strongly reactive to CB and PV neuropils, while the striosomes exhibit low reactivity, especially in the dorsal Cd. Therefore, we provide a detailed description morphology and distribution of striatal interneuron populations in a model as a valuable tool for studying neurodegenerative pathogenesis, progression and treatment strategies.

COMPARISON BETWEEN C-REACTIVE PROTEIN AND SERUM LACTATE IN EVALUATION OF SEPSIS INDUCED ACUTE KIDNEY INJURY AND SEPSIS WITHOUT ACUTE KIDNEY INJURY

Background One of the main causes of death in critically ill patients is sepsis. In Intensive care unit half of the patients develop Acute Kidney Injury, and has high mortality rates. In assessment of sepsis there are multiple markers in which C-reactive protein, serum lactate are commonly used. C-reactive protein is known biomarker of inammation and involved in several immunological functions. Aim To correlate C-reactive protein and Serum lactate in evaluation of sepsis induced acute kidney injury and compare these parameters in sepsis without acute kidney injury. Materials &amp; Methods This is a Cross-sectional study which will be done at Mahatma Gandhi Medical College and Research Institute, Pondicherry. This study comprises of 52 patients out of which Group 1 are 26 patients having sepsis with AKI and Group 2 are 26 patients are having sepsis without AKI. Complete case history including the socio-demographic data and other medical conditions of the patient was recorded and &amp; the collected blood samples were recorded for CRP, Serum lactate as well as other parameters. Results Mean age of subjects in sepsis with AKI and sepsis without AKI are almost similar, with 55.8 % being females. 90% of sepsis with AKI group and 76% of sepsis without AKI group has CRP of &gt;10 and 42% of sepsis with AKI group and 34% of sepsis without AKI group has Lactate of &gt;2. Conclusion CRP and Serum lactate are markers of inammation. CRP has been used to monitor the course of infection and inammatory diseases. CRP has not only a biochemical marker of inammation but also an active modulator of the inammatory response.

Aldolase B attenuates clear cell renal cell carcinoma progression by inhibiting CtBP2.

Aldolase B (ALDOB), a glycolytic enzyme, is uniformly depleted in clear cell renal cell carcinoma (ccRCC) tissues. We previously showed that ALDOB inhibited proliferation through a mechanism independent of its enzymatic activity in ccRCC, but the mechanism was not unequivocally identified. We showed that the corepressor C-terminal-binding protein 2 (CtBP2) is a novel ALDOB-interacting protein in ccRCC. The CtBP2-to-ALDOB expression ratio in clinical samples was correlated with the expression of CtBP2 target genes and was associated with shorter survival. ALDOB inhibited CtBP2-mediated repression of multiple cell cycle inhibitor, proapoptotic, and epithelial marker genes. Furthermore, ALDOB overexpression decreased the proliferation and migration of ccRCC cells in an ALDOB-CtBP2 interaction-dependent manner. Mechanistically, our findings showed that ALDOB recruited acireductone dioxygenase 1, which catalyzes the synthesis of an endogenous inhibitor of CtBP2, 4-methylthio 2-oxobutyric acid. ALDOB functions as a scaffold to bring acireductone dioxygenase and CtBP2 in close proximity to potentiate acireductone dioxygenase-mediated inhibition of CtBP2, and this scaffolding effect was independent of ALDOB enzymatic activity. Moreover, increased ALDOB expression inhibited tumor growth in a xenograft model and decreased lung metastasis in vivo. Our findings reveal that ALDOB is a negative regulator of CtBP2 and inhibits tumor growth and metastasis in ccRCC.

The Basolateral Amygdala Sends a Mixed (GABAergic and Glutamatergic) Projection to the Mediodorsal Thalamic Nucleus.

The medial prefrontal cortex receives converging inputs from the mediodorsal thalamic nucleus (MD) and basolateral amygdala (BLA). Although many studies reported that the BLA also projects to MD, there is conflicting evidence regarding this projection, with some data suggesting that it originates from GABAergic or glutamatergic neurons. Therefore, the present study aimed to determine the neurotransmitter used by MD-projecting BLA cells in male and female rats. We first examined whether BLA cells retrogradely labeled by Fast Blue infusions in MD are immunopositive for multiple established markers of BLA interneurons. A minority of MD-projecting BLA cells expressed somatostatin (∼22%) or calretinin (∼11%) but not other interneuronal markers, suggesting that BLA neurons projecting to MD not only include glutamatergic cells, but also long-range GABAergic neurons. Second, we examined the responses of MD cells to optogenetic activation of BLA axons using whole-cell recordings in vitro Consistent with our immunohistochemical findings, among responsive MD cells, light stimuli typically elicited isolated EPSPs (73%) or IPSPs (27%) as well as coincident EPSPs and IPSPs (11%). Indicating that these IPSPs were monosynaptic, light-evoked EPSPs and IPSPs had the same latency and the IPSPs persisted in the presence of ionotropic glutamate receptor antagonists. Overall, our results indicate that the BLA sends a mixed, glutamatergic-GABAergic projection to MD, which likely influences coordination of activity between BLA, MD, and medial prefrontal cortex. An important challenge for future studies will be to examine the connections formed by MD-projecting glutamatergic and GABAergic BLA cells with each other and other populations of BLA cells.SIGNIFICANCE STATEMENT The mediodorsal thalamic nucleus (MD) and basolateral amygdala (BLA) send convergent projections to the medial prefrontal cortex. Although many studies reported that the BLA also projects to MD, there is conflicting evidence as to whether this projection is glutamatergic or GABAergic. By combining tract tracing, immunohistochemistry, optogenetics, and patch clamp recordings in vitro, we found that BLA neurons projecting to MD not only include glutamatergic cells, but also long-range GABAergic neurons. Differential recruitment of these two contingents of cells likely influences coordination of activity between the BLA, MD, and medial prefrontal cortex.

Clinical outcomes of a large-scale, partnership-based regional food prescription program: results of a quasi-experimental study

BackgroundFood prescription programs are gaining interest from funders, policy makers, and healthcare payers as a way to provide value-based care. A small body of research suggests that such programs effectively impact health outcomes; however, the quality of existing studies is variable, and most studies use small samples. This study attempts to address these gaps by utilizing a quasi-experimental design with non-equivalent controls, to evaluate clinical outcomes among participants enrolled in a food prescription program implemented at scale.MethodsWe completed a secondary analysis of participant enrollment and utilization data collected between May 2018 and March 2021, by the Houston Food Bank as part of its multi-institution food prescription program. Enrollment data was obtained from 16 health care partners and redemption data from across 40 food pantries in Houston, Texas. Our objective was to assess if program participation impacted multiple cardio-metabolic markers. Exposure was defined as any visit to a food pantry after receipt of prescription. Linear and logistic regression models were used to estimate change in outcomes by exposure status and number of food pantry visits.ResultsExposed patients experienced a −0.28% (p = 0.007) greater change in HbA1c than unexposed patients, over six months. Differences across exposure categories were seen with systolic blood pressure (−3.2, p < 0.001) and diastolic blood pressure (−2.5, p = 0.028), over four months. The odds of any decline in HbA1c (OR = 1.06 per visit, p < 0.001) and clinically meaningful decline in HbA1c (OR = 1.04 per visit, p = 0.007) showed a linear association with visit frequency.ConclusionsOur study of a large food prescription program involving multiple health care and food pantry sites provides robust evidence of a modest decline in HbA1c levels among participants. These results confirm that food prescription programs can continue to be effective at scale, and portend well for institutionalization of such programs.

Utility of monocyte HLA-DR and rationale for therapeutic GM-CSF in sepsis immunoparalysis.

Sepsis, a heterogeneous clinical syndrome, features a systemic inflammatory response to tissue injury or infection, followed by a state of reduced immune responsiveness. Measurable alterations occur in both the innate and adaptive immune systems. Immunoparalysis, an immunosuppressed state, associates with worsened outcomes, including multiple organ dysfunction syndrome, secondary infections, and increased mortality. Multiple immune markers to identify sepsis immunoparalysis have been proposed, and some might offer clinical utility. Sepsis immunoparalysis is characterized by reduced lymphocyte numbers and downregulation of class II human leukocyte antigens (HLA) on innate immune monocytes. Class II HLA proteins present peptide antigens for recognition by and activation of antigen-specific T lymphocytes. One monocyte class II protein, mHLA-DR, can be measured by flow cytometry. Downregulated mHLA-DR indicates reduced monocyte responsiveness, as measured by ex-vivo cytokine production in response to endotoxin stimulation. Our literature survey reveals low mHLA-DR expression on peripheral blood monocytes correlates with increased risks for infection and death. For mHLA-DR, 15,000 antibodies/cell appears clinically acceptable as the lower limit of immunocompetence. Values less than 15,000 antibodies/cell are correlated with sepsis severity; and values at or less than 8000 antibodies/cell are identified as severe immunoparalysis. Several experimental immunotherapies have been evaluated for reversal of sepsis immunoparalysis. In particular, sargramostim, a recombinant human granulocyte-macrophage colony-stimulating factor (rhu GM-CSF), has demonstrated clinical benefit by reducing hospitalization duration and lowering secondary infection risk. Lowered infection risk correlates with increased mHLA-DR expression on peripheral blood monocytes in these patients. Although mHLA-DR has shown promising utility for identifying sepsis immunoparalysis, absence of a standardized, analytically validated method has thus far prevented widespread adoption. A clinically useful approach for patient inclusion and identification of clinically correlated output parameters could address the persistent high unmet medical need for effective targeted therapies in sepsis.

Expansion microscopy reveals subdomains in C. elegans germ granules.

Light and electron microscopy techniques have been indispensable in the identification and characterization of liquid-liquid phase separation membraneless organelles. However, for complex membraneless organelles such as the perinuclear germ granule in C. elegans, our understanding of how the intact organelle is regulated is hampered by (1) technical limitations in confocal fluorescence imaging for the simultaneous examination of multiple granule protein markers and (2) inaccessibility of electron microscopy. We take advantage of the newly developed super resolution method of expansion microscopy (ExM) and in situ staining of the whole proteome to examine the C. elegans germ granule, the P granule. We show that in small RNA pathway mutants, the P granule is smaller compared with WT animals. Furthermore, we investigate the relationship between the P granule and two other germ granules, Mutator foci and Z granule, and show that they are located within the same protein-dense regions while occupying distinct subdomains within this ultrastructure. This study will serve as an important tool in our understanding of germ granule biology and the biological role of liquid-liquid phase separation.