👤 Sara K Orlowski

Pulmonary large-cell neuroendocrine carcinoma (LCNEC) is a rare lung malignancy characterized by an aggressive clinical course and an unfavorable prognosis. Next-generation sequencing (NGS) has revealed that LCNECs exhibit molecular features resembling either small-cell lung carcinoma (SCLC-like LCNEC) or non-small cell lung carcinoma (NSCLC-like LCNEC). This study aimed to characterize the incidence of actionable gene variants in a retrospective cohort of LCNEC patients using a targeted NGS approach. Microscopic diagnosis was established according to the 2021 World Health Organization (WHO) classification using a standard immunohistochemical (IHC) panel. In total, 216 LCNEC tumor samples were analyzed for molecular variants in 17 genes using the RNA-based Archer FusionPlex Lung NGS assay (Integrated DNA Technologies, USA) and the MiSeq platform (Illumina, USA)-an algorithm utilized for routine NSCLC diagnosis. Overall, 46 variants were identified in 46/216 (21.3%) tumor samples, with 28/216 (13%) LCNECs harboring at least one actionable molecular variant potentially targetable by registered or investigational agents. Show less

Drought and water scarcity, exacerbated by global warming, are enormous threats to global food sustainability and security. Poultry, in particular, are highly impacted by adverse environmental stressors. As nutrient absorption and intestinal integrity are critical for growth and performance, understanding the impact on the broiler gastrointestinal tract is highly relevant. Here, we examined the effect of chronic cyclic heat stress (HS) on the jejunal expression profile of tight-junction, gap-junction, adherens, and desmosome genes in the 4th generation of broiler lines divergently selected for low (LWE)- and high-water efficiency (HWE). Male HWE and LWE broilers (n = 240/line) were allotted to 12 environmental chambers (2 floor pens/chamber, 6 chambers/line, 20 birds/pen) and were exposed to cyclic HS (36°C for 9h/day from 9:00 am to 6:00 pm) or thermoneutral conditions (25°C) from day 29 to 49 of age in a 2 × 2 factorial design. Growth performance and mortality were recorded. At day 49, jejunal tissues were collected for molecular analyses using real-time quantitative PCR and immunoblot. Jejunal gene expression of multiple gut integrity factors were higher (P < 0.05) in the HWE as compared to the LWE lines, including claudin 22 (CLDN22), -34, occluding (OCDN), zona-occludin-2 (ZO-2), gap junction alpha1 (GJA1), GJA3, GJC1, and cadherin 1 (CDH1). CLDN8, -20, -25, -4, GJC2, and GJD2 were also greater (P < 0.05) in HWE, but were additionally downregulated (P < 0.05) during HS. Conversely PALS1-associated tight junction protein (PATJ) and desmocollin 1 (DSC1) mRNAs were significantly downregulated in the HWE as compared to the LWE broilers. Significant interactions between the line and environment were seen in CLDN1, where the expression was decreased in the LWE but increased in the HWE in HS. Additionally, CLDN15 and -16 genes were greatest in the HWE under TN conditions, while catenin alpha 2 (CTNNA2) was highest in the HWE during HS. Overall, the jejunal expression profile of key genes associated with intestinal barrier integrity likely contributes to the water efficiency phenotype and the response of these birds to HS. Show less

Heat stress (HS) has long posed a significant challenge to the poultry industry due to its adverse effects, such as depressed feed intake, decreased growth performance, and increased water consumption. Water efficiency (WE, conversion of water intake into body weight gain), although often neglected, is a key economic and production trait that is significantly affected by HS. Recently, we selected two broiler lines for high WE (HWE) and low WE (LWE) and showed a differential hypothalamic expression of genes involved in water homeostasis regulation. As the gut also plays a significant role in water absorption, the present study aimed to determine the effect of chronic HS on duodenal barrier integrity in LWE and HWE broilers. Male HWE and LWE chicks (240 chicks/line) were individually wing-banded for line identification, weighed, and placed in 12 controlled environmental chambers (2 pens/chambers). On day 29, birds were subjected to thermoneutral conditions (TN, 25 °C) or cyclic HS conditions (HS, 36 °C for 9 h/day from 9:00 a.m. to 6:00 p.m.) (120 birds/line/environment) for 3 weeks. On day 49, duodenal tissues were collected for histological, biochemical, and molecular analyses. Hematoxylin and eosin (H&E) staining revealed that HS significantly reduced villus height in the duodenum. Further analysis using qPCR showed that the mRNA expressions of intestinal barrier integrity-related genes, including claudins (CLDN1, 4, 5, 8, 16, and 22), PALS1-associated tight junction protein ( Show less

Lysosomes and related precursor organelles robustly build up in swollen axons that surround amyloid plaques and disrupted axonal lysosome transport has been implicated in worsening Alzheimer's pathology. Our prior studies have revealed that loss of Adaptor protein-4 (AP-4) complex function, linked primarily to Spastic Paraplegia (HSP), leads to a similar build of lysosomes in structures we term "AP-4 dystrophies". Surprisingly, these AP-4 dystrophies were also characterized by enrichment of components of APP processing machinery, β-site cleaving enzyme 1 (BACE1) and Presenilin 2. Our studies examining whether the abnormal axonal lysosome build up resulting from AP-4 loss could lead to amyloidogenesis revealed that the loss of AP-4 complex function in an Alzheimer's disease model resulted in a strong increase in size and abundance of amyloid plaques in the hippocampus and corpus callosum as well as increased microglial association with the plaques. Interestingly, we found a further increase in enrichment of the secretase, BACE1, in the axonal swellings of the plaques of Alzheimer model mice lacking AP-4 complex compared to those having normal AP-4 complex function, suggestive of increased amyloidogenic processing under this condition. Additionally, the exacerbation of plaque pathology was region-specific as it did not increase in the cortex. The burden of the AP-4 linked axonal dystrophies/AP-4 dystrophies was higher in the corpus callosum and hippocampus compared to the cortex, establishing the critical role of AP-4 -dependent axonal lysosome transport and maturation in regulating amyloidogenic amyloid precursor protein processing. Show less

Lysosomes and related precursor organelles robustly build up in swollen axons that surround amyloid plaques and disrupted axonal lysosome transport has been implicated in worsening Alzheimer's pathology. Our prior studies have revealed that loss of Adaptor protein-4 (AP-4) complex function, linked primarily to Spastic Paraplegia (HSP), leads to a similar build of lysosomes in structures we term "AP-4 dystrophies". Surprisingly, these AP-4 dystrophies were also characterized by enrichment of components of APP processing machinery, β-site cleaving enzyme 1 (BACE1) and Presenilin 2. Our studies examining whether the abnormal axonal lysosome build up resulting from AP-4 loss could lead to amyloidogenesis revealed that the loss of AP-4 complex function in an Alzheimer's disease model resulted in a strong increase in size and abundance of amyloid plaques in the hippocampus and corpus callosum as well as increased microglial association with the plaques. Interestingly, we found a further increase in enrichment of the secretase, BACE1, in the axonal swellings of the plaques of Alzheimer model mice lacking AP-4 complex compared to those having normal AP-4 complex function, suggestive of increased amyloidogenic processing under this condition. Additionally, the exacerbation of plaque pathology was region-specific as it did not increase in the cortex. The burden of the AP-4 linked axonal dystrophies/AP-4 dystrophies was higher in the corpus callosum and hippocampus compared to the cortex, establishing the critical role of AP-4 -dependent axonal lysosome transport and maturation in regulating amyloidogenic amyloid precursor protein processing. Show less

High environmental temperature has strong adverse effects on poultry production, welfare, and sustainability and, thereby, constitutes one of the most challenging stressors. Although colossal information has been published on the effects of heat stress on poultry productivity and gut health, the fundamemntal mechanisms associated with heat stress responses and intestinal barrier function are still not well defined. The aim of the present study was, therefore, to determine the effects of acute (2 h) heat stress on growth performance, gut integrity, and intestinal expression of heat shock and tight junction proteins in slow- (broilers of the 1950's, ACRB), moderate- (broilers of 1990's, 95RAN), rapid-(modern broilers, MRB) growing birds, and their ancestor wild jungle fowl (JF). Heat stress exposure significantly increased the core body temperature of 95RAN and MRB chickens by ~0.5-1°C, but not that of JF and ACRB compared to their counterparts maintained at thermoneutral conditions. Heat stress also depressed feed intake and increased serum fluorescein isothiocyanate-dextran (FITC-D) levels ( Show less