bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanisms involved in the disruption of the intestinal barrier during Necrotizing Enterocolitis (NEC)
Authors: Bein Amir , Schwartz Betty
Published: June 2021

Objectives and Methods

Characterize cellular mechanisms involved in development of NEC. Assessments were conducted in in vitro systems measuring differential response of mature intestinal epithelial cells (IEC6) versus non mature cells (FHs74Int – embryonic cells) to NEC associated stress.

Establish how NEC‐associated stress perturbs intestinal tissue functionality. Evaluations carried out in an ex‐vivo novel organ culture model mimicking NEC developed for these studies.

Compare events of abovementioned in vitro and ex vivo NEC models to evaluations obtained in intestinal specimens from premature healthy and NEC affected infants.

Results

Occludin was down regulated in mature cells exposed to NEC (hypoxia and LPS); in embryonic cells only minor alternations occurred. Embryonic cells expressed TLR4 but were devoid of junction structures. NHE1 was down regulated and a shift of β‐catenin from the cytosol to the nucleus was observed in both cell lines exposed to hypoxia. Transcription and translation of ZO1 was significantly down regulated in organ culture model exposed to NEC conditions were applied to, similarly to intestinal specimens from NEC affected infants.

Conclusions

NEC affects tight junctions, Wnt‐pathway and functional intestinal proteins.
Disruption of the epithelial barrier upon Shigella flexneri infection
Authors: Maldonado-Contreras Ana , Mumy Karen , McCormick Beth
Published: June 2021

Shigella is one of the most communicable bacterial pathogens and causes the disease shigellosis. Shigella invades the colonic mucosa from the basolateral surface, but how Shigella gets to the basolateral domain is incompletely understood. We have shown that S. flexneri alters the cell‐cell junction complex facilitating paracellular passage of the organism across the epithelial barrier. To understand the mechanism by which this bacterium alters the intestinal epithelial junctions, we focused on SepA, a bacterial protein associated with tissue invasion and destruction upon infection. To test the hypothesis that SepA is involved in altering the intestinal epithelial barrier function, polarized monolayers of human intestinal epithelia or xenotransplanted human intestine were infected with wild‐type S. flexneri or an isogenic SepA mutant strain. We found that absence of SepA lead to a less severe disturbance on the intestinal barrier, which correlate with not only a decrease in bacterial invasion, but also significant reduction of neutrophils transepithelial migration. Functionally, we found SepA to elicit cleavage of the intracellular domain of E‐cadherin, which we speculate contributes to the observed cell‐cell junction permeability, and in turns permits S. flexneri access to the basolateral domain to invade and trigger the intense inflammatory response, a hallmark of this disease.

NIH
Immune System Adaptation to Rapid Changes in Bacteremia
Authors: Londono Diana , Diego Cadavid
Published: June 2021

Analogous to sepsis, relapsing fever (RF) borreliosis is a rapidly progressive and at times fatal infection of blood and tissues caused by different Borrelia species. RF offers a valuable opportunity to study the adaptation of the mammalian immune system to rapid changes in pathogen load due to the rapid appearance and disappearance of large numbers of bacteria from the circulation The purpose of the present study was to investigate how the immune system adapts to the rapid changes in pathogen load that take place before, during, and after bacteremia in immunocompetent mice. The course of bacteremia varies depending of the initial inoculum and the genetic background. The kinetics on specific antibody production depends of the initial inoculum. The anti‐inflammatory cytokines response is fundamental at the time of high pathogen load. The chemokine responses are different on each period of bacteremia but are command by CCL2, CXCL10 and CXCL13. Cells from the innate immune system play an important role during the first peak bacteremia in the acute infection. The results identified the key molecular and cellular immunological events that play a leading role during the various phases of bacteremia in this model.
Plasma gelsolin enhances lung macrophage host defense against bacterial pneumonia in mice
Authors: Yang Zhiping , Chiou Terry , Lee Po-Shun , Stossel Tom , Kobzik Lester
Published: June 2021

Plasma gelsolin (pGSN) functions as part of the ‘extracellular actin scavenging system’ but its potential to improve host defense against infection has not been studied. Mice (C57BL/6, male) were pre‐treated with recombinant human plasma gelsolin (rhu‐pGSN, 400 mg/kg, s.c.) or vehicle 2 h before infection with Strep. pneumoniae (serotype 3, 150K CFU, i.n.) and evaluated 24 h later. rhu‐pGSN caused enhanced bacterial clearance (% bacterial left alive in lungs, mean±SD: control, 88 ± 38 vs. rhu‐pGSN‐treated, 10 ± 5; n ≥ 13 mice/grp) and reduced acute inflammation (% BAL PMNs: control, 30 ± 11 vs. rhu‐pGSN‐treated, 8 ± 3). Similar results were seen with aerosolized rhu‐pGSN. In vitro, rhu‐pGSN rapidly improved lung macrophage uptake and killing of bacteria. rhu‐pGSN caused an activating phosphorylation (ser1177) of macrophage nitric oxide synthase type III (NOS3), which has important bactericidal functions in the AM. rhu‐pGSN failed to enhance bacterial killing in NOS3−/− macrophages. pGSN binds sphingosine‐1‐phosphate (S1P), and may enhance its delivery to the S1P1 receptor. Both S1P and the S1P mimic, FTY720P, enhanced macrophage uptake and killing of bacteria, and activated macrophage NOS3 as well. Plasma gelsolin enhances innate resistance to pneumococcal pneumonia via lung macrophage NOS3, and is a potential immunomodulator for improving lung host defense against bacterial pneumonia. Support: NIH ES00002 ES11008
Phospholipase A2 is involved in S. pneumoniae‐elicited PMN transepithelial migration
Authors: Bhowmick Rudra , McCormick Beth A. , Leong John M.
Published: June 2021

Alveolar infiltration of neutrophils (PMNs) characterizes Streptococcus pneumoniae lung infection. Using in vitro lung epithelium and human PMNs, we have shown that S. pneumoniae‐induced PMN transepithelial migration is mediated by the PMN chemoattractant hepoxilin A3 (HXA3), an eicosanoid metabolized from arachidonic acid (AA). Release of AA from membrane phospholipids is the rate‐limiting step in eicosanoid generation. We studied the role of phospholipase A2 (PLA2), which generates AA, on S. pneumoniae‐elicited PMN transpeithelial migration. We found that pneumococcal infection resulted in release of AA from lung epithelial cells. This event was partially inhibited by pan‐ PLA2 inhibitors ONO‐RS‐082 and ACA, and also by diacylglycerol (DAG) lipase inhibitor. Pan‐PLA2 inhibitors significantly reduced S. pneumoniae‐induced PMN transmigration, but the DAG lipase inhibitor had no effect. We found that, both cytosolic PLA2 (cPLA2) and secretory PLA2 (sPLA2) are important for generation of free AA and inhibition of these enzymes lead to dramatic reduction in PMN migration. We observed that S. pneumoniae infection caused increase in the phosphorylation of cPLA2, suggesting a mechanism whereby S. pneumoniae activates cPLA2 and sPLA2 to generate free AA that may be converted to HXA3 which ultimately mediate PMN transmigration. This research is supported by institutional funds of Professor Leong.
Redefining cleft lip and palate
Authors: Vieira Alexandre Rezende
Published: June 2021

Cleft lip and palate is the most common craniofacial birth defect and its etiology, in regards to both genetic and environmental influences, has been focus of many reports in the literature. Information regarding research on long term outcomes, genes and their interactions with other genes, and gene‐environment interactions are compiled to provide the audience with a critical and up to date overview on the current knowledge of the etiology of these defects. Recent epidemiological evidence strongly suggests individuals born with clefts have a shorter life spam and may have a higher incidence of cancer and psychological disorders. IRF6 was shown to be an important contributor to cleft lip and palate, but the functional variant leading to the defect has not been defined yet. A likely scenario is that IRF6 interacts with other genes. Inactivation of MSX1 and genes in the FGF family were shown to lead to cleft lip and palate as well. In addition, missense mutations in a number of candidate genes may cause cleft lip and palate, but definitive evidence regarding the biological consequences of these mutations is yet to be unraveled. The latest effort in gene identification and the associations between isolated cleft lip and palate and the loci harboring IRF6 (1q32) and 8q24.21 will be highlighted as well as the latest insight from more sophisticated phenotypical characterization and the inclusion of covariables related to the environment in the analysis of genetic variation. Maternal cigarette smoking increases the risk of a baby born with clefts, in particular when the mother carries the GSTT1‐null variants. It is possible that the underlying mechanism invloved in smoking is hypoxia, and hypoxia could also provide an explanation to why there is a higher incidence of some craniofacial anomalies (cleft lip, microtia, and hemifacial microsomia) at sites 2,000 meters above the sea level. The latest approaches in craniofacial research include the analysis of several additional phenotypical features of the population with the goal to increase statistical power of genetics studies.
The present and the future of stem cell therapies
Authors: Giacin Kenneth
Published: June 2021

StemCyte is a leading Umbilical Cord Blood Stem Cell Therapy Company with one of the largest and most racially diverse Public Cord blood inventories in the world. We have been providing a life saving cellular therapy product since 2001 and have delivered high quality, HLA matched umbilical cord blood stem cells for more than 1600 transplants to 240+ of the world's leading Transplant Centers in over 30 countries. StemCyte has operations in the US, Taiwan and India and has embarked on a very deliberate strategy over the past 8 years to enhance our base business by consistently growing our revenue, year over year, while supporting research and being engaged in new clinical indications for stem cells sourced from umbilical cord blood. This presentation “The Present and Future of Stem Cell Therapies from the perspective of a Public Umbilical Cord Blood Bank CEO”; .chronicles the advances in UCB Clinical life saving applications, the current indications being explored by StemCyte and others and what we believe to be the potential uses of this rich source of adult stem cells while understanding the regulatory environment as well as the need to remain focused and sustain our ongoing business objectives.
Mechanisms underlying antinociception exerted by endomorphin in the spinal dorsal horn
Authors: Li Yun-Qing
Published: June 2021

Background

Endomorphin (EM) is a new opioid peptide and considered as the endogenous ligand for the mu‐opioid receptor (MOR) with high affinity. EM has similar effect in pain inhibition as morphine, but with much less side effects. In the dorsal root ganglion (DRG) and superficial layers (laminae I and II) of the spinal dorsal horn, many substance P (SP), EM and MOR immunpositive neurons and dense fibers and terminals are found, respectively. Based on these phenomena, we proposed a hypothesis that EM binding to its autoreceptor MOR on the pre‐synaptic buttons will inhibit further release of the noxious neurotransmitters, such as SP.

Aims

To prove whether the hypothesis is true or not, the antinociceptive effects of EM through pre‐synaptic mechanism in the superficial layers of the spinal dorsal horn were investigated.

Methods

Behavioral observation, whole‐cell patch clamp recording, immunofluorescent histochemistry and post‐embedding immunohistochemical staining methods were used.

Results

(1) EM pre‐treatment through intrathecal administration could dose‐dependent inhibit the primary mechanical allodynia induced by subcutaneous injection of complete Freund's adjuvant (CFA). This effect might be reversed by antagonists of neurokinin 1 receptor, and attenuated by SP or co‐administration of SP and EM, respectively. (2) EM remarkably reduced the frequency instead of the amplitude of miniature EPSCs in lamina II. This effect was antagonized by MOR antagonist beta‐FNA. EM could also inhibit the EPSCs induced by stimulating of the attached dorsal root. The average amplitude of the residual EPSCs increased during high concentration of SP being perfused. (3) SP, EM and MOR were co‐localized in the DRG neurons. SP and EM were observed within the same large dense‐coated granular synaptic vesicles in the pre‐synaptic buttons, whereas MOR was localized on the pre‐synaptic membrane of the pre‐synaptic buttons in which SP and EM were found to co‐exist in the same vesicles.

Conclusions

SP and EM are co‐localized in the pre‐synaptic button, of which MOR can be found on the pre‐synaptic membrane. EM inhibits excitatory synaptic transmission and decreases the release of SP from primary afferent terminals in the superficial layers of the spinal dorsal horn through activating MOR on primary afferent terminals to exert their analgesic effects.
mRNA decay in mammals
Authors: Maquat Lynne E.
Published: June 2021

In mammals, two different messenger ribonucleoproteins (mRNPs) serve as templates for protein synthesis. Newly synthesized CBP80‐CBP20 (CBC)‐bound mRNPs initially undergo a pioneer round of translation (Maquat et al., 2010). One purpose of this round is to ensure the quality of gene expression, as exemplified by nonsense‐mediated mRNA decay (NMD). NMD functions to eliminate mRNAs that prematurely terminate translation, and also contributes to proper gene control, and it targets CBC‐bound mRNPs (Sato et al., 2008; Isken et al., 2008). CBC‐bound mRNPs are remodeled to eIF4E‐bound mRNPs as a consequence of the pioneer round of translation and also independently of translation (Sato & Maquat, 2009). eIF4E‐bound mRNPs support the bulk of cellular protein synthesis and are the primary targets of mRNA decay mechanisms that conditionally regulate gene expression, such as Staufen1‐mediated mRNA decay (SMD) (Gong et al., 2009). Mechanistic aspects of NMD will be discussed, including how CBP80, which is acquired by the 5′ caps of newly synthesized transcripts, promotes NMD at multiple steps by promoting specific mRNP rearrangements (Hwang et al., 2010). Mechanistic aspects of SMD will also be described, including how Staufen1‐binding sites form not only by intramolecular base‐pairing within an mRNA 3′‐untranslated region but also by intermolecular basepairing between the Alu element of an mRNA 3‐untranslated region and a partially complementary Alu element within a long noncoding RNA (Gong & Maquat, 2011).
Consumption of capsaicin and capsiate potentiate glucose‐stimulated insulin secretion in 90% pancreatectomized diabetic rats
Authors: Park Sunmin , Kwon Dae Young , Kim Min Jung , Yang Hye Jeong
Published: June 2021

Red peppers and red pepper paste have anti‐obesity and anti‐inflammatory effects in animals and humans. These actions are related to the capsaicin in red pepper. We investigated whether the oral consumption of capsaicin and capsiate, a nonpungent capsaicin analogue, altered glucose homeostasis in 90% pancreatectomized (Px) diabetic rats. Diabetic rats were divided into 3 groups: 1) capsaicin, 2) capsiate or 3) dextrose (control). Rats were provided with a high fat diet (40 energy % fat) containing assigned components (0.03% capsaicin, capsiate, or dextrose) for 8 weeks. CPA and CPI reduced body weight, visceral fat, and serum leptin levels without modulating caloric intake in diabetic rats, compared to the control. CPA and CPI improved glucose tolerance. In comparison to the control, CPA and CPI potentiated first and second phase insulin secretion at hyperglycemic clamp. Both also increased β‐cell mass by increasing proliferation and decreasing apoptosis of β‐cells through potentiating insulin/IGF‐1 signaling. However, only CPI, not CPA, enhanced hepatic insulin sensitivity during euglycemic hyperinuslinemic clamp. This was related to enhanced pAkt→PEPCK and pAMPK signaling. CPA alone reduced triglyceride storage through activating pAMPK. In conclusion, CPA and CPI improve glucose homeostasis in different manners in diabetic rats, however, CPI may be better for alleviating diabetic symptoms.
Flavan‐3‐ol isolated from rhizome of Drynaria fortunei (Kunze) J. Sm. exerts osteoprotective effects via its actions on osteoblastogenesis and osteoclastogenesis
Authors: Wong Man-Sau , Wong Ka-Chun , Dong Xiaoli , Law Man-Chun , Chan Tak-Hang
Published: June 2021

(‐)‐Epiafzelechin (FRD) is a flavan‐3‐ol found in rhizome of Drynaria fortunei (Kunze) J. Sm., a Chinese herb commonly used for management of bone diseases. Previous study suggested FRD could stimulate osteoblastic functions. Our study aims to determine if FRD act on osteoblastogenesis and osteoclastogenesis, and if its actions in bone cells involve the activation of Estrogen Receptors (ER) using pre‐osteoblast MC3T3‐E1, pre‐osteoclast RAW 264.7 as well as osteoblast‐like UMR 106 cells. FRD could increase alkaline phosphatase activity and secretory collagen in MC3T3‐E1 cells.

FRD could reduce viability of RAW 264.7 cells and prevent the formation of osteoclastic cells, as observed by a reduction of Tartrate‐resistant acid phosphatase activity. Co‐incubation of ER antagonist (ICI 182780) with FRD abolished its positive effects on UMR 106 cell growth. FRD has a weak binding affinity to ER‐alpha and beta and could only weakly activate ER‐alpha, but not ER‐beta, mediated Estrogen Response Element (ERE)‐dependent transcriptional activity in UMR 106 cells.

In conclusion, our results showed that FRD could stimulate osteoblastogenesis and inhibit osteoclastogenesis and its positive effects in bone cells appeared to be ER dependent.

Funding support: RGC General Research Fund [POLYU 563209] and research studentship from the Hong Kong Polytechnic University
Effects of high‐fat diet on intestinal permeability and inflammation‐associated tumorigenesis
Authors: Kim Min-Young , Bae Yun-Jung , Bak Youn-Kyung , Kim Jin , Sung Mi-Kyung
Published: June 2021

High‐fat diet induces “chronic low‐grade” inflammation and oxidative stress. Intestinal barrier plays an important role and its disruption leads to the increased permeability implicated pathogenic changes in several diseases. The aim of this study was to investigate the effects of high‐fat diet on intestinal permeability and carcinogenesis. APCmin+/− mice were fed with normal diet(ND) or high fat diet(HD), to examine intestinal permeability and tumorigenesis. Although there was no significant difference in body weight, the total tumor number was higher in HD group than that of ND group. Plasma total antioxidant capacity was lower in HD group, while 8‐OHdG level was higher. Protein expressions of ZO‐1, claudin‐1 and occludin were lower in HD group suggesting intestinal permeability was increased. Plasma MPO and IL‐6 levels were higher in HD group than those of ND group. CD14, the only LPS receptor, plasma level and colonic TLR4 mRNA expression were increased in HD group than ND group. These results suggest that high‐fat diet increases oxidative stress and inflammation induced by endotoxemia disrupting intestinal gap junction proteins which increase membrane permeability and accelerating tumorigenesis. [This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education, Science and Technology (2010‐0000918) and the SRC program(Center for Food&Nutritional Genomics:2011‐0000918) of the National Research Foundation(NRF) of Korea funded by MEST.]

Grant Funding Source : Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education,Science and Technology (2010‐ 0000918) and the SRC program(Center for Food&Nutritional Genomics:2011‐0000918) of the National Research Foundation(NRF) of Korea funded by MEST
Children's perceptions of obesity and its relation to nutrition and physical activity
Authors: Economos Christina D. , Bakun Peter J. , Herzog Julia Bloom , Dolan Peter R. , Lynskey Vanessa M. , Sharma Shanti , Nelson Miriam E.
Published: June 2021

Data indicate that about one third of children in the US are overweight/obese (ov/ob). Understanding children's perceptions of obesity and related socio‐behavioral factors is important in crafting child‐receptive interventions to address this epidemic. In 2010 an online Harris Interactive Youth Query omnibus survey among 1224 children age 8–18 yrs in the US was commissioned to capture children's views. Though 46% of children knew that one third of US children are ov/ob another 46% believed that it was half or more. When asked to categorize themselves, only 28% reported being ov/ob compared to 35% among 6–18 yr olds nationally, underscoring the shift in cultural weight norms. 80% of self‐categorized ov/ob children worry about their weight compared to 31% of normal weight children. 92% of children felt it was important to not be overweight at their age and that it leads to teasing at school, inability to play sports well, and overweight and illness in adulthood. Children focused on eating habits (especially sugar intake) as the main reasons for childhood ov/ob. They did not report physical activity as a main reason. Children also focused on food more than physical activity when asked what adults need to know to help people their age achieve a healthy weight. The contemporary knowledge and views of children captured here should be addressed if interventions to reverse the childhood obesity epidemic are to be successful. RWJF
Dietary Supplements service members are using in Afghanistan and why they use them
Authors: Carvey Christina E , McGraw Susan M , Lieberman Harris R
Published: June 2021

Military service in Afghanistan is a stressful experience and service members may use Dietary supplements (DS) to cope with demands of deployment to combat. In 2010, we conducted an anonymous, voluntary survey of DS intake, demographic/lifestyle factors, and reasons for use in a convenience sample of service members in Afghanistan. A total of 260 questionnaires were completed (78% Army; 78% male). Most service members (72%) used at least one DS a week, with 25% using 5 or more per week. Among males, the most popular DS were multi‐ or megavitamins/minerals (MVI/Mega) (46%), protein powders/drinks (44%), and nitric oxide boosting DS (43%). Males most often used vitamins, minerals and antioxidants (AO) to promote general health (67, 48, and 75% of MVI/Mega, calcium, and AO users, respectively). Males frequently used protein powders/drinks, creatine, and amino acids to increase muscle strength (58, 67, 46%, respectively) and/or to enhance performance (19, 15, 21%, respectively). Nitric oxide DS were used to increase energy (41%) and muscle strength (35%). As expected, males consumed weight/fat loss DS primarily to lose weight (65%), but 25% cited increased energy as a benefit of weight/fat loss DS. Deployed service members regularly use DS, and many use multiple DS concurrently; DS are used for many reasons, some of which have little to no scientific justification. (Support: US Army Medical Research & Materiel Command)
Association of the Parental Feeding Practices and Type and Home Availability of Sugar‐Sweetened Beverages and Fruit Juice of Preschool Children from Low Income Families
Authors: Lora Karina , Quesada Catalina , Wakefield Dorothy , Ferris Ann
Published: June 2021

This study evaluated the association of parental feeding practices with home availability and children's reported portion size (PS) of sugar‐sweetened beverages (SSBs): juice drinks, soda, and fruit juice (FJ) for 300 poor parents of 3–5 y. old children. Participants provided demographic and anthropometric data, completed the Preschooler Feeding Questionnaire (PFQ) (Baughcum et al, 2001) and an inventory of beverages in the home. PFQ Factor analysis retained eight factors for 31 questions. Multivariate regression models tested if PFQ factors predicted child's daily PS of SSBs and FJ while controlling for demographic variables. Child's reported daily PS (oz) were 16.2±15.4 fruit juice, 13.8± 12.0 juice drinks, and 1.43±3.2 soda and were correlated with home availability. Higher structure during feeding interactions was associated with lower reported consumption of SSBs by the child. Latino parents were significantly more child‐centric, used more rewards, and more likely to feed older children inappropriate foods than African‐American parents. Being a young parent (19–25 y) and SNAP participant was positively associated with home availability of juice drinks and child's reported PS. Parents’ education was negatively associated with child's soda PS (P< .05). Fewer SSBs were available in the homes of parents with controlled feeding practices.

Grant Funding Source : USDA/NRI 2008‐55215‐19071
Characterization of flavanol and procyanidin absorption and metabolism in humans: identification of potential bioactive metabolites
Authors: Ottaviani Javier Ignacio , Momma Tony Y. , Keen Carl L. , Schroeter Hagen
Published: June 2021

Accumulating data suggest a causal relationship between diets rich in flavanols (F) and procyanidins (P) and cardiovascular health in humans. Thus, it becomes important to understand the absorption, distribution, metabolism and excretion (ADME) of these food constituents. F and P encompass a complex group of phytonutrients, including 4 F stereoisomers, and various P, consisting of F oligomers of different chain lengths. In this study, we assessed metabolism and the systemic concentrations of F stereoisomers and P in humans, following the intake of these compounds in a well‐characterized food matrix. The results show that the ADME of F is stereoisomer‐specific. Moreover, F stereochemistry and metabolism represented major determinants for the vasodilation activity of these compounds in vivo. Compared to F, P reached negligible levels in circulation. Contrary to previous suppositions, P did not contribute to the systemic presence of F via depolymerization in the gastrointestinal tract. However, 5‐(3,4‐dihydroxyphenyl)‐γ‐valerolactone, a gut microbiome‐related catabolite, was detected after the intake of F and P. Taken together, our results argue for a more comprehensive discrimination between the F and P content in foods, which may result in the identification of clearer epidemiological outcomes, and a better understanding of the putative nutritional and health‐promoting properties of F and P in general.

Grant Funding Source: Mars, Inc
The immunological cellular and biochemical contents of breastmilk respond to maternal or infant infections
Authors: Hassiotou Foteini , Metzger Philipp , Trengove Naomi , Lai Ching Tat , Hartmann Peter , Filgueira Luis
Published: June 2021

Breastmilk contains immunological components that protect the infant. Among those, leukocytes have been variably present in breastmilk. To examine whether this variability may be related to the health status of the mother/infant dyad, mature breastmilk was collected from 20 dyads longitudinally, while they were healthy and while they had an infection. Leukocyte content was measured by flow cytometry via the pan‐leukocyte marker CD45. Breastmilk from healthy dyads had 0–1.5% of leukocytes (0–1,365 immune cells/mL breastmilk). During an infection of the mother and/or the infant, leukocyte content increased to up to 94% of total cells (2,594,982 leukocytes/mL breastmilk), with the majority of leukocytes being activated T‐cells that responded to viral peptides if stimulated in vitro, with corresponding proliferation and cytokine secretion. Interestingly, leukocyte content returned to baseline levels (≤1.5%) upon recovery from the infection. Similar responses were observed for immunoglobulins and lactoferrin. Despite their higher concentrations in mothers weaning or very late into lactation, leukocyte content remained low (0–1.5%) under healthy conditions. It is concluded that infection induces a rapid mechanism of leukocyte movement into breastmilk, followed by a biochemical response. Unless induced by infection, leukocytes are either absent or present in very small amounts in mature breastmilk.

Grant Funding Source : Women and Infants Research Foundation, Medela AG
The preventive effect of melatonin and/or exercise on cartilage destruction in collagenase‐induced osteoarthritis rats
Authors: Kim Hyunsoo , Hong Yunkyung , Hong Yonggeun
Published: June 2021

Osteoarthritis (OA) is the major cause of disability in the adult population. HIF‐2a is a crucial catabolic regulator of arthritic cartilage destruction. The purpose of this study is to evaluate the prophylactic effect of melatonin and/or exercise on the degenerative change in cartilage of collagenase‐induced OA rats. Melatonin (MT) was injected twice/day subcutaneously. The treadmill exercise (Ex) was conducted for 1‐hr/day(5 days/week). The width of affected knee joints was reduced by co‐intervention at day 28 after collagenase injection (P<.05). MT+Ex reduced EPAS1 as well as MMP13, whereas it increased ADAMTS4 level at day 28 (P<.05). There were ill‐defined cell layer, abnormally condensed cell morphology, and cartilage surface erosion in the OA+Veh group. However, it was reorganized in the OA+MT+Ex group. Not only bone surface density(BS/BV) but also trabecular number(Th.N) was significantly increased in OA+Veh as well as OA+MT group. However, it was recovered to normal level in OA+MT+Ex group. Taken together, melatonin with treadmill exercise effectively inhibits the initiation of OA induction suggesting prophylactic effect on degenerative OA progression.
Innate immune signaling plays a crucial role in the pathogenesis of avascular osteonecrosis of the femoral head
Authors: Okazaki Shunichiro , Nagoya Satoshi , Tateda Kenji , Katada Ryuichi , Mizuo Keisuke , Watanabe Satoshi , Yamashita Toshihiko
Published: June 2021

Objectives

Pathogenesis of avascular osteonecrosis of the femoral head (ONF) still remains unclear. We hypothesized that innate immune signaling via toll‐like receptor (TLR) play any role in pathogenesis of ONF. We found that administration of ligands of TLR with steroid treatment induces ONF. The present study verified the hypothesis and evaluated effect of steroid in the development of the ONF.

Methods

Male Wistar rats (n=120) were randomly divided six groups. Each group rats were given saline or ligands of TLR on day 1 and saline or methylprednisolone on day 2. Animals were sacrificed 1, 3, 7 or 14 days after last injection. Histopathological, biochemical analyses were then performed.

Results

ONF was observed in groups given ligands of TLR and methylprednisolone treatment. Although the activity of IRF7 did not change, the activity of NF‐κB was decreased after methylprednisolone injection.

Conclusions

The present study shows that the innate immunity signaling pathway via the TLR, especially MyD88 dependent pathway may play a crucial role in pathogenesis of ONF.
Development of a new chick chorioallantoic membrane model for human pancreas adenocarcinoma
Authors: CASTRONOVO Vincent , GONZALEZ Arnaud , DELVENNE Philippe , PEULEN Olivier
Published: June 2021

Pancreatic cancer is the fourth leading cause of cancer death in developed countries. Early‐stage pancreatic cancer is usually clinically silent, and disease only becomes apparent after the tumor invades surrounding tissues or metastasizes to distant organs. One of the urgent challenges in pancreas cancer research is to set up model systems allowing the development and testing of new therapies as well as the discovery of new diagnostic markers. The chick chorioallantoic membrane (CAM) has been successfully used to produce tumors from various cancer cell lines. Here, using BxPC3 human pancreatic cells we were able to produce within 7 days post‐grafting 30 mm3vascularized tumor on CAM. Histology analysis revealed that the cancer cells formed tumors compatible with PanIN‐3 grade human malignant adenocarcinomas. Immunohistochemistry revealed that the malignant cells were positive for carcinoembryonic antigen and cytokeratin‐7, and ‐19. Functional blood vessels were numerous between pancreatic islets. Interestingly, we were able to demonstrate that two biomarkers recently identified in our lab from human pancreas cancer tissues, latent‐transforming growth factor beta‐binding protein 2 (LTBP‐2) and transforming growth factor beta‐induced (TGFBI) were found to be also expressed in the BxPC3. The possibility to perfuse the tumor allows to expect this model to be used to detect new biomarkers as well as inject and test drugs.

This work was support by the Fond Léon‐Fredericq (ULg) and FNRS.
Are your students ready for A & P? Developing an effective tool to identify students at risk for failure in introductory science courses
Authors: Gultice Amy Denise , Kallmeyer Robert , Witham Ann
Published: June 2021

Introductory science courses, such as Anatomy and Physiology, are gateway courses to professional degree programs in nursing, radiation technology, and other fields. At the University of Cincinnati Blue Ash College, Anatomy and Physiology I and Fundamentals of Biology I have failure/withdrawal rates of ~35%. In order to identify students at risk for failure, an online survey that collected information regarding students’ high school grades, current college status, personal responsibilities, and math and reading skills was administered to 200 students. Analysis of the data demonstrates statistically significant differences in the academic preparation of passing versus failing students. These results led to a comprehensive study of the college records of more than 2,000 students from a 5‐year period. The analysis shows statistically significant differences between passing and failing students in the following categories: age, high school GPA, college GPA, number of college hours earned, and placement test scores in English and math. Logistic regression analysis using the historical data produces a model with factors that predict student success in these courses. Evidence‐based advising that communicates this information to students could encourage underprepared students to delay entry into gateway courses until they develop the necessary skills, potentially increasing student success.
Acceptance and usefulness of an audience response system in teaching an undergraduate pharmacology class
Authors: Sekar M. Chandra
Published: June 2021

At all levels of education, teachers are encouraged to promote the concept of active learning in their classrooms and curriculums. Use of an audience response system encourages greater participation of the audience and should therefore stimulate learning.

Senior students, enrolled in the fourth year of their six‐year Pharm D program at the University of Findlay, were required to bring “clickers” for each of my classes in the cardiovascular disease module. While students were not graded for their answers, positive incentives in the form of participation points were awarded.

Forty‐eight of the fifty‐four students completed the anonymous Google Docs survey, the results of which are presented below:

Question
Response %

SA
A
D
SD
NA

Promoted learning
73
23
0
0
0

Improved concentration
40
54
6
0
0

Class more interesting
46
48
2
0
0

Identified difficult to understand concepts
52
44
4
0
0

Wrong answer explanation helped me to learn
38
56
2
4
0

Results clearly indicate that over 90% of the students believed clicker use in the class made the class more interesting and promoted learning.
BNIP3 and OGD‐induced oligodendrocyte death: Role in white matter injury in cerebral ischemia
Authors: Guan Teng , Li Chengren , Kong Jiming
Published: June 2021

We sought to identify potential role of BNIP3, a proapoptotic member of the Bcl‐2 family of death‐regulating proteins that promote programmed cell death, in Oxygen–glucose deprivation (OGD) treated primary cultures of rat oligodendrocyte lineage cells (OLs). OGD induced cytotoxic injury to various maturational stages of OLs. Survival assays revealed that oligodendrocyte precursor cells (OPCs) and later‐stage precursors were highly vulnerable, whereas more mature OLs were relatively resistant. Exposure of OLs to OGD resulted in a significant increase in BNIP3 expression. Knockdown of BNIP3 with shRNA reduced OGD‐triggered OL apoptosis. Thirty minutes ischemic preconditioning (IPC) of later‐stage precursors and more mature OLs down‐regulated the BNIP3 expression and shown cytoprotection. Interestingly, OPCs were more susceptible to OGD after IPC pretreatment, concurrent with highly BNIP3 expression. These studies unveil BNIP3 as an important mediator in OLs death and reveal a novel mechanism of action of IPC on stage‐specific OLs cultures, which may contribute to a better understanding of white matter injury in cerebral ischemia.
Immune synapse malformation: Role for CAV1 and PAK1
Authors: Gilling Christine Elizabeth , Mittal Amit K , Joshi Shantaram S
Published: June 2021

Caveolin‐1 (CAV1) a scaffold molecule, is a member of the tolerogenic signature and is significantly differentially expressed in chronic lymphocytic leukemia (CLL). CAV1 controls transmembrane signaling and has been hypothesized to regulate actin polymerization. Additionally, impaired immune synapse (IS) formation has been reported in CLL. Therefore the objective of this report was to identify the molecular basis of IS malformation in CLL. We hypothesized that IS malformation is caused by dysregulation of cytoskeletal molecules associated with CAV1. Ingenuity pathway analyses of differentially expressed cytoskeletal molecules in CLL cells revealed direct association of ARPC5, CALD1, CASK, DSTN, PFN2, STMN1, VASP, and WAS with CAV1. Further, high expression of CAV1 and DSTN were clinically correlated with a poor prognosis (p=0.047 and p=0.007, respectively). CAV1 knockdown in Raji and Jurkat cells, a model for IS formation, resulted in decreased IS formation with decreased expression of F‐actin (involved in actin polymerization) and PAK1 (associated with cell protrusions). Together, our results suggest a role for CAV1 in regulating immune synapse formation by regulating F‐actin and PAK1 molecules.

Grant Funding Source : Supported by the CLL Foundation, Houston, TX and the NIH INBRE grant P20 RR016469.
Tubulobulbar Complexes Formed In Sertoli Cells In Primary Cultures Associate With Intercellular Junction Elements
Authors: Du Min , Roskelley Calvin , Nicholls Peter , Stanton Peter , Deng Wanyin , Finlay Brett , Vogl Wayne
Published: June 2021

In the seminiferous epithelium, unique double membrane structures termed tubulobulbar complexes (TBCs) develop in association with basal junction complexes. These actin‐related structures are proposed to internalize intercellular junctions during the movement of spermatocytes from basal to adluminal compartments of the epithelium. To study the role of basal TBCs in junction turnover, we use a primary Sertoli cell culture system in which TBCs develop in the vicinity of junction complexes between Sertoli cells. We immunoprobed cultured Sertoli cells for the intercellular junction elements Claudin‐11, Connexin‐43 and Nectin‐2 and discovered rod‐like structures that protrude into Sertoli cells from the cell periphery. These protrusions resemble TBCs that form in the intact seminiferous epithelium. These protrusions also co‐localize with F‐actin, which is known to associate with the proximal tubular part of TBCs and can be used as a marker for TBCs. Significantly, cultured Sertoli cells that express GFP‐tagged Claudin‐11 and Connexin‐43 extend similar protrusions into neighboring non‐transfected Sertoli cells. Our results are consistent both with the conclusion that junction elements from neighboring Sertoli cells are found together within TBCs, and with the hypothesis that tubulobulbar complexes may play a role in the turnover of intercellular junctions. Supported by an NSERC Discovery Grant to AWV.

Grant Funding Source : NSERC
Bacterial LPS Influences Receptor Activator NF Kappa B and Fractalkine Receptors Expression In Arterial Smooth Muscle Cells Derived From Limbs Amputated For Peripheral Arterial Disease
Authors: Zamin Rasheeda , Norman Paul , Filgueira Luis
Published: June 2021

Bacterial infections may trigger peripheral arterial disease (PAD) through their lipopolysaccharide (LPS) molecules by changing expression of CX3CR1 and RANK, which are associated with atherosclerosis and arterial calcification.

Arterial samples were harvested from 9 patients undergoing lower limb amputation for PAD. We initially identified the presence of bacterial (chlamydia and mycoplasm) antigens/LPS and inflammatory markers (CX3CR1 and RANK) in tissue samples using immunofluorescence staining. Secondly, the effect of LPS was tested in vitro (10ng/ml) using proliferating primary human arterial smooth muscle cells (pASMC). The expression of CX3CR1 and RANK post‐activation, with or without LPS, was assessed on 2 different pASMC cell lines using flow cytometry.

All samples showed evidence of infection. RANK and CX3CR1 were expressed in all arterial samples. Flow data showed that LPS down‐regulated RANK expression on pASMC (0.52–0.62 reduction in standardized mean intensity), while CX3CR1 expression was up regulated (1.27–1.75 increment) compared to non‐activated pASMC.

Our studies suggest that LPS may induce inflammatory changes seen in PAD lesions. Down‐regulation of RANK may play a role in calcification of infected arteries, while up regulation of CX3CR1 may contribute to the atherosclerotic changes.
The Atlas of Human Cardiac Anatomy: A free‐access educational website
Authors: Iaizzo Paul A , Bateman Michael G , Quill Jason L , Rolfes Christopher D , Howard Stephen A , Eggum Julianne H , Goff Ryan P , Eggen Michael D , Williams Gary W , Hill Alexander J
Published: June 2021

The University of Minnesota's Visible Heart® Laboratory has developed a novel free‐access educational website highlighting both functional and static human cardiac anatomy: The Atlas of Human Cardiac Anatomy (http://www.vhlab.umn.edu/atlas/). This website contains freely downloadable footage from over 100 human hearts, with associated text describing these anatomies, and with various related educational tutorials. To collect these images and videos, human hearts (deemed non‐viable for transplant) were obtained from LifeSource (MN) or the U of MN's Bequest Program, as gifts for research from the organ donors and their families. To honor these donations, all obtained/developed educational information has and will be gifted back for the training of interested individuals: which may range from students in K‐12, undergraduates, post‐graduates, cardiovascular residents or fellows, physicians, or patients themselves. This library of heart images and videos, includes those obtained with videoscopes, fiberscopes, ultrasound, fluoroscopy, MRI, and/or CT: from perfusion‐fixed specimens and/or from reanimated specimens using Visible Heart® methodologies (Hill et al., 2001). “The Atlas” is a living website that is updated on a monthly basis with new educational information, images, video footage, and/or requested materials. This project was funded by NIH training grant T32AR007612 and Medtronic, Inc.
Creation, Implementation, and Evaluation of a Neuroscience Curriculum for Grades 4–6
Authors: Rider Kristina , Dao Kim , Puder Barb
Published: June 2021

Anatomical sciences education requirements for grades K‐12 have recently been updated at state and national levels. At the local school level, many school systems are unable to fulfill the anatomical sciences education requirements due to financial constraints and/or lack of anatomical knowledge by the classroom teacher. Current research indicates that a neuroscience curriculum has not been developed and implemented in the academic instruction of students in grades K‐12. This prospective study involved developing a four‐week neuroscience curriculum targeted to students in grades 4–6 and determining if the curriculum was effective in outlining neuroscience concepts. The curriculum was presented to the 5th grade classrooms of a pre‐selected Oakland, CA area elementary school. During each one hour weekly session, the students were presented a neuroscience topic which was reinforced with interactive hands‐on activities and take home assignments. To assess the effectiveness of the curriculum, anonymous pre and post tests were administered and statistical significance was achieved using paired sample T –tests. Upon analysis of the data, the neuroscience curriculum allows students to retain neuroscience concepts and supports part of the anatomical sciences requirements currently mandated by state and national agencies.

Grant Funding Source : none
Cadaver dissection videos as an effective lab instruction tool for medical and other graduate level health professional students
Authors: Bee Mary Tracy , Montante James M , McAuley Bob
Published: June 2021

Human anatomy laboratory remains to be one of the most challenging courses in medical school. Students are required to perform high quality dissections, even though most of them do not have previous experience and struggle with learning by merely reading a lab manual. To make this task less daunting we produced a series of anatomy dissection videos to visually and audibly teach students the landmarks to focus on and how to dissect. Videos are approximately ten minutes in duration and illustrate dissection techniques and important anatomical relationships. Student feedback of the videos was overwhelmingly positive with 68% increased satisfaction from a group of students utilizing the dissection videos versus a control group. Student comments include “I have greater confidence and understanding of the dissections” and “I couldn't understand what they described in the book, but the video helped it all make sense.” Furthermore, dissection quality of the students viewing the videos was significantly better than the control group (p<0.01). We discuss the process of producing the videos and advocate the use of dissection videos to help guide students in human anatomy dissection.
Clinical Cases and TBL in Developmental and Gross Anatomy Course
Authors: Mavrych Volodymyr , Bolgova Olena , Seetharama Rao
Published: June 2021

The application of clinical cases studies and team‐based learning (TBL) as a component of a developmental and gross anatomy course in the medical school was evaluated. We adopted TBL in teaching first semester students and modified classical TBL to fit our curriculum. The core components of TBL were instituted in 5 clinical cases (CC) sessions and 3 question and answers (Q&As) sessions in the course. Students were surveyed to assess their preference. Respondents (n 76; 83% response) were asked to rate the TBL activity using scale 1–5 (strongly disagree – strongly agree). 74.7% of students admitted that CC sessions gave them an opportunity to integrate Anatomy and Clinical Medicine but only 23.8% of them prefer this activity over the in‐class didactic teaching. 65.1% of respondents acknowledged that CC sessions help them to understand better clinical importance of different anatomical structures and 30.1% thought that they are more confident working with X‐rays, CTs and MRIs. 70.7% of our students feel that TBL activity improves their communication skills and helps them to become a member of a team, 50.8% of respondents believe that this activity helps in overcoming the fear of talking in front of auditoria. 54.2% feel that Q&As sessions help in better understanding of Gross Anatomy material through clinical cases integration and admitted that discussion within the team during the answering is really useful and productive.