bluuebunny/crossref_metadata_2025 huggingface dataset:
Do nutrition rating systems promote a healthy diet? An evaluation of the Overall Nutritional Quality Index (ONQI) and risk of chronic disease
Authors: Chiuve Stephanie , Sampson Laura , Willett Walter
Published: June 2021

Nutrition rating systems rank foods based on nutrient composition and aim to guide consumers in making healthier food choices. These rating systems have not been evaluated with respect chronic disease prevention. The NuVal™ Nutritional Scoring System is based on the Overall Nutritional Quality Index (ONQI) algorithm that incorporates over 30 dietary components, and recently was endorsed by the American College of Preventive Medicine. We examined whether the ONQI score predicted major chronic disease risk prospectively. A total of 62,287 healthy women from the Nurses’ Health Study and 42,559 healthy men from Health Professionals Follow‐Up Study were followed from 1986–2006. Dietary data were collected from questionnaires at baseline. Major chronic disease included incident CVD, cancer, diabetes and non‐trauma death. We documented 20,005 cases of chronic disease in women and 13,554 cases in men. The multivariate relative risk (RR) of chronic disease, comparing the highest to lowest quintile, was 0.91 (95%CI: 0.87–0.95) in women and 0.88 (95%CI: 0.83–0.93) in men. In women, the ONQI score was associated with lower risk of CVD (22% in women and men) and diabetes (15% in women, 18% in men), but not cancer. The ONQI score was inversely associated with all‐cause mortality (p‐trend<0.01). The NuVal™ Nutritional Scoring System may help consumers target foods that contribute to a healthy diet. Funded by NIH grants.
Effect of Mulberry leaves extract on lipid metabolism in SD rats fed high fat diet
Authors: Choi Bong Kyoum , Kim Yeon Kyoung , Kim Ji Yeon , Kwon Oran
Published: June 2021

The aim of this study was to examine the effect of mulberry leaves extract(MLE) on lipid metabolism in 5 week‐old Sprague‐dawley rats(SD, n=32) fed normal or high fat diet(HF, 60% energy as fat). After obesity induction for 16 weeks, animals were fed different two experimental diets containing 0% or 5% (w/w) MLE for 6 weeks. Plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity and total protein and albumin concentrations were not different among groups. Liver total lipid and TG concentrations were significantly lower in 5% MLE group than 0% MLE group in normal diet groups(p<0.05). And 5% MLE group in high fat diet group was significantly lower than 0% MLE group in high fat diet group in total cholesterol concentration of liver and feces (p<0.05). But, plasma total lipids, TG, total cholesterol and HDL cholesterol concentrations were not significant among groups. In conclusion, MLE intake decreased liver total lipid and TG in normal diet groups and that decreased total cholesterol of liver and feces in high fat diet groups.
Differential effects of ginsenosides Rb1 and Rg3 on adipogenic and myogenic differentiation
Authors: Lee Ok Hwan , Hong Hee‐Do , Kim Yong Ook , Kim Young‐Cheul
Published: June 2021

Ginsenosides Rb1 and Rg3, the main bioactive constituents in fresh and steamed Panax ginseng, have been shown to exert protective effects against hyperglycemia and diabetes. In the present study, we tested the effects of Rb1 and Rg3 on adipogenic and myogenic gene expression and differentiation using 3T3‐L1 and C2C12 cells. The changes in levels of lipid, mRNA expression of key adipogenic and myogenic transcription factors were determined using semi‐quantitative RT‐PCR. Compared to control, both ginsenosides Rb1 and Rg3 enhanced differentiation of 3T3‐L1 preadipcytes in parallel with increased PPARγ mRNA, a key transcription factor for adipogenic determination. This correlated with increased expression of aP2 mRNA, a PPARγ‐regulated late marker gene. Moreover, ginsenosides Rg3 dose‐dependently stimulated myogenesis by increasing expression of both myogenin, a critical myogenic transcription factor, and MHC1, a marker gene for myotube formation. However, Rb1 was without effects on both myogenic marker gene expression and differentiation. These results indicate that the anti‐diabetic activities of ginseng may be attributable for Rb1 and Rg3 through a mechanism that involves activation of PPARγ in addition to the beneficial effect of Rg3 in stimulating myogenesis.
Fanconi Anemia Group J Mutation Abolishes its DNA Repair Function by Uncoupling DNA Translocation from Helicase Activity
Authors: Brosh Robert Michael , Sommers Joshua , Suhasini Avvaru , Leonard Thomas , Deakyne Julianna , Mazin Alexander , Shin‐ya Kazuo , Kitao Hiroyuki , Wu Yuliang
Published: June 2021

Fanconi anemia (FA) is a genetic disease characterized by bone marrow failure and cancer. FANCJ, one of thirteen genes linked to FA, encodes a DNA helicase proposed to operate in homologous recombination repair and replicational stress response. The FANCJ disease‐causing A349P mutation resides immediately adjacent to a highly conserved cysteine of the Iron‐Sulfur domain. Given the clinical severity of the A349P allele, we investigated its effect on the functions of FANCJ. Purified recombinant FANCJ‐A349P protein had reduced iron and was defective in coupling ATP hydrolysis and translocase activity to unwinding forked duplex or G‐quadruplex DNA substrates. The FANCJ‐A349P allele failed to rescue cisplatin or telomestatin sensitivity of a FA‐J null cell line as detected by cell survival or ƒ×‐H2AX foci formation. Expression of FANCJ‐A349P in a wild‐type background exerted a dominant negative effect, indicating that the mutant protein interferes with normal DNA metabolism. The ability of FANCJ to transduce force production from ATP hydrolysis to catalytic strand separation or destabilize protein (e.g., RAD51) bound to DNA is required for its role in DNA repair.
Chronic IL‐22Fc administration induces liver hepcidin expression and relative iron deficiency anemia.
Authors: Rottman James Brian , Smith Carole L , Li Hongyan , Rose Mark J , Maxwell Joseph R
Published: June 2021

Interleukin‐22 (IL‐22), a member of the IL‐10 cytokine family, is produced by activated TH17 and NK cells and binds to a specific heterodimeric receptor expressed on epithelium of the skin, respiratory and digestive tracts and hepatocytes. In vitro, IL‐22 stimulates hepatoma cell lines to express α1‐antichymotrypsin, haptoglobin, serum amyloid A, and in vivo administration stimulates LPS binding protein expression by the liver. Treatment of C57BL/6 mice with IL‐22Fc for either 9 days (200 μg I.P. SID) or 4 weeks (150 μg I.P., 3 times per week) induced hypochromic anemia associated with a decreased serum iron concentration, suggesting relative iron deficiency. Given that IL‐22 is also known as “hepatocyte stimulating factor” and iron balance is partially controlled by hepatic expression of the protein hepcidin, we measured serum hepcidin levels by mass spectrometer analysis. Animals treated with IL‐22 had elevated serum hepcidin levels as compared to controls. Previously reported to be elevated in chronically inflamed tissues, IL‐22 may participate in the anemia of chronic inflammatory disease through hepcidin‐induced alterations of iron metabolism.
The liver regeneration of land snail Helix lucorum
Authors: Dzidziguri Diana , Chiladze Natalia , Modebadze Irina , Mchedlidze Tamar , Tumanishvili Teimuraz , Kudryavtsev Boris
Published: June 2021

Study of regenerative peculiarities in organisms standing on the various steps of phylogenesis is important in order to establish the common biological meaning and basic pattern of regeneration.

The aim of the work was to study the regeneration characters of the mollusc liver so called hepatopancreas which is the analogue of vertebrate liver.

Experiments were carried out on a land snail Helix lucorum using the experimental model elaborated by us. The transcriptional activities within the liver cell nuclei were studied using by test‐system of RNA‐synthesis. To evaluate the mitotic activity of the liver tissue 5–7 μm paraffin sections were used under the light microscope.

The rhythmic character of RNA‐synthesis was revealed in the liver cells of intact snail during the day. The live resection induced the growth control genes activation like in rodents. The peak of transcriptional activity of these genes was appeared at 9th hour after the operation. Thereby the first peak of mitotic activity within the liver tissue was observed at 27th hour after the operation. The liver mass was regenerated after a week from the resection. The quantitative correlation of the main cell types (digestive, calcium and excretory ‐ 8:1:1) within the liver lobes remained stable.

On the basis of our results we conclude that the initial stages of reparative processes carry the similar character in phylogenetically distant organisms.
The inhibitory effect of chimeric decoy oligodeoxynucleotide in the regulation of transcription factors NF‐¥êB and Sp1 in an animal model of liver cirrhosis
Authors: Park Kwan‐Kyu , Kim Kyung‐Hyun , Park Ji‐Hyun , Kim Soo‐Jung , Lee Woo‐Ram
Published: June 2021

Liver fibrosis is a process of healing and scarring in response to chronic liver injury. Following injury an acute inflammation response takes place resulting in moderate cell necrosis and extracellular matrix damage. To develop a novel therapeutic approach in hepatic fibrogenesis, we examined the simultaneous suppression of the transcription factor NF‐κB and Sp1, which regulate acute inflammation and continuous deposition of extracellular matrix in liver fibrosis. In this study, we employed chimeric decoy oligodeoxynucleotide containing the consensus sequences of both NF‐κB and Sp1 binding sites, to suppress these transcription factors simultaneously. Treatment of chimeric decoy oligodeoxynucleotide reduced the activity of hepatic stellate cells in vitro, and decreased the expression of fibrotic and proinflammatory gene responses in a mouse model of liver fibrosis. These results suggest that chimeric decoy oligodeoxynucleotide strategy might be the potential therapeutic application to prevent liver fibrosis.
Protective effects of Red/Near Infrared Radiation on Murine Cardiac Ischemia/Reperfusion Injury
Authors: Spagnolli Ester , Anderson R. Rox , Hamblin Michael R. , Nagasaka Yasuko , Bloch Kenneth D. , Zapol Warren M.
Published: June 2021

Red/Near infrared radiation (R/NIR) has been reported to reduce myocardial infarct (MI) size in rabbits after ischemia/reperfusion (I/R) injury (Lohr N.L, JMCC 2009). The precise mechanisms responsible for the cardioprotective effects of R/NIR are incompletely understood. We sought to identify the most beneficial wavelengths in the R/NIR light spectrum for limiting murine cardiac I/R injury. Anesthetized and ventilated mice underwent left coronary artery ligation for 60 min followed by reperfusion. R/NIR was applied for three min at the beginning of reperfusion. Area at risk (AAR) and MI size were measured at 24h of reperfusion.

We investigated several wavelengths at varying levels of irradiance. A 19% decrease of MI/AAR was observed in wild mice when hearts were irradiated with red light (690 nm) at 0.06 W/cm2 of irradiance for 3 min at the time of reperfusion (59 ± 9% control group, n=7, vs 47 ± 7% irradiated group, n=6, P<0.05).

The cardioprotective effect of light appears related to both the specific wavelength and the level of irradiance across the broad spectrum of R/NIR radiation. By defining the specific wavelengths that provide maximal cardioprotection, genetically altered mice will further provide the precise mechanisms underling the cardioprotective effect of R/NIR.

This work was supported by NIH grant R01AI050875 and laboratory royalties from licensed patents.
S1P triggers cerebral vessel constriction through the store‐operated calcium entry pathway
Authors: Hopson Kristen Park , Truelove Jessica , Wang Yumei , Waeber Christian
Published: June 2021

Objective

To characterize signaling pathways by which sphingosine‐1‐phosphate (S1P) modulates vascular tone.

Methods

The role of store‐operated calcium entry (SOCE) in S1P‐induced Ca2+ entry in vascular smooth muscle cells (VSMCs) and vessel constriction was assessed using ratiometric Ca2+ measurements, quantitative RT‐PCR and isolated arteries.

Results

In VSMCs, S1P triggered STIM1 puncta formation, and SOCE activated by S1P was inhibited by 2‐aminoethoxydiphenylborate (2‐APB), diethylstilbestrol and gadolinium. In VSMCs prepared from S1P3 receptor knockout mice, the initial calcium rise triggered by S1P was abolished, but SOCE was still present suggesting that S1P acts via multiple signaling pathways. S1P‐induced SOCE was larger in proliferative than in contractile VSMCs, correlating with increases in STIM1, Orai1, S1P1 and S1P3 receptor mRNA. STIM1, S1P1 and S1P3 receptor mRNA expression were larger in freshly isolated brain vessels than in aorta. These data, and the fact that 2‐APB inhibits S1P‐induced cerebral artery constriction, suggest that SOCE modulates S1P‐induced vasoconstriction in vivo.

Conclusion

These data support a role for SOCE in S1P induced calcium entry and vessel constriction. The relationship between increased S1P signaling and SOCE in proliferative VSMCs suggests a role of S1P in disorders associated with vascular proliferation, such as atherosclerosis or vasospasm.
The defensive effect of nitric oxide on the pathogenesis of steroid‐induced femoral head osteonecrosis
Authors: Kenji Tateda , Okazaki Shunichiro , Watanebe Satoshi , Katada Ryuichi , Mizuo Keisuke , Nagoya Satoshi , Yamashita Toshihiko , Matsumoto Hiroshi
Published: June 2021

Objectives

Femoral head osteonecrosis(ON) is observed in patients treated with steroids. However, the pathogenesis of ON remains unclear. Nitic oxide(NO) is known to be related to the pathogenesis of osteonecrosis. Last year we established a rat model with ON by injecting lipopolysaccharide(LPS) and MP, and this was associated with a disruption of the innate immune system. In the present study, we investigated the effects of NO on pathogenesis of ON in our rat model.

Methods

Male Wistar rats were injected intravenously with 2mg/kg LPS on days 0 and 1 and intramascularly with 20mg/kg MP on days 2, 3 and 4, and then intraperitoneally with 50mg/kg NG‐nitro‐L‐arginine methyl ester(L‐NAME), an NO synthase inhibitor, on days 5,6 and 7 in group A, and on days 11,12 and 13 in group B. The animals were sacrificed 2 weeks after the last MP injection. Blood cytokines and other factors were performed by multiplex analysis system using Luminex.

Results

ON was observed in 67% of group A, not observed in group B. The plasma AST and ALT concentrations were increased in group A and B. The plasma IFN‐gamma concentrations were significantly higher in group A and B compared with control. The plasma IL‐17 concentrations were significantly higher in group B than control, while IL‐17 levels in group A were not different from those in control.

Conclusions

Inhibition of NO production just after MP injection caused ON in the rat model. IL‐17 levels were decreased in the rats with ON indicating that NO and IL‐17 may have defensive effects on pathogenesis of ON.
Prevalence of Heparin Platelet Factor 4 Antibody in Hip Fracture Patients Undergoing Orthopaedic Surgery Treated with Heparin versus Enoxaparin and Their Relevance to Serum Amyloid A
Authors: Griffin Justin , Litinas Evangelos , Zhu He , Hoppensteadt Debra , Lassen Michael , Hopkinson William , Fareed Jawed
Published: June 2021

Orthopaedic surgical interventions often result in the activation of hemostatic and inflammatory processes contributing to the pathophysiology of thrombotic and vascular complications especially in the elderly patient. Anticoagulants represent a standard of care for the management of postoperative thrombosis. Nonfunctional HIT antibodies are also generated in these patients, which may contribute to adverse outcomes. More recently, serum amyloid A (SAA) has also been implicated as an inflammatory mediator after surgical interventions. To investigate the relative prevalence of HIT antibodies and their subtypes, as well as the circulating level of SAA blood samples from elderly patients treated with heparin and enoxaparin (n=50 in each group) were collected at one day prior to and on 1, 3, 5, 7 postoperative days. Circulating levels of HIT antibodies were measured using GTI and Hyphen methods. The subtyping of the antibodies in terms of IgG, A, and M were also studied using the Hyphen kits. The SAA was measured using a commercially available ELISA method (Abazyme, Needham, MA). Patients treated with unfractionated heparin showed a greater prevalence of anti‐heparin platelet factor 4 antibodies as compared to enoxaparin. The heparin group also showed a greater prevealence of the IgG antibodies. Interestingly, SAA levels were increased in both the heparin and enoxaparin treated groups after surgery which persisted in both groups on day 5.
Improvements in Methods for Detection of Properties of High Density Lipoprotein
Authors: Yousefian Omid , Hough Greg , Hama Susan
Published: June 2021

Objective

We have developed an assay in which monocyte migration through filters, determines the level of inflammatory properties of low density lipoprotein (LDL) and high density lipoprotein (HDL) protective capacity. Automated monocyte enumeration demonstrated this property of HDL.

Methods

Cultured artery wall cells were exposed to either media alone, media plus LDL, or media plus LDL and HDL. The media from the treated cells was assayed in a monocyte chemotaxis assay. Image‐Pro Plus 5 software was used to automate the quantification of migrated monocytes. The accuracy of the data obtained from automated counting of images with 20X objective was compared with manual counting and tested by a two‐tailed paired t‐test. An integrated tool, Macro, was used to perform the automated enumeration on several images with 4X objective (the entire field).

Summary of Results

HDL, when presented to co‐cultures of artery wall cells together with LDL, significantly reduced the increased monocyte transmigration due to LDL alone. The statistical results, comparing the manual and automatic enumeration, demonstrated accuracy and precision (p=0.02, df=11, α= 0.05).

Conclusion

The results demonstrate that automated counting can replace manual counting, which should result in higher‐throughput of measurement of monocyte chemotaxis being used to determine the level of HDL proinflammatory or anti inflammatory properties.
Hematologic analysis of students in the region of Ferronikel smelter in Gllogovc
Authors: Bislimi Kemajl , Gashi Agim M , Bajraktari Ismet D , Grapci Linda , Orana Ilirjana , Halili Fetah M
Published: June 2021

The experiment involved students (N = 20) of the same age group (18–20 year old) from region polluted by smelter of Ferronikeli in Gllogovc and students from Gjakova (N = 20), located far from the source of pollution. In blood was investigated the number of red blood cells, the concentration of hemoglobin, hematocrit values, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) and leukocyte formula. The analysis were done with devices SYSMEX KX21N.

Statistical processing and analysis of preliminary results showed that compared to control group, in blood of students from Gllogovc the number of leucocytes is higher, whereas the number of platelets, red blood cells, the hematocrit values, hemoglobin concentration and MCV, MCH, MCHC are lower.

Leukocyte differential analysis of students in the contaminated region shows an increase in the number of lymphocytes, reduction of MXD (mixed forms of leucocytes) and neutrophils.
Morphometric evaluation of masticator muscles after experimental administration of a carcinogen
Authors: Loureiro Marco , Farinha Rodrigo , Rebelo Diana , Cabrita António Silvério
Published: June 2021

In developed countries, over 80% of the cancers in the oral cavity are attributable in part to tobacco smoking. This study evaluates the morpho‐histologic effects in masticator muscles of 7,11‐dimethylbenzanthracene (DMBA), a carcinogen present in tobacco smoke, being administered in a solution by gavage. Sixteen Wistar rats were used, randomly distributed in two groups. The control group had no manipulation. The test group had an experimental administration of DMBA by gavage 3 times in a week, in a dosage of 25 mg/kg. After two weeks, the animals were sacrificed and fragments from the masseter and temporal muscles were collected and analysed in a morphometric program, ImageJ®.

At morphometric evaluation, the masseter muscle from the control group had an average area of muscle fibers (A) of 10704±6835 for a 95% confidence interval (CI), while the test group had an A of 9503±4283 for a CI of 95%. The temporal muscle from control group had an A of 8103±2398 for a CI of 95%, while the test group had an A of 16282±7315 for a CI of 95%.
Increase of the antidepressant action of desipramine caused by caffeine and amphetamine
Authors: VILLAFAÑA SANTIAGO , ROBLES‐MOLINA EVELYN , MILLAN DANIEL , HONG ENRIQUE , HUANG FENGYANG
Published: June 2021

Desipramine is an antidepressive agent widely used which acts through the inhibition of the recapture of noradrenaline and serotonin, on the other hand, central stimulants such as caffeine and amphetamine help to release noradrenaline and serotonin, for this reason the combination of these agents with desipramine could increase the antidepressant response to desipramine, The aim of this work was to evaluate the antidepressive response of desipramine when its combine with caffeine and amphetamine in the forced swimming model. Male mice were used they received different doses of desipramine, caffeine, amphetamine, desipramine‐caffeine and desipramine‐amphetamine. The results showed that all agents produce immobility time decreased. When performing dose combinations without antidepressant effect of caffeine and amphetamine with desipramine was observed an increase in antidepressant response to desipramine, presenting only the combination of desipramine‐amphetamine on motor activity changes. These results suggest that drugs that promote the release of noradrenaline and serotonin could be used in combination with antidepressants to increase its effect, especially caffeine.
The histone acetyltransferase inhibitor garcinol blocks estradiol‐induced enhancement of object memory consolidation
Authors: Frick Karyn Morrione , Zhao Zaorui , Fan Lu
Published: June 2021

We recently showed that histone acetylation and DNA methylation in the dorsal hippocampus (DH) are critical for the potent estrogen, 17β‐estradiol (E2), to enhance object recognition memory (Zhao et al., PNAS, in press). However, the necessity of histone acetylation in mediating the effects of E2 on memory is unknown. The present study examined whether intrahippocampal infusion of the histone acetyltransferase inhibitor garcinol blocks E2 effects on memory and epigenetics. Ovariectomized female mice were trained in object recognition, and then immediately infused bilaterally into the DH with vehicle, E2 (5 μg/side), garcinol (1 ng/side), or E2 plus garcinol. After 48 h, garcinol blocked the memory‐enhancing effects of E2; this dose of garcinol did not impair memory on its own. Two weeks later, mice were infused again and killed 30 min later for measurement of epigenetic alterations in the DH. Garcinol reversed the E2‐induced increase in acetylation of histone H3 and decrease in expression of histone deacetylase 2 protein. Protein levels of DNA methyltransferase (DNMT) enzyme subtypes 1, 3A, and 3B were not altered by E2 or garcinol 30 min after infusion, but may be 4 h after infusion, at which point E2 increases DNMT3B protein levels. These findings suggest that histone acetylation is critical for the beneficial effects of E2 on object memory consolidation in females. Sponsored by NIH grant AG022525 to KMF and Yale.
Evidence for the role of PPAR‐β/δ in the development of spinal cord injury
Authors: Esposito Emanuela , Paterniti Irene , Mazzon Emanuela , Genovese Tiziana , Galuppo Maria , Di Paola Rosanna , Bramanti Placido , Kapoor Amar , Thiemermann Christoph , Cuzzocrea Salvatore
Published: June 2021

Evidence suggests a biological role for peroxisome proliferator activated receptor (PPAR)‐β/δ in the pathogenesis of a number of diseases. The aim of the present study was to evaluate the contribution of PPAR‐β/δ in the secondary damage following experimental spinal cord injury (SCI) in mice. Spinal cord trauma was induced by the application of vascular clips to the dura via a four‐level T5–T8 laminectomy. SCI resulted in severe trauma characterized by oedema, neutrophil infiltration, production of inflammatory mediators, tissue damage, and apoptosis. The high affinity PPAR‐β/δ agonist GW0742 (0.3 mg/kg, i.p.) 1 and 6 h after the SCI significantly reduced: (1) the degree of spinal cord inflammation and tissue injury, (2) neutrophil infiltration, (3) nitrotyrosine formation, (4) pro‐inflammmaory cytokines, (5) NF‐κB activation, (6) iNOS expression and (6) apoptosis (TUNEL staining, FasL, Bax and Bcl‐2 expression). Moreover, GW0742 significantly ameliorated the recovery of locomotory function (evaluated by motor score). In order to elucidate whether the protective effects of GW0742 are related to activation of the PPAR‐β/δ, we investigated the function of PPAR‐β/δ antagonist, GSK0660, on the protective effects of GW0742. GSK0660 (1 mg/kg i.p. 30 min prior to GW0742)significantly blunted the protective effect of the PPAR‐β/δ agonist. Our results demonstrate that GW0742 treatment reduces the development of inflammation and tissue injury associated with spinal cord trauma.
Secreted APP and amyloid beta quantification in SH‐SY5Y cell media using high sensitivity AlphaLISA kits
Authors: Hohenauer Krystyna , Anani Shihadeh , Marcil Alexandre , Roby Philippe , Parent Stephane , Dahan Sophie
Published: June 2021

The aggregation and accumulation of amyloid beta peptides is one of the pathological hallmarks of Alzheimer's disease (AD). Amyloid beta is produced by sequential proteolytic cleavage of Amyloid Precursor Protein (APP) by beta‐ and gamma‐secretase, which are plausible molecular targets for AD treatment. Thus, identification of modulators of these secretases could lead to new therapeutics for this prevalent and debilitating disease. SH‐SY5Y are human neuroblastoma cells that express APP and the various secretases that cleave this membrane‐bound protein into sAPPβ, sAPPα and amyloid beta peptides. We have developed new highly sensitive and specific AlphaLISA kits that allow the detection of endogenous amounts of these various APP metabolites. Specific inhibitors of alpha‐, beta‐ and gamma‐secretase were used to modulate the relative activities of these enzymes. Application of these inhibitors resulted in characteristic signature levels of product sAPPα, sAPPβ, Aβ 1–40 and Aβ x‐40 (defined as the combined amount of both Aβ 1–40 and Aβ 17–40) from SH‐SY5Y cell culture supernatants. These results show that the new AlphaLISA kits have sufficient sensitivity and specificity to study this pathway, even in wild‐type cells that do not overexpress APP or BACE. The Z′‐factor obtained for these cell‐based secretase assays was greater than 0.6, indicating reproducible and robust assays for HTS. In summary, these new AlphaLISA detection kits can accurately quantitate endogenous APP cleavage products. This will in turn result in a better understanding of the APP degradation pathway and facilitate the identification of novel therapeutic drugs for the treatment of Alzheimer's disease.
The effect of an experimental model of affective disorder on the expression of GluR2 in rat brain
Authors: Rosa Maria Luiza Nunes Mamede , Iyomasa Melina Mizusaki
Published: June 2021

Glutamatergic hypoactivity may contribute to cognitive dysfunction in schizophrenia. Isolation rearing of rats from weaning has been used as experimental model of affective disorders like schizophrenia. In this study we evaluated the changes in the expression of GluR2 AMPA receptor in hippocampus, amygdala and entorhinal cortex (EC) induced by isolation rearing. In two groups of Wistar rats (n=6/each) the pups remained with their mothers (6/mother) until weaning (21 days) when they were randomly allocated to one of two conditions: 1) grouped, housed 3/cage and handled 3 times/week; 2) isolated, housed individually and handled once/week. After 10 weeks all animals were deeply anaesthetized (50ml/kg of Urethane 25%), perfused and their brains removed. 40‐μm sections were used for immunohistochemistry. The immunopositive cells (IC) were counted bilaterally, in 3 sections/rat, in hippocampus, amygdala and EC. Data were compared by t‐test (p<0.05). Isolation rearing induced a significant decrease in GluR2‐IC in hippocampus. This reduction was 52% in the hillus (p<0.001) and 29% in CA1 (p=0.002). No change was found in CA3, amygdala or EC. The results suggest that isolation rearing induces changes in GluR2 AMPA receptor in hippocampus similar to those reported for postmortem human brains with schizophrenia, contributing with additional evidence for using isolation of rats from weaning as an animal model of schizophrenia. Grants from FAPESP (05/01501‐7) and FPA.
NAD+ blocks H2O2‐induced HMGB1 translocation of PC12 cells and decreases survival of microglia
Authors: Chen Heyu , Han Jin , Shao Jiaxiang , Xia Weiliang , Ying Weihai
Published: June 2021

We have reported that NAD+ administration can profoundly decrease ischemic brain injury. However, the mechanisms underlying the protection remain unclear. Because inflammation plays a crucial role in ischemic brain damage, we hypothesized that NAD+ may decrease ischemic brain injury partially by affecting the inflammation. Because HMGB1 translocation outside of the nucleus has been suggested as a key signal activating inflammatory responses, we determined the effects of NAD+ on the HMGB1 translocation in PC12 cells. Our study has suggested that NAD+ treatment could decrease the H2O2‐induced HMGB1 translocation. Our study using microglial BV2 cell lines has also found that NAD+ treatment can significantly decrease the number of surviving microglial cells. Treatment of relatively high density (80–90%) microglia cultures with 0.1 mM or 1 mM NAD+ for 24 hrs decreased the number of surviving cells by 30% and 50%, respectively. Treatment of relatively low density (50–60%) microglia cultures with 0.01 mM or 0.1 mM NAD+ decreased the number of surviving cells by 30% and 40%, respectively. These observations suggest that NAD+ administration may attenuate ischemic brain injury partially by inhibiting inflammatory reactions. (Supported by a key basic research grant from Shanghai Municipal Scientific Committee (to W.Y.), and a National 973 grant #2010CB834306 (to W.Y.)).
Synthesis of some new derivates of [(heteroaryl‐2‐ylimino)‐ethyl]‐benzopyran‐2ones and their antimicrobial activity
Authors: Selmani Pranvera Breznica , Hoti Ramiz , Mulliqi‐Osmani Gjyle , Damoni‐Rexha Dafina , Gashi Fatbardh
Published: June 2021

Novel derivates of 3‐[1‐(pyridin‐2‐ylimino)‐ethyl]‐benzopyran‐2‐ones 3(a–c) and 3‐[1‐(pyrimidin‐2‐ylimino)‐ethyl]‐benzopyran‐2‐ones 3(d–e) are synthesized by condensation reactions of 3‐acetyl‐benzopyiran‐2‐one 2 with corresponding heterocyclic amines 2(a–e). These products are synthesized by catalytic reactions using zinc chloride, in reflux conditions. Products are characterized on the basis of their spectroscopic data. Antimicrobial activity of products 4(a–e) against S. aureus, E. coli and Candida albicans are investigated measuring inhibition zones around disks marked previously with corresponding compounds. Results are shown for a considerable activity of products against these microorganisms
Redox Sensitive MR Contrast Agent
Authors: Matsumoto Ken‐ichiro , Nakanishi Ikuo , Anzai Kazunori
Published: June 2021

Nitroxyl radicals (also known as nitroxide) have been used as in vitro and in vivo redox probes in fields of electron paramagnetic resonance (EPR) spectroscopy and imaging. Nitroxyl radicals are paramagnetic species, which also can be detected using T1‐weighted MRI. Reactive oxygen species (ROS) react with nitroxyl radicals to lose paramagnetism. The time course of in vivo MRI intensity induced by a nitroxyl radical can give tissue redox information, which is the so called the redox imaging technique. Employing a membrane permeable nitroxyl radical, redox information in tissues could be available. A brief theory of redox imaging techniques and applications of redox imaging techniques are introduced.
Pharmacodynamic differences between biosimilar and branded enoxaparins with reference to their immunogenic profile
Authors: Ramacciotti Eduardo , Rao Gundu , Hoppensteadt Debra , Jeske Walter , Fareed Jawed , Walenga Jeanine M.
Published: June 2021

Low molecular weight heparins are complex mixtures of sulfated oligosaccharides produced by depolymerization of porcine mucosal heparin. The depolymerization process inflicts a variety of structural modifications which are process‐dependent. The objective of this study was to compare branded enoxaparin (Lovenox, Sanofi‐Aventis) with Cutenox (Gland Pharma) in healthy human volunteers for pharmacodynamic equivalence. Individual groups of volunteers (n=110) were administered test agent at doses of 40 mg OD SC for 10 days and blood samples were collected at baseline and after 1, 3, 7, 10 and 14 days. Hemostatic parameters such as aPTT, anti‐Xa, anti‐IIa and TFPI release were measured. In addition, anti‐heparin‐PF4 antibodies were measured using two commercially available assays (GTI and Hyphen Biomedical). The antibody subtype (IgG, IgA, IgM) was determined using an assay from Hyphen Biomedical. No significant differences in the various hemostatic parameters were noted. The prevalence of positive antibody titers was different between assays, but not between treatment groups. The titer of anti‐heparin‐PF4 IgG antibodies was higher following Cutenox treatment (0.37 ± 0.21) compared to following Lovenox treatment (0.21 ± 0.11). The relative proportion of IgG positive samples was also different in the two groups. These studies suggest that Cutenox produces a different immunogenic profile than Lovenox.
Thalidomide: A Boon to Dermatology? Current Indications and Concerns
Authors: Kumar Uma Senthil , Rajasekaran Senthil Kumar
Published: June 2021

Thalidomide is a drug with a complex past. Initially marketed as a sedative and anti‐emetic for use in pregnancy, thalidomide was linked to embryologic defects and was withdrawn from the market. However, it has since made a comeback for the U.S. FDA‐approved use in the treatment of erythema nodosum leprosum (ENL).

This presentation analyses the publications made between 2000 and 2009 obtained by searching the pubmed database using key words “thalidomide and dermatology and uses”.

Thalidomide's effect on immune system include; suppression of the production of IgM antibodies inhibition of tumour necrosis factor‐α, inhibition of IL‐8, IL‐12, enhancement of IL‐2, IL‐4, IL‐5, inhibition of cytokine‐induced NF‐κB gene expression and decrease of B lymphocytes in the spleen.

Currently, thalidomide is known to be effective in unresponsive dermatological conditions such as actinic prurigo, adult Langerhans cell histiocytosis, aphthous stomatitis, Behçet's syndrome, graft‐versus‐host disease, cutaneous sarcoidosis, erythema multiforme, Kaposi sarcoma, lichen planus, lupus erythematosus, melanoma, prurigo nodularis, pyoderma, gangrenosum and uraemic pruritus.

While there are undoubted benefits with use of thalidomide in the field of dermatology, the more serious adverse drug reactions such as teratogenecity, peripheral neuropathy and the relatively uncommon side‐effects like neutropenia, mood changes, thromboembolic complications, such as deep vein thrombosis and pulmonary embolism must be watched for. As most of the dermatological conditions require a long term administration of thalidomide, these adverse drug reactions must be thoroughly screened for. Further studies on the exact mechanism of action of thalidomide may help us in identifying the potential therapeutic benefits of this drug even further.
Cell‐based screening for discovery of prostaglandin (PG) E1 agonist and PGE2 antagonist from dietary TMC herbs
Authors: Chillar Annirudha J , Mohite Anita J , Ruan Ke‐He
Published: June 2021

Cell‐based screening for discovery of prostaglandin (PG) E1 agonist and PGE2 antagonist from dietary TMC herbs

Annirudha J Chillar, Anita J Mohite, Ke‐He Ruan

Centre for Experimental Therapeutics and Pharmacoinformatics, Department of Pharmacological and Pharmaceutical sciences, College of Pharmacy, University of Houston, and Houston, Texas 77004

The prostaglandin (PG) E1 (PGE1) and PGE2 acting through EP receptors (EP1, EP2, EP3 and EP4) are involved in a variety of biological functions. PGE1 is a potent anti‐inflammatory molecule whereas PGE2 is inflammatory and cancer mediators. Little information is available for the dietary herbs with PGE1 agonist or PGE2 antagonist properties. In this study, a HEK293 cell line expressing EP1 receptor stably was created by gene transferring and G418 screening. A highly sensitive fluorescence calcium signaling mediated by the EP1 receptor expressed in the cell line was created and used as indicator for identification of the dietary TMC herbs with PGE1 agonist and PGE2 antagonist activities. Six herbs that demonstrated PGE2 antagonist activity inhibited the PGE2 signal and seven herbs with PGE1 agonist activity, mimicked PGE1 signaling were identified by a wide range screening started from 96 TMC dietary herbs. In conclusion, the established cell line stably expressing EP1 receptor can be used as a cell‐based target screening for discovery of PGE1 agonist and PGE2 antagonist. Many anti‐inflammatory and anti cancer ingredients are present in the dietary TMC herbs, which have potential to be further developed into preventive and therapeutic interventions against inflammation and cancers.

Funded by NIH grant‐ R01HL56712 (For KHR); RO1HLO79389 (For KHR).
Decreased expression of Cdc25a in breast stroma of diethylstilbestrol treated female ACI rats
Authors: Taylor LaShaundra , Newell Oneil , Darling‐Reed Selina , Soliman Karam , Hudson Alicia , Elie Yasmeen
Published: June 2021

Diethylstilbestrol (DES), a known human carcinogen and synthetic nonsteroidal estrogen has been shown to cause pituitary, breast and cervix cancers. DES has been shown to produce mammary tumors and double strand breaks in rats and induce breast cancer in humans. Diallyl sulfide (DAS), a garlic organosulfur compound (OSC) has been shown to inhibit chemical carcinogenesis in animals. OSCs like DAS are known to decrease proliferation of cancer cells by delaying cell cycle progression. The effect of DAS on the proliferation of stroma breast tissue by DES is not fully elucidated. This study will determine the proliferative properties of DAS in breast stroma of DES induced female ACI rats by examining Cdc25a and ATM expression. Female ACI rats were treated for 6 months with corn oil (control), 10mg/kg DES and/or .01mg/g DAS. RNA was isolated from breast tissue using trizol and converted to cDNA using Stratagene first strand cDNA synthesis reaction. Gene expression profiles were determined using Super Array pathway finder and confirmed with real time RT‐PCR. Cdc25a and ATM expression in the DES treated group is decreased significantly compared to the control group. Cdc25a shows an increase in the fold change in the DES/DAS group as compared to the control. We conclude that the decreased Cdc25a expression in the DES treated group may induce cell cycle arrest while the co‐treated group is approaching the level of the control.

Funded by RCMI Grant #2G12RR03020‐24
Diallyl Sulfide inhibits Diethystilbestrol induced Breast Tumor in Female ACI rats
Authors: Newell Oneil G , Taylor LaShaundra , Hudson Alicia , Darling‐Reed Selina , Reams Renee , Oriakiu Ebenezer , Reuben Jayne
Published: June 2021

Previous studies in this lab have shown that diallyl sulfide (DAS) inhibits DNA adduct formation, lipid peroxidation and modulate detoxification and DNA repair genes and enzymes in breast tissue of diethylstilbesterol (DES) treated female ACI rats. These findings are important since estrogen and estrogenic compounds such as DES have been shown to induce breast cancer in humans and animals. In addition, scientists have postulated that DAS, a constituent of garlic, is an effective cancer prevention agent. In this study, we hypothesize that the release of inteleukin 6 by DES induced cancerous cells leads to activation of the JAK‐ Stat pathway which can increase cell proliferation and inhibit apoptosis. DAS may modulate these changes. To prove this hypothesis, 6 week old female ACI rats were implanted with Control (silicon implant alone) and DES (50mg/kg) silicon pellets were implanted s.c. In addition, these animals were fed normal chow or DAS (as co‐treatment) containing chow ad libitum for six months. At sacrifice, representative mammary tissueand all palpable mammary tumors were removed for genetic and histopathologicalanalysis. Genetic analysis was done by microarray analysis and validated using PCR array. Our data show significant increase in interleukin 6 gene expression in cancerous cells (DES group) and a significant decrease in the DAS co‐treatment group. Furthermore, a significant modulation of genes associated with the JAK/STAT pathway was observed. Animals co‐treated with DAS were free of tumor. For the first time our lab was able to show that DAS prevents DES induced breast tumors in female ACI rats.

Funded by RCMI Grant #2G12RR03020‐24
Intraocular transplantation of umbilical cord mesenchymal stem cells rescues retinal functional loss in light‐induced retinopathy
Authors: Lee Yih‐Jing , Lim Yah‐Teem , Tien Lu‐Tai
Published: June 2021

Apoptosis of photoreceptor is the final event that leads to degeneration of retina, such as retinitis pigmentosa and age‐related macular degeneration, which often lead to blindness. At present, there are no effective vision‐restoring treatments for such disorders. Stem cell is a potent resource for treating several degenerative diseases including retinal disorders. Among them, the umbilical cord mesenchymal stem cell (UMSC) is a noncontroversial and inexhaustible source for stem cell transplant therapies. The purpose of this study is to investigate the fate of UMSC transplanted into retinas of animal models of light‐induced retinopathy. The rescue effect of UMSC intraocular transplantation was examined functionally by measuring the electroretinogram (ERG). Retinas were harvested for immunohistochemistry and histology analysis by measuring the thickness of the outer nuclear layer (ONL). The data showing 7 days exposure caused significant reductions in a‐wave and b‐wave in ERG. After UMSC transplantation, the amplitude of a‐wave and b‐wave in UMSC‐treated eyes are larger than that measured from non‐treated eyes. The ONL of treated retina was thicker than non‐treated eyes. The transfected UMSCs differentiated into several different kinds of retinal cells. The results indicate that transplantation of UMSCs recovering both function and morphology of retina.
Use of an excretory and secretory antigen of Fasciola hepatica for the immunodiagnosis in fasciolasis and a potential target for immunoprophylaxis
Authors: Gaudier José Francisco , Espino Ana M.
Published: June 2021

Fasciola hepatica, is a prevalent and economically important disease in the husbandry industry. A 17KDa protein termed Fh4.26 (Q5I5Y3) was recently identified by means of successive screenings of a cDNA library previously prepared from F. hepatica adult worms using a rabbit anti‐F. hepatica excretion and secretion (E/S) antigens serum and a serum from rabbit with 4 week of F. hepatica infection. Fh4.26 preserves two DM9 Domains of unknown function, this repeats are conserved in some methyltransferases protein. Recombinant Fh4.26 protein was purified by affinity chromatography. The immunodiagnostic potential of the antigen rFh4.26 was assessed by the antibody detection Enzyme‐Linked ImmunoSorbent Assay and Western blot using a large panel of sera from animals infected with the liver fluke F. hepatica. The assay was highly sensitive and revealed that animals infected with F. hepatica develop antibodies against Fh4.26 at early and late stages of infection. Confocal microscopy analysis demonstrated that Fh4.26 is localized on vitellaria cells of the liver fluke. Fh4.26 could be one of the antigens responsible for this maintenance which make it an attractive target for immunoprophylaxis or chemotherapy. This study was approved by IACUC #7870104.
Chronic Treatment with an orally active formulation of Angiotensin‐(1‐7) improves autonomic control in SHR
Authors: Casali Karina Rabello , Bertagnolli Mariane , Pinto G , Dias L D , Rigatto Katya Vianna , Schaan Beatriz , Irigoyen Maria Claudia , Santos Robson
Published: June 2021

Angiotensin‐(1–7) (ANG‐(1‐7)) seems to have an important role in cardiovascular regulation. In hypertension, autonomic control and ANG(1‐7) are altered. Heart rate (HRV) and blood pressure variability (BPV) are important predictors of cardiovascular risk and provide information about cardiac and vascular autonomic modulation. In this work we investigated the effect of chronic treatment with orally active formulation of (HPâCD/Ang‐(1‐7)) on autonomic modulation in spontaneously hypertensive rats (SHR). Treated group (AG, n=8) received, equivalent to 30 ug/Kg Ang‐(1‐7), by gavage once a day for 10 weeks. The control group (CG, n=8) received tap water. Blood pressure was recorded and stationary sequences of 250 beats were randomly chosen. Frequency domain analysis of HRV and BPV was performed with an autoregressive model. Systolic blood pressure, as well as BPV and vascular sympathetic modulation, were reduced after treatment. With respect to cardiac autonomic modulation, although the treatment did not cause a reduction in HR, ANG(1‐7) caused an increase in HRV, showing an improvement in the cardiac autonomic control. The sympathetic‐vagal balance was reduced in the CG. Our results show that the oral treatment with ANG(1‐7) was able to improve relevant cardiovascular parameters which are altered in hypertension, suggesting the possibility of a novel pharmacological intervention in hypertension therapy.
Oral Tetrahydrobiopterin Treatment Prevents Accelerated Atherosclerosis Caused by Oscillatory Shear Stress
Authors: Li Li , Rezvan Amir , Chen Wei , Jo Hanjoong , Harrison David G
Published: June 2021

Tetrahydrobiopterin (BH4) is a critical cofactor for NO synthesis by endothelial NO synthase (eNOS). BH4 synthesis is regulated by phosphorylation of the rate‐limiting enzyme, GTP cyclohydrolase I (GTPCH‐1). We previously found that laminar, but not oscillatory shear stress stimulates GTPCH‐1 phosphorylation and markedly increases BH4 levels. A recent study showed that disturbed blood flow in vivo induced by partial carotid ligation results in endothelial dysfunction and accelerated atherosclerosis in ApoE−/− mice. We have now found that oscillatory shear in vivo is associated with reduced GTPCH‐1 phosphorylation and markedly lower levels of BH4. We therefore aimed to test the hypothesis that atherosclerosis induced by oscillatory shear is due to low levels of BH4 and eNOS uncoupling, and that increasing BH4 will prevent atherosclerosis. We found that oral BH4 treatment at 10 mg/kg/day in drinking water virtually eliminated atherosclerotic lesion formation in ApoE−/− mice after 3 weeks of ligation. Furthermore, BH4 treatment preserved the decline of NO production induced by oscillatory shear. We also showed that BH4 treatment abolished the marked increase in superoxide production induced by disturbed flow. Thus, BH4 treatment appears to prevent eNOS uncoupling and atherosclerosis in this model of rapid lesion development caused by disturbed flow. Supported by HL39006 and an AHA predoctoral fellowship.