bluuebunny/crossref_metadata_2025 huggingface dataset:
Conditioned Media from Small‐sized Human Umbilical Cord Blood‐Derived Mesenchymal Stem Cells Suppresses a‐MSH Induced Melanogenesis
Authors: Oh Hyun ah , Kim Ji Hyun , Kim Ju yeon
Published: November 2023

Human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSCs) has been extensively investigated in terms of cell therapy. Especially, small‐sized hUCB‐MSCs have multiple biological effects in a variety of diseases. Previously, conditioned media from human umbilical cord blood‐derived mesenchymal stem cells (hUCB‐MSC‐CM) inhibited melanin synthesis via microphthalmia‐associated transcription factor (MITF) degradation by ERK pathway activation. Base on this, we investigated these melanogenesis activity to vary according to the cell size, type of MSCs. In this study, we show that the effect of conditioned media derived from small sized hUCB‐MSCs (small‐sized hUCB‐MSC‐CM) in melanocytes. Treatment of small‐sized hUCB‐MSC‐CM strongly inhibited the alpha‐melanocyte stimulating hormone (α‐MSH) induced hyperpigmentation in B16F1 cells and Melan‐a cells. On the other hand, conditioned media derived from bone marrow‐MSCs (BM‐MSC‐CM), adipocyte‐MSCs (AD‐MSC‐CM) almost not reduced melanin production. We also found that small‐sized hUCB‐MSC‐CM inhibits tyrosinase activity and down‐regulates melanogenic enzymes in melanocytes. These results small sized hUCB‐MSC‐CM efficiently suppressed melanin synthesis in melanocytes and melanoma cells.
β‐hydroxy‐β‐methylbutyrate (HMB) production in human subjects
Authors: Walker Dillon , Thaden John , Engelen Marielle , Deutz Nicolaas
Published: November 2023

Rationale

HMB is produced from the amino acid leucine in the body. Studies in sheep/pigs have demonstrated that HMB production accounts for 2–10% of Leucine turnover, but HMB production in humans is unknown. In contrast, HMB supplementation to individuals promotes maintenance of muscle mass when subjected to muscle wasting conditions. Therefore, we hypothesize that in subjects with low muscle mass, whole body HMB production is reduced. We first developed a method to measure whole body HMB production and the conversion of Leucine to HMB in humans.

Method

We assessed the whole body production of leucine and HMB by providing a single intravenous pulse dose of L‐[U‐13C6]Leucine and [3,4, methyl‐13C3]HMB in human subjects in the post absorptive state and sampled blood over a 3 hour period. Plasma enrichment of [3,4, methyl‐13C3]HMB was determined by GC/MS/MS methods and plasma enrichment of L‐[U‐13C6]Leucine was determined by LC/MS/MS. Data are summarized as mean (SE) in μmol/kg ffm/hr. Fitting of enrichment curves, ANOVA and student's t‐tests were done by GraphPad Prism 6.

Results

The decay of the plasma HMB and Leucine enrichments were strictly two exponential. We determined HMB production to be 0.28 μmol/kg ffm/hr and leucine turnover to be 100 μmol/kg ffm/hr. Only 0.28% of the total leucine turnover is converted to HMB.

Conclusion

Whole body HMB production is much lower in humans than reported production in large animals.
Multiplexed Micronutrient Array for Population Surveillance
Authors: Lillis Lorraine , Barney Becky , Brindle Eleanor , Peck Roger , Tyler Abby , Lyman Chris , Boyle David
Published: November 2023

Objective

Deficiencies of vitamin A, iron and iodine affect a significant portion of the world's population and are often associated with adverse health outcomes, particularly among pregnant women and children. Efforts to characterize deficiency patterns have been hampered by a lack of measurement tools appropriate for large‐scale use. Since many of these micronutrients are not easily measured directly, reliable proxy biomarkers indicative of deficiency status have been identified and widely adopted. Inflammation or infection biomarker levels must also be measured, as inflammation affects the levels of vitamin A and iron status biomarkers. Furthermore, malaria infection is known to deplete iron levels, thus screening for malaria is also recommended. We previously developed a prototype multiplex immunoassay for the simultaneous measurement of five biomarkers relevant to assessing vitamin A, iron status and inflammation; retinol binding protein, soluble transferrin receptor, ferritin, alpha‐1‐acid glycoprotein and C‐reactive protein. Here we present an improved version of the immunoassay which has also been expanded to include measurement of the biomarkers for iodine deficiency (thyroglobulin) and malarial parasitemia (Histidine Rich Protein II).

Method

Using affordable technology from Quansys Biosciences, antibodies are coated in seven discrete regions of the well of a microtiter plate and the seven analytes are assayed in a single volume of sample. A control standard was developed for the assay that reflected the clinical range of each biomarker being assayed. Assay performance was evaluated by comparing multiplex and conventional assay results for plasma from 72 US volunteers.

Results

The new multiplex immunoassay and established conventional assay methods showed high correlation for all analytes tested (Average 0.77, p<.0001), improving on the values observed with the original 5‐plex plate. Use of a control standard specially designed for the multiplex assay in place of a commercially available standard allowed for more accurate quantitation of each analyte. The assay was also validated against the available WHO reference standards.

Conclusions

This 7‐plex micronutrient assay has excellent potential for use as a cost effective tool for population surveillance of vitamin A, iron and iodine deficiencies as well as malaria infectivity rates.

Support or Funding Information

This research is supported by the Bill and Melinda Gates Foundation.
Portion size selection of pizza and modeled impact on nutritional intake in the US adult population
Authors: Masset Gabriel , Vlassopoulos Antonis , Mölenberg Famke , Lehmann Undine
Published: November 2023

Introduction

In the last decades, food and beverage portion sizes have increased together with obesity prevalence. Portion size reduction has been identified as a cost‐effective strategy to reduce energy intake of the general population. Pizza is a major contributor to diets in the US, with approximately an eighth of the population consuming pizza on any given day.

Objective

To model the effect of selecting the RACC serving size for pizza (140g) on the nutritional intake of NHANES 2011–12 adult participants.

Methods

Dietary intakes of 4948 participants aged 19+ years were derived from the first 24hr‐recall. Nutritional intakes (energy (E), protein (P), total fat (TF), saturated fat (SFA), sodium (Na), and total sugars (S)) were compared between three scenarios: Baseline, Observed intakes as reported in the 24‐hour recall; RACC1, All reported pizza consumption occasions set at 140g (RACC); RACC2, All pizza consumption reported above 140g was set to 140g. Intake was analyzed in the whole population and among pizza consumers only. All analyses were conducted using the ‘survey’ package of R.

Results

Among the target sample, 11% of participants reported consuming pizza. Average and median pizza portion size was 235g and 194g, respectively; 62% of declared serving sizes were above the RACC. In the whole population, RACC1 and RACC2 scenarios were associated with a 1.5 and 2.2% reduction in energy intake (−33kcal and −39kcal), respectively, which was translated in reductions of other nutrients (not shown). Among pizza consumers, forcing a 140g serving size (RACC1 scenario) led to a significant reduction of energy intake (−301kcal or 12%), and thereby of all the other nutrients (Table). In the RACC2 scenario, the reduction in energy intake was strengthened (−356kcal or 14%, Table).

Conclusion

Shifting the portion size of a single food group led to relevant reduction in energy, saturated fat and sodium intake in this representative sample of the US adult population, demonstrating the potential for portion size selection to be a key lever within dietary advice and guidelines.

Support or Funding Information

This study was funded by Nestec SA.

Average daily intakes of US pizza consumers aged 19 and above (n=459, NHANES 2011–12, day 1), at Baseline and in RACC1 and RACC2 protion size selection scenarios

Baseline
RACC 1
RACC 2

Nutrient
Mean
SE
Mean
SE

Mean
SE

Energy (kcal)
2558
58.9
2257
55.8
*
2202
57.3
*

Protein (g)
92.5
2.05
79.2
1.86
*
76.9
1.88
*

Total fat (g)
98.7
2.91
85.1
2.79
*
82.6
2.90
*

Saturated fat (g)
34.9
1 12
29.4
1.07
*
28.4
1.12
*

Sodium (mg)
4309
130
3669
120
*
3542
124
*

Total sugars (g)
124
3.07
120
3.01
*
120
3.01
*

RACC1, All reported pizza consumption occasions set at 140g (RACC); RACC2, All pizza consumption reported above 140g was set to 140g.

Pizza consumption was defined as having reporting pizza at least once during the 24\hr recall.

pttest vs. Baseline < 0.001
Nutritional Status in People Living With HIV and Trends in HIV Prevalence Using NHANES (1999–2012)
Authors: Thuppal Sowmy , Bailey Regan
Published: November 2023

Background

Under‐nutrition is an independent predictor of mortality in people living with Human Immunodeficiency Virus (PLHIV). Introduction of the highly active antiretroviral therapy (ART) has improved the nutritional status in PLHIV but has not completely eliminated malnutrition.

Aim

To understand the current nutritional status in PLHIV and to study the trends in prevalence of HIV status using NHANES (1999–2012).

Methods

NHANES 1999–2012 was used to estimate the prevalence of PLHIV in U.S. adults (n=16,083; ages 19–15) and for calculating selective biochemical parameters and biomarkers. Two 24‐hour dietary recalls were used to estimate energy adjusted mean values for micro‐ and macro‐nutrients; height and weight were used to calculate BMI.

Results

The overall prevalence of HIV infection in U.S. adults was 0.44% (CI: 0.26–0.75) in 1999–2002, 0.53% (CI: 0.35–0.78) in 2003–2006, and started to decline thereafter to 0.25% (95% CI: 0.12–0.51) in the 2011–2012. HIV prevalence was highest in non‐Hispanic blacks followed by Mexican Americans and non‐Hispanic whites across all survey years. The mean age of PLHIV ranged from 35.4 years to 49.6 years. The body mass index (BMI) was lower in men and higher in women living with HIV compared to respective negative controls. Energy expenditure was significantly lower in women living with HIV compared to men living with HIV. Macronutrient intake was similar between PLHIV and HIV negative controls in both men and women. All the micronutrients were similar between PLHIV and controls except the levels of vitamin E was significantly lower in women living with HIV compared to the HIV negative controls. Micronutrient and macronutrient intake were similar between men and women living with HIV, except women had greater BMI and lower energy expenditure, while men had higher dietary fat intake. Women living with HIV had insufficient levels of vitamin D (37 nmol/L) and significantly lower levels of RBC folate (509 nmol/L) and Serum Folate (19 nmol/L). Mean triglycerides in men living with HIV was (410 mg/dL). There were no differences in the electrolyte levels and other biochemical markers compared to negative controls.

Conclusions

With the initiation of ART the nutrient intake in PLHIV has improved and is similar compared to HIV negative population. Women living with HIV have lower levels of vitamin D and folic acid. They have higher BMI and lower energy expenditure compared to men in spite of consuming similar quantities of micro‐ and micro‐nutrients, warranting life‐style modifications.
Household Food Insecurity but not participation in a Nutrition Assessment and Counselling Support (NACS) program is significantly associated with current BMI in People Living with HIV/AIDS (PLWHA) in the Greater Accra Region of Ghana
Authors: Sackey Joachim , Aryeetey Richmond , Zhang Fang Fang , Rogers Beatrice , Wanke Christine
Published: November 2023

Background

Ghana has about 230,000 adults living with HIV. Nutrition plays a key role in the management of HIV. The NACS program developed by USAID's (U.S. Agency for International Development) FANTA (Food and Nutrition Technical Assistance) project seeks to enhance access to nutrition services for PLWHA in developing countries, including Ghana.

Objectives

To determine whether Ghanaian PLWHA participating in NACS differ from non‐participants by age, sex, BMI, and food insecurity.

To determine factors associated with BMI among this group of PLWHA.

Methods

Seventy‐seven adult PLWHA participating in NACS at three district hospitals in Accra and 75 not participating in NACS, in three comparable district hospitals in the same region were recruited into this study in 2015. Participants’ height and weight were measured using a stadiometer and digital scale, respectively. Food insecurity was assessed using the Household Food Insecurity Access Scale. Analysis of covariance was used to evaluate baseline characteristics by participation in NACS, and the association between food insecurity and BMI.

Results

Among 152 participants (mean age=39.9yrs; 84% female), 26.5% were overweight (BMI=25–29 kg/m2) and 20.5% obese (BMI ≥30 kg/m2). Over a third (36.2%) were classified severely food insecure by the guidelines of the HFIAS. Participants in NACS did not differ from those who were not in NACS in age, sex, food insecurity and BMI. Amongst all participants, food secure PLWHA had a BMI 4.2 kg/m2 higher compared to severely food insecure (p=0.0002) after multivariable adjustments.

Conclusion

Food insecurity is associated with a lower BMI in this group of PLWHA. Interventions to improve BMI among PLWHA in this region should consider including components which will improve food security.

Future steps

Participants will be followed for the next 6 months with measurements repeated at 3 and 6 months to prospectively assess the impact of the NACS program on their BMI.

Support or Funding Information

Brown/Tufts/Miriam Fogarty AIDS International Training and Research Program (2D43TW000237)
Novel Association Of Estimated Glomerular Filtration Rate With NKAIN2 Gene Variants In Hispanic Children
Authors: Chittoor Geetha , Haack Karin , Laston Sandra , Mehta Nitesh R , Cole Shelley A , Comuzzie Anthony G , Butte Nancy F , Voruganti V. Saroja
Published: November 2023

Background

Obesity has been associated with reduced kidney function and increased chronic kidney disease risk, in both adults and children.

Objective

To identify genetic loci contributing to the variation in estimated glomerular filtration rate (eGFR) using genome‐wide association (GWAS) approach in 789 Hispanic children of the Viva La Familia Study (VFS).

Methods

We conducted a GWAS using SOLAR after accounting for familial relationships. Empirical thresholds for genome‐wide significant and suggestive associations were set at p <1 × 10−7, and p <1 × 10−6, respectively. The eGFR was estimated by the ‘Bedside Schwartz Equation’, which is specially developed for children.

Results

Mean± SD values of serum creatinine and eGFR were 1.79 ± 0.97 mg/dL and 39.3 ± 14.7 mL/min/1,73m2, respectively, and were significantly different (p <3×10−15) between obese (2.06 ± 1.09; 35.64 ± 14.45) and non‐obese children (1.52 ± 0.75; 43.02 ± 13.82). eGFR, adjusted for age, age*sex, and BMI z‐scores, was significantly heritable (51%, p <1×10−16). The GWAS identified rs2689886 (p <9 ×10−8, minor allele [G] frequency [MAF] of 0.42) of Na+/K+ Transporting ATPase Interacting 2 (NKAIN2 located on chromosome 6) to be strongly associated with eGFR. This variant also showed suggestive evidence of association with serum creatinine. NKAIN2 is a membrane enzyme that energizes the Na‐pump and evidence suggests that it has a significant role in thermogenesis, energy balance, and weight regulation. Other NKAIN2 gene variants that showed suggestive evidence of association with eGFR include rs1973961 and rs2689887, p <5×10−7, MAF (A) of 0.42 and 0.48. These NKAIN2 variants were not in LD, thus signifying that a number of functional variants may exist in NKAIN2 influencing variation in eGFR. Suggestive associations with eGFR were also observed for gene variants (rs10954465, rs10279045, and rs2347709) in LOC102724947 located on chromosome 7.

Conclusions

Children and adolescents, particularly obese, of the VFS exhibit significantly lower eGFR. Additionally, for the first time, we have shown the association of the NKAIN2 gene with eGFR and serum creatinine in Hispanic children warranting future functional studies pertaining to this gene and renal function in pediatric cohorts.

Support or Funding Information

R01DK092238
Trends in Obesity and Non‐Communicable Chronic Diseases in China and Projected Future Situation for 2030
Authors: Zhu Jinzhou , Zhang Yong , Xue Hong , Wang Limin , Wang Yaogang , Wang Youfa
Published: November 2023

Background

China, the world's second largest economy, has experienced rapid economic development and many social and environmental changes including people's lifestyle and disease patterns in the past 40 years.

Methods

Based on nationally representative data, we studied trends of obesity and non‐communicable chronic diseases (NCDs) in China and projected future situation for 2030.

Results

The past two decades witnessed an increase in overweight and obesity in Chinese adult (aged ≥ 18 y). During 1992–2012, based on WHO standard of body mass index (BMI)≥25, the combined prevalence of overweight and obesity increased from 14.6 to 32.3%, while it increased from 20.0 to 42.0% based on Chinese standard of BMI≥24. Assuming the future trends would be similar as what observed in the past, we projected it will reach 51.2% and 61.5% in 2030 by WHO and Chinese BMI cut‐points, respectively. In school age children (7–18 y), the prevalence of obesity increased from 0.1% in 1985 to 5.0% in 2010, and will reach 9.7% by 2030. In terms of NCDs, between 1993 and 2008, the morbidity of cardiovascular diseases increased from 31.4 to 85.5‰, and will be 153.8‰ by 2030, while diabetes increased from 1.9 to 10.7‰, and will reach 22.3‰. Although the proportion of mortality attributable to major NCDs presented no remarkable change during 1990–2012, that due to malignant tumors increased from 18 to 22%, and cardio‐/cerebral‐vascular diseases increased from 27 to 36%. The proportion of respiratory diseases decreased from 21 to 12%. Alarmingly, the percentages of awareness, treatment and control of hypertension and diabetes remain very low in China. The figures for hypertension were 46.5%, 41.1% and 13.8% in 2012, increased from 26.6%, 12.2%, and 2.9% in 1991. The figures for diabetes increased from 28.5%, 24.7% and 9.4% in 2000 to 36.1%, 33.4% and 11.6% in 2012.

Conclusions

In the past two decades China saw a steady increase in obesity and NCDs, but awareness, treatment and control of related diseases like hypertension and diabetes remain very low. Future effective national programs and policies are needed to fight the epidemic.

Support or Funding Information

Funding: NIH, U54HD070725
Cost efficiency of nutrient delivery to primary school recipients of National Dietary Services Limited (NDSL) lunches in Trinidad
Authors: Nichols Selby , Francis Marlon
Published: November 2023

Background

In Trinidad, the National Schools Dietary Services Limited (NSDSL) uses contracted caterers to provide over one hundred and forty thousand meals daily to its recipients. This government sponsored programme cost taxpayers approximately 2 40 million TT6 dollars annually. The programme aims to meet approximately one third the nutrient needs at of its recipients, encourage regular school attendance and stimulate the production of local agricultural produce through its inclusion on planned menu cycles.

Objectives

We investigated the cost efficiency of various menus to deliver carbohydrates, proteins, fats, vitamin A, iron, calcium and fiber to recipients.

Methodology

Menus served on the NSDSL were randomly selected and standardized recipes were analyzed using Food Works College Edition dietary analysis software. Cost of the production and delivery of one meal was determined by adding the cost of each meal paid to the caterer plus the cost in administrative overheads incurred by NDSL per meal served. We used a cut‐off of 25% and greater of meal components not consumed as our definition of wastage. We multiplied this fraction be the cost producing the particular menu component and divided by the total major macronutrient level to determine a cost per unit of the nutrient wasted.

Results

Our results suggest that that chicken was superior to fish, processed meats and legumes in delivering the protein component of the meals. While bread was superior to rice and rice mixed with vegetables, and cassava in delivering the carbohydrate component of the diet. Menus with legumes and vegetables as part of the meal were better than menus that included one or none of these components in delivering fiber to recipients.

Conclusion

Our results suggest that certain menus are better than other at delivering key nutrients in this school meal programme. These finding should be considered in planning menus that give good value per unit of nutrient delivered.

Support or Funding Information

Not Applicable
The Prevalence of Obesity in Alaska Native Children: Results from the Children's Healthy Living Program
Authors: Power Julianne Mary , Philip Jacques , Bersamin Andrea
Published: November 2023

The prevalence of obesity among American Indian and Alaska Native (AI/AN) children is significantly higher than among children of other ethnic groups. Health statistics presented under the broad heading AI/AN, however, obscure important differences in obesity prevalence that correspond to wide geographic and cultural differences. Our objectives were to estimate and compare the prevalence of obesity among 2–8 year old Alaska Native and non‐ native Alaskan children. Participants were recruited at a range of public events and pre‐schools as part of the Children's Healthy Living Program for Remote Underserved Minority Populations of the Pacific Region (CHL). We measured the height and weight of each child using a standardized protocol (N = 476). Body mass index (BMI) was calculated and converted into BMI Z‐scores based on age and sex using the United States Center for Disease Control (CDC) 2000 growth charts. Parents or guardians self‐reported child behaviors related to obesity, as well as child race, which was categorized as AN (n=104), White (n=315), or Mixed Race (AN and White, n=57) for analysis. Linear mixed models were used to evaluate the association between BMI Z‐score and race. Generalized linear mixed models were used to estimate the odds ratio of a binary outcome (overweight + obese vs. healthy weight) by race. Both models (n=379 because of missing values in covariates) included a random intercept for household group because up to three children could be sampled from each of 252 households. All models were adjusted for sex, age, child sleep time, household income, birth weight and sampling region. Forty‐nine percent of AN children were overweight or obese compared to 32.1% of White children and 29.8% of Mixed Race children. We found a positive association between BMI Z‐score and being AN vs White (β=0.34, p=0.018) but not for being Mixed Race vs. White. Similarly, AN children, but not Mixed Race children, were more likely to be overweight or obese compared to White children (OR=2.71, CI=[1.25, 5.86]). Alaska Native children appear to be at increased risk of being overweight or obese compared to White children and public health efforts should focus on understanding the causes of this difference so we can more effectively design culturally relevant interventions to reduce this trend.
Platinum Resistance in High‐Grade Serous Ovarian Cancer: an Intratumoral Collagen Fiber Implication
Authors: Pavanello Marina , de Andrade Natal Rodrigo , Ramalho Susana , Vassallo José , de Angelo Andrade Liliana Aparecida , Derchain Sophie , Sarian Luis Otávio
Published: November 2023

Introduction

Overall survival of women with high‐grade serous ovarian cancer (HGSOC) is highly related with platinum resistance. Parameters of collagen can be used as potential new prognostic biomarkers, since their relation with tumor growth, invasion and metastasis has been demonstrated. These parameters are (1) quantity which is the amount of fibers deposited, (2) uniformity which refers to how collagen fibers are closely deposited side by side and (3) organization refers to how the fibers are arranged.

Aim

To evaluate the association of intratumoral collagen fiber parameters with platinum resistance in HGSOC patients.

Methods

35 women with HGSOC, 14 platinum resistant and 21 platinum sensitive were included. Their samples of paraffin embedded human were sectioned (4 μm) and stained with hematoxylin and eosin. Three intratumoral regions of interest were analyzed by second harmonic generation microscopy, in a Zeiss LSM 780‐NLO confocal (Carl Zeiss AG, Germany). The images were captured (1024×1024 format; 40×/1.3 NA oil) with an excitation wavelength of 890 nm, a pulse length of approximately 100 fs, and emission filter centered at 445 nm with a 20 nm bandwidth (Semrock, Rochester, New York) (Figure 1). Each image was divided in 16 regions (256 × 256) and analyzed in an independently way, using the ImageJ (NIH, Bethesda, USA). Intratumoral collagen fiber parameters were compared considering platinum resistance status using t‐tests. Sensitivity and specificity were analyzed using receiver operator characteristics (ROC) curves.

Results

There was a significant association between platinum resistance and thick collagen fiber quantity (p=0.04) and a trend of association with thin collagen fiber quantity (p=0.06). Further, intratumoral thick and thin collagen fiber uniformity also were associated with platinum resistance (p=0.02 and p=0.02, respectively). There was no association between intratumoral thick and thin collagen fiber organization with platinum resistance (p=0.29 and p=0.27, respectively). ROC curves demonstrated the highest area under the curve using thin collagen fiber uniformity as parameters, in this case it was found 47.6% of sensibility and 92.9% of specificity to determine platinum resistance (Figure 2).

Conclusion

Our preliminary results are encouraging since there was an association between intratumoral thick and thin collagen fiber quantity and uniformity with platinum resistance status in women with HGSOC.

Support or Funding Information

Partially financed by Cnpq number 306583/2014‐3.
PRKCA, PTPN1 and ZYX Are Possible Markers to Target CD133+ GBM Stem Cells
Authors: Ozkanca Seyma , Bal Merve Gulsen , Yukselten Yunus , Yildiz Dilara Akcora , Ugur Hasan Caglar , Demircan Kadir , Sunguroglu Asuman
Published: November 2023

Introduction

Glioblastoma Multiforme (GBM) accounts for more than %75 of malignant brain tumors. GBM stem cells (GSCs) have a crucial role in tumor formation, development, chemo‐resistance and recurrence. CD133, the first discovered cell surface marker for GBM, is a pentaspan transmembrane glycoprotein. In this study, it is aimed to investigate new therapeutic targets within cell adhesion molecules to select and eliminate CD133+ GSCs effectively.

Methods

Ten primary GBM cells were isolated from freshly obtained GBM tissue samples and they were cultured. CD133+ and CD133− cells were seperated by MACS method via anti‐CD133 (AC133) antibody from those GBM primary cells and cell lines. Following RNA isolation from CD133 positive and negative cells, cDNA synthesis was performed. Finally, by applying micro array protocol for cell adhesion molecules, mRNA expression levels of 88 genes were detected. Proper results were collected and analysed statistically.

Results

According to the results of microarray; it has been found that CD133+ cells express 8,73 fold PRKCA, 2,17 fold PTPN1 and ZYX 5,29 fold when they are compared with CD133− cells.

Conclusions

PRKCA (Protein Kinase C, Alpha) is a serine‐threonine kinase that regulates PI3K‐Akt pathway. This gene has been reported to have a role in proliferation, apoptosis, differentiation, migration and adhesion, angiogenesis and therefore tumorigenesis. It also activates MAPK pathway, which leads to cell proliferation.

PTPN1 (Protein Tyrosine Phosphatase, Non‐receptor Type 1) is a tyrosine phosphatase. It dephosphorylates kinases like EGFR, JAK2 and TYK2. By inhibiting MAPK pathway PTPN1 promotes cell proliferation.

ZYX (Zyxin) is a zinc‐binding phosphoprotein that concentrates at focal adhesions and along the actin cytoskeleton. It has a proline rich domain which may interact with SH3 domains of proteins that have a role in signaling pathways. Moreover, ZYX has a role in cell migration. High expressions of zyxin might mean higher motility of malignant tumors and therefore tumor initiation, progression and metastasis.

To conclude, since PRKCA, PTPN1 and ZYX are related with tumorigenesis and malignant cell motility, and are highly expressed on GBM CD133+ cells, it seems that PRKCA, PTPN1 and ZYX may be essential markers to target CD133+ GBM stem cells.

Support or Funding Information

This research has been supported by The Scientific and Technological Research Council of Turkey (No:114S189)
Functional Analysis of the Synthetic Compatible Solute Homoectoine in the Stabilization of Intestinal Epithelial Barrier During Inflammation
Authors: Ochoa Karla Fabiola Castro , Robles Hilda Vargas , Madrid Alí Francisco Citalán , Schnoor Michael , Galinski Erwin A.
Published: November 2023

Background and Aim

Inflammatory bowel disease (IBD) is a multifactorial disorder that comprises Ulcerative Colitis (UC) and Crohn's disease (CD) affecting millions of people worldwide with alarmingly increasing incidences every year. Dysfunction of the intestinal epithelial barrier has been associated with the pathogenesis of IBD. IBD therapies include anti‐inflammatory drugs that enhance intestinal barrier function but they frequently have adverse side effects. Compatible solutes such as bacterial ectoines have been shown to stabilize cell membranes and proteins. Here, we test if homoectoine (4,5,6,7‐tetrahydro‐2‐methyl‐1H‐(1,3)‐diazepine‐4‐carboxylic acid), a synthetic derivative of ectoine, prevents excessive intestinal epithelial permeability during inflammation.

Methods

Confluent Caco‐2 cells in transwell filters were incubated with the inflammatory cytokines TNF‐α and IFN‐γ with or without parallel administration of homoectoine. In order to observe changes in epithelial permeability, transepithelial electrical resistance (TEER) was measured every 24 hours. Paracellular flux of 4kDa FITC‐dextran was measured when differences in TEER were observed. For in vivo experiments, homoectoine was administered orally to C57BL/6 mice in parallel to the induction of colitis with dextran sulfate sodium (DSS) during 7 days. To determine the clinical severity of colitis, the disease activity index (DAI) was evaluated and hematoxylin/eosin stainings of colon tissues were analyzed. Intestinal epithelial permeability was determined by means of an Evans Blue assay.

Results

In vitro assays showed that homoectoine protects against the increase of epithelial permeability induced by pro‐inflammatory cytokines such as TNF‐α and IFN‐γ as indicated by increased TEER. We also observed that cells receiving homoectoine were able to recover faster from inflammation. However, paracellular flux of macromolecules was not improved significantly. In vivo, homoectoine treatment alone did not affect the DAI suggesting that homoectoine has no negative effects on healthy mice. Importantly, homoectoine treatment in colitic mice significantly reduced the DAI score with greater effects on intestinal bleeding and stool consistency. Moreover, intestinal epithelial permeability was reversed to control levels in the group of colitic mice receiving homoectoine. Histologically, homoectoine reduced edema formation, leukocyte influx and tissue damage induced by DSS.

Conclusions

Homoectoine has protective effects on the epithelial barrier during inflammation and may thus serve in the future as diet supplementation in IBD patients to reach or extend phases of remission.

Support or Funding Information

CONACyT Grants; 179895, 207268 and 233395
Allosteric HSP70 Family Inhibitors Block Proinflammatory Signaling and Toxicity in an In Vitro Non‐Alcoholic Fatty Liver Disease Model
Authors: Khachatoorian Ronik , Ganapathy Ekambaram , Shao Hao , Gestwicki Jason E , French Samuel W
Published: November 2023

Endoplasmic reticulum stress is associated with steatosis of the liver and progression to steatohepatitis in a significant number of patients. This can ultimately lead to cirrhosis and hepatocellular carcinoma. In this study, we sought to determine the effect of a number of allosteric heat shock protein (HSP) 70 family inhibitors (AHIs) on lipid‐induced inflammation and injury in hepatocytes. Briefly, Huh7 cells were treated with 300 μM palmitic acid and immediately treated with a number of AHIs or DMSO as negative control. Then, 72 hours after treatment, the cells were harvested and the levels of phospho‐JNK and total JNK were determined through Western analyses. The results indicated that a subset of the AHIs significantly blocked activation of JNK. Further MTT assays showed the same AHIs completely blocked lipotoxicity. These results implicate HSP70 family members in fatty acid‐mediated inflammation and toxicity and raise the possibility of utilizing AHIs as therapeutic agents.

Support or Funding Information

NIH R01DK090794, SWF NIH NS059690, JEG
High‐molecular‐weight kininogen and its cleavage product bradykinin are critical in the pathogenesis of autoantibody‐induced arthritis
Authors: Wu Yi
Published: November 2023

High‐molecular‐weight kininogen (HK) is a major component of plasma kallikrein‐kinin system (KKS), the activation of which mediates the cleavage of HK to release bradykinin (BK). BK is a nonapeptide with potent proinflammatory and vasoactive activities. Although patients with rheumatoid arthritis (RA) exhibits the activation of the KKS and the cleavage of HK in plasma, the role of HK in arthritis pathogenesis has never been characterized. In this study we characterized the phenotype of our newly‐generated HK‐deficient (Kng1−/−) mice in arthritis. To induce autoantibody‐induced arthritis, two groups of Kng1+/+ and Kng1−/− mice were injected intraperitoneally with anti‐type II collagen antibody cocktail (4 mg) on day 0 and lipopolysaccharide (LPS, 50 μg) on day 3. Although wild‐type (WT) mice developed severe arthritis, the Kng1−/− mice exhibited significantly attenuated disease severity as evaluated by joint diameter change and clinical scores (p<0.0001). Compared with WT mice, Kng1−/− mice displayed reduced synovial hyperplasia and cartilage erosions, and decreased protein and mRNA levels of IL‐1b and IL‐6 in joint tissue (p<0.001). BK level and kallikrein activity in Kng1−/− plasma were significantly lower than that of WT mice. Incubation with Collagen‐anti‐collagen complex mediated BK production in human plasma. In consistence with the phenotype of Kng1−/−mice, the mice lacking BK receptors also displayed a reduced arthritis severity, as well as a decrease in IL‐1b and IL‐6 production in joint tissue. Our results the first genetic evidence demonstrating an important role of plasma HK in the pathogenesis of arthritis, and the blockade of HK cleavage to prevent BK production might be a novel therapeutic strategy for RA treatment.
Profiles and biological analysis of lncRNAs in RAW264.7 macrophages infected by space mutagenized Escherichia coli after simulated microgravity
Authors: PEI Xuefeng , ZHANG Xiaoyi , YAO Jing , LIU Rong , WANG Jingyu , YUAN Min , YUAN Ming
Published: November 2023

Objective

To explore the difference expression of long non‐coding RNAs(LncRNAs) of RAW264.7 macrophages infected by space mutagenized Escherichia coli(E.coli) after simulated microgravity.

Methods

RAW264.7 macrophages were randomly divided into four groups. G1 and G2 were cultured in normal gravity for 72 hours. G3 and G4 were cultured in simulated microgravity for 72 hours, then G2 and G4 were infected for 6 hours by space mutagenized E.coli in Shenzhou 10 spacecraft, G1 and G3 were cultured for 6 hours in normal culture media. Total RNA was extracted from the cells in four groups. The profile of lncRNAs were detected with SBC‐lncRNA chips and the significant difference expression of lncRNAs were screened (fold change≥2; P≤0.05). The significant signaling pathways were enriched from the different expression of gene in G2 vs G2 and G4 vs G1. The Pathway‐Network was constructed based on the pathways with significant differences to get the key signaling pathway. The gene network graph was constructed based on the expression values of G2 and G4 to screen the key genes in the coexpression network.

Results

Differently expressed lncRNAs were detected as follows(fold change≥2, P≤0.05): there were 5843 lncRNAs detected totally in G2 vs G1, in which there were 2140 up‐regulated lncRNAs and 3703 down‐regulated lncRNAs. G4 vs G1 detected totally 7187 lncRNAs, in which 3230 lncRNAs were up‐regulated and 3957 lncRNAs were down‐regulated. G4 vs G2 detected totally 31 lncRNAs, in which there were 27 up‐regulated and 4 down‐regulated lncRNAs. The Csf3, Ripk2, A130077B15Rik were recognized as key genes in coexpression network. The most different expression gene was Csf3, which only existed in G2 but not existed in G4, and related with PI3K‐Akt signaling pathway, Cytokine‐Cytokine receptor interaction and Jak‐STAT signaling pathway. Ripk2 is related with immunologic function and can induce cell apoptosis by activating the NF kB signaling pathway. A130077B15Rik, as a LncRNA, might be associated with immunity and inflammation.

Conclusion

Our work explored the difference expression of LncRNAs in RAW264.7 macrophages infected by space mutagenized E. coli after simulated microgravity, which established the basis for the research of LncRNAs mechanisms of bacterial pathogenicity in spaceflight.

Support or Funding Information

Funded by National Basic Research Programs of China, No.2014CB744404
Muscle function recovery after acute lung injury is impaired in aged mice, linked in part, to metabolic alterations attributed to MuRF1 by quantitative targeted metabolomics analysis of acyl‐carnitines
Authors: Ilaiwy Amro , Parry Traci L. , Bain James R. , Muehlbauer Michael J. , Willis Monte S. , Files Clark
Published: November 2023

Most survivors of acute lung injury suffer from substantial physical complications months after the initial lung injury resolves, and persistent muscle weakness is a very common sequelae of lung injury. Using a mouse model of LPS‐induced lung injury [acute lung injury (ALI) mice], we found that old (20 month) ALI mice failed to recover skeletal muscle function 10 days after injury compared to young (6 month) ALI control mice. Non‐targeted GC‐MS analysis of old ALI gastrocnemius muscle vs. vehicle treated controls identified 18 significantly different metabolites, including the long chain fatty acid linoleic acid (1.75 fold increased), indicative of altered linoleic acid metabolism. Our laboratory recently identified that muscle ring finger protein 1 (MuRF1) inhibits fatty acid oxidation in vivo via mono‐ubiquitination of peroxisome proliferator‐activated receptor alpha. Moreover, we've identified that MuRF1−/− mice are resistant to LPS‐induced muscle atrophy. Western analysis of old and young ALI muscles for MuRF1 found significantly greater expression of MuRF1 3 days following lung injury in old ALI mice, leading us to hypothesize that old ALI muscles had inhibited fatty acid oxidation. We next performed targeted metabolomics analysis of fatty acid intermediates (acyl‐carnitines) and amino acids of the affected muscles. At day 10, we identified 13 significantly altered acyl carnitines (by t‐test, p<0.05) compared to vehicle treated controls (0.27–1.14 fold). We also identified that total long chain acyl‐carnitines were almost decreased by half, paralleling the acyl‐carnitine profile recently published in MuRF1−/− hearts. Together, these studies demonstrate for the first time age‐associated susceptibility of ALI‐induced muscle wasting and characterize the metabolic alterations in fatty acid metabolism, suggesting MuRF1's role in regulating fatty acid metabolism, which may contribute to impaired skeletal muscle functional recovery.

Support or Funding Information

This work was supported by the National Institutes of Health (R01HL104129 to M.W.), the Leducq Foundation Transatlantic Networks of Excellence (to M.W.), the Claude D. Pepper Older Americans Independence Center (P30AG21332 to D.C.F and O.D.), and the American Thoracic Society Foundation (D.C.F.).
Novel lymphoid targeted prodrugs of the glutamine antagonist DON for the treatment of hematological malignancies
Authors: Jancarik Andrej , Rais Rana , Alt Jesse , Tenora Lukáš , Krecmerova Marcela , Cheng Chih , Pommier Elie , Rojas Camilo , Powell Jonathan , Majer Pavel , Slusher Barbara
Published: November 2023

Objective

The glutamine antagonist 6‐diazo‐5‐oxo‐l‐norleucine (DON) has shown robust efficacy in both lymphoma and leukemia animal models and exploratory clinical studies. DON broadly blocks glutamine utilizing reactions critical for the synthesis of nucleic acids/proteins and the generation of alpha‐ketoglutarate for energy metabolism. Unfortunately its severe toxicity hampered its development. Using a prodrug approach, we aimed to synthesize lymphoid‐targeted prodrugs of DON designed to circulate intact in blood and preferentially activate inside PBMCs affording reduced systemic exposure and improved tolerability, while maintaining its robust efficacy.

methods

Synthetic efforts for DON prodrugs were particularly challenging due to DON's inherent instability. We found it was imperative to mask both the carboxylate as well as the amine functionality of DON to achieve sufficient chemical stability. We concealed the carboxylate and amine with various alkyl esters (e.g. methyl, ethyl and isopropyl), amino acids (e.g. leucine, alanine), and other bulkier promoeities used in FDA approved drugs (e.g. POC, POM, ODOL). We then evaluated the prodrugs for in vitro plasma stability across multiple species (mouse, dog, monkey, pig, human). Ex vivo whole blood experiments were further conducted to determine selective conversion in PBMCs. Lastly in vivo experiments using pigs were carried out and PBMC to plasma ratios were calculated.

results

Our novel DON prodrugs showed species specific plasma stability with rodents metabolizing the prodrug more rapidly compared to higher species. Of the various modifications on the carboxylate, the isopropyl ester afforded significant lymphoid‐targeting in pigs, monkeys, dog, and humans. In human whole blood ex vivo studies several of the isopropyl ester containing prodrugs delivered DON into PBMCs versus plasma with a >30‐fold differential, in contrast to DON which delivered a small percentage (<10%) of drug to PBMCs. Lastly in vivo studies in pigs confirm this PBMC selectivity with favorable pharmacokinetics.

conclusions

We have successfully synthesized lymphoid‐targeted DON prodrugs which deliver >30‐fold more DON to lymphoid cells in human whole blood versus plasma. Comparative efficacy and toxicity studies are ongoing in lymphoma animal models. These novel prodrugs may pave a path for the development of a new and safer therapeutic for hematological malignancies.
Challenges and solutions to developing an automated high‐throughput/high‐content screening platform for the ‘neglected’ schistosome parasite
Authors: Caffrey Conor , Chen Steven , Suzuki Brian , Singh Rahul , Arkin Michelle
Published: November 2023

Automated high‐throughput (HT) and/or high‐content (HC) screening platforms are now well‐established in early drug discovery, including for many parasitic organisms causing ‘neglected tropical diseases.’ However, for the helminth (worm) parasites responsible for a range of globally prevalent and debilitating infections, such systems are still in their infancy. Using the schistosome, a flatworm parasite that infects over 200 million people, as a particularly challenging example, we developed an HTS/HCS platform to quantify the parasite's responses to chemical insult. I will highlight the hurdles encountered to standardize the preparation and handling of the parasite, and how we interfaced the parasite with an automated instrumentation environment. I will then discuss the approaches taken regarding image acquisition, object segmentation and tracking, and feature extraction. We tested the platform with seven anti‐schistosomal drugs to measure a range of static and dynamic parasite responses as a function of time and concentration. We designed a user interface to visualize and interrogate the data, which are maintained in a customized database. We combined the high‐dimensional data into a single metric output suitable for primary first‐pass screening: data from screening campaigns of high‐value small molecule collections will be presented. The new platform increases throughput, improves rigor and will support the identification of drugs, drug targets and mechanisms of action.

Support or Funding Information

Supported by NIH R01AI089896 and R21AI107390.
Modulation of GRP78‐CHOP‐JNK‐Caspase12 pathway by Chinese herb medicine in atrophied soleus muscle induced by tail suspension
Authors: Zhang Shu , Wu Shiwen , Yuan Ming , Wang Jingyu , Yang Honghui
Published: November 2023

Objective

Microgravity could induce a series of changes on the physiological state in humans and animals, among which skeletal muscle atrophy is striking. The countermeasure of skeletal muscle atrophy mainly relies on the physical exercise which need expensive and heavy special apparatus in space. However, complementary medicine such as Chinese herb medicine may have distinctive effects on the countermeasure of skeletal muscle atrophy induced by microgravity or simulated microgravity. This study was carried out to test the effects of Xuefuzhuyu capsules(XFZY), an Chinese medicine known to improve the blood supply of tissue, on the skeletal muscle atrophy induced by simulated microgravity.

Methods

Rats fed the XFZY were tail‐suspended for 30 days(XFZY group), and the muscle mass and cross‐sectional area (CSA) were then compared with those of the suspended rats(TS group) and control rats(Con group). The protein expression and phosphorylation level in soleus muscle were measured by Western blot.

Results

Treatment with the XFZY attenuated the loss of muscle mass and CSA induced by tail suspension in rats. Furthermore, western blot analysis showed that the phosphatidyl‐inositol 3‐kinase/Phospho‐AKT were both down‐regulated after 30d tail suspension, which were reversed by treatment of XFZY. Meanwhile, Tai suspension increased the expression of GRP78, CHOP, phospho‐JNK level and Caspase 12 in soleus muscle, and XFZY alleviated the increased protein expression in soleus muscle induced by tail suspension.

Conclusion

Our results suggested that the XFZY can partially reverse the effects of tail suspension on soleus muscle and maybe target on the endoplasmic reticulum stress (ERS) system by regulating GRP78‐CHOP‐JNK‐Caspase12 pathway.