Sphingosine 1-Phosphate and Apolipoprotein M Levels and Their Correlations with Inflammatory Biomarkers in Patients with Untreated Familial Hypercholesterolemia

High-density lipoprotein (HDL)-bound apolipoprotein M/sphingosine 1-phosphate (ApoM/S1P) complex in cardiovascular diseases serves as a bridge between HDL and endothelial cells, maintaining a healthy endothelial barrier. To date, S1P and ApoM in patients with untreated heterozygous familial hypercholesterolemia (HeFH) have not been extensively studied. Eighty-one untreated patients with HeFH and 32 healthy control subjects were included in this study. Serum S1P, ApoM, sCD40L, sICAM-1, sVCAM-1, oxLDL, and TNFα concentrations were determined by ELISA. PON1 activities were measured spectrophotometrically. Lipoprotein subfractions were detected by Lipoprint. We diagnosed FH using the Dutch Lipid Clinic Network criteria. Significantly higher serum S1P and ApoM levels were found in HeFH patients compared to controls. S1P negatively correlated with large HDL and positively with small HDL subfractions in HeFH patients and the whole study population. S1P showed significant positive correlations with sCD40L and MMP-9 levels and PON1 arylesterase activity, while we found significant negative correlation between sVCAM-1 and S1P in HeFH patients. A backward stepwise multiple regression analysis showed that the best predictors of serum S1P were large HDL subfraction and arylesterase activity. Higher S1P and ApoM levels and their correlations with HDL subfractions and inflammatory markers in HeFH patients implied their possible role in endothelial protection.     

Compressibility and flow properties of a cohesive limestone powder in a medium pressure range

The most important design parameters for roller presses can be referred to flow characteristic of bulk materials. Usually the flow properties are measured in the low stress range 1–50 kPa at the shear rate about 1 mm/min. But this does not fit the stressing conditions in the roller press. Press shear cell was used for shear tests with cohesive limestone powder from Gummern in the so-called medium pressure range 50–1000 kPa.     

Dietary fat composition and tocopherol requirement: III. Quantitative studies on the relationship between dietary linoleate and vitamin E

The present study has involved biologically titrating linoleate vs. vitamin E using the male rat as the indicator. In the first of the titration studies, the dietary tocopherol level was held constant, while in the second study the linoleate intake was held constant. The investigation was conducted with male rats since these have a much higher linoleate requirement than females. By first depleting such animals of their stores of essential fatty acids by feeding a fat-free diet from weaning, a sensitive test organism was provided. These animals have an immediate need for linoleate during the repletion periods. If an imbalance between linoleate and vitamin E content existed in any of the dietary regimens, such an imbalance would have been more likely noted in test animals actively metabolizing the ingested linoleate. Based upon various nutritional and biochemical indices, the amount of tocopherol ordinarily included in the basic diets fed to our rats, 0.01% as dl-alphatocopheryl acetate, was adequate even when the diet provided up to 5% linoleate; an amount corresponding to ca. 12% of the total calories and providing a ratio of linoleate to the tocopherol of ca. 500:1. In the reverse biological titration with all test diets now providing the constant level of 5% linoleate, ratios of linoleate to vitamin E were satisfactory even in a ratio of as much as 2500:1 (or 0.4 mg gram of vitamin E per polyunsaturated fatty acid). The control animals continued on the fat-free diet indicated that there is a need for added tocopherols even in the absence of linoleate according to a number of biochemical indices. Based upon a number of accepted bioanalytical approaches, the minimum requirement for linoleate by the fat-depleted male rat was found to be between 100–200 mg/day or ca. 1–2% of the caloric intake. Although the fatty acid composition of tissue lipid fractions is markedly affected by the amount of linoleate in the diet, dietary tocopherol supplements have little effect on these values.     

Regional Differences in the Permeability Barrier of the Skin—Implications in Acantholytic Skin Diseases

The chemical milieu, microbiota composition, and immune activity show prominent differences in distinct healthy skin areas. The objective of the current study was to compare the major permeability barrier components (stratum corneum and tight junction (TJ)), investigate the distribution of (corneo)desmosomes and TJs, and measure barrier function in healthy sebaceous gland-rich (SGR), apocrine gland-rich (AGR), and gland-poor (GP) skin regions. Molecules involved in cornified envelope (CE) formation, desquamation, and (corneo)desmosome and TJ organization were investigated at the mRNA and protein levels using qRT-PCR and immunohistochemistry. The distribution of junction structures was visualized using confocal microscopy. Transepidermal water loss (TEWL) functional measurements were also performed. CE intracellular structural components were similarly expressed in gland-rich (SGR and AGR) and GP areas. In contrast, significantly lower extracellular protein levels of (corneo)desmosomes (DSG1 and CDSN) and TJs (OCLN and CLDN1) were detected in SGR/AGR areas compared to GP areas. In parallel, kallikrein proteases were significantly higher in gland-rich regions. Moreover, gland-rich areas were characterized by prominently disorganized junction structures ((corneo)desmosomes and TJs) and significantly higher TEWL levels compared to GP skin, which exhibited a regular distribution of junction structures. According to our findings, the permeability barrier of our skin is not uniform. Gland-rich areas are characterized by weaker permeability barrier features compared with GP regions. These findings have important clinical relevance and may explain the preferred localization of acantholytic skin diseases on gland-rich skin regions (e.g., Pemphigus foliaceus, Darier’s disease, and Hailey–Hailey disease).     

Dietary fat composition and tocopherol requirement: II. Nutritional status of heated and unheated vegetable oils of different ratios of unsaturated fatty acids and vitamin E

In studies conducted on male and female rats and involving evaluation of growth, reproductive and lactation performances and of lipid peroxidation, no evidence could be found for the need for added vitamin E (a-tocopherol) over and above that naturally present as tocopherols in the vegetable oils investigated. These oils are in common usage in industry, i.e., liquid nonhydrogenated cottonseed oil, a lightly hydrogenated cottonseed oil and a hydrogenated soybean oil shortening. The ratio of polyunsaturates to total tocopherol in the test oils varied from 640:1 to 9:1. Even those oils obtained from a commercial frying operation after a steady state had been attained contained sufficient vitamin E to meet dietary requirements. Results of in vitro peroxide hemolysis tests conducted on the red blood cells of the test animals did not correlate well with biological performance.     

Janus Kinase Inhibitors Improve Disease Activity and Patient-Reported Outcomes in Rheumatoid Arthritis: A Systematic Review and Meta-Analysis of 24,135 Patients

Pain, fatigue, and physical activity are major determinants of life quality in rheumatoid arthritis (RA). Janus kinase (JAK) inhibitors have emerged as effective medications in RA and have been reported to exert direct analgesic effect in addition to reducing joint inflammation. This analysis aims to give an extensive summary of JAK inhibitors especially focusing on pain and patient reported outcomes (PRO). MEDLINE, CENTRAL, Embase, Scopus, and Web of Science databases were searched on the 26 October 2020, and 50 randomized controlled trials including 24,135 adult patients with active RA met the inclusion criteria. JAK inhibitors yielded significantly better results in all 36 outcomes compared to placebo. JAK monotherapy proved to be more effective than methotrexate in 9 out of 11 efficacy outcomes. In comparison to biological disease-modifying antirheumatic drugs, JAK inhibitors show statistical superiority in 13 of the 19 efficacy outcomes. Analgesic effect determined using the visual analogue scale and American College of Rheumatology (ACR) 20/50/70 response rates was significantly greater in the JAK group in all comparisons, and no significant difference regarding safety could be explored. This meta-analysis gives a comprehensive overview of JAK inhibitors and provides evidence for their superiority in improving PROs and disease activity indices in RA.     

Effect of varying levels of dietary protein on tumor development and lipid metabolism in rats exposed to aflatoxin

Reports in the literature concerning the relationship of protein nutrition to aflatoxicosis are contradictory. In an attempt to elucidate this relationship more clearly, we have examined the effects of low, normal, and high protein-containing diets on tumor incidence and development, as well as on several biochemical indices, in rats which have been exposed to low levels of aflatoxin in a “chronic” rather than “acute” situation. In our study, male weanling rats were place for 3 months on otherwise adequate diets containing either 8, 22, or 30% casein with and without aflatoxin B₁ at 1.7 ppm. Half of the animals in each group received diets which were further supplemented with the amino acid, cystine, at 0.6% of the diet. (Sulfur-containing amino acids are the most limiting amino acids in casein, and the addition of cystine to the diet serves to improve the biological quality of the protein source.) After 3 months the animals were fed control diets without aflatoxin until they were killed at 1 year. Weight gain was markedly decreased and liver weights increased in response to aflatoxin in all groups except those on the low protein diets, where aflatoxin had no effect on these protein diets, where aflatoxin had no effect on these indices. No tumors were found in the livers of rats fed the low protein, aflatoxin-supplemented diet. In the other groups, the severity of the liver involvement increased progressively with increased protein levels in the diet. When cystine was included in the diet, tumors were observed also in the animals fed the low protein diet; furthermore, the livers of those animals on “normal” and high protein diets were much more severely involved than were the livers of animals on non-cystine supplemented diets. Plasma cholesterol levels were increased in response to aflatoxin when the diets containing 22 and 30% protein were fed and when cystine was included in the 8% protein diet. Liver cholesterol levels were increased in response to aflatoxin in all groups except in those receiving the low protein diets. Among these latter animals, aflatoxin administration had no effect on liver cholesterol values. Changes as a result of aflatoxin administration were also observed in the fatty acid composition of sterol esters, triglycerides, and phospholipids of liver and tumor tissue.     

Structural Characterization of Daunomycin-Peptide Conjugates by Various Tandem Mass Spectrometric Techniques

The use of peptide-drug conjugates has generated wide interest as targeted antitumor therapeutics. The anthracycline antibiotic, daunomycin, is a widely used anticancer agent and it is often conjugated to different tumor homing peptides. However, comprehensive analytical characterization of these conjugates via tandem mass spectrometry (MS/MS) is challenging due to the lability of the O-glycosidic bond and the appearance of MS/MS fragment ions with little structural information. Therefore, we aimed to investigate the optimal fragmentation conditions that suppress the prevalent dissociation of the anthracycline drug and provide good sequence coverage. In this study, we comprehensively compared the performance of common fragmentation techniques, such as higher energy collisional dissociation (HCD), electron transfer dissociation (ETD), electron-transfer higher energy collisional dissociation (EThcD) and matrix-assisted laser desorption/ionization–tandem time-of-flight (MALDI-TOF/TOF) activation methods for the structural identification of synthetic daunomycin-peptide conjugates by high-resolution tandem mass spectrometry. Our results showed that peptide backbone fragmentation was inhibited by applying electron-based dissociation methods to conjugates, most possibly due to the “electron predator” effect of the daunomycin. We found that efficient HCD fragmentation was largely influenced by several factors, such as amino acid sequences, charge states and HCD energy. High energy HCD and MALDI-TOF/TOF combined with collision induced dissociation (CID) mode are the methods of choice to unambiguously assign the sequence, localize different conjugation sites and differentiate conjugate isomers.     

Analysis of mir-9 Expression Pattern in Rat Retina during Postnatal Development

It is well established that miR-9 contributes to retinal neurogenesis. However, little is known about its presence and effects in the postnatal period. To expand our knowledge, miRNA-small RNA sequencing and in situ hybridization supported by RT-qPCR measurement were carried out. Mir-9 expression showed two peaks in the first three postnatal weeks in Wistar rats. The first peak was detected at postnatal Day 3 (P3) and the second at P10, then the expression gradually decreased until P21. Furthermore, we performed in silico prediction and established that miR-9 targets OneCut2 or synaptotagmin-17. Another two microRNAs (mir-135, mir-218) were found from databases which also target these proteins. They showed a similar tendency to mir-9; their lowest expression was at P7 and afterwards, they showed increase. We revealed that miR-9 is localized mainly in the inner retina. Labeling was observed in ganglion and amacrine cells. Additionally, horizontal cells were also marked. By dual miRNA-in situ hybridization/immunocytochemistry and qPCR, we revealed alterations in their temporal and spatial expression. Our results shed light on the significance of mir-9 regulation during the first three postnatal weeks in rat retina and suggest that miRNA could act on their targets in a stage-specific manner.     

Effect of sterculic acid upon aflatoxicosis in rats fed diets containing saturated and unsaturated fat

Investigations on trout have shown that the cyclopropenoid fatty acids, which occur naturally in small amounts in unrefined cottonseed oil, may act as powerful cocarcinogens when fed in conjunction with aflatoxin. Attempts at confirming these findings in mammals, i.e. rats, have been inconclusive. In this study, the effects of sterculic acid and aflatoxin upon lipid metabolism and tumor formation in male rats have been examined using basal diets containing either saturated or unsaturated fat to which the following additions were made: (A) basal diet (no supplements); (B) aflatoxin B₁ at 1.7 ppm; (C) sterculic acid at 210 ppm; and (D) aflatoxin B₁ at 1.7 ppm, plus sterculic acid at 210 ppm. The rats consumed these diets for 3 months and, thereafter, were fed the unsupplemented basal diet until sacrifice 9 months later. Growth was depressed in rats in groups B, C, and D, but no synergistic inhibition was observed, regardless of the fat source. Liver wt doubled in response to aflatoxin; however, only when the diet contained unsaturated fat did sterculic acid, in combination with aflatoxin, exaggerate the increase in liver wt (a reflection of the more severe liver pathology observed in these rats). In the animals fed the saturated fat diet, aflatoxin administration to animals fed the control or sterculic acid supplemented diets resulted in marked increases in plasma cholesterol levels; the unsaturated fat diets, supplemented with aflatoxin, evoked a slight increase in plasma cholesterol content which was nullified by sterculic acid supplementation.