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Calcium carbonate-enriched pumpkin affects calcium status in ovariectomized rats

2023, Wawrzyniak, Natalia, Gramza-Michałowska, Anna, Kurzawa, Paweł, Kołodziejski, Paweł Antoni, Suliburska, Joanna

AbstractCalcium carbonate (CaCO3)-enriched pumpkin may serve as a good source of calcium for patients diagnosed with osteoporosis. In this study, we aimed to determine the effect of CaCO3-enriched pumpkin on Ca status in ovariectomized rats. The study included 40 female Wistar rats divided into five groups (n = 8). One group was fed with a standard diet (control group), while the other four groups were ovariectomized and received a standard diet (control ovariectomized group), or a diet containing CaCO3-enriched pumpkin, alendronate, or both. The nutritional intervention lasted 12 weeks, and then the rats were euthanized. Tissue and blood samples were collected and assessed for the levels of total Ca, estradiol, parathyroid hormone, and procollagen type I N propeptide. In addition, a histological analysis was performed on femurs. The results of the study suggest that CaCO3-enriched pumpkin can increase Ca content in femurs and improve bone recovery in ovariectomized rats. Furthermore, enriched pumpkin contributes to Ca accumulation in the kidneys, and this effect is more pronounced in combination with alendronate.

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Impact of dietary salicylates on angiogenic factors and biochemical parameters in a rat model of preeclampsia

2025, Suliburska, Joanna, Cholik, Rafsan Syabani, Karaźniewicz-Łada, Marta, Wronka, Dorota, Karlik, Anna, Waśkiewicz, Agnieszka, Skrypnik, Katarzyna, Kołodziejski, Paweł, Cieślak, Adam, Przybył, Łukasz

Background The pathophysiology of preeclampsia involves impaired cytotrophoblastic invasion, placental ischemia, inflammation, and angiogenic imbalance. Prophylactic low-dose aspirin can reduce the risk of preeclampsia and fetal growth restriction in high-risk women. This study evaluated the effect of dietary salicylates on the development of preeclampsia in rats treated with L-NAME (NG-nitro-L-arginine-methyl ester). Methodology Pregnant Sprague-Dawley rats were randomly assigned to six groups and treated with dietary salicylates at two dose levels (1 and 10 mg/kg diet) or aspirin (doses adjusted to dietary salicylates). Preeclampsia was induced by administering L-NAME in drinking water from gestational days 6–19. Results Neither dietary salicylates nor aspirin, at either dose, affected blood pressure in L-NAME-treated rats. The lower dose of dietary salicylates significantly reduced urinary albumin levels. Both interventions prevented an increase in the sFlt/PLGF ratio and mitigated histopathological placental changes in preeclamptic rats. The higher dose of aspirin reduced placental VEGFR2 protein levels. Conclusion Dietary salicylate supplementation does not provide clear preventive effects against preeclampsia.

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Cord Blood Spexin Level in Mothers with Obesity—Forecast of Future Obesity?

2023, Wojciechowska, Malgorzata, Kołodziejski, Paweł, Pruszyńska-Oszmałek, Ewa, Leciejewska, Natalia, Krauss, Hanna, Checinska-Maciejewska, Zuzanna, Sassek, Maciej, Rekas-Dudziak, Anna, Bernatek, Malgorzata, Skrzypski, Marek, Wilczak, Maciej

Spexin (SPX) is a peptide that plays an important role in the regulation of food intake and body weight (BW) by the effect on carbohydrate-lipid metabolism. However, the role of SPX in fetal life, in children, and in adolescent metabolism is limited. Therefore, we decided to check whether obesity affects the concentration of SPX in the mother’s peripheral blood (MB) and umbilical cord blood (UCB). Using MB and UCB sera on the day of delivery obtained from 48 women (24 non-obese and 24 obese) and commercially available Elisa kits and colorimetric assays, we determined changes in SPX and the relationship between SPX concentration and other metabolic and anthropometric markers (body weight and BMI) on the day of delivery and in children at the age of 36 months. We found lower concentrations of SPX in MB (p < 0.05) and UCB (p < 0.01) derived from obese women (BMI > 30) and a moderate linear correlation (r = 0.4429; p < 0.01) between SPX concentrations in MB and UCB. We also noted that the concentration of SPX is not correlated with the child’s body weight on the day of birth (r = −0.0128). However, there is a relationship between SPX at birth and body weight at 3 years of age (r = −0.3219; p < 0.05). Based on the obtained results, it can be assumed that spexin is one of the factors modulating the child’s metabolism already in the fetal period and can be considered a potential marker of future predisposition to obesity. However, confirmation of this thesis requires additional research.

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MOTS-c modulates pancreatic islet function in rats and pigs in vitro

2025, Bień, Jakub, Pruszyńska-Oszmałek, Ewa, Kołodziejski, Paweł, Leciejewska, Natalia, Szczepankiewicz, Dawid, Grzęda, Emilia, Sassek, Maciej

Abstract MOTS-c is a promising regulator of metabolism and energy homeostasis. While its effects have been studied in cell lines, our team aimed to investigate its influence on more complex structures—specifically, isolated pancreatic islets. We used two animal models: the rat, which is commonly studied, and the pig, which shares greater physiological similarities with humans. This study assessed the expression and secretion of insulin and glucagon, the expression of their receptors, cell viability, and cell death following MOTS-c treatment of the islets. Additionally, we examined how MOTS-c secretion is affected by different incubation media, such as the presence of free fatty acids, pancreatic hormones, and different glucose concentrations. The results indicate that MOTS-c impacts pancreatic islet physiology by, for example, reducing insulin and glucagon secretion and enhancing cell viability. Notably, the effects differed between the two species, which may be attributed to anatomical differences in their pancreatic islets or structural variations in rat and pig MOTS-c. These facts may lead to the conclusion that if MOTS-c may be helpful in human medicine, the pig model should be considered another valuable choice.

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Effect of a high-fat high-fructose diet on the composition of the intestinal microbiota and its association with metabolic and anthropometric parameters in a letrozole-induced mouse model of polycystic ovary syndrome

2024, Pieczyńska-Zając, Joanna Maria, Malinowska, Anna Maria, Pruszyńska-Oszmałek, Ewa, Kołodziejski, Paweł Antoni, Drzymała-Czyż, Sławomira, Bajerska, Joanna

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Special Issue: Lipid Metabolism, Adipogenesis and Fat Tissue Metabolism: Gene Regulation

2023, Skrzypski, Marek, Kołodziejski, Paweł

Lipid metabolism is pivotal in controlling energy homeostasis [...]

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The Effects of Peruvian maca (Lepidium meyenii) Root Extract on In Vitro Cultured Porcine Fibroblasts and Adipocytes

2025, Loba-Pasternak, Weronika, Aksoy, Mehmet Onur, Stuper-Szablewska, Kinga, Szwajkowska-Michałek, Lidia, Kołodziejski, Paweł Antoni, Szczerbal, Izabela, Nowacka-Woszuk, Joanna

Peruvian maca (Lepidium meyenii) is a plant known for its nutritional and medicinal properties whose use as a supplement in animal diets has attracted much interest. We studied the effects of powdered maca root extract on the growth potential of in vitro cultured porcine cells prior to its use as an additive in animal nutrition. Fibroblast cell viability (MTT), cell proliferation (BrdU), and apoptosis level (TUNEL) were measured for a range of extract doses (0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 7.0, and 10 mg/mL). Transcript levels of CCND1, MCM2, and PCNA genes as molecular markers of cell proliferation were also determined. Next, the effects of maca extract at 2 and 5 mg/mL on in vitro induced adipogenesis were evaluated over eight days of differentiation. The transcript levels of three adipocyte marker genes (CEBPA, PPARG, and FABPB4) were measured at days 0, 4, and 8 of adipose differentiation, and lipid droplet accumulation (BODIPY staining) was also noted. No cytotoxic effect was detected on fibroblast cell viability, and the inhibitory concentration (IC50) value was determined to be IC50 > 10 mg/mL. Doses of maca extract above 3 mg/mL decreased cell proliferation. The transcript level decreased in concentrations above 5 for the MCM2 and PCNA genes. For the CCND1 gene, the transcript level decreased when the greatest maca dose was used. In the in vitro adipogenesis experiment, it was found that the rate of lipid droplet formation increased on day 4 of differentiation for both doses, while decreased lipid droplet formation was observed on day 8 for 5 mg/mL of maca extract. Significant changes were seen in the mRNA level for CEBPA and PPARG on days 4 and 8, while the transcript of FABP4 increased only on day 8 at 2 mg/mL dose. It can be concluded that the addition of Peruvian maca in small doses (<3 mg/mL) has no negative effect on porcine fibroblast growth or proliferation, while 2 mg/mL of maca extract enhances adipocyte differentiation.

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LEAP2 in Physiology—A Narrative Review

2025, Sosinski, Oskar, Pruszyńska-Oszmałek, Ewa, Leciejewska, Natalia, Sassek, Maciej, Kołodziejski, Paweł Antoni

Liver Enriched Antimicrobial Peptide 2 (LEAP2) is a fascinating peptide that has gained significant attention since its discovery in 2003. Initially identified as an antimicrobial peptide, LEAP2 has more recently been found to play a key role in the regulation of energy metabolism. One of the most notable functions of LEAP2 is its interaction with the ghrelin hormone, which is known for stimulating hunger. LEAP2 acts as an inhibitor of ghrelin, thereby reducing food intake and influencing energy balance. The physiological roles of LEAP2 extend beyond appetite suppression. Studies have shown that LEAP2 has an impact on insulin secretion, suggesting its potential involvement in glucose metabolism and possibly insulin sensitivity, which is crucial in managing conditions like type 2 diabetes. Moreover, LEAP2 levels appear to fluctuate based on factors such as gender, developmental stage, and even interventions like bariatric surgery, which is known for its role in managing obesity and diabetes. Given these findings, LEAP2 shows potential as a therapeutic target, particularly for addressing obesity and metabolic diseases such as type 2 diabetes. Its ability to influence food intake and energy balance makes it a promising candidate for further research into therapies aimed at weight regulation and glycemic control. In the future, LEAP2 could become an important agent in the development of treatments aimed at curbing obesity and its associated metabolic disorders.

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Research Project

LEAP2 jako modulator metabolizmu tkanki tłuszczowej oraz trzustki w cukrzycy typu 2

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Combination of emulsifier and xylanase in triticale-based broiler chickens diets

2023, Wiśniewska, Zuzamma, Kołodziejski, Paweł Antoni, Pruszyńska-Oszmałek, Ewa, Konieczka, Paweł, Kinsner, Misza, Górka, Paweł, Flaga, Jadwiga, Kowalik, Kinga, Hejdysz, Marcin, Kubiś, Marta, Jarosz, Łukasz S., Ciszewski, Artur, Kaczmarek, Sebastian

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Insect fat influences broiler performance, meat quality, and the cecal microbiota similarly to plant oils rather than animal fats

2025, Aslam, Muhammad Rumman, Kierończyk, Bartosz, Rawski, Mateusz, Szymkowiak, Piotr, Stuper-Szablewska, Kinga, Kołodziejski, Paweł, Mikuła, Robert, Dankowiakowska, Agata, Józefiak, Damian

Abstract We evaluated the addition of Hermetia illucens larvae (BSFL) fat to broiler diets compared to that of soybean oil (SO), rapeseed oil (RO), palm oil (PO), palm kernel fatty acid distillate (PKFD), poultry fat (PF), pig lard (PL), and beef tallow (BT) on performance, meat quality, and cecal microbiota abundance. BSFL addition reduced the feed conversion ratio compared to PO, PF, and BT addition and improved nutrient digestibility at 14 days and 28 days of age. BSFL addition enhanced broiler immune responses compared with SO, PO, and PL by increasing IgA levels; compared to PO by increasing IgG levels; and compared to PL by increasing IgY and IL-6 levels. Moreover, broiler free triiodothyronine levels increased when BSFL was added. Reduced drip loss after 7 days was observed in the RO, PO, PF, PL, and BT groups, while the salt-induced water uptake in the PL group was greater than that in the BSFL group. Compared with the SO group, BSFL addition increased C18:3 and overall n-3 FAs in breast meat. BSFL fat in broiler diets leads to outcomes that are more similar to those of plant oils than to those of animal fats, particularly regarding nutrient digestiblity and immune response.

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Ostarine does not enhance the metabolic effect of exercise in obese rats

2023, Leciejewska, Natalia, Pruszyńska-Oszmałek, Ewa, Sassek, Maciej, Głowacki, M., Lehmann, T., Rekas-Dudziak, A., Nogowski, Leszek, Nowak, Krzysztof W., Kołodziejski, Paweł

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GIP_HUMAN [22–51] Peptide Encoded by the Glucose-Dependent Insulinotropic Polypeptide (GIP) Gene Suppresses Insulin Expression and Secretion in INS-1E Cells and Rat Pancreatic Islets

2023, Pusch, Emily, Krążek, Małgorzata, Wojciechowicz, Tatiana, Sassek, Maciej, Kołodziejski, Paweł, Strowski, Mathias, Nowak, Krzysztof W., Skrzypski, Marek

GIP_HUMAN [22–51] is a recently discovered peptide that shares the same precursor molecule with glucose-dependent insulinotropic polypeptide (GIP). In vivo, chronic infusion of GIP_HUMAN [22–51] in ApoE−/− mice enhanced the development of aortic atherosclerotic lesions and upregulated inflammatory and proatherogenic proteins. In the present study, we evaluate the effects of GIP_HUMAN [22–51] on insulin mRNA expression and secretion in insulin-producing INS-1E cells and isolated rat pancreatic islets. Furthermore, we characterize the influence of GIP_HUMAN [22–51] on cell proliferation and death and on Nf-kB nuclear translocation. Rat insulin-producing INS-1E cells and pancreatic islets, isolated from male Wistar rats, were used in this study. Gene expression was evaluated using real-time PCR. Cell proliferation was studied using a BrdU incorporation assay. Cell death was quantified by evaluating histone-complexed DNA fragments. Insulin secretion was determined using an ELISA test. Nf-kB nuclear translocation was detected using immunofluorescence. GIP_HUMAN [22–51] suppressed insulin (Ins1 and Ins2) in INS-1E cells and pancreatic islets. Moreover, GIP_HUMAN [22–51] promoted the translocation of NF-κB from cytoplasm to the nucleus. In the presence of a pharmacological inhibitor of NF-κB, GIP_HUMAN [22–51] was unable to suppress Ins2 mRNA expression. Moreover, GIP_HUMAN [22–51] downregulated insulin secretion at low (2.8 mmol/L) but not high (16.7 mmol/L) glucose concentration. By contrast, GIP_HUMAN [22–51] failed to affect cell proliferation and apoptosis. We conclude that GIP_HUMAN [22–51] suppresses insulin expression and secretion in pancreatic β cells without affecting β cell proliferation or apoptosis. Notably, the effects of GIP_HUMAN [22–51] on insulin secretion are glucose-dependent.

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Implementation of a metabolizable energy regression model for black soldier fly larvae fat in broiler chicken diets: effect on growth performance, nutrient digestibility, and selected physiological indices

2024, Kierończyk, B., Kaczmarek, S. A., Hejdysz, M., Szymkowiak, P., Aslam, M. R., Rawski, M., Kołodziejski, P. A., Mikuła, R., Stuper-Szablewska, K., Józefiak, D.

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Effect of Feeding Dried Apple Pomace on Ruminal Fermentation, Methane Emission, and Biohydrogenation of Unsaturated Fatty Acids in Dairy Cows

2023, Gadulrab, Khaled, Sidoruk, Pola, Kozłowska, Martyna, Szumacher, Małgorzata, Lechniak, Dorota, Kołodziejski, Paweł Antoni, Pytlewski, Jarosław, Strzałkowska, Nina, Horbańczuk, Jarosław Olav, Jóźwik, Artur, Yanza, Yulianri Rizki, Irawan, Agung, Patra, Amlan Kumar, Cieślak, Adam

Industrial fruit by-products are now being utilized as animal feeds for several reasons. They may substitute the conventional cereal feeds, and also offer economic and environmental benefits. One of the most important industrial fruit by-products is apple pomace, which can be used as a source of energy in the ration of ruminant species, including dairy cattle. The aim of the present study was to evaluate the effect of feeding dried apple pomace to dairy cattle on ruminal fermentation, fatty acid concentration, microbial populations, and methane production. The experiment lasted 64 days and was conducted with 4 cannulated commercial dairy cows. The control animals received a standard diet, while the experimental animals was fed a standard diet supplemented with 150 g/kg DM dried apple pomace. Ruminal fluid samples were collected at three different time intervals. The samples were obtained at 0-, 3-, and 6-h post-feeding. The ruminal fluid was used to assess the ammonia concentration, pH, volatile fatty acids (VFA), long-chain fatty acids (FA), microbial population. A number of ruminal fermentation variables changed as a result of the addition of dried apple pomace to the standard diet. Ruminal pH slightly increased (p < 0.01) while the ammonia concentration decreased (p < 0.01) by 46%. There was a significant decrease in total protozoa count (p < 0.01) and an increase (p < 0.01) in total volatile fatty acids. In addition, there was a decline in methane emission (p = 0.05) by 8% due to dried apple pomace feeding. To sum up, this study demonstrated a positive effect of 150 g/kg DM dietary dried apple pomace on ruminal metabolism including a decrease in ammonia concentration and methane emissions, alongside with an increase in total ruminal VFAs, higher nutrient digestibility, and milk production. Also, beneficial changes to the ruminal fatty acid profile resulting from reduced biohydrogenation were observed although a decreased content of the C18:2 cis 9 trans 11 isomer was also noticed. The dietary inclusion of DAP can serve as a valuable, sustainable, and environmentally friendly dietary component for dairy cows.

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Neopterin Concentration in Umbilical Cord Blood as a Reflection of Maternal Insulin Resistance—A Pilot Study

2025, Chęcińska-Kopeć, Aleksandra, Pruszyńska-Oszmałek, Ewa, Checinska-Maciejewska, Zuzanna, Rekas-Dudziak, Anna, Wojciechowska, Małgorzata, Ślósarz, Piotr Paweł, Krauss, Hanna, Szymanowski, Krzysztof, Kołodziejski, Paweł

Background/Objective: Insulin resistance (IR) during pregnancy, even in women with normal body mass index (BMI), may affect maternal and fetal metabolic and immune status. This study aimed to evaluate neopterin (NPT), leptin, insulin, and ghrelin concentrations in maternal blood (MB) and umbilical cord blood (CB) in normoglycemic women with and without IR, all with normal BMI. Methods: Peripheral and cord blood was collected from 36 Caucasian women with term, uncomplicated vaginal deliveries. The participants were classified into control (n = 16; age = 30.81 ± 4.875 years) and IR (n = 20; age = 31.95 ± 4.979 years) groups based on a professional medical diagnosis. Anthropometric parameters were recorded, and metabolic/hormonal markers were measured using ELISA and RIA. Results: NPT concentrations in CB were significantly higher in the IR group (p < 0.05), correlated positively with MB NPT levels (r = 0.3809, p < 0.05). A significantly higher concentration of both insulin and leptin was observed in the MB of women with IR compared to the control group (p < 0.0001), whereas in CB, only insulin concentration was significantly higher in the IR group than in healthy controls (p < 0.05). Ghrelin levels did not differ between the groups. Conclusions: Insulin resistance in non-obese pregnant women is associated with increased NPT concentration in CB, which may suggest fetal immune activation. However, defining the role of NTP as a metabolic “messenger” between mother and child requires further study.

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Effects of isoflavone and probiotic intake on calcium transport and bone metabolism biomarkers in female rats

2023, Harahap, Iskandar Azmy, Kuligowski, Maciej, Schmidt, Marcin, Kołodziejski, Paweł, Suliburska, Joanna

AbstractCalcium is essential for maintaining bone health as it contributes to bone formation, remodeling, strength, and density. This study investigated the effect of isoflavones and probiotics on calcium transporters' gene expression, serum calcium levels, and bone metabolism biomarkers in healthy female rats. Forty‐eight female Wistar rats were classified into six groups. Bone metabolism biomarkers (pyridinoline, deoxypyridinoline, parathyroid hormone, and osteocalcin) and serum calcium levels were measured by enzyme‐linked immunosorbent assay (ELISA) and atomic absorption spectroscopy (AAS), respectively. Gene expression of calcium transporters (Trpv5 and Trpv6) was evaluated in duodenum and jejunum tissue samples using quantitative polymerase chain reaction (qPCR). Trpv5 and Trpv6, epithelial calcium channels, play a crucial role in calcium transport and homeostasis in the body. The study consisted of a1‐week adaptation period for the rats to adjust to the controlled conditions, followed by an 8‐week intervention phase. The daidzein and genistein group showed a significant increase in the gene expression of the Trpv6 transporter in the duodenum and a marked decrease in serum pyridinoline levels compared to the control group. The tempeh and soybean groups showed a significant decrease in the gene expression of the Trpv5 calcium transporter in the jejunum. However, no significant influence of the Lactobacillus acidophilus diet on calcium transport and bone metabolism biomarkers was observed in the L. acidophilus group. The correlation analysis showed a significant positive relationship between serum calcium, bone metabolism biomarkers, and calcium transporters. In conclusion, our study demonstrates that the daidzein and genistein diet improves calcium transport in the duodenum and reduces pyridinoline serum concentrations, while tempeh and soybean diets reduce calcium transport in the jejunum. However, the combination of daidzein, genistein, and L. acidophilus did not demonstrate a synergistic effect on calcium transport and bone metabolism, suggesting that further investigations are needed to elucidate their potential interactions.

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Effect of clinically uneventful carotid endarterectomy on pentraxin 3 concentration: relationships between cognitive functions, local inflammation, and duration of brain ischemia

2025, Rękas-Dudziak, Anna Renata, Rojek, Joanna, Blacha, Anna, Pruszyńska-Oszmałek, Ewa, Kołodziejski, Paweł, Matuła, Przemysław

IntroductionThe pentraxins are a superfamily of proteins characterized by the presence of a pentraxin domain at the C-terminal. One of them is pentraxin 3 (PTX3), which is of great interest to scientists, primarily due to its function as an endogenous modulator of the inflammatory response. Local inflammation is a common phenomenon during various types of surgery and procedures, and its causes, among others, are local ischemia or tissue irritation. One such procedure is carotid plaque removal by carotid endarterectomy (CEA), which is performed to reduce the risk of stroke.Material and methodsThe purpose of this study was to investigate the effect of CEA on PTX3 concentration and the relationship between cognitive functions and the duration of brain ischemia in patients. We evaluated the relationships using blood collected from patients undergoing CEA, the Mini-Mental State Examination (MMSE) questionnaire, and commercially available ELISA tests.ResultsResults: We found that the concentration of PTX3 increased after surgery (from 0.59 ± 0.11 to 1.29 ± 0.29 ng mL<sup>–1</sup>; <i>P</i> < 0.01). Also, a decrease in the MMSE score was observed 24 hours after the procedure (<i>P</i> < 0.01). However, just one month after the procedure, it significantly increased compared to the day of admission to the hospital (<i>P</i> < 0.01). Our results did not demonstrate any interactions between the tested parameters. It can be concluded that neither the changes in PTX3 concentration following the procedure nor the timing of these changes directly contribute to the temporary deterioration of cognitive functions observed after CEA.ConclusionsPTX3 increases independently of the duration of cerebral hypoxemia/ hypercapnia after clamping the carotid artery. This rise is an inflammatory marker that occurs independently within the central nervous system.

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Cleft Lip and Palate in Four Full-Sib Puppies from a Single Litter of Staffordshire Bull Terrier Dogs: An Anatomical and Genetic Study

2023, Ruszkowski, Jakub Jędrzej, Nowacka-Woszuk, Joanna, Nowak, Tomasz, Rozynek, Jędrzej, Serwańska-Leja, Katarzyna, Gogulski, Maciej, Kołodziejski, Paweł Antoni, Świtoński, Marek, Zdun, Maciej, Szczerbal, Izabela

Cleft lip and palate (CLP) is a well-known congenital defect in dogs, characterized by abnormal communication between the oral and nasal cavities. Its incidence rate is high and affects all dog breeds. The etiology of CLP is thought to be multifactorial, caused by both genetic and environmental factors. In this study, four puppies out of seven from a single litter of Staffordshire Bull Terrier dogs with craniofacial abnormalities were anatomically and genetically examined. Classical anatomical preparation, dyed-latex-injection of the arterial vessels, and cone-beam computed tomography were used. The puppies showed variations in their observable abnormalities: three of them had a complete cleft of the palate on both sides, while one puppy had a cleft on the right side only. Cytogenetic analysis showed a normal diploid chromosome number (2n = 78,XX or 78,XY) in the studied animals. Known genomic variants of CLP were examined in the ADAMTS20, DLX6, and MYH3 genes, but no mutations were identified. Further studies are needed to identify the breed-specific genetic variants associated with canine CLP.

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MOTS-c Impact on Muscle Cell Differentiation and Metabolism Across Fiber Types

2025, Leciejewska, Natalia, Pruszyńska-Oszmałek, Ewa, Kołodziejski, Paweł Antoni, Szczepankiewicz, Dawid, Nogowski, Leszek, Sassek, Maciej

Background/Aims: MOTS-c belongs to a group of mitochondrial peptides involved in metabolic processes in the body. This peptide has garnered increasing attention since its discovery in 2015 because of its potential to ameliorate metabolic parameters in animals with diabetes or insulin resistance. MOTS-c is involved in muscle metabolism; however, little is known about its role in fiber differentiation. Materials: We conducted a study to explore the effect of MOTS-c on cellular processes using the C2C12 and L6 cell lines, representing different metabolic types of muscle fibers. The research methods were real-time PCR, Western blot, and lipid accumulation measurement. Results: >Notably, our investigations revealed that MOTS-c increased the survival of C2C12 cells at doses of 10 and 100 nM (p<0.01) and stimulated the phosphorylation of extracellular signal-regulated kinase within 5 min of incubation (p<0.05). Remarkably, these effects were not observed in L6 cells; however, both cell lines showed a reduced rate of proliferation. Furthermore, MOTS-c promotes the differentiation of C2C12 cells by increasing the expression of muscle regulatory factors, but it does not produce such an effect in L6 cells. Additionally, cells were treated with physiological concentrations of free fatty acids and MOTS-c, unveiling an augmentation in lipid accumulation observed in L6 cells and a decrease in lipid accumulation in C2C12 cells. Conclusion: In conclusion, our findings have suggested a diverse response to MOTS-c depending on the type of muscle fibers, particularly in the domains of survival, cell differentiation, and lipid accumulation.