Increased Amount of Polyunsaturated Fatty Acids in the Intestinal Contents of Patients with Morbid Obesity.

INTRODUCTION: Obesity is associated with disturbed gut microbiota homeostasis that translates into altered intestinal and blood metabolite profiles. The long-chain fatty acid (LCFA) may be absorbed in the intestine, but until now, their composition in intestinal contents of patients with obesity has not been studied. The aim of the present study was to verify whether obesity is related to any changes in fecal LCFA content and whether intestinal LCFA content may be associated with the health status of patients with obesity. METHODS: The fatty acid composition has been studied in stool samples obtained from 26 patients with morbid obesity and 25 lean subjects by gas chromatography-mass spectrometry. The dietary habits were assessed using the Food Frequency Questionnaire (FFQ-6). RESULTS: Our results show for the first time that lean subjects and patients with obesity differ in their stool LCFA profiles. The levels of most n-3 polyunsaturated fatty acids (PUFAs) and n-6 PUFAs were significantly higher in fecal samples from people with obesity than in those from lean controls. CONCLUSIONS: Based on the current knowledge, we have defined three hypotheses that may explain proving the cause-and-effect relationships observed differences in fecal LCFA profiles between patients with obesity and lean subjects. They may be related to alterations in fat digestion and/or LCFA absorption and diet. However, proving the cause-and-effect relationships requires further research.

Specific Needs of Patients and Staff Reflected in the Design of an Orthopaedic and Rehabilitation Hospital-Design Recommendations Based on a Case Study (Poland).

This article presents results of the research conducted with the use of participatory methods by the Faculty of Architecture, Poznan University of Technology on architectural design of healthcare facilities. The studies concerned the needs of patients and hospital staff in an orthopaedic and rehabilitation hospital. Preferences and expectations of all the users of healthcare facilities should be considered as early as at the stage of planning and designing. The hospital profile and the type of its users predetermine the architectural design in the area of the building functions, its internal circulation and interior design. Participation of the user in the process of compiling design recommendations is a chance for a designer to confront the original assumptions with expectations and to adjust relevant solutions to factual needs of the users. This study, undertaken in a regional setting, provided an in-depth exploration of staff's experiences of hospital space to indicate possible spatial improvements. METHODS: The research was conducted on the basis of a case study of a renowned regional orthopaedic and rehabilitation hospital in Poznan, Poland. Rapid assessment methods and questions were examined to describe current approaches and synthesize results. Semi-structured interviews and thematic analysis identified staff and patient's experiences. RESULT: Participation of hospital staff and patients resulted in design recommendations of high utility value. It was found that the two groups to a certain extent proposed similar recommendations; however, certain proposals submitted by the two groups were totally opposite. CONCLUSION: the research highlighted the importance of the active research methodology that engages the researcher/expert in the action and critical reflection process. Such a methodology can successfully underlie the formulation of accurate recommendations.

Pro-Inflammatory Profile of Adipokines in Obesity Contributes to Pathogenesis, Nutritional Disorders, and Cardiovascular Risk in Chronic Kidney Disease.

Obesity is a disease which leads to the development of many other disorders. Excessive accumulation of lipids in adipose tissue (AT) leads to metabolic changes, including hypertrophy of adipocytes, macrophage migration, changes in the composition of immune cells, and impaired secretion of adipokines. Adipokines are cytokines produced by AT and greatly influence human health. Obesity and the pro-inflammatory profile of adipokines lead to the development of chronic kidney disease (CKD) through different mechanisms. In obesity and adipokine profile, there are gender differences that characterize the male gender as more susceptible to metabolic disorders accompanying obesity, including impaired renal function. The relationship between impaired adipokine secretion and renal disease is two-sided. In the developed CKD, the concentration of adipokines in the serum is additionally disturbed due to their insufficient excretion by the excretory system caused by renal pathology. Increased levels of adipokines affect the nutritional status and cardiovascular risk (CVR) of patients with CKD. This article aims to systematize the current knowledge on the influence of obesity, AT, and adipokine secretion disorders on the pathogenesis of CKD and their influence on nutritional status and CVR in patients with CKD.

Intestinal Microbiota as a Contributor to Chronic Inflammation and Its Potential Modifications.

The gut microbiota is a crucial factor in maintaining homeostasis. The presence of commensal microorganisms leads to the stimulation of the immune system and its maturation. In turn, dysbiosis with an impaired intestinal barrier leads to accelerated contact of microbiota with the host's immune cells. Microbial structural parts, i.e., pathogen-associated molecular patterns (PAMPs), such as flagellin (FLG), peptidoglycan (PGN), lipoteichoic acid (LTA), and lipopolysaccharide (LPS), induce inflammation via activation of pattern recognition receptors. Microbial metabolites can also develop chronic low-grade inflammation, which is the cause of many metabolic diseases. This article aims to systematize information on the influence of microbiota on chronic inflammation and the benefits of microbiota modification through dietary changes, prebiotics, and probiotic intake. Scientific research indicates that the modification of the microbiota in various disease states can reduce inflammation and improve the metabolic profile. However, since there is no pattern for a healthy microbiota, there is no optimal way to modify it. The methods of influencing microbiota should be adapted to the type of dysbiosis. Although there are studies on the microbiota and its effects on inflammation, this subject is still relatively unknown, and more research is needed in this area.

Short-Term Effect of One-Anastomosis Gastric Bypass on Essential Fatty Acids in the Serum of Obese Patients.

One-anastomosis gastric bypass is a promising type of bariatric surgery, but it may lead to a deficiency in important nutrients, such as fatty acids. The short-term effects of one-anastomosis gastric bypass on serum fatty acids have not been studied thus far. Therefore, the aim of this study was to determine the effect of one-anastomosis gastric bypass on serum fatty acid composition two weeks after surgery. This study included 38 patients who underwent one-anastomosis gastric bypass as surgical treatment for morbid obesity. Serum fatty acid composition was analyzed before and two weeks after surgery using gas chromatography-mass spectrometry. We observed a decrease in essential polyunsaturated fatty acids (p < 0.001 for linolenic acid and p < 0.001 for linoleic acid) and odd-chain fatty acids (p = 0.004) in the serum of obese patients shortly after a one-anastomosis gastric bypass. Considering the benefits of the aforementioned fatty acids for human health, the implementation of a fatty-acid-rich diet or the use of supplementation may be recommended for patients immediately after one-anastomosis gastric bypass.

A prospective evaluation of the influence of three bariatric procedures on insulin resistance improvement. Should the extent of undiluted bile transit be considered a key postoperative factor altering glucose metabolism?

INTRODUCTION: Insulin resistance (IR), the essential step in development of type 2 diabetes mellitus (T2DM), resolves quickly after bariatric surgery, but the effectiveness depends on the type of the procedure. Although the long-term influence on IR improvement is well documented, the mechanisms of the ultra-fast response after restrictive and bypass procedures require explanation. AIM: To determine IR evolution from the initial preparative period to 6 months after the operation, exposing the rapid postoperative response while comparing the 3 bariatric methods, with the belief that the metabolic effect may be correlated with anatomical combinations. MATERIAL AND METHODS: From January to December, 2013, a cohort of severely obese, insulin resistant individuals recruited to the prospective study underwent laparoscopic sleeve gastrectomy (SG-30), Roux-en-Y gastric bypass (RYGB-30) and one-anastomosis gastric bypass (OAGB-30). Main laboratory parameters of glucose metabolism were evaluated in fasting patients preoperatively, 4 days and 1, 3 and 6 months after surgery. RESULTS: Within the whole observation period the most significant improvement in homeostasis model assessment for IR (HOMA-IR) was observed in the first 4 days after each operation. The decrease of HOMA-IR was higher (p < 0.0001) in gastric bypass groups than in patients after SG (-41%). The difference between bypass groups favors OAGB over RYGB (63 vs. -56%, p = 0.0489). CONCLUSIONS: Among all bariatric management factors, operation type is the most important in IR improvement. The significant difference in response after SG vs. RYGB and OAGB supports the concept of metabolic competence of duodeno-jejunal exclusion. Altered bile flow after duodeno-jejunal exclusion may be responsible for enhanced glucose metabolism improvement.

Fasting and postprandial acyl and desacyl ghrelin levels in obese and non-obese subjects.

INTRODUCTION: The potentially differential roles of both forms of ghrelin in obesity are undefined, and little is known about desacyl ghrelin's (DAG) regulation by meals. We aimed to assess changes in acyl ghrelin (AG) and DAG in response to mixed-meal consumption in obese and non-obese subjects. MATERIAL AND METHODS: Venous blood for plasma glucose, AG and DAG assays were collected in both groups after an overnight fast and two hours after the consumption of a standard 300 kcal-mixed meal (Nutridrink, Nutricia). RESULTS: Mean fasting values of both AG and DAG were significantly lower in the obese individuals. On the other hand, among non-obese controls, the mean postprandial DAG levels did not change and AG levels decreased, whereas in obese individuals the mean DAG levels after a mixed-meal diminished and AG levels were unchanged. CONCLUSIONS: It is necessary to distinguish between the desacylated and acylated forms of ghrelin, as we have shown differential postprandial AG and DAG responses in obese and non-obese individuals. Whether targeting changed proportions between AG and DAG could be a successful strategy in obesity treatment remains a question for future studies.

Obesity and diet awareness among Polish children and adolescents in small towns and villages.

BACKGROUND: In addition to genetic predispositions and environmental factors, healthy lifestyle education is very important for children and adolescents. The purpose of this research was to estimate the number of overweight and obese children and adolescents from small towns and villages and to find out an association between health awareness in children and the risk of becoming overweight or obese. METHODS: The research was conducted in 1,515 healthy children aged 6-18 years from small towns and villages in Poland. Overweight was diagnosed when BMI for age and sex was over the 90th percentile; obesity--when it was over the 95th percentile. The study consisted of a lifestyle interview and anthropometrical measurements. The lifestyle interview was conducted with the use of an anonymous questionnaire form and included questions about food frequency, diet habits and physical activity. The research was analysed using the SAS System for Windows, release 8.02. RESULTS: Overweight status was diagnosed in 9.0% and obesity in 5.1% of respondents. Excess body mass was statistically more frequently diagnosed in girls than in boys aged 14-18 years. Girls of this age group significantly more frequently chose wholemeal bread, smoked sausages, meat and poultry as products that are believed to keep them fit. Older children substantially more often indicated that stress, smoking cigarettes, consuming fatty meat, sweets, being obese, and a lack of physical activity are factors that damage health. Boys spent more time in front of a computer or TV than girls; in the older group of children, the phenomenon even intensified. CONCLUSION: Awareness of healthy lifestyle behaviour is not sufficient to maintain optimal body mass. Knowledge about proper eating habits is better among girls than among boys, especially in the older age groups. However, in older groups, there was less physical activity due to spending more time in front of TV or the computer. High percentage of obese/overweight children and insufficient knowledge of nutrition may consequently result in increased risk of cardio-vascular diseases in adult population.

The universe of Hsp90.

Abstract Molecular chaperones are key components in the maintenance of cellular homeostasis and survival, not only during stress but also under optimal growth conditions. Among the ATP-dependent chaperones, heat shock proteins (Hsp90) proteins play a special role. While Hsp90s can interact with unfolded and misfolded proteins, their main (and in eukaryotic cells essential) function appears to involve interactions with a limited number of protein clients at late steps of maturation or in "alter-native" conformations for regulating their stability and activity. Because Hsp90 clients are hubs of diverse signaling networks and participate in nearly every cellular function, Hsp90s interconnect many regulatory circuits and link them to environmental impacts. The availability and activity of Hsp90 may thus influence complex physiological and pathophysiological processes, such as differentiation, development, aging, cancer, neurodegeneration, and infectious diseases. Furthermore, through homeostatic effects on differentiation and development, Hsp90s act as capacitors of phenotypic evolution. In this review, we discuss recent insights in the structure and chaperone cycle of Hsp90s, the mechanisms underlying Hsp90 binding to clients, and potential reasons why client proteins specifically require the assistance of Hsp90s. Moreover, the current views on Hsp90-cochaperone interactions and regulation of Hsp90 proteins via posttranslational modifications are summarized. The second half of this article is devoted to the role of Hsp90 proteins in health and disease, aging, and evolution.

CHIP participates in protein triage decisions by preferentially ubiquitinating Hsp70-bound substrates.

The E3 ubiquitin ligase CHIP (C-terminus of Hsc70-interacting protein) is believed to be a central player in the cellular triage decision, as it links the molecular chaperones Hsp70/Hsc70 and Hsp90 to the ubiquitin proteasomal degradation pathway. To better understand the decision process, we determined the affinity of CHIP for Hsp70 and Hsp90 using isothermal titration calorimetry. We analyzed the influence of CHIP on the ATPase cycles of both chaperones in the presence of co-chaperones and a substrate, and determined the ubiquitination efficacy of CHIP in the presence of the chaperones. We found that CHIP has a sixfold higher affinity for Hsp90 compared with Hsc70. CHIP had no influence on ADP dissociation or ATP association, but reduced the Hsp70 cochaperone Hdj1-stimulated single-turnover ATPase rates of Hsc70 and Hsp70. CHIP did not influence the ATPase cycle of Hsp90 in the absence of co-chaperones or in the presence of the Hsp90 cochaperones Aha1 or p23. Polyubiquitination of heat-denatured luciferase and the native substrate p53 was much more efficient in the presence of Hsc70 and Hdj1 than in the presence of Hsp90, indicating that CHIP preferentially ubiquitinates Hsp70-bound substrates.

Insights into the conformational dynamics of the E3 ubiquitin ligase CHIP in complex with chaperones and E2 enzymes.

The dimeric E3 ubiquitin ligase CHIP binds with its tetratricopeptide repeat (TPR) domain the C-terminus of molecular chaperones Hsp70 and Hsp90 and with its U-box region E2 ubiquitin-conjugating enzymes. By ubiquitinating chaperone-bound polypeptides, CHIP thus links the chaperone machinery to the proteasomal degradation pathway. The molecular mechanism of how CHIP discriminates between folding and destruction of chaperone substrates is not yet understood. Two recently published crystal structures of mouse and zebrafish CHIP truncation constructs differ substantially, showing either an asymmetric assembly or a symmetric assembly with a highly ordered middle domain. To characterize the conformational properties of the intact full-length protein in solution, we performed amide hydrogen exchange mass spectrometry (HX-MS) with human CHIP. In addition, we monitored conformational changes in CHIP upon binding of Hsp70, Hsp90, and their respective C-terminal EEVD peptides, and in complex with the different E2 ubiquitin-conjugating enzymes UbcH5a and Ubc13. Solution HX-MS data suggest a symmetric dimer assembly with highly flexible parts in the middle domain contrasting both the asymmetric and the symmetric crystal structure. CHIP exhibited an extraordinary flexibility with a largely unprotected N-terminal TPR domain. Formation of a complex with intact Hsp70 and Hsp90 or their respective C-terminal octapeptides induced folding of the TPR domain to a defined, highly stabilized structure with protected amide hydrogens. Interaction of CHIP with two different E2 ubiquitin-conjugating enzymes, UbcH5a and Ubc13, had distinct effects on the conformational dynamics of CHIP, suggesting different roles of the CHIP-E2 interaction in the ubiquitination of substrates and interaction with chaperones.

Spatially and kinetically resolved changes in the conformational dynamics of the Hsp90 chaperone machine.

The highly conserved 90 kDa heat shock protein (Hsp90) chaperones use ATP to regulate the stability and activity of many signalling molecules like protein kinases and transcription factors. Studies using crystallography, electron microscopy and small-angle X-ray scattering yielded controversial results for the conformational states that these dimeric multidomain proteins assume while progressing through the ATPase cycle. To better understand the molecular mechanism of Hsp90 proteins, we studied the conformational dynamics of the Escherichia coli homologue HtpG in solution using amide hydrogen exchange mass spectrometry (HX-MS) and fluorescence spectroscopy. A conformation-sensitive fluorescent probe allowed to elucidate the ATPase cycle of HtpG. Continuous-labelling and pulse-labelling HX-MS experiments revealed major ATP-induced conformational changes throughout the protein that do not occur simultaneously, but progress surprisingly slow from the immediate nucleotide-binding site towards the N terminus and the middle domain. The conversion between the different conformational states is rate limiting for ATP hydrolysis, and the nucleotide-coordinating residue, Glu34, is important for the rate constant of conversion. Our findings, for the first time, allow to kinetically resolve changes in the conformational dynamics of individual structural elements of Hsp90.

Wernicke's syndrome after sleeve gastrectomy.

We report a case of Wernicke's encephalopathy after sleeve gastrectomy, which had been complicated by stomach wall edema and aggravated by dietary non-compliance. Despite intense parenteral nutrition, thiamine deficiency became clinically evident. It suggests that nutritional preparations used were unable to cover the increased thiamine requirement. After intense thiamine supplementation, gradual improvement occurred during the 6 months after the diagnosis, without permanent cognitive impairment. Clinicians involved in postoperative management of bariatric surgery patients must consider Wernicke's syndrome in hyper-emetic patients, who show unclear neurological deterioration. Early diagnosis and treatment can instantly improve the patient's condition without permanent sequelae.

The use of molecular markers in apple breeding for disease resistance.

Molecular markers have become a useful tool simplifying and speeding up breeding work. They are also helpful in the genetic analysis of complex agronomic traits. The investigations concern the use of already available SCAR markers for molecular analysis of breeding materials. The markers are the apple powdery mildew resistance gene Pl2, originating from Malus zumi, and the apple scab resistance gene Vf, from M. floribunda 821. The clone U 211 was found to be highly resistant to mildew under field conditions and transmitted a high level of resistance to the majority of its progeny. The presence of the Vf gene conferring resistance to scab was confirmed by molecular analysis. As the molecular markers for the Pl2 gene are not present in the DNA of U 211, it is probable that the clone U 211 is a new source of mildew resistance.