Functional metagenomic analysis of carbohydrate degrading enzymes from the human gut microbiota

Abstract The gut microbiota is a complex and diverse microbial community that is adapted to a carbohydrate-rich ecosystem. Plant cell wall components (cellulose, hemicelluloses and pectins), resistant starch and various oligosaccharides reach the colon by escaping digestion in the upper gastrointestinal tract. Fermentation of these dietary carbohydrates by the gut microbiota has well-recognised beneficial effects on host health. The microbial community in the human gut requires specific enzymes to efficiently degrade these carbohydrates. In this project, a culture-independent approach based on functional screening of genomic and metagenomic libraries using Escherichia coli and Lactococcus lactis as heterologous expression hosts, was used to isolate novel genes encoding glycoside hydrolase (GH) enzymes. The study identified several active GH enzymes involved in the breakdown of dietary polysaccharides such as starch, cellulose, xylan and β-glucan, recovered from the E. coli metagenomic library. The bioinformatic analysis of the insert from positive clones showed the presence of ORFs with the similarity to enzymes from GH families 13, 43 and 51 encoded by dominant bacterial genera from the human colon (Bacteroides sp., Roseburia sp., Ruminococcus sp.). A group of clones encoding potentially novel GH enzymes was also identified, emphasising the importance of functional-based study. One highly active clone was detected during screening of the L. lactis metagenomic library and showed fibrolytic activity on cellulose-, lichenanand xylan-containing plates. The insert contained a partial gene with the GH9 catalytic domain and identity to the protein from Coprococcus eutactus ART55/1. Further functional analysis established the fibrolytic activity of selected Coprococcus species. Moreover, several active clones were isolated from the Ruminococcus sp. 80/3 genomic library which encoded protein with the similarity to enzymes from GH families 2, 3 and 5. In this work, the traditional approach of expression in E. coli was complemented by using an alternative host – L. lactis. While this did not improve the screening efficiency in terms of number of recovered clones, differences in gene expression and protein export between E. coli and L. lactis were noted during this study which highlights the benefits of using different heterologous hosts in functional metagenomic approaches

The reaction of the WIG stock market index to changes in the interest rates on bank deposits

Determination of the relationship between the money market and capital market is particularly important from the point of view of taking a decision on the location of investment capital. It may help to forecast future states. This study seeks to determine the relationship of the interest rate on deposits in zloty with the WIG stock index and the volume of turnover on the Warsaw Stock Exchange. Analysis of correlation and VAR models are used. Analysis of long-term correlation indicates a negative relationship between the interest rate on deposits in banks and the value of the WIG stock-index. However, this may be spurious. The dependence between these variables may be more complex and should rather be seen as short term. It seems that in general the impact of an increase in interest rates on the value of the WIG index is negative in the short term, just as in the long term. In addition, in the short term these variables can move in the same direction. The results obtained in the research are consistent with results obtained for other national markets. This applies in particular to the relatively weak, negative correlation described above.interest rate on deposits, WIG index, correlation, causality

Nursing care of a patient in terminally ill cancer - case study

The aim was to show the nurses and hospice care as a member of the team over the patient in a hospice for terminally ill cancer resident inthe house. Nurse choosing to work with patients who die must know that not only medical knowledge will be needed in carrying out its duties but alsoa lot of love, patience and above all humility toward life and death. Characterized range of palliative care and its residents. The paper presents adescription and policies for hospices and home desktops and volunteers working in them. Material - the subject of research - the case of a patient withpancreatic cancer he cultivated to the last moments of life. Test method - method of individual cases, which applies to nursing, is a qualitativedescription of the passage of reality and its interpretation. Used to collect all kinds of information about the case, in particular about the familysituation, social, environmental, material and existential, psychological, physical or other person examined. This is a descriptive study undertaken bytype of research problem, which requires the use of methods relating to a single case or small their number.Research problems:1. How long the patient has symptoms associated with cancer diagnosed?2. Have there been cases of death from cancer in the family sick?3. Have you previously was treated oncological?4. When the first signs of disease?5. Does the patient know about his condition?6. Are they symptomatic treatment introduced hospice doctor brings relief from symptoms?7. Is recommended to take the product (Megace) improves the appetite, gives the desired effect?8. Is your oxygen therapy brought relief in breathing?9. How effective were notmorphotic painkillers?10.Do you sick of the constant care exercised a family?11. Is the hospice team visits took place at regular intervals or whenever the call of the family?12. Is the patient dying in dignity, surrounded by kin?The goal of the research work presented, a description of the patient (the subject of research), research problems and the method ofresearch, technology and research tools, describes the results of research. Presented in a special way and meaningful roles of a nurse who has thecare of the patient in the terminal stage of cancer, staying at home. The problem of mental, spiritual, and emotional problems of the patient and theenvironment. Based on the literature of the subject and based on scientific study, the following conclusions:1. In view of the cancer patient who is not eligible for treatment at the forefront the problem of cancer pain and other ailments of thedigestive and respiratory systems, and greatly reduced physical fitness.2. The patient was the biggest problem was lack of appetite and emaciation of the body related to it which greatly limited its self-care, andin a later stage, prevented completely.3. Above the patient in the terminal who will remain in the home should be planned nursing care consulting and communicating withmembers of her family, educating them in implementing the active care, care and assist.4. The nurse takes action on dying nursing care aimed at improving the quality of life, eliminating pain, psychological comfort, eliminationof other ailments associated with the disease, assist

Conditions of agriculture compared to economic growth in selected countries

Purpose: In the past years, agriculture has been undergoing large transformations. It has become more modern, but its share in the GDP growth has been diminishing. The question of the connection between the condition on the agriculture market and the general economic condition seems fundamental. Design/Methodology/Approach: Based on the added value of agriculture and the Gross Domestic Product in 1992-2017 in the United States, Great Britain, France, and Poland, the connection was determined between these variables. Correlative and cointegration research was carried out. Findings: Based on the research results, conclusions may be drawn about a poorer and less stable increase in agriculture in Europe as compared to the United States. Practical Implications: Grounds for the statement were found that disturbances in agriculture may have a negative impact on the entire economy. This is particularly evident in Europe. Hence, it is recommended to examine the current policy of the EU. Originality/value: Research shows how important the implications of a single branch of the economy are for the entire economy. Moreover, it provides grounds for remodeling EU policy towards market deregulation following the US pattern.peer-reviewe

Living with diabetes - case study

Introduction. Diabetes is a chronic disorder of metabolism. It is caused by impaired insulin secretion and action. There are type 1 diabetes,type 2 diabetes, gestational diabetes and other specific types of diabetes. Risk factors for this disease include age (risk increases after 45 years ofage), obesity, low physical activity, hypertension, lipid disorders genetics. Symptoms of the disease are frequent urination, excessive thirst, weakness,lethargy, emaciation, fungal infections, furunculosis. Patients have elevated blood glucose levels. Complications of diabetes can be divided into acuteand chronic. Complications include acute hyperglycemia and hypoglycemia. Chronic complications is microangiopathy (retinopathy, nephropathy,neuropathy) and macrovascular disease (myocardial infarction, stroke, peripheral vascular disease). Diabetes occurs more frequently, is an importantsociological and social problem. Her diagnosis always causes anxiety, uncertainty, sense of danger. Ill try to find answers to many questions.Objective. The aim of this study was to determine the quality of life for patients with diabetes, diagnosis of care problems, patienteducation in terms of information essential for living with chronic disease.Materials. Material - the subject of research was a woman aged 64 suffering from diabetes type II. Patient presented information aboutdiabetes, we got to know her health problems and their solutions.Methods. A Study of the individual case. Techniques: Observation, interview, examination of documents.Research problems posed in the work are:What are the health problems common in patients with diabetes?The extent to which a patient with diabetes is capable of self-care and self-control?As the nurse prepares the patient to change and lifestyle modification in chronic disease that is diabetes?The paper presents a range of activities including patient self-care and self-control.Results. The patient reported the following health problems relating to diabetes: the lack of insulin, inability to use the meter, the lack ofsufficient knowledge about the complications of diabetes, skin abrasion on the foot and clammy skin between the toes, being overweight.Conclusion. Patient is a walking, self-contained. All nursery operations performed alone. With the support of her daughter is trying toovercome all the difficulties associated with the disease. She uses medical care as a medical indication, every 6-8 weeks in primary exercise controlstudy of blood glucose levels, use of medical visits, always ordered the hypoglycemic drugs, insulin, diabetic diet, was informed by a nurse for furthercare and self-control. The paper presents a range of activities including patient self-care and self-control.The patient was informed of the need for lifestyle changes such as diet, avoidance of drugs, the constant control of blood glucose(glucometer training will be offered), a small regular exercise, etc.First of all making clear to patient that diabetes is not cited, you can live with this disease, do not fall apart. The proper operation oftreatment with a conscious discipline of the patient, can maintain good performance in life and personal life satisfaction.She learned to use the same meter and examine the level of blood sugar at a specified time. She is aware of the problems associated withthe disease. He is aware that only applies to the stringent requirements and restrictions, will allow it to avoid future complications. Despite anincurable disease is trying to discern the meaning of life and the meaning of their sacrifices. He believes in the fact that a positive attitude to thedisease is part of the success on the road to quality of life

Cdc42 protein acts upstream of IQGAP1 and regulates cytokinesis in mouse oocytes and embryos

AbstractCdc42 and Rac1 Rho family GTPases, and their interacting protein IQGAP1 are the key regulators of cell polarity. We examined the role of Cdc42 and IQGAP1 in establishing the polarity of mouse oocyte and regulation of meiotic and mitotic divisions. We showed that Cdc42 was localized on the microtubules of meiotic and mitotic spindle and in the cortex of mouse oocytes and cleaving embryos. IQGAP1 was present in the cytoplasm and cortex of growing and fully-grown oocytes. During maturation it disappeared from the cortex and during meiotic and mitotic cytokinesis it concentrated in the contractile ring. Toxin B inhibition of the binding activity of Cdc42 changed the localization of IQGAP1, inhibited emission of the first polar body, and caused disappearance of the cortical actin without affecting the migration of meiotic spindle. This indicates, that in maturing oocytes accumulation of cortical actin is not indispensable for spindle migration. In zygotes treated with toxin B actin cytoskeleton was rearranged and the first and/or subsequent cytokinesis were inhibited. Our results indicate that Cdc42 acts upstream of IQGAP1 and is involved in regulation of cytokinesis in mouse oocytes and cleaving embryos, rather than in establishing the polarity of the oocyte

Shifts in Female Facial Attractiveness during Pregnancy

It has been proposed that women’s physical attractiveness is a cue to temporal changes in fertility. If this is the case, we should observe shifts in attractiveness during pregnancy—a unique physiological state of temporal infertility. The aim of this study was to examine how women’s facial attractiveness changes during the subsequent trimesters of pregnancy and how it compares to that of nonpregnant women. Sixty-six pictures of pregnant women (22 pictures per trimester) and 22 of nonpregnant women (a control group) were used to generate four composite portraits, which were subsequently assessed for facial attractiveness by 117 heterosexual men. The results show considerable differences between facial attractiveness ratings depending on the status and progress of pregnancy. Nonpregnant women were perceived as the most attractive, and the attractiveness scores of pregnant women decreased throughout the course of pregnancy. Our findings show that facial attractiveness can be influenced by pregnancy and that gestation, even at its early stages, affects facial attractiveness. Considerable changes in women’s physiology that occur during pregnancy may be responsible for the observed effects

Mutational analysis in podocin-associated hereditary nephrotic syndrome in Polish patients: founder effect in the Kashubian population

Hereditary nephrotic syndrome is caused by mutations in a number of different genes, the most common being NPHS2. The aim of the study was to identify the spectrum of NPHS2 mutations in Polish patients with the disease. A total of 141 children with steroid-resistant nephrotic syndrome (SRNS) were enrolled in the study. Mutational analysis included the entire coding sequence and intron boundaries of the NPHS2 gene. Restriction fragment length polymorphism (RFLP) and TaqMan genotyping assay were applied to detect selected NPHS2 sequence variants in 575 population-matched controls. Twenty patients (14 %) had homozygous or compound heterozygous NPHS2 mutations, the most frequent being c.1032delT found in 11 children and p.R138Q found in four patients. Carriers of the c.1032delT allele were exclusively found in the Pomeranian (Kashubian) region, suggesting a founder effect origin. The 14 % NPHS2 gene mutation detection rate is similar to that observed in other populations. The heterogeneity of mutations detected in the studied group confirms the requirement of genetic testing the entire NPHS2 coding sequence in Polish patients, with the exception of Kashubs, who should be initially screened for the c.1032delT deletion

Ten-year trends in epidemiology and outcomes of pediatric kidney replacement therapy in Europe : data from the ESPN/ERA-EDTA Registry

Background For 10 consecutive years, the ESPN/ERA-EDTA Registry has included data on children with stage 5 chronic kidney disease (CKD 5) receiving kidney replacement therapy (KRT) in Europe. We examined trends in incidence and prevalence of KRT and patient survival. Methods We included all children agedPeer reviewe

Genetic Drivers of Kidney Defects in the DiGeorge Syndrome

Background The DiGeorge syndrome, the most common of the microdeletion syndromes, affects multiple organs, including the heart, the nervous system, and the kidney. It is caused by deletions on chromosome 22q11.2; the genetic driver of the kidney defects is unknown. Methods We conducted a genomewide search for structural variants in two cohorts: 2080 patients with congenital kidney and urinary tract anomalies and 22,094 controls. We performed exome and targeted resequencing in samples obtained from 586 additional patients with congenital kidney anomalies. We also carried out functional studies using zebrafish and mice. Results We identified heterozygous deletions of 22q11.2 in 1.1% of the patients with congenital kidney anomalies and in 0.01% of population controls (odds ratio, 81.5; P=4.5×10(-14)). We localized the main drivers of renal disease in the DiGeorge syndrome to a 370-kb region containing nine genes. In zebrafish embryos, an induced loss of function in snap29, aifm3, and crkl resulted in renal defects; the loss of crkl alone was sufficient to induce defects. Five of 586 patients with congenital urinary anomalies had newly identified, heterozygous protein-altering variants, including a premature termination codon, in CRKL. The inactivation of Crkl in the mouse model induced developmental defects similar to those observed in patients with congenital urinary anomalies. Conclusions We identified a recurrent 370-kb deletion at the 22q11.2 locus as a driver of kidney defects in the DiGeorge syndrome and in sporadic congenital kidney and urinary tract anomalies. Of the nine genes at this locus, SNAP29, AIFM3, and CRKL appear to be critical to the phenotype, with haploinsufficiency of CRKL emerging as the main genetic driver. (Funded by the National Institutes of Health and others.)