Convenient broad-host-range unstable vectors for studying stabilization cassettes in diverse bacteria

Background: Low-copy-number vectors of potential wide application in biotechnology need to encode stabilization modules ensuring their stable inheritance. The efficiency of stabilization may vary depending on the plasmid host so a thorough analysis of stabilization functions is required before use. Results: To facilitate such analysis highly unstable, mobilizable, broad-host-range (BHR) vectors based on RK2 replicon were constructed. The vectors are suitable for testing of various stabilization functions, including plasmid and chromosomal partitioning cassettes encoding ParB homologues capable of spreading on DNA. The xylE or lacZ reporter systems facilitate easy monitoring of plasmid segregation. Conclusion: The range of BHR vectors with different reporter cassettes and alternative mobilization systems expands their application in diverse bacterial species

Effects of genistein on insulin pathway-related genes in mouse differentiated myoblast C2C12 cell line: evidence for two independent modes of action

Introduction. Genistein (plant isoflavone) is a well-known anti-cancer drug with estrogenic-like properties. Genistein also regulates sugar and lipid metabolism; thus, it has anti-diabetic properties. The aim of the study was to evaluate in vitro effects of genistein on glucose transport, fatty acids oxidation, activation of PKB, and expression of genes related to insulin pathway in differentiated myoblast C2C12 mouse cell line. Material and methods. Differentiated myoblast C2C12 mouse cell line was used to assess the effects of different genistein concentrations on glucose transport and fatty acids oxidation measured by radioactivity technique, activation of PKB, and expression of selected genes related to insulin signaling pathway (IR-a, IR-b, IRS-1, PKB, GLUT-4, PP2A, SH-PTP2) at the mRNA and protein levels. Cells were incubated with various concentrations of genistein under standard conditions for 0–48 hours.Results. Genistein in low concentrations (0.1–1 μM) significantly increased glucose transport and decreased fatty acids oxidation in C2C12 cells after 48 h of incubation. High concentration of genistein (50 μM) had the opposite effect. Genistein stimulated PKB phosphorylation during the first 5–10 minutes of incubation. There was no significant impact on the protein expression of selected genes (IR-a, IR-b, IRS-1, PKB, GLUT-4, PP2A-Ca, ER-a and ER-b) after 48 h treatment. We observed inverse correlation between genistein concentration and the expression of SH-PTP2 protein. Genistein affected the expression pattern of mRNAs for genes related to the insulin pathway, however, not the expression of the encoded proteins. Conclusions. The results of this study showed that depending on the concentration and time of incubation genistein significantly affects glucose and lipid metabolism and at low concentration modifies expression patternof a few genes in C2C12 cells

Pitted keratolysis – a frequently misdiagnosed, mild, infectious disorder of soles

Introduction . Pitted keratolysis (PK) is a mild infectious skin disorder caused by Corynebacterium spp., Kytococcus sedentarius or Dermatophilus congolensis . These bacteria produce enzymes that digest keratin, causing superficial lesions in the plantar surface. The disease is predominantly observed in young men. Objective . Pitted keratolysis despite the characteristic presentation of skin lesions is often misdiagnosed. In this article we aimed to remind readers of its clinical aspects and treatment by presenting a typical PK case. Case report. A 35-year-old man was admitted to the dermatological clinic due to skin lesions on both soles. In the physical examination we found multiple crateriform pits, associated with hyperhidrosis and malodour diagnosed as PK. Remission of lesions was observed after treatment with oral erythromycin. Conclusions . The differential diagnosis of plantar skin lesions should include PK. Due to typical clinical manifestation the diagnosis is based on physical examination

Intense pustular reaction during treatment of actinic keratosis with ingenol mebutate

Introduction. Actinic keratosis is a common diagnosis in dermatological practice. Lesions usually appear on senile skin or skin exposed to sunlight. As they can progress to squamous cell carcinoma, all lesions should be properly treated. Ingenol mebutate is one of the newest topical agents. Objective. To present a severe site reaction during treatment of actinic keratosis with ingenol mebutate gel. Case report. An 85-year-old patient was treated with ingenol mebutate for an actinic keratosis lesion on the head. After the third day of treatment a massive local reaction was observed, with numerous pustular lesions. It was treated with a topical antibacterial agent. Complete remission of actinic keratosis was observed one month later. Conclusions. Treatment of actinic keratosis with ingenol mebutate carries the risk of local adverse effects, including severe pustular reaction. Patients should always be informed about possible side effects of the treatment and their transient nature

Influence of protein deficient diet, vitamin B₂ supplementation and physical training on serum composition of polyunsaturated fatty acids (PUFAs) in rats

Introduction Prolonged shortages of protein in the diet significantly alter the composition and content of polyunsaturated fatty acids (PUFA) in tissues and body fluids. One of nutritional factors which may reduce negative effects of protein malnutrition might be vitamin B2 due to its influence on lipids metabolism. Objective The aim of the study was to investigate the influence of low protein (LP) diet enriched with vitamin B2 on the content and composition of PUFA in the blood serum of rats treated with dosed physical exercise. Material and Methods The experiment was carried out for 3 months on 72 growing male Wistar rats divided into 5 groups. Animals were fed ad libitum on a diet with an energy value of 350 kcal/100 g, in which 4.5% of the energy was provided by protein. In the control diet, 20% of the energy was provided by protein. Two groups were fed the diet enriched with vitamin B2. The two groups of tested animals were trained for 5 days a week. Results LP diet caused a decrease in α-linolenic acid (ALA) after 30 days, and a decrease in docosahexaenoic acid (DHA) after 60 days of experiment, compared with rats fed the control diet. After 60 and 90 days of the experiment, a significant decrease was noted in arachidonic acid (AA) in serum of trained rats, compared with sedentary rats fed the LP diet. Physical activity increased LA (mainly on day 30), EPA (on day 90) and reduced AA content (on day 90) in serum of rats fed the LP diet. B2 supplementation in the trained LP group did not change the EPA and AA dependence; however, there was a decrease in LA content in comparison to the non-supplemented trained group. Conclusions Results of this study suggest that all investigated factors (protein deficiency, physical exercise and supplementation of vitamin B2) have significant impact on PUFA composition of serum in rats

Influence of protein deficient diet, vitamin B₂ supplementation and physical training on serum composition of polyunsaturated fatty acids (PUFAs) in rats

Introduction Prolonged shortages of protein in the diet significantly alter the composition and content of polyunsaturated fatty acids (PUFA) in tissues and body fluids. One of nutritional factors which may reduce negative effects of protein malnutrition might be vitamin B2 due to its influence on lipids metabolism. Objective The aim of the study was to investigate the influence of low protein (LP) diet enriched with vitamin B2 on the content and composition of PUFA in the blood serum of rats treated with dosed physical exercise. Material and Methods The experiment was carried out for 3 months on 72 growing male Wistar rats divided into 5 groups. Animals were fed ad libitum on a diet with an energy value of 350 kcal/100 g, in which 4.5% of the energy was provided by protein. In the control diet, 20% of the energy was provided by protein. Two groups were fed the diet enriched with vitamin B2. The two groups of tested animals were trained for 5 days a week. Results LP diet caused a decrease in α-linolenic acid (ALA) after 30 days, and a decrease in docosahexaenoic acid (DHA) after 60 days of experiment, compared with rats fed the control diet. After 60 and 90 days of the experiment, a significant decrease was noted in arachidonic acid (AA) in serum of trained rats, compared with sedentary rats fed the LP diet. Physical activity increased LA (mainly on day 30), EPA (on day 90) and reduced AA content (on day 90) in serum of rats fed the LP diet. B2 supplementation in the trained LP group did not change the EPA and AA dependence; however, there was a decrease in LA content in comparison to the non-supplemented trained group. Conclusions Results of this study suggest that all investigated factors (protein deficiency, physical exercise and supplementation of vitamin B2) have significant impact on PUFA composition of serum in rats

Effect of Vitamin D Treatment on Dynamics of Stones Formation in the Urinary Tract and Bone Density in Children with Idiopathic Hypercalciuria

Vitamin D supplementation in patients with urolithiasis and hypercalciuria is considered to be unsafe. We analyzed the impact of vitamin D supplementation on selected health status parameters in children with idiopathic hypercalciuria. The study included 36 children with urolithiasis resulting from excessive calcium excretion. The level of calcium and 25(OH)D (hydroxylated vitamin D - calcidiol) in serum, urinary calcium excretion and the presence of stones in urinary tract were assessed prospectively. Blood and urine samples were collected at the time when the patient was qualified for the study and every three months up to 24 month of vitamin D intake at a dose of 400 or 800 IU/day. At time zero and at 12, and 24 months of vitamin D supplementation, densitometry was performed. Supplementation with vitamin D caused a statistically significant increase in the concentration of 25(OH)D in serum. There were no significant changes in calcium concentration in serum, excretion of calcium in urine but also in bone density. There was no significant increase in the risk of formation or development of stones in the urinary tract. Supplementation with vitamin D (400–800 IU/day) in children with idiopathic hypercalciuria significantly increases 25(OH)D concentration, does not affect calciuria, but also does not improve bone density

Different effects of feeding pregnant and lactating mice Rhodiola kirilowii aqueous and hydro-alcoholic extracts on their serum angiogenic activity and content of selected polyphenols

Angiogenesis plays an important role in many physiological processes, among them the formation of tissues and organs during embryogenesis. A lot of medicinal plants exhibit angiomodulatory properties. This creates the need for a thorough check of whether the plant extracts that we would like to give to pregnant women in order to increase their resistance to bacterial or viral infection will have negative effects on angiogenesis, and consequently on fetal development. This paper seeks to investigate the effect of serum of pregnant and nursing Balb/c mice that received aqueous (RKW) or hydro-alcoholic (RKW-A) R. kirilowii extracts (20 mg/kg), or epigallocatechin (0.2 mg/kg), on the in vitro proliferation and migration of mouse endothelial cell line HECa10. Of the 15 identified polyphenols in the extracts by HPLC, 8 were present in the sera. Chemical analysis revealed higher salidroside, kaempferol, chlorogenic acid, bFGF and VEGF concentration in RKW-A sera than in the sera of RKW group of mice. RKW-A and EGC sera did not affect migration of endothelial cells, however we noted some increase of migrating cells after RKW-sera treatment. RKW and EGC sera did not affect proliferation of endothelial cells. Sera of mothers from RKW-A group impaired the proliferation of endothelial cells in comparison to other groups. These data allow us to assume that Rhodiola kirilowii hydro-alcoholic extract (RKW-A) is potentially able to modulate pre- and post- natal angiogenesis what might influence the development of organs in progeny. Sera of RKW mothers have not harm the proliferation of endothelial cells, despite they also contain antiangiogenic catechins and salidroside. This suggests the existence in RKW-A extract and in RKW-A sera of some other, as yet unidentified substances influencing endothelial cells proliferation

The long-term effect of a protein-deficient-diet enriched with vitamin B6 on the blood parameters in unexercised and exercised rats

International audienceProtein undernutrition affects inter alia blood parameters and immune system. These negative effects may be improved by the addition of vitamin B6 to the diet. Here, we evaluated the effect of vitamin B6 supplementation and physical exercise in rats fed a protein-deficient diet. Rats were divided into six groups exercised or unexercised-control, protein deficient, protein deficient with vitamin B6 supplementation. Sixty days of protein malnutrition caused significant changes in the rat body weight, haematological parameters (mainly red blood cells parameters), immunological parameters (NK and NKT cells) and biochemical parameters (total protein and albumin concentration and activity of AST). The rat exercise did not intensify the negative effects of protein malnutrition. The vitamin B6 supplementation in protein-deficient groups significantly improved body weight and red blood cell parameters. These results indicate that vitamin B6 should be considered as a valuable supplement for individuals suffering from protein malnutrition