Fecal microbiota transplantation - new possibility to influence the results of therapy of cancer patients.

BACKGROUND: The intestinal microbio-me is essential for the function of the human body, it affects not only metabolism and digestion, but also the immune and neurobehavioral systems. The composition of the human intestinal microbio-me has been of interest to many scientific teams around the world in recent years, aided by the rapid development of molecular genetics methods. Intestinal microbio-me imbalance (so-called dysbio-sis) can help develop several pathological conditions such as autoimmune diseases or can be involved in the process of carcinogenesis. Microbio-me research in oncology has so far focused most on the effect of intestinal microbio-me composition on the effectiveness of checkpoint inhibitors. Differences in the relative proportions of individual bacterial strains and the overall microbio-me diversity in patients treated with checkpoint inhibitors appear to be related to the efficacy of this therapy. Many projects are currently studying the possibility of manipulating the composition of the intestinal microbio-me, especially by means of fecal microbial transplantation (FMT). Two published clinical studies have confirmed that it is possible to overcome resistance to checkpoint inhibitor therapy in malignant melanoma with this method and to re-establish a clinical response after FMT. One of the problems of this effort is the significant diversity in the composition of the microbio-me in different populations. Therefore, knowledge of the microbial composition in a particular population is of key importance. The Department of Oncology of the 1st Faculty of Medicine at Charles University and the General University Hospital in Prague is part of this effort, where a program to investigate intestinal microbio-me composition in patients with non-small cell lung cancer, renal cell carcinoma and malignant melanoma during checkpoint inhibitor therapy has been running for several years. PURPOSE: The aim of the publication is to demonstrate the current information and the importance of fecal transplantation in oncology and also to present our currently ongoing research project.

Future of lung cancer treatment.

The evolution of lung cancer treatment is an example of new perspectives in clinical oncology. Genomically determined targeted therapy of non-small cell lung cancer (NSCLC) is developing very rapidly with the gradual identification of new target structures and the concomitant development of innovative drugs are a great promise for the future. The historical development of systemic treatment of NSCLC is a model example of the path to accurate (precise) treatment. The innovation of the treatment has led to the shift from (non-targeted) cytostatic treatment to targeted therapy and immunotherapy. The targeted treatment and immunotherapy with checkpoint inhibitors have led to breakthrough prolongation of survival in patients with advanced NSCLC. According to a recent European Society for Medical Oncology (ESMO) recommendation, NSCLC is therefore one of the diagnoses where an examination using the next-generation sequencing panel should be performed as a standard.

The importance of interictal electroencephalography in paroxysmal states.

INTRODUCTION: Electroencephalography (EEG) is a non-invasive investigation method playing an important role in differential diagnostics of seizures. In this article authors point out to its importance, but also limitations. MATERIAL AND METHODS: Native interictal EEG findings were evaluated in inpatients after solitary unprovoked epileptic seizures (n=84), patients with sporadic epileptic seizures (n=179), patients with "chronic" epilepsy (n=324), outpatients with epilepsy (n=300), patients with syncope (n=100), patients with neurocardiogenic syncope (n=70), patients with migraine (n=100) and patients with tetanic syndrome (n=100). EEG findings were evaluated as normal or abnormal and abnormal findings were further divided into epileptic and non-epileptic, focal and generalized. RESULTS: In native EEG, epileptic manifestations were registered in 14.29 % of patients after solitary unprovoked epileptic seizures, in 25.7 % of patients with sporadic epileptic seizures, in 37.34 % of patients with chronic epilepsy and in 32 % of outpatients with epilepsy. Interictal EEG abnormalities (epileptiform and non-epileptiform) in non-epileptic diagnoses were at least registered in patients with syncope, but also in this group abnormal findings occurred in 30 % of them. We registered epileptiform abnormalities in 5 % of patients with migraine, in 4 % of patients with tetanic syndrome and in 2 % of patients with syncope. CONCLUSION: The diagnosis of epilepsy and non-epileptic seizures is a only a clinical diagnosis. EEG is a very important investigational method in this group of patients, but still only additional (Tab. 4, Fig. 2, Ref. 14).

Individual response of growing pigs to lysine intake.

Nitrogen (N) balance experiment was carried out to study the individual and group response of growing pigs to lysine (Lys) intake. A series of 15 purified diets limiting in Lys were fed sequentially to nine pigs (initial body weight, 39 kg) during a 15-day experiment. Lysine concentration ranged from 50% to 140% of its assumed requirement. N retention in individual pigs was related to Lys intake using linear-plateau (LP) and quadratic-plateau (QP) models. No difference in the closeness of fit between the models was found (p = 0.72). There were significant differences between the slopes of individual regression lines in the LP model (p = 0.018) and between the individual plateau values in both models (p < 0.0001). The breakpoint values in the QP model were greater than those of the LP model (p = 0.027), while the plateau values estimated by both models were similar (p = 0.32). In the LP model, no relationship was found between the slopes and plateau values (p = 0.67, r = 0.17) or between the slopes and breakpoint values (p = 0.55, r = -0.23), thus suggesting that pigs with higher protein deposition rate do not utilize lysine more efficiently. However, there was a close positive correlation between breakpoint and plateau values (p = 0.0005, r = 0.92). Based on LP and QP models, the mean daily requirement of Lys for a 47-kg gilt was estimated to be 20.8 and 24.2 g (0.79 and 0.92 g/MJ metabolizable energy), respectively, with coefficients of variation of approximately 10%. Marginal efficiency of Lys utilization derived from the LP model was 0.67 and was dependent on the range of input data selected. Lysine disappearance was a curvilinear function of Lys intake, indicating that Lys catabolism is not directly related to Lys intake. The closeness of fit of exponential, saturation kinetics or four-parameter logistic models applied to data set of all pigs was similar to that of LP or QP models.

Individual response of growing pigs to sulphur amino acid intake.

Two N balance experiments were conducted to study the individual response of growing pigs to limiting amino acid (AA) intake. Series of fifteen diets with increasing concentration of sulphur amino acids (SAA, Expt 1) or methionine in the presence of excess cystine (Expt 2) were fed sequentially to nine pigs during a 15-day experimental period. The concentration of the AA under test ranged from 50% to 140% of the requirement while other essential AA were given in a 25% excess relative to the limiting AA. N retention was related to the limiting AA intake using rectilinear and curvilinear models. In Expt 1, the quadratic-plateau model fitted the individual data significantly better (p = 0.01) than the linear-plateau model. No difference was found between the two models in Expt. 2, presumably due to the sparing effect of excess cystine on methionine utilization. Exponential, saturation kinetics or four-parameter logistic models fitted to data for all pigs showed that their goodness of fit was similar to those of quadratic-plateau or linear-plateau models. Significant differences (p < 0.05) were found between individual plateau values for N retention within each experiment while the slopes of the regression lines did not significantly differ either in Expt 1 (p = 0.07) or Expt 2 (p = 0.45). There was a positive correlation between the slope and plateau values of the linear-plateau model in Expt 1 (r = 0.74, p = 0.02) but no significant correlation was found in Expt 2 (r = -0.48, p = 0.13). Marginal efficiencies of SAA and methionine utilization derived from the linear-plateau model were 0.43 and 0.65 respectively. Based on linear-plateau and quadratic-plateau models, daily requirements of SAA and methionine for a 50 kg pig were estimated to be 13.0 and 5.9 g and 14.3 and 6.1 g respectively.

Efficiency of amino acid utilization in the growing pig at suboptimal levels of intake: branched-chain amino acids, histidine and phenylalanine + tyrosine.

Five balance experiments on growing pigs were carried out to study the marginal efficiency of utilization of isoleucine, leucine, valine, histidine and phenylalanine + tyrosine and to estimate their requirements for maintenance. Purified diets based on casein and crystalline amino acids as the sole source of N contained graded levels of each amino acid, corresponding to protein accretion rates of 0, 33, 66, 99 and 132 g/day, respectively. All other essential amino acids were given in a 30% excess. N retention increased linearly (p < 0.01) as the dietary concentration of the limiting amino acid increased. Based on linear regression equations relating amino acid deposition in body protein to amino acid intake, marginal efficiencies of ileal digestible amino acid utilization were calculated to be isoleucine - 0.81, leucine - 0.81, valine - 0.82, histidine - 1.17 and phenylalanine + tyrosine - 0.67. Extrapolating the regression equations to zero N retention, the daily requirements of amino acids for N equilibrium were estimated to be (mg/kg0.75) isoleucine - 18, leucine - 33, valine - 23, histidine - 14 and phenylalanine + tyrosine - 43.

Efficiency of amino acid utilization in the growing pig at suboptimal levels of intake: lysine, threonine, sulphur amino acids and tryptophan.

A series of N balance experiments using growing pigs was conducted to study the efficiency of utilization of lysine, threonine, sulphur amino acids and tryptophan and to estimate their maintenance requirements. Purified diets based on casein and crystalline amino acids as the sole source of N contained graded levels of each amino acid, corresponding to expected protein accretion rate of 0, 33, 66, 99 and 132 g/day, respectively. N retention increased linearly (p < 0.01) as the dietary concentration of the limiting amino acid increased. Based on linear regression equations relating amino acid deposition in body protein to amino acid intake, marginal efficiencies of ileal digestible amino acid utilization were calculated to be lysine 0.91, threonine 0.83, sulphur amino acids 0.85 and tryptophan 0.66. Extrapolating the regression equations to zero N retention, the daily requirements of amino acids for N equilibrium were estimated to be (mg/kg(0.75)) lysine 39, threonine 49, sulphur amino acids 46 and tryptophan 16.

Effect of nonessential amino acids on nitrogen retention in growing pigs fed on a protein-free diet supplemented with sulphur amino acids, threonine and tryptophan.

Two N balance experiments were conducted on growing pigs to study the effect of essential and nonessential amino acids added to a protein-free diet on N retention. In Expt. 1, the addition of sulphur amino acids, threonine and tryptophan to a protein-free diet at levels two times the maintenance requirements reduced (p > 0.1) daily N loss from -131 to -108 mg/kg(0.75). A further addition of nonessential amino acids equivalent to 250 mg N/kg(0.75)/d resulted in a marked increase (p < 0.01) in daily N retention to 28 mg/kg(0.75). In Expt. 2, nonessential amino acids were added to a protein-free diet supplemented with sulphur amino acids, threonine and tryptophan at levels corresponding to 100, 200 and 300 mg N/kg(0.75)/d. N retention increased linearly as dietary nonessential N increased. The slope of the best-fit regression line indicated that the marginal efficiency of nonessential N utilization for protein accretion was 0.26. The results suggest that nonessential amino acids may be a limiting factor for the re-utilization of amino acids released by body protein breakdown or that they may serve as precursors for de novo synthesis of amino acids by gut microorganisms, thus contributing to the amino acid requirements of the pig.

Bacterial inoculum enhances keratin degradation and biofilm formation in poultry compost.

Native microbial populations can degrade poultry waste, but the process can be hastened by using feather-degrading bacteria. Strains of Bacillus licheniformis and a Streptomyces sp. isolated from the plumage of wild birds were grown in a liquid basal medium and used to inoculate feathers in compost bioreaction vessels. Control vessels had only basal medium added to the feathers, litter and straw. Temperature, ammonia, carbon and nitrogen were monitored for 4 weeks. Scanning electron microscopy of the feather samples showed more complete keratin-degradation, more structural damage, and earlier microbial biofilm formation on inoculated feathers than on uninoculated feathers. A diverse community of aerobic bacteria and fungi were cultured early, but declined rapidly. Thermophilic B. licheniformis and Streptomyces spp. were abundant throughout. Enteric gram-negative bacteria, (e.g., Salmonella, E. coli) originally found on waste feathers were not recovered after day 4. Vessel temperatures reached 64-71 degrees C within 36 h and stabilized at 50 degrees C. When tumble-mixed at day 14, renewed activity peaked at 59 degrees C and quickly dropped as available carbon was used. Feathers soaked in an inoculum of B. licheniformis and Streptomyces degraded more quickly and more completely than feathers that were not presoaked. Inoculation of feather waste could improve composting of the large volume of feather waste generated every year by poultry farms and processing plants.