Pavlovian backward conditioned inhibition in humans: summation and retardation tests

Two experiments using human participants investigated whether a Pavlovian backward inhibitory treatment (nonreinforced trials in phase 1 followed by reinforced trials in phase 2; i.e., AX- followed by A+) produces a stimulus which can pass summation and retardation tests for inhibition. The rationale for conducting these experiments was that previous demonstrations of Pavlovian backward inhibition informed participants about the nature of the outcome before starting the experiment. According to some theoretical views, this is a potential confound. In the present experiments we used a predictive task in which participants had no knowledge about the outcome until phase 2, when reinforcement occurred. The results of Experiment 1 (summation test) and Experiment 2 (retardation test) provide a clear demonstration of backward conditioned inhibition.     

Oligosaccharide transfer from lipid sugar intermediates to endogenous protein(s) of rat liver microsomal subfractions

Summary The subcellular localization and characterization of some of the components involved in the glycosylation of asparagine type glycoproteins was attempted using dolichyl diphosphate [¹⁴c]mannose oligosaccharide as precursor of the glycosylation reaction in vitro. Isolated rough and smooth microsomel fractions were able to carry out the transfer of the carbohydrate moiety from lipid oligosaccharide to endogenous protein acceptors. The protein glycosylating activity remained practically the same after stripping the vesicles from their ribosomes or partially releasing their vesicular content. Isolation of polysomes from rough microsomes after glycosylation has taken place, reveals that a large proportion of mannose labeled glycoproteins is in the membranous fraction. The remaining labeled glycoproteins co-sediment with the polysomal fraction. If the isolation is carried out before glycosylation only the membranous fraction shows enzyme activity, whereas the polysomes alone are not able to carry out glycosylation. All these results taken together indicate that the protein glycosylating enzyme is a structural component of the rough and smooth microsomes of rat liver.     

Preniche as missing link of the metastatic niche concept explaining organ-preferential metastasis of malignant tumors and the type of metastatic disease

Here we attempt to supplement the metastatic niche concept with a stage of “preniche” that determines the site of development of a premetastatic niche and of a subsequent metastasis. The “preniche” includes all cellular and molecular events in the site of a prospective metastasis preceding the entrance of myeloid progenitor cells. The “preniche” integrates an activation of vascular endothelium of the microcirculatory vessels of target organs in the site of a future metastasis under conditions of chronic persistent productive inflammation that can be induced by cytokines from the primary tumor and independently of it. The endothelium activation is responsible for adhesion and clustering of the recruited myeloid progenitor cells and also for the retention of cells of malignant tumors. The preniche easily arises in organs enriched with organspecific macrophages (lungs, liver, brain, etc.) where the endothelium is predisposed for intensive recruiting of myeloid progenitor cells of macrophages, especially under conditions of inflammation. Introduction of the “preniche” concept allows us to avoid difficulties associated with the development of the metastatic niche concept, especially concerning the problem of organ-preferential localization of metastases, and to make some predictions for experimental verification and potential approaches for preventing metastasizing in some oncologic patients.     

Vaginal lactic acid bacteria in the mare: evaluation of the probiotic potential of native Lactobacillus spp. and Enterococcus spp. strains

Lactic acid bacteria (LAB) are important members of the human vaginal microbiota and their presence is considered beneficial. However, little is known about native vaginal bacteria in other animal species such as the horse. The aim of this work was to quantify the vaginal lactic acid bacteria and lactobacilli of mares and to establish if selected equine vaginal lactic acid bacteria, particularly Lactobacillus and Enterococcus spp. strains, could exhibit potential as probiotics. The vaginal lactic acid bacteria and lactobacilli of 26 mares were quantified by plate counts. Five strains (three Lactobacillus spp. and two Enterococcus spp.) were characterised and adhesion to vaginal epithelial cells, antimicrobial activity and ability to form biofilms were evaluated. Lactic acid bacteria were recovered from the 26 samples and lactobacilli counts were detected in 18 out of 26 mares (69%). Probiotic properties tested in this study varied among the isolates and showed promising features for their use as equine probiotics.     

In vitro activity of potential probiotic Lactobacillus murinus isolated from the dog

Aims:  The aim of this study was to isolate and identify Lactobacillus spp. isolates from faeces of a healthy dog, and to characterize their potential as probiotics in order to evaluate their possible use as probiotics for dogs.
Methods and Results:  An in vitro approach was used to characterize the isolates as potential probiotics including the evaluation of pH and bile salts tolerance, production of antimicrobial substances, biofilm formation on glass and polystyrene surfaces, aggregation ability and adhesion to canine intestinal mucus. The isolates survived to different pH and bile salts conditions, inhibited the in vitro growth of Escherichia coli and Clostridium perfringens , and adhered to glass and intestinal mucus.
Conclusions:  The properties shown by these isolates may indicate that they could colonize and persist in the gastrointestinal tract and induce beneficial effects to the host.
Significance and Impact of Study:  The evaluation of native canine isolates and future exprimental feeding assays may be useful tools to develop probiotics to improve animal health and reduce the risk of gastrointestinal disorders.     

Safety assessment and enteric colonization ability of a native canine Lactobacillus murinus strain

The aim of this work was to assess the safety and survival in the canine gastrointestinal tract of a native Lactobacillus murinus strain using an in vivo approach. A spontaneous rifampicin-resistant strain generated from a native Lactobacillus murinus strain isolated from dog feces was used in an in vivo administration protocol performed in dogs. The parental strain had been previously characterized in our laboratory, showing interesting probiotic-related properties. The rifampicin-resistant mutant generally exhibited similar properties to the parental strain. When it was administered to healthy dogs, this strain was able to survive in the canine gut and diverse physiological parameters of the animals did not differ from those assessed in non-treated ones. The present in vivo study using a native canine Lactobacillus murinus strain allowed us to conclude that the assessed strain transiently persisted in the canine gut and is safe for administration in dogs. Results of this assay can contribute to the development of new strategies for health promotion in dogs. Future studies will be carried out to evaluate the potential probiotic strain in vivo in dogs under different clinical conditions.     

Evaluation of native potential probiotic bacteria using an in vitro ruminal fermentation system

Rumen microbiota provides an important source of protein to grazing animals and produces volatile fatty acids (VFA), the main energy source for ruminants generated by fibre fermentation. Probiotics can be used to modulate rumen fermentation, and native microbiota is a source of potentially useful microorganisms. In this work, ruminal bacterial strains were isolated and subsequently identified, and their potential to modify fermentation patterns with wheat straw, microcrystalline cellulose and oat xylan as substrates was assessed by in vitro gas production and VFA fermentation patterns. Four of the isolates were identified as Pseudobutyrivibrio ruminis and two corresponded to new members of the Lachnospiraceae family. The addition of one P. ruminis (strain 50C) and one Lachnospiraceae (strain 21C) to the fermentation system which used wheat straw as the substrate significantly increased total VFA concentration without altering the total gas produced in one case and showed a decrease in total gas production in the other. All bacterial strains induced higher butyric acid concentrations with the three substrates (up to 31 mM in the case of Lachnospiraceae 21C incubated with oat xylan and 25 mM in microcrystalline cellulose fermenters to which P. ruminis 50C had been added) compared to the control, which had concentrations of <1 mM. Analysis of the fermentation products suggested that the addition of probiotics to the fermentation system had the potential to induce metabolic shifts that would result in better energy yields. These results show that native bacteria have promising features as fermentation modulators, thereby justifying further research to assess their use as probiotics for ruminants.     

Collateral presentation of antigens as physiological prototype for lymph node metastases

The formation of lymphogenic metastases remains enigmatic. In particular, the much more pronounced predilection of carcinomas than of sarcomas to metastasizing into regional lymph nodes is an unsolved problem. We suggest that this difference could be due to the ability of epitheliocytes for a hypothetical process termed by us “collateral presentation of antigens”. Under conditions of infection of epithelium with intracellular pathogens or during inflammation, epithelial cells acquire a special receptor phenotype, undergo the epithelial-mesenchymal transition, and migrate along lymphatic vessels into lymph nodes where they present antigen to immunocytes. The collateral presentation of antigens can be of significant biological importance in the case of insufficient classical pathway of antigen presentation (by dendritic cells) or on disturbance in the death mechanisms of the infected cells. Depending on conditions of induction of the epithelial-mesenchymal transition and on possible ability of epitheliocytes to express MHC II with co-stimulating molecules, two pathways, “container-mediated” and “MHC II-dependent”, of antigen presentation in lymph nodes resulting in development of immunogenesis or anergy of immunocytes are supposed to exist. All pathways of delivery of the epithelial cells into lymph nodes and of antigen presentation by epitheliocytes terminate by death of these cells. The lymphogenic metastasizing realizes the same mechanism under conditions of tumor disease; however, this is not associated with cell death, but they actively colonize the lymph node. The proposed hypothesis allows us to explain the metastasizing of sarcomas into lymph nodes. The main prerequisite for lymphogenic metastasizing seems to be related with the mesenchymal-epithelial transition of sarcoma cells promoting their involvement in the presentation of antigens.     

Types of immune-inflammatory responses as a reflection of cell–cell interactions under conditions of tissue regeneration and tumor growth

Inflammatory infiltration of tumor stroma is an integral reflection of reactions that develop in response to any damage to tumor cells including immune responses to antigens or necrosis caused by vascular disorders. In this review, we use the term “immune-inflammatory response” (IIR) that allows us to give an integral assessment of the cellular composition of the tumor microenvironment. Two main types of IIRs are discussed: type 1 and 2 T-helper reactions (Th1 and Th2), as well as their inducers: immunosuppressive responses and reactions mediated by Th22 and Th17 lymphocytes and capable of modifying the main types of IIRs. Cellular and molecular manifestations of each IIR type are analyzed and their general characteristics and roles in tissue regeneration and tumor growth are presented. Since inflammatory responses in a tumor can also be initiated by innate immunity mechanisms, special attention is given to inflammation based on them. We emphasize that processes accompanying tissue regeneration are prototypes of processes underlying cancer progression, and these processes have the same cellular and molecular substrates. We focus on evidence that tumor progression is mainly contributed by processes specific for the second phase of “wound healing” that are based on the Th2-type IIR. We emphasize that the effect of various types of immune and stroma cells on tumor progression is determined by the ability of the cells and their cytokines to promote or prevent the development of Th1- or Th2-type of IIR. Finally, we supposed that the nonspecific influence on the tumor caused by the cytokine context of the Th1- or Th2-type microenvironment should play a decisive role for suppression or stimulation of tumor growth and metastasis.     

Intratumor heterogeneity: Nature and biological significance

Intratumor heterogeneity inherent in the majority of human cancers is a major obstacle for a highly efficient diagnosis and successful prognosis and treatment of these diseases. Being a result of clonal diversity within the same tumor, intratumor heterogeneity can be manifested in variability of genetic and epigenetic status, gene and protein expression, morphological structure, and other features of the tumor. It is most likely that the appearance of this diversity is a source for the adaptation of the tumor to changes in microenvironmental conditions and/or a tool for changing its malignant potential. In any case, both processes result in the appearance of cell clones with different undetermined sets of hallmarks. In this review, we describe the heterogeneity of molecular disorders in various human tumors and consider modern viewpoints of its development including genetic and non-genetic factors of heterogeneity origin and the role of cancer stem cells and clonal evolution. We also systematize data on the contribution of tumor diversity to progression of various tumors and the efficiency of their treatment. The main problems are indicated in the diagnosis and therapy of malignant tumors caused by intratumor heterogeneity and possible pathways for their solution. Moreover, we also suggest the key goals whose achievement promises to minimize the problem of intratumor heterogeneity and to identify new prognostic, predictive, and target markers for adequate and effective treatment of cancer.