Circulating subpopulations of non-cytotoxic ILCs in diffuse large B-cell lymphoma.

Non-cytotoxic innate lymphoid cells (ILCs) have been added to the list of immune cells that may contribute to the tumor microenvironment. Elevated levels of total ILCs and their subgroups have been reported in peripheral blood and tissue samples from patients with solid tumors, but their frequency in non-Hodgkin lymphomas, particularly diffuse large B-cell lymphoma (DLBCL), has not been clearly established. This study examined frequency and subset distribution in newly diagnosed DLBCL patients (nodal and extra-nodal) and compared it with blood specimens from healthy donors. The percentage of total ILCs (Lin - CD127+) was assessed by flow cytometry, as well as the four ILC subsets, defined as ILC1 (Lin - CD127 + cKit - CRTH2-), ILC2 (Lin - CD127 + cKit+/- CRTH2+), ILCp NCR- (Lin - CD127 + cKit + CRTH2- NKp46-) and NCR + ILC3 (Lin - CD127 + cKit + NKp46+). In the studied group of patients (n = 54), significantly lower levels of circulating total ILCs, ILC1, and ILCp NCR- were observed compared to the control group (n = 43). Similarly, there was a statistically significant decrease in the median frequency of NKp46 + ILC3 cells in lymphoma patients. Analysis of the ILC2 subpopulation showed no significant differences. The correlation of the distribution of individual subpopulations of ILCs with the stage and location of the tumor was also demonstrated. Our results suggest that circulating ILCs are activated and differentiated and/or differentially recruited to the lymph nodes or tumor microenvironment where they may be involved in antitumor defense. However, our observations require confirmation in functional studies.

Limited Long-Term Impact of Insect Venom Immunotherapy on the Micro-RNA Landscape in Whole Blood.

OBJECTIVE: To perform a genome-wide characterization of changes in microRNA (miRNA) expression during the course of venom immunotherapy (VIT). METHODS: miRNA was isolated from the whole-blood of 13 allergic patients and 14 controls, who experienced no allergic reaction upon stings by honeybees and wasps. We analyzed 2549 miRNAs from the whole blood of these patients prior to VIT and 12 months after the start of VIT. The results for differential expression obtained on a microarray platform were confirmed by quantitative real-time PCR. Out of the 13 patients, 8 had a negative allergic reaction with VIT, thus indicating that this approach was successful. RESULTS: By comparing time points before and 12 months after ultrarush VIT, correlation analysis and principal component analysis both indicated a limited effect of VIT on the overall miRNA expression pattern. Volcano plot analysis based on raw P values revealed few deregulated miRNAs, most of which were increasingly expressed after VIT as compared with before VIT. Based on the 50 most altered miRNAs, no clear clustering was observed before or after VIT. CONCLUSIONS: Our results indicate an overall reduced effect of VIT on the miRNA expression pattern in whole blood.

Limited long-term impact of insect venom immunotherapy on the micro-RNA landscape in whole blood

Objective: To perform a genome-wide characterization of changes in microRNA (miRNA) expression during the course of venom immunotherapy (VIT). Methods: miRNA was isolated from the whole-blood of 13 allergic patients and 14 controls, who experienced no allergic reaction upon stings by honeybees and wasps. We analyzed 2549 miRNAs from the whole blood of these patients prior to VIT and 12 months after the start of VIT. The results for differential expression obtained on a microarray platform were confirmed by quantitative real-time PCR. Out of the 13 patients, 8 had a negative allergic reaction with VIT, thus indicating that this approach was successful. Results: By comparing time points before and 12 months after ultrarush VIT, correlation analysis and principal component analysis both indicated a limited effect of VIT on the overall miRNA expression pattern. Volcano plot analysis based on raw P values revealed few deregulated miRNAs, most of which were increasingly expressed after VIT as compared with before VIT. Based on the 50 most altered miRNAs, no clear clustering was observed before or after VIT. Conclusions: Our results indicate an overall reduced effect of VIT on the miRNA expression pattern in whole blood

Objetivo: Realizar la caracterización genómica de los cambios en la expresión de microARN (miARN) en el curso de ITV (inmunoterapia con veneno). Métodos: Los microARNs se analizaron en la sangre total de 13 pacientes alérgicos y 14 controles sin reacción alérgica a las picaduras de abejas y avispas. Se analizaron 2549 miRNAs diferentes en la sangre total de estos pacientes antes de la ITV y 12 meses después del inicio de la ITV. Los resultados de expresión diferencial obtenidos en la plataforma de microarrays se confirmaron mediante PCR cuantitativa a tiempo real (qRT-PCR). De los 13 pacientes, se confirmó que ocho tenían una reacción alérgica negativa tras la ITV, lo que indicó una ITV exitosa. Resultados: Al comparar los resultados de microRNAs, previa IT y 12 meses después de la ITV, la correlación y el análisis de componentes principales indican un efecto limitado de la ITV en el patrón de expresión general de miARN. El análisis de Volcano basado en los valores de P crudos, reveló la existencia de pocos miRNAs desregulados estando la mayoría de ellos sobre-expresados tras la ITV en comparación con la previa. Utilizando los 50 miRNAs que más se alteraban, no se observó una agrupación clara en función del tiempo, es decir, pre y post-ITV. Conclusiones: Nuestros resultados indican que la ITV tiene poco efecto en el patrón de expresión de miARN en sangre completa

Comparative lipidomic analysis of synovial fluid in human and canine osteoarthritis.

OBJECTIVE: The lipid profile of synovial fluid (SF) is related to the health status of joints. The early stages of human osteoarthritis (OA) are poorly understood, which larger animals are expected to be able to model closely. This study examined whether the canine groove model of OA represents early OA in humans based on the changes in the lipid species profile in SF. Furthermore, the SF lipidomes of humans and dogs were compared to determine how closely canine lipid species profiles reflect the human lipidome. METHODS: Lipids were extracted from cell- and cellular debris-free knee SF from nine donors with healthy joints, 17 patients with early and 13 patients with late osteoarthritic changes, and nine dogs with knee OA and healthy contralateral joints. Lipid species were quantified by electrospray ionization tandem mass spectrometry (ESI-MS/MS). RESULTS: Compared with control canine SF most lipid species were elevated in canine OA SF. Moreover, the lipid species profiles in the canine OA model resembled early OA profiles in humans. The SF lipidomes between dog and human were generally similar, with differences in certain lipid species in the phosphatidylcholine (PC), lysophosphatidylcholine (LPC) and sphingomyelin (SM) classes. CONCLUSIONS: Our lipidomic analysis demonstrates that SF in the canine OA model closely mimics the early osteoarthritic changes that occur in humans. Further, the canine SF lipidome often reflects normal human lipid metabolism.

Safety of Ultrarush Venom Immunotherapy: Comparison Between Children and Adults.

BACKGROUND: The ultrarush protocol is an attractive approach in the buildup phase of venom immunotherapy (VIT-UR). However, the degree of risk of VIT-UR in children remains unknown. The objective of this study was to compare the safety of VIT-UR in children and adults. METHODS: We performed a study based on prospectively gathered medical records of children and adults with hymenoptera venom allergy treated with VIT-UR in 3 allergy centers in Poland. RESULTS: The study population comprised 134 children (mean [SD] age, 12.6 [3.7] years; males, 70.1%) and 207 adults (mean age, 42.4 [14.0] years; males, 47.8%). The number of children in the subgroups of bee venom (BV) allergy and wasp venom (WV) allergy were comparable, although sensitization to WV was more predominant in the adult group (70.1%). Skin reactivity to both venoms was more common in children than in adults (P < .001); however, children had higher concentrations of total IgE and specific IgE to BV (both P < .001). Systemic allergic reactions (VIT-SARs) occurred in 6.2% of the patients (3.7% in children and 7.7% in adults; nonsignificant). In adults, SARs occurred more frequently in patients treated with BV than WV extracts (21.4% vs 2.6%; P < .001). The same pattern was observed in children (7.2% vs 0%; P = .058). However, VIT-SARs to BV were less frequent in children than in adults (P = .034). Similarly, no significant relationship was noted between children and adults receiving WV VIT (2.6% vs 0%; nonsignificant). The severity of VIT-SAR did not differ between children and adults. CONCLUSIONS: VIT-UR is safer in children. Age below 18 is not a risk factor for VIT-SARs.

Therapeutic antitumor potential of endoglin-based DNA vaccine combined with immunomodulatory agents.

Therapy targeting tumor blood vessels ought to inhibit tumor growth. However, tumors become refractory to antiangiogenic drugs. Therefore, therapeutic solutions should be sought to address cellular resistance to antiangiogenic therapy. In this regard, reversal of the proangiogenic and immunosuppressive phenotype of cancer cells, and the shift of the tumor microenvironment towards more antiangiogenic and immune-stimulating phenotype may hold some promise. In our study, we sought to validate the effects of a combination therapy aimed at reducing tumor blood vessels, coupled with the abrogation of the immunosuppressive state. To achieve this, we developed an oral DNA vaccine against endoglin. This antigen was carried by an attenuated Salmonella Typhimurium and applied before or after tumor cell inoculation into immunocompetent mice. Our results show that this DNA vaccine effectively inhibited tumor growth, in both the prophylactic and therapeutic settings. It also activated both specific and nonspecific immune responses in immunized mice. Activated cytotoxic T-lymphocytes were directed specifically against endothelial and tumor cells overexpressing endoglin. The DNA vaccine inhibited angiogenesis but did not affect wound healing. In combination with interleukin-12-mediated gene therapy, or with cyclophosphamide administration, the DNA vaccine resulted in reduced microvessel density and lowered the level of Treg lymphocytes in the experimental tumors. This effectively inhibited tumor growth and prolonged survival of the treated animals. Polarization of tumor milieu, from proangiogenic and immunosuppressive, towards an immunostimulatory and antiangiogenic profile represents a promising avenue in anticancer therapy.

Impact of different dimethyl sulphoxide concentrations on cell recovery, viability and clonogenic potential of cryopreserved peripheral blood hematopoietic stem and progenitor cells.

BACKGROUND AND OBJECTIVES: The procedure of autologous hematopoietic stem/progenitor cell transplantation requires cryopreservation. Addition of DMSO is necessary to secure the viability of such cells, but this solvent is potentially toxic to stem cells' recipient. 10% DMSO solution is used by the majority of transplant centres. The aim of our study was to test if DMSO concentration might be reduced without negative impact on cell recovery and clonogenicity. MATERIALS AND METHODS: Samples were prospectively collected from 20 patients. Small volumes of leukapheresis products were frozen with different cryoprotective mixtures, containing 10%, 7·5%, 5% and 2·5% DMSO, respectively. The quality of cryoprotective mixtures was evaluated based on recovery, viability and clonogenic potential of hematopoietic stem cells after defreezing. RESULTS: Reduction in DMSO concentration to 7·5% or lower was associated with decreased recovery of nucleated cells. In contrast, the number of colonies was highest for 7·5% DMSO with significant differences when compared to 10% DMSO solution. CONCLUSION: Reduction in DMSO concentration from 10% to 7·5% may have favourable impact on hematopoietic recovery after autologous transplantation. The findings require confirmation in a clinical setting.

Biological maturity at birth, the course of the subsequent ontogenetic stages and age at menarche.

The main aim of the study was to assess the influence of biological maturity at birth on growth processes in the subsequent years and during puberty in girls. The material of this study comes from the outpatient clinic cards and cross-sectional research on girls from the province of Wielkopolska in Poland. It includes data of 527 girls. The influence of perinatal maturity on body weight in the later stages of ontogeny was determined with the use of the Kruskal-Wallis test and the Mann-Whitney U test. In order to determine the relationship between perinatal maturity and age at menarche, the survival analysis module was used. The results show a diverse influence of perinatal maturity on the values of body weight achieved in later years of life. The indicated predictive factors included both birth weight and gestational age. In the examined girls menarche occurred between the 10th year and the 17th year of life (X¯=12.87, s=1.26; Me=13 years). The comparison showed a significant variation in age at menarche depending on the length of pregnancy (log-rank χ(2)(2)=27.068, p<0.0001) and birth weight (log-rank χ(2)(2)=23.241, p<0.0001). There was no variation in maturation of the examined girls conditioned by the occurrence of intra-uterine growth retardation (log-rank χ(2)(2)=2.046, p>0.05). Remote prognoses as to the postnatal development of preterm-born children and/or children with low birth weight indicate adverse influence of these variables on age at menarche. Perinatal biological maturity of a newborn conditions the course of postnatal development.

Biological status of adult patients with cystic fibrosis.

Cystic fibrosis (CF) is the most common autosomal disorder in the Caucasian population. The main goal of the study was to assess the biological condition of adult patients with CS. Data of 90 CF patients aged 18-31 were considered. The biological condition was determined by the measurement of somatometric traits and the nutritional status. The results show a considerable physical retardation and a poor nutritional status of the studied patients. Nearly 45% of the patients showed symptoms of malnutrition, ranging from slight undernutrition to emaciation. The results, however, show a considerable variability of data among the CF patients compared with the healthy population. A significant relationship between the type of mutation and nutritional status was demonstrated.

Spermatocyte-specific expression of constitutively active heat shock factor 1 induces HSP70i-resistant apoptosis in male germ cells.

Spermatocytes, the most sensitive male germ cells to heat-induced apoptosis, do not respond to hyperthermia by inducing heat shock proteins (HSPs), including HSP70i, which has been previously shown to confer resistance to apoptosis in somatic cells. To dissect the mechanism of heat-induced apoptosis and to determine if we could protect spermatocytes by expressing HSP70i, we engineered transgenic mice that express in spermatocytes constitutively active heat shock transcription factor (HSF)1. Such HSF1 expression did not lead to transcription of inducible Hsp70 genes, but instead induced caspase-dependent apoptosis that mimicked heat shock-induced death of spermatogenic cells. Both mitochondria-dependent and death receptor-dependent pathways appear to be involved in such HSF1-induced apoptosis: the levels of Bcl-2 family proteins became increased, p53 protein accumulated and expression levels of caspase-8 and death-receptor-interacting proteins (including Fas-associated death domain protein and TNF receptor associated death domain protein) became elevated. Surprisingly, the constitutive spermatocyte-specific expression of HSP70i in double-transgenic males did not protect against such HSF1-induced apoptosis.

[Evaluation of the character of morphologic traits in persons with obstructive sleep apnea]. / Ocena charakteru cech morfologicznych osób z zespolem obturacyjnego bezdechu sennego.

The aim of the study was carried out to show the anthropometric analysis of patients with obstructive sleep apnoea syndrome and to answer the question about the relations between the degree of SAS and obesity. The research has begun since May 1993 in an interdisciplinary team. The study was carried out in a group of 40 men diagnosed as SAS in Sleep Apnoea Unit of Department of Pulmonary Diseases. The anthropometric analysis consists of basic somatometric measurements. The relations between obesity and the degree of apnoeas was determined by analysis of variance and the model of single and multiple regression. The obtained results demonstrate the dependence between the grade of apnoeas pathology during sleep and the measurements of upper body parts. The slope of the line B not equal to 0 indicates that the intensity of SAS increases in patients "commonly considered as obese". Obesity is an important factor leading to disturbances in respiratory ventilation. An important development of fatty tissue of the neck can cause pressure changes and can induce adipose degeneration. An increased fatty thickness of the thorax is a factor which can lead to the aggravation of symptoms.