Targets for Ibrutinib Beyond B Cell Malignancies.

Ibrutinib (Imbruvica™) is an irreversible, potent inhibitor of Bruton's tyrosine kinase (BTK). Over the last few years, ibrutinib has developed from a promising drug candidate to being approved by FDA for the treatment of three B cell malignancies, a truly remarkable feat. Few, if any medicines are monospecific and ibrutinib is no exception; already during ibrutinib's initial characterization, it was found that it could bind also to other kinases. In this review, we discuss the implications of such interactions, which go beyond the selective effect on BTK in B cell malignancies. In certain cases, the outcome of ibrutinib treatment likely results from the combined inhibition of BTK and other kinases, causing additive or synergistic, effects. Conversely, there are also examples when the clinical outcome seems unrelated to inhibition of BTK. Thus, more specifically, adverse effects such as enhanced bleeding or arrhythmias could potentially be explained by different interactions. We also predict that during long-term treatment bone homoeostasis might be affected due to the inhibition of osteoclasts. Moreover, the binding of ibrutinib to molecular targets other than BTK or effects on cells other than B cell-derived malignancies could be beneficial and result in new indications for clinical applications.

Effects of IL-17 on erythroid progenitors growth: involvement of MAPKs and GATA transcription factors.

Interleukin-17 is Th17 cell cytokine implicated in regulation of hematopoiesis and inflammation. Besides promoting granulopoiesis, we have previously shown that IL-17 also affects erythropoiesis stimulating the development of early erythroid progenitors, BFU-E, but suppressing, at least partly via p38 MAPK, the growth of late stage erythroid progenitors, CFU-E. The aim of the present study was to investigate the involvement of other MAPKs, JNK and ERK1/2, as well as GATA transcription factors, in IL-17-mediated effects on murine bone marrow erythroid progenitors. Data obtained by use of specific MAPKs inhibitors indicated that MEK1/2-ERK1/2 MAPK signaling mediates IL-17-induced CFU-E inhibition, as well as that JNK and/or MEK1/2-ERK1/2 MAPKs activation underlies IL-17-induced stimulation of BFU-E growth. Furthermore, Western blot analyses demonstrated no effect on early hematopoiesis transcription factor, GATA-2, and enhanced expression level of erythroid-specific factor GATA-1 in murine bone marrow cells after IL-17 stimulation, which in light of previous reports that GATA-1 overexpression inhibits erythroid differentiation, could be related to IL-17-mediated inhibition of CFU-E growth. Although, no contribution for p38, JNK and ERK MAPKs in IL-17-induced GATA-1 expression was shown, data obtained using specific inhibitors pointed to the role of JNK and MEK1/2-ERK1/2 in GATA-1 downregulation. Overall, obtained data gave an insight into the mechanisms by which IL-17 exerts its effects on erythropoiesis, implying the involvement of JNK and ERK MAPKs, as well as GATA-1, in IL-17-regulated growth of erythroid progentors.

Establishment and initial characterization of SOX2-overexpressing NT2/D1 cell clones.

SOX2, a universal marker of pluripotent stem cells, is a transcription factor that helps control embryonic development in vertebrates; its expression persists in neural stem/progenitor cells into adulthood. Considering the critical role of the SOX2 transcription factor in the regulation of genes required for self-renewal and pluripotency of stem cells, we developed and characterized SOX2-overexpressing NT2/D1 cell clones. Using Southern blot and semi-quantitative RT-PCR, we confirmed integration and expression of exogenous SOX2 in three NT2/D1 cell clones. Overexpression of the SOX2 gene was detected in two of these clones. SOX2 overexpression in NT2/D1 cell clones resulted in altered expression of key pluripotency genes OCT4 and NANOG. Furthermore, SOX2-overexpressing NT2/D1 cell clones entered into retinoic acid-dependent neural differentiation, even when there was elevated SOX2 expression. After 21 days of induction by retinoic acid, expression of neural markers (neuroD1 and synaptophysin) was higher in induced cell clones than in induced parental cells. The cell clone with SOX2 overexpression had an approximately 1.3-fold higher growth rate compared to parental cells. SOX2 overexpression did not increase the population of cells undergoing apoptosis. Taken together, we developed two SOX2-overexpressing cell clones, with constitutive SOX2 expression after three weeks of retinoic acid treatment. SOX2 overexpression resulted in altered expression of pluripotency-related genes, increased proliferation, and altered expression of neural markers after three weeks of retinoic acid treatment.

The effect of interleukin-17 on hematopoietic cells and cytokine release in mouse spleen.

To evaluate whether the response of hematopoietic cells to interleukin-17 (IL-17) depends on the tissue microenvironment in which hematopoiesis occurs, the influence of recombinant mouse IL-17 on spleen hematopoietic cells and cytokine release was assessed in normal mice in vitro and in vivo. In vitro, IL-17 did not significantly affect the growth of granulocyte-macrophage (CFU-GM) and erythroid (BFU-E and CFU-E) derived colonies. A single injection of IL-17 in vivo exhibited stimulatory effects on hematopoietic cells from both granulocytic and erythroid lineages. The increased number of metamyelocytes 48 h after treatment imply to the IL-17-induced stimulation of granulopoiesis. The number of BFU-E was increased at 24 h, while the number of CFU-E increased 6 h and 24 h after treatment. Since the same treatment in the bone marrow decreased the number of CFU-E, it may be concluded that the local microenvironment plays an important role in IL-17-mediated effects on CFU-E. IL-17 increased the release of IL-6 both in vitro and in vivo, but showed tendency to suppress the constitutive secretion of IL-10 by spleen cells. Our results suggest the complexity of target cell response and interplay of secondary induced cytokines by IL-17 in different hematopoietic organs.

In vivo effects of interleukin-17 on haematopoietic cells and cytokine release in normal mice.

In order to gain more insight into mechanisms operating on the haematopoietic activity of the T-cell-derived cytokine, interleukin-17 (IL-17) and target cells that first respond to its action in vivo, the influence of a single intravenous injection of recombinant mouse IL-17 on bone marrow progenitors, further morphologically recognizable cells and peripheral blood cells was assessed in normal mice up to 72 h after treatment. Simultaneously, the release of IL-6, IL-10, IGF-I, IFN-gamma and NO by bone marrow cells was determined. Results showed that, in bone marrow, IL-17 did not affect granulocyte-macrophage (CFU-GM) progenitors, but induced a persistant increase in the number of morphologically recognizable proliferative granulocytes (PG) up to 48 h after treatment. The number of immature erythroid (BFU-E) progenitors was increased at 48 h, while the number of mature erythroid (CFU-E) progenitors was decreased up to 48 h. In peripheral blood, white blood cells were increased 6 h after treatment, mainly because of the increase in the number of lymphocytes. IL-17 also increased IL-6 release and NO production 6 h after administration. Additional in vitro assessment on bone marrow highly enriched Lin- progenitor cells, demonstrated a slightly enhancing effect of IL-17 on CFU-GM and no influence on BFU-E, suggesting the importance of bone marrow accessory cells and secondary induced cytokines for IL-17 mediated effects on progenitor cells. Taken together, these results demonstrate that in vivo IL-17 affects both granulocytic and erythroid lineages, with more mature haematopoietic progenitors responding first to its action. The opposite effects exerted on PG and CFU-E found at the same time indicate that IL-17, as a component of a regulatory network, is able to intervene in mechanisms that shift haematopoiesis from the erythroid to the granulocytic lineage.

Regulation of the SOX3 gene expression by retinoid receptors.

Sox3/SOX3 gene is considered to be one of the earliest neural markers in vertebrates. Despite the mounting evidence that Sox3/SOX3 is one of the key players in the development of the nervous system, limited data are available regarding the transcriptional regulation of its expression. This review is focused on the retinoic acid induced regulation of SOX3 gene expression, with particular emphasis on the involvement of retinoid receptors. Experiments with human embryonal carcinoma cells identified two response elements involved in retinoic acid/retinoid X receptor-dependent activation of the SOX3 gene expression: distal atypical retinoic acid-response element, consisting of two unique G-rich boxes separated by 49 bp, and proximal element comprising DR-3-like motif, composed of two imperfect hexameric half-sites. Importantly, the retinoic acid-induced SOX3 gene expression could be significantly down-regulated by a synthetic antagonist of retinoid receptors. This cell model provides a solid base for further studies on mechanism(s) underlying regulation of expression of SOX3 gene, which could improve the understanding of molecular signals that induce neurogenesis in the stem/progenitor cells both during development and in adulthood.

Combined effect of IL-17 and blockade of nitric oxide biosynthesis on haematopoiesis in mice.

AIM: The study was undertaken to extend our investigation concerning both the in vivo activity of interleukin (IL)-17 and the specific role of nitric oxide (NO) in IL-17-induced effects in the process of haematopoiesis. METHODS: CBA mice were simultaneously treated with IL-17 and/or nitric oxide synthase (NOS) inhibitor, l-NAME, for 5 days and changes within various haematopoietic cell lineages in bone marrow, spleen and peripheral blood were analysed. RESULTS: Findings showed that administration of both IL-17 and l-NAME stimulated increase in net haematopoiesis in normal mice. IL-17-enhanced myelopoiesis was characterized by stimulation of both femoral and splenic haematopoietic progenitor cells and morphologically recognizable granulocytes. Additionally, IL-17 induced alterations in the frequency of erythroid progenitor cells in both bone marrow and spleen, accompanied with their mobilization to the peripheral blood. As a consequence of these changes in the erythroid cell compartments, significant reticulocytosis was observed, which evidenced that in IL-17-treated mice effective erythropoiesis occurred. Exposure of mice to NOS inhibitor also increased the number of both granulocyte-macrophage and erythroid progenitors in bone marrow and spleens, and these alterations were followed by the mobilization of erythroid progenitors and elevated content of reticulocytes in peripheral blood. The specific role of NO in IL-17-induced haematopoiesis was demonstrated only in the IL-17-reducing effect on bone marrow late stage erythroid progenitors, CFU-E. CONCLUSION: The results demonstrated the involvement of both IL-17 and NO in the regulation of haematopoietic cell activity in various haematopoietic compartments. They further suggest that IL-17 effects are differentially mediated depending on the haematopoietic microenvironments.

[Epidermolysis bullosa hereditaria dystrophica: diagnostic problems]. / Epidermolysis bullosa hereditaria dystrophica: dijagnosticki problem.

A female patient at the age of 8 years was presented in this paper. The patient was affected by an exceptionally rare disease. A complete clinical picture of disease together with its complication on the esophagus was set forth. According to the literature available a contemporary classification of disease was made and therefore, this case was put into one of the possible varieties of disease.

Medico-legal and ethical approach to the legal interruption of pregnancy after aspiration biopsy of the chorionic villi and early amniocentesis.

PIP: According to article 191 of the 1974 Yugoslavian Constitution, men have the right to decide freely on the birth of a child. According to the Law of conditions and proceedings for permitting abortion (1978), pregnant women are allowed to make the decision to abort freely up to the 10th week of pregnancy. Between the 10th and 20th week there are medical, eugenic, and moral indications for abortion; medical indication being disease or a high risk to the other of the mother; eugenic indications being heredity deformities in the fetus or heavy abnormalities; and moral indications being pregnancies caused by the commission of a crime like rape or incest. In the region of Vojvodina, population 2 million, there were 25,000 live births and twice as many abortions. The traditions of Yugoslavia are that fetuses have a right to life including a high quality of life free from disease and genetic abnormalities. Doctors are relied upon to take the greatest responsibility in terms of doing the ethical things for a pregnant woman and the fetus. Modern medical technology allows doctors and patients to have ever greater amounts of information so that an informed decision can be made about the health and well being of the mother and the fetus.^ieng

Frequency estimation of the gene alleles for genodermatoses in the population of Vojvodina.

Genodermatoses are hereditary skin disorders or anomalies. The authors had investigated the frequency of genes, species and clinical forms of hereditary dermatoses in Vojvodina including age and sex distribution. The experimental group was selected by random sampling of the diseased of genodermatosis and/or genetic diseases at the Clinic of Dermatology and Venereology and Institute for Children and Adolescents in Novi Sad. The experimental group included 152 cases, and that made simultaneously the experimental and positive control group in relation to the diseased population with genodermatoses and/or genetic diseases in Vojvodina. In the investigation we applied the following methods: family history taking including genealogy; dermatoglyphic examination; screening tests in medical genetics; cytogenetic analysis of patient's karyotype; histopathological analysis of the material obtained by skin biopsy; dermatovenereological, genetic and dysmorphologic examination of skin diseases by analysis of dysmorphologic signs on the skin using a special computer program--POSSUM (Pictures of Standard Syndromes and Undiagnosed Malformations). Application of non-parametric statistics and Log-linear analysis, revealed that there is no statistically significant difference between the experimental group and the group with genetic diseases in population of Vojvodina. The obtained incidence of genodermatoses and/or genetic diseases was computed by "Hardy-Weinberg's principle". These methods of genetic population investigations give possibilities for valid incidence estimation of the diseases and frequency of the gen allele's for genodermatoses and/or genetic disorders and syndromes in defined population. Our results of investigation of the incidence genodermatoses in our's population showed significantly increased values in relation to literature data for the same hereditary disorders.

[Minor malformation score in congenital cleft of the lip and palate]. / Minor malformacioni skor kod urodenih rascepa usne, vilice i nepca.

The authors analyzed minor malformation score in 63 children (36 boys and 27 girls) with congenital cleft of the lip and palate in order to evaluate purposefulness of its application as a screening method in detecting various major malformations. The results obtained revealed extremely increased minor malformation score in our patients and confirmed that an increased minor malformation score is associated with major malformation, i.e. clefts of the lip and palate. Therefore, it could be used as a screening method for detection of major malformations.

[Morbidity and mortality in adolescents in Vojvodina]. / Morbiditet i mortalitet adolescenata U SAP Vojvodini.

Adolescents. The aim of this study was to analyze the morbidity and mortality of adolescents in SAP Vojvodina. For the analysis, data was used from the statistical yearbooks on the health protection of the population and data from individual statistical reports. The period from 1980 to 1986 was observed. In order to more easily follow the morbidity and mortality, and for a better insight into the pathology, the adolescents were divided into 2 age groups; the younger one from 10 to 14 years of age, and the older one from 15 to 19. Adolescents in Vojvodina mostly ail from respiratory diseases, illnesses of the digestive tract, and infective diseases, but a significant role in the morbidity is also taken by injuries and poisoning. Adolescents of the age group from 10 to 14 years, get sick more often. In SAP Vojvodina 156 adolescents die annually. The highest mortality is due to injuries and poisoning (16.36/1,000,000), and after that comes the mortality due to neoplasms (5.84/1,000,000), and the diseases of the respiratory and circulatory system (3.5/1,000,000). Mortality is higher in the adolescent group from 15 to 19 years of age. With this study only a partial insight was achieved into the pathology of the adolescent age group. In order to attain a full insight, it is absolutely necessary to actively and prospectively follow the health state of adolescents and the conditions of environment.

[Multicystic dysplastic kidney. Therapeutic dilemmas and personal experience]. / Multicisticni displazicni bubreg. Terapijske dileme i nase iskustvo.

A multicystic dysplastic kidney (MCDK) is one of the most frequent causes of abdominal mass in the neonate. Prenatal echography permits early and frequent diagnostics. It is a nonfamilial disease without associated cystic disease of the pancreas, liver or lungs. Indications for elective surgery are clear when there is a symptomatic disease. However, treatment of asymptomatic patients is controversial. A rising number of authors prefer nonsurgical approach, leaving MCDK intact with a close follow-up of patients for possible severe complications (malignancy, hypertension, infection, pain, rupture). We treated 15 patients with MCDK from 1984 to 1994. Diagnosis was passed antenatally in 7 (47%) patients, accidentally in 2, and based on the presence of abdominal mass in 6 patients. Two patients had renal failure due to the abnormal contralateral kidney. Nine patients were operated on and 6 were treated nonsurgically. The risk of complications associated with nonsurgical treatment, easiness of efficient surgery at the age of 3-6 months, avoiding stress in the child and family due to long-term follow-up, all suggest operative treatment. We suggest to parents both operative and nonoperative options, explaining the risk and danger of both. We believe that nephrectomy is the best solution in a child with MCDK who is growing, develops hypertension, with uncertain diagnosis or when adequate follow-up is impossible.

[Skin changes in patients with Lyme borreliosis]. / Kozne promene u obolelih od Lyme borreliosis.

In introduction some clinical characteristics of Erythema migrans, Borrelia lymphocytoma and acrodermatitis chronica atrophica has been described. The importance of atypical forms of Erythema migrans and the difficulties in differential diagnosis of cutaneous manifestation has been stressed. In a prospective, and partly retrospective investigation of 1292 persons with tick bites, signs of Lyme borreliosis have been found in 18.96%. Number of such persons seen in dermatology wards is rising, and 18.2% of these are children less than 15 years of age. Patients seen in dermatology are mostly women (56.5%:43.5%). Nearly half of the patients with Erythema migrans did not known that they had a tick bite (42.5%). Lyme borreliosis was manifested mainly as Erythema migrans, 89% of patients. Borrelia lymphocytoma was encountered in 2%, and Acrodermatitis chronica atrophicans in 0.4% of patients, significantly less than in other reports. Sclerotic skin lesions were found in 4.1% of patients, and some macular and urticarial lesions were recorded. An incubation period generally less than three weeks preceded to skin manifestations, but in some patients this period could not be recorded. Skin lesions were located on lower extremities in 50.4% of patients, trunk in 25.5%, and upper extremities in 10.5% of patients. In 87% of patients skin lesions lasted less than three weeks. Symptoms were present in 62% of patients. Seropositivity to Borrelia burgdorferi has been found in 10.2% of patients, mostly three weeks after the tick bite.

[Hereditary and congenital defects in children in the regions of Indija-Stara Pazova and Pancevo-Kovin]. / Urodene i prirodene mane u dece regiona Indija-Stara Pazova i Pancevo-Kovin.

An analysis of frequency and range of hereditary and congenital defects was performed in all infants born in the period from January 1 to December 31, 1986 in the Ward for Neonates at the Department of Gynecology and Obstetrics in Pancevo and in the out-patient maternity home in Indija and Kovin. In Pancevo 2.559 (1.286 m. and 1.273 f.) newborn infants were born, i.e. there were 2.314 (1.178 m. and 1.136 f.) mature children and 245 (108 m. and 187 f.) premature children. In Indija 367 (180 m. and 187 f.) newborn infants were born, i.e. there were 355 (174 m. and 181 f.) mature children and 12 (6 m. and 6 f.) premature children. In Kovin 146 (65 m. and 81 f.) newborn infants were born, i.e. 150 (82 m. and 68 f.) mature and 10 (1 m. and 9 f.) premature children. A total 3.072 (1.531 m. and 1.541 f.) newborn infants in all three places were born. In all three places there was a total of 2.805 (1.417 m. and 1.387 f.) mature and 267 (115 m. and 152 f.) premature children. In Pancevo 68 children were born with defects (40 m. and 28 f.), i.e. 2.66% (3.11% m. and 2.20% f.). In Indija only one defect was noted in one female child (0.53% related to female newborns).(ABSTRACT TRUNCATED AT 250 WORDS)