Encouraging effect of autologous bone marrow aspirate concentrate in rehabilitation of children with cerebral palsy.

OBJECTIVE: In this study, we used autologous bone marrow aspirate concentrate (BMAC) transplantation to treat children with cerebral palsy (CP) to improve their motor and cognitive functions. PATIENTS AND METHODS: Forty-two patients with CP received BMAC. The transplantation of stem cells via the intrathecal route includes three BMAC applications. The patients' examination was before the injection of stem cells, with follow-ups on 1, 3, 6, and 12 months after the injections. The assessments included the gross motor function classification scale, the Ashworth scale, and the Learning accomplishment profile-diagnostic scale. RESULTS: This study included 42 patients with CP who received three BMAC intrathecal administrations. A personalized home rehabilitation program was designed and included for each patient in the study. After the treatment, we observed a reduction of spasticity in 58% of patients and significant cognitive improvement in 35% of patients. CONCLUSIONS: The outcome of this study indicates that stem cell therapy and personalized training can improve the development of children with CP. The crucial goal of this therapeutic intervention is to substitute injured tissue with new tissues by activating the regenerative capacity of stem cells.

Serum cytokine profile of laryngeal squamous cell carcinoma patients.

OBJECTIVES: This study aimed to evaluate serum cytokine concentrations in healthy individuals and laryngeal squamous cell carcinoma patients. METHODS: A total of 59 laryngeal squamous cell carcinoma patients and 44 healthy controls were included. Multiplex analysis of interleukins 2, 4, 5, 6, 10, 12, 13 and 17 and interferon-gamma with respect to the presence of laryngeal carcinoma, tumour-node-metastasis T stage, nodal involvement and larynx subsite was performed. RESULTS: Statistical analysis revealed no difference in serum cytokine levels between patients and healthy controls. The serum interleukin-12 concentration was significantly higher in patients with early (T1-2) than in those with late (T3-4) stage disease and without nodal involvement (p < 0.05). Serum interleukin-10 levels were significantly higher in T3-4 stage than in T1-2 stage patients (p < 0.05). Additionally, serum interleukin 10, 12 and 13 concentrations (p < 0.05) and interleukin-6 concentration (p < 0.01) were significantly higher in patients with T1-2 stage supraglottic vs glottic tumours. CONCLUSION: Serum cytokines level cannot be used as laryngeal squamous cell carcinoma markers. Progression from T1-2 to T3-4 stage is followed by decreased serum interleukin-12 levels and increased interleukin-10 levels. Nodal involvement is associated with lower serum interleukin-12 levels. In patients with early stage tumours, serum interleukin 6, 10, 12 and 13 concentrations are significantly higher in those with supraglottic vs glottic tumours.

An outbreak of tick-borne encephalitis associated with raw goat milk and cheese consumption, Croatia, 2015.

The aim of this report is to emphasize the risk of acquiring TBE by the consumption of raw milk and dairy products. In April-May 2015, we registered the first outbreak of tick-borne encephalitis in Croatia in seven members out of ten exposed persons who consumed raw goat milk or cheese from the same supplier. Infection was confirmed by TBEV enzyme-linked immunosorbent assay (ELISA) in all patients. None had been vaccinated nor had observed a tick bite.

Effects of Preoperative Clarithromycin Administration in Patients with Nasal Polyposis.

OBJECTIVE: In recent years, various investigators have shown considerable interest in the use of macrolide antibiotics for treatment of chronic rhinosinusitis and nasal polyposis. The aim of this study was to evaluate the clinical effects of preoperative long-term, low-dose clarithromycin administration in patients with nasal polyposis. METHODS: Eighty nasal polyp patients (42 non-atopic and 38 atopic) were included in this prospective, non-placebo controlled investigation and randomized equally to either the combined clarithromycin-surgical or surgical group. Forty patients received 500 mg of clarithromycin daily for eight weeks, and, after evaluation, they were treated by functional endoscopic sinus surgery (FESS). The other 40 patients were treated only surgically. The nasal symptom scores and endoscopic scores after macrolide treatment/surgical treatment, and after six and 12 months of follow-up were evaluated. RESULTS: After clarithromycin therapy, we found improvement in symptom scores in 25/40, and improvement in endoscopic scores in 19/40 patients. We found no significant difference in nasal symptom score between allergic and non-allergic patients regarding the outcome to macrolide (p = 0.352) or surgical treatment (p = 0.396). When we compared differences between endoscopic scores at the time points of 12 months and six months postoperatively (ESt12 minus ESt6), we found statistically lower differences in the clarithromycin-surgery group than in the surgery group (p = 0.006). CONCLUSION: Preoperative clarithromycin administration postponed nasal polyp relapse after FESS. Allergies have no influence on the clinical efficacy of clarithromycin therapy and on the efficacy of FESS.

Influence of allergy on clinical, immunological and histological characteristics of nasal polyposis.

OBJECTIVES: Although polyps seem to be a manifestation of the chronic inflammation of nasal/paranasal sinuses mucosa in both allergic and non-allergic subjects, the pathogenesis of nasal polyposis remains unknown. The aim of this prospective study was to compare the clinical characteristics of nasal polyposis in non-allergic and allergic patients, to compare the cytokine levels in nasal secretions in atopic and non-atopic nasal polyp patients and to correlate these levels with eosinophil counts in nasal polyp tissue specimens. METHODS: This study included thirty patients with nasal polyposis (13 atopic and 17 non-atopic) requiring surgical treatment. Nasal secretion samples were collected from the nasal cavities of all 30 subjects a few days before the surgical treatment. The levels of tumour necrosis factor-alpha (TNF-alpha), tumour necrosis factor-beta (TNF-beta), interleukin (IL)-1beta, IL-2, IL-12, interferon-gamma (IFN-gamma), IL-4, IL-5, IL-6, IL-10, and IL-8 were measured using the flow cytometric method. Each of the 30 patients was staged clinically according to nasal symptom score, endoscopic score and Lund-Mackay computed tomography (CT) score. All these patients had undergone sinus surgery. Eosinophils were counted in hematoxylin- and eosin-stained sections of all nasal polyp samples. RESULTS: Our results showed that allergy does not modify the symptoms, or the endoscopic and CT findings, of nasal polyposis. We found significantly higher concentrations of IL-4 (p < 0.01), IL-5 (p < 0.05), IL-6 (p < 0.05) and TNF-beta (p < 0.05) in nasal secretions of allergic nasal polyp patients than in non-allergic ones. Eosinophil counts were significantly higher in tissues of atopic patients' polyps than in non-atopic subjects (p < 0.01). No correlation was observed between cytokine levels and eosinophil counts. CONCLUSION: Non-atopic and atopic patients' polyps have different immunological patterns. Our results showed that the presence of Th2 cytokines was a more significant feature in allergic patients with nasal polyposis than in non-allergic patients.

Effect of long-term, low-dose clarithromycin on T helper 2 cytokines, eosinophilic cationic protein and the 'regulated on activation, normal T cell expressed and secreted' chemokine in the nasal secretions of patients with nasal polyposis.

BACKGROUND: Little is known about the effects of macrolides on the cytokines and chemokines that modulate the function of eosinophils in nasal polyposis. METHODS: Twenty-two non-allergic and 18 allergic patients with nasal polyps were administered clarithromycin 500 mg/day (single oral dose) for eight weeks. We measured the nasal secretion levels of the T helper 2 (also known as Th2) cytokines interleukin 4, 5 and 6, the 'regulated on activation, normal T cell expressed and secreted' (also known as RANTES) chemokine, and the eosinophilic cationic protein, before and after treatment. RESULTS: After clarithromycin treatment, we found reduced levels of the 'regulated on activation, normal T cell expressed and secreted' chemokine in samples from both non-allergic and allergic patients (p < 0.05). Clarithromycin treatment decreased the levels of eosinophilic cationic protein only in non-allergic patients (p < 0.05), and decreased the level of interleukin 6 only in allergic patients (p < 0.05). Decreased levels of the 'regulated on activation, normal T cell expressed and secreted' chemokine were associated with a reduction in polyp size both in non-allergic and allergic patients. CONCLUSION: Clarithromycin has a strong anti-inflammatory effect in nasal polyposis, but has different immunomodulatory effects in allergic and non-allergic nasal polyposis patients.

Correlation between cytokine levels in nasal fluid and eosinophil counts in nasal polyp tissue in asthmatic and non-asthmatic patients

Background/ Aims: Concentrations of mediators in nasal secretions could reflect the inflammatory status of the nasal mucosa and evolution of sinus disease. So, the aim of our study was to evaluate local immune reaction by measuring crucial Th1, Th2 and inflammatory cytokines innasal fluid samples of patients with nasal polyps (NP), and to correlate them to clinical, radiological findings and to the degree of eosinophil infiltration of polyp tissue. Therefore, in our study we compared the cytokine levels in nasal fluid of asthmatic and non-asthmatic patients with nasal polyposis, the eosinophil counts in NP tissues of these patients, and we correlated cytokine levels with eosinophil counts in NP tissue specimens. Material and methods: Thirty patients with nasal polyposis (NP) (15 asthmatic and 15 non asthmatic) were included in this prospective study. Nasal secretion samples were collected from nasal cavities of all subjects. The levels of 11 cytokines (TNF- , TNF- , IL-1 , IL-2, IL-4,IL-5, IL-6, IL-8, IL-10, IL-12, and IFN- ) were measured using commercial flow cytometric kit. Eosinophils were counted in haematoxylin-and-eosin-stained NP sections. Results: The concentrations of Th2 cytokines IL-5, IL-6, IL-10, and Th1 cytokine IFN- were significantly higher in patients with NP and asthma compared with non-asthmatic subjects. A positive correlation was found between IL-6 and TNF- levels in nasal fluid and eosinophil counts in polyp tissue in non-asthmatic subjects. In asthmatic NP patients, we found positive correlation between level of IL-6 and eosinophil counts and negative correlation between IFN- level and number of eosinophils in NP tissue specimens. Conclusion: Our results showed that these patients with similar clinical findings had significantly different mediator profiles in their nasal secretions, implying clear differences in pathogenesis of their NP (AU)

Correlation between cytokine levels in nasal fluid and eosinophil counts in nasal polyp tissue in asthmatic and non-asthmatic patients.

BACKGROUND/AIMS: Concentrations of mediators in nasal secretions could reflect the inflammatory status of the nasal mucosa and evolution of sinus disease. So, the aim of our study was to evaluate local immune reaction by measuring crucial Th1, Th2 and inflammatory cytokines in nasal fluid samples of patients with nasal polyps (NP), and to correlate them to clinical, radiological findings and to the degree of eosinophil infiltration of polyp tissue. Therefore, in our study we compared the cytokine levels in nasal fluid of asthmatic and non-asthmatic patients with nasal polyposis, the eosinophil counts in NP tissues of these patients, and we correlated cytokine levels with eosinophil counts in NP tissue specimens. MATERIAL AND METHODS: Thirty patients with nasal polyposis (NP) (15 asthmatic and 15 non-asthmatic) were included in this prospective study. Nasal secretion samples were collected from nasal cavities of all subjects. The levels of 11 cytokines (TNF-α, TNF-ß, IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, and IFN-γ) were measured using commercial flow cytometric kit. Eosinophils were counted in haematoxylin-and-eosin-stained NP sections. RESULTS: The concentrations of Th2 cytokines IL-5, IL-6, IL-10, and Th1 cytokine IFN-γ were significantly higher in patients with NP and asthma compared with non-asthmatic subjects. A positive correlation was found between IL-6 and TNF-α levels in nasal fluid and eosinophil counts in polyp tissue in non-asthmatic subjects. In asthmatic NP patients, we found positive correlation between level of IL-6 and eosinophil counts and negative correlation between IFN-γ level and number of eosinophils in NP tissue specimens. CONCLUSION: Our results showed that these patients with similar clinical findings had significantly different mediator profiles in their nasal secretions, implying clear differences in pathogenesis of their NP.

Proinflammatory cytokines (IL-1beta and TNF-alpha) and chemokines (IL-8 and MIP-1alpha) as markers of peri-implant tissue condition.

Analysis of peri-implant crevicular fluid (PICF) offers a non-invasive means of studying the host response in peri-implant disease and may provide an early indication of patients at risk for active disease. This study examined the PICF levels of interleukin-1beta (IL-1beta), tumour necrosis factor alpha (TNF-alpha), interleukin-8 (IL-8) and macrophage inflammatory protein-1alpha (MIP-1alpha) in patients with non-manifesting inflammation, early and late stages of mucositis. The study group comprised 90 adult healthy volunteers with endosseal titanium implants inserted. Samples were taken from peri-implant sulcus using a filter paper technique. Implant tissues were categorized clinically as healthy, early mucositis or advanced mucositis. Clinical manifestations were determined by: gingival index and bleeding on probing, plaque index and radiographic analyses. Cytokine concentrations were assesed using commercially available enzyme-linked immunosorbent assay kits. Patients from the control group (healthy patients) have significantly lower concentrations of IL-1beta, TNF-alpha, IL-8 and MIP-1alpha in PICF compared with both groups with mucositis. Positive correlation was noted in the control group between IL-1beta and TNF-alpha and between MIP-1alpha and IL-8 in the group with early mucositis. The results suggest that cytokines could be prognostic markers of implant failure.

Production of interleukin-8 in vitro by mononuclear cells isolated from human periapical lesions.

INTRODUCTION: Interleukin-8 (IL-8) is an important mediator of inflammation. However, little is known about its production in chronic dental periapical lesions and this was the main aim of this work. METHODS: Inflammatory cells were isolated from clinically different periapical lesions and analyzed by morphological criteria. The mononuclear cells were isolated, phenotypically analyzed by immunocytochemistry and cultivated in vitro. IL-8 was measured in culture supernatants of these periapical lesion mononuclear cells (PL-MNC) using a microbeads fluorescence assay. RESULTS: We found a relatively high production of IL-8 in 19 out of 21 periapical lesions included in the study. The level of IL-8 and the proportion of neutrophil granulocytes were significantly higher in the group of symptomatic lesions, compared to the asymptomatic lesions, but there was no statistically significant correlation between these parameters. According to the predominance of CD3(+) T cells and Ig(+)/CD19(+) B cells and plasma cells, lesions were divided into T-type and B-type lesions, respectively. The levels of IL-8 were significantly higher in the culture supernatants of PL-MNC in the T-type lesions and were positively correlated with the proportion of macrophages/dendritic cells (CD11c(+) cells) and CD4(+) T cells. Such a correlation was not shown in B-type lesions. CONCLUSION: These results suggest that PL-MNC are a significant source of IL-8, which is probably an important chemokine for the migration and function of different cell types at the site of chronic inflammation.

The cleft palate patient: a challenge for prosthetic rehabilitation--clinical report.

Although cleft palate patients are not regularly seen in general dental practice, their number is not negligible. Prosthodontic treatment of such patients requires good planning that takes into account all remaining teeth and roots, deformation of maxillary segments, residual palatal defects, and the disproportion between the maxillary and mandibular alveolar ridge. With the aim to provide satisfactory function and esthetics and alleviation of the deformities, the authors describe prosthetic therapy of a cleft palate patient using root copings, attachments, telescope and cone crowns, and a metal-base partial prosthesis. The patient's mastication, phonation, and esthetics were improved. Successful results can be best achieved through the judicious use of appropriate treatment modalities tempered by clinical experience and creativity.

Skin lesions--an indicator of disease activity in systemic lupus erythematosus?

Cutaneous manifestations have great diagnostic value for systemic lupus erythematosus (SLE). In this study we tried to establish a correlation between lupus erythematosus LE-specific and LE-nonspecific cutaneous lesions and disease activity measured by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Sixty-six patients with SLE were evaluated. They were divided into three groups having: (1) only LE-specific lesions (38 or 58.46%); (2) only LE-nonspecific lesions (4 or 6.15%); and (3) both types of lesions (23 or 35.38%). Results were analyzed using the Student t-test. Patients with LE-nonspecific skin manifestations had significantly increased disease activity compared to those with only LE-specific lesions. The number of different skin lesion types also correlated with disease activity. It was significantly increased in a group with three different types of lesion, either specific or nonspecific. Patients with only one type of lesion had mild disease. An intermediate disease activity was found in the group with two different lesion types. Lupus-specific skin manifestations serve primarily as an important diagnostic clue. In conclusion, patients with LE-nonspecific lesions have significantly more active SLE than those with LE-specific lesions and may therefore require more intensive therapy and disease monitoring.

[Detection of B-lymphocyte clonality in samples of salivary gland tissue in patients with primary Sjogren syndrome]. / Detekcija klonalnosti B limfocita u uzorcima tkiva pljuvacnih zlezda bolesnika sa primarnim Sjegrenovim sindromom.

Intensive lymphoplasmocytic infiltration with atrophy of glandular tissue structures is the dominant patohistological feature found in exocrine glands of patients with Sjögren syndrome (SS). The infiltrates consist of T and B lymphocyte clusters that make the structures resembling germinal centers, and numerous plasmocytes that are secreting imunoglobulines locally, including autoantibodies. By applying the polymerase chain reaction (PCR) in our study we have shown the existence of dominant B cell clone in salivary glands samples of 4 out of 6 patients with SS, in the absence of clinical, routine laboratory, and patohistological signs of the lymphoma. B lymphocyte clones were detected upon the amplification of gene segment that encoded variable heavy chain immunoglobulin CDR3 region. Finding of single, dominant B lymphocyte clone could be of predictive significance, because these patients are predisposed to non-Hodgkin lymphoma (NHL) for which there is an assumption that it originates out of salivary glands from one of the clusters of proliferating B lymphocytes.

[Detection of K-ras and p-53 oncogene mutations in a patient with acute lymphoblastic leukemia before and after bone marrow transplantation]. / Detekcija mutacija u K-ras i p-53 onkogenima kod obolelog od akutne limfoblastne leukemije pre i posle transplanticije kostine srzi.

We present a case of 22 years old male patient, who was submitted to singenic transplantation of hematopoietic cells originating from the bone marrow in the remission phase of the diagnosed acute lymphoblastic leukemia (ALL). The bone marrow sample was donated by his healthy twin brother. The pretransplantation and transplantation phases were regular. We analyzed the presence of K-ras and p-53 point mutations in our patient with ALL and for the first time we had the opportunity to analyze the samples from two monozygotic twins. DNA was isolated from the peripheral blood mononuclear cells (PBMNC) by the standard procedure, of the patient with ALL before and after bone marrow (BM) transplantation and of his clinically healthy twin brother. Samples were subjected to PCR amplification of K-ras exons 1 and 2 and p-53 exons 5, 6, 7 and 8. In PBMNC of the patient with ALL before BM transplantation, mutations were observed in exon 1 of K-ras and exon 8 of p-53 gene. These mutations were found neither in PBMNC sample of his twin brother, nor in PBMNC of the patient with ALL after BM transplantation. In the p-53 exons 5, 6 and 7 and exon 2 of K-ras, there were no mutations in any analyzed samples. Detected mutations in K-ras and p-53 genes could be a part of larger genetic abnormalities and the obtained results had shown the possibility of using DNA mutational changes in the follow-up of the success of BM transplantation. The molecular disease marker that was found by this method was also significant for the detection of minimal residual disease at the molecular level.

The effect of tooth loss on the TM-joint articular eminence inclination.

The inclination between the posterior wall of the articular eminence and the referential plane (Frankfurt line) was measured on 137 dry skull specimens (78 with complete dental arches and 59 edentulous). Both left and right joints were measured using direct craniometric method. The results were compared with respect to the loss of teeth, side and sex. A large range of measured values of the eminence inclination shows great individual differences, so that the mean values can have only orientational significance. The mean value of eminence inclination for all measured specimens was 61.9 angular degree. The difference between the two groups established upon dental status is very small and without any statistical significance (P > 0.05). The right joint shows a slightly steeper eminence inclination compared to the left one, but without any statistical significance (P > 0.05). Asymmetry between the right and left joint appears almost as a rule, while maximum measured differences reach up to 33 degrees, with mean absolute difference of 6.9 degrees. The group of edentulous specimens shows a greater symmetry and less differences between the left and right joint, which indicates that different bimechanical conditions in the joint, due to tooth loss might lead to remodellation of the articular eminence. The eminence inclination was significantly steeper in male specimens in all subgroups (P < 0.001), which confirms sexual dimorphism.

Bond strengths of two dental bonding systems.

A new bonding system named Kevloc has been introduced. It is based on acrylization of the metal surface with the goal of preventing the occurrence of a marginal gap between the metal and the resin. The purpose of this investigation was to determine the values of the shear bond strength achieved using the Kevloc technique on Ag-Pd (Auropal SE) and Co-Cr (Basil S) alloys and to compare them with those obtained with the OVS technique. The shear bond strengths were measured with the Smitz-Schulmayer shear test in a universal testing machine for polymer materials. A microscope image analyzer was used to measure the thickness of bonding layers and to reveal the possible occurrence of the marginal gap with both techniques. No marginal gap was detected with either technique. Kevloc provided better results than OVS only in a group of specimens tested after polymerization. Immersion in water and thermocycling reduced the initially high bond strength values of Kevloc specimens, whereas the bond strength values of OVS specimens remained unchanged regardless of which aging treatment was used. Microscopic examination did not reveal the existence of the marginal gap for either bonding system. According to the results obtained, it can be concluded that the Kevloc bonding system does not provide better shearing bond strength than the OVS bonding system.

Donor leukocyte infusion--the effect of mutual reactivity of donor's and recipient's peripheral blood mononuclear cells on hematopoietic progenitor cells growth.

Donor leukocyte infusions are an effective therapy for patients who relapse with leukemia after bone marrow transplantation. We report the case of 14-year-old boy who relapsed 34 months after sibling donor bone marrow transplant for Philadelphia-positive chronic myeloid leukemia. Subsequently, he received three infusions of donor mononuclear cells (DMNC) harvested in steady state hematopoiesis and one G-CSF mobilized-peripheral blood mononuclear cells (PBMC) infusion. Simultaneously, test named as--"Test of Mixed Progenitors" (TMP) was performed for the assessment whether the outcome of donor leukocyte infusion treatment could be predicted. Prior to DMNC infusions, the CFU-GM and BFU-E colony assays were performed for donor's and recipient's PBMC individually, as well as for the mixture of these cells at 1:1 ratio. The cells were plated either directly in the semisolid medium or after 24 h preincubation treatment. Significantly lower values for CFU-GM derived colonies were determined in TMP in comparison to the CFU-GM values obtained for the recipient's cells. The reduced number of CFU-GM was determined both in TMP performed without preincubation treatment, app. 80% and after the 24 h preincubation, app. 55%. The reduced number of BFU-E derived colonies (app. 44%) was observed only related to recipient's cells and after the preincubation treatment of the cells. The patient did not develop GVHD and currently (40 months after the first infusion). He remained well in complete hematological, cytogenetic, molecular and clinical remission, which was the most direct evidence of the GVL effect. The novel in vitro TMP test in which the specific contribution of donor's leukocytes to the growth of recipient's hematopoietic precursor cell growth was determined, correlated with the clinical outcome.

Prosthetic rehabilitation of a warfare facial trauma.

This case report presents the prosthetic treatment of a warfare facial trauma patient with the aim of providing satisfactory function, esthetics, and alleviation of deformities. Veneer crowns with rests, root coping, and metal-base partial prostheses were used for the patient therapy and treatment.

Bond strengths of silicoated and acrylic resin bonding systems to metal.

STATEMENT OF PROBLEM: The Kevloc bonding system is based on the bond of acrylic resin with the metal surface, in contrast to the Silicoater system, which covers the metal surface with a SiOx-C layer. Bonding prevents the occurrence of the marginal gap between the metal and the resin, which causes weaker bond strengths and discoloration. PURPOSE: This investigation examined the bond strength values achieved by using the Kevloc technique on Ag-Pd and Co-Cr alloys, and compared them with those obtained by the Silicoater technique. After artificial aging, shear testing of the specimens was performed. MATERIAL AND METHODS: A microscope image analyzer measured the thickness of bonding layers to reveal any marginal gap. RESULTS: No marginal gap was noticed for either technique. The Kevloc technique provided better results than Silicoater regardless of alloy used. Immersion in water (7.64 to 8. 05 MPa) and thermocycling (6.12 to 6.95 MPa) reduced the initial bond strength values (12.52 to 12.94 MPa), but the breakdown occurred between the opaque layers or between the opaque and the resin, thus giving a cohesive type of failure. CONCLUSION: The dental alloy used did not affect bond strengths and artificial aging caused reduction in bond strengths. The Kevloc bonding system exhibited higher bond strengths than Silicoater, but the marginal gap was not seen for either bonding system.