Effect of Annona muricata aqueous leaf extract on reactive oxygen and nitrogen species.

OBJECTIVE: Annona muricata extracts are used in traditional medicine due to their significant biological effects. Verification and elucidation of their mechanisms is beneficial in terms of the usefulness of these extracts in everyday life or in the context of disease treatment or prevention. MATERIALS AND METHODS: The effectiveness of the extract was assessed from dried A. muricata leaves available for direct consumption. It is targeted against reactive oxygen and nitrogen species such as superoxide (O2•-), hydroxyl (•OH), nitric oxide (NO) radicals, and peroxynitrite anion (ONO2-) at concentrations of 5, 10, 25, 50, 100 µg.ml-1. RESULTS: No significant inhibitory activity was measured against O2·- at the assessed concentrations of the extract. Conversely, substantial antioxidant properties were found towards ·OH. Moreover, very efficient uptake was recorded at low concentrations of the extract, 5 µg.ml-1 (53.91%) and 10 µg.ml-1 (45.3%). The antioxidant effect decreased with increasing concentration. By indirect determination of NO oxidation derivatives it was found that, as the extract concentration increased, the nitrite concentration decreased. In contrast, even at low concentrations, the extract causes an increase in the peroxynitrite concentration. CONCLUSIONS: The results themselves show that the effects of A. muricata leaf extract are mainly mediated by the activity against •OH, as well as the consequences of increased ONO2- formation.

Expression of selected inflammatory proteins and metalloproteinases in periodontitis.

OBJECTIVE: Periodontitis is a chronic inflammatory disease caused by microbial dental plaque which leads to the destruction and loss of supporting tissues of the tooth. Microbial plaque alone, however, is not enough to cause the disease. The body's response plays an important role, in which an imbalance between the pro-inflammatory and anti-inflammatory effects of cytokines leads to an inflammatory reaction. PATIENTS AND METHODS: We detected changes in mRNA expression and protein levels of MIP-1α, and metalloproteinases (MMP-2, MMP-9) contributing to cascades in the initiation and progression of inflammatory bone resorption and destruction of periodontal soft tissues in patients with aggressive (AP) or chronic (CP) forms of periodontitis in comparison with healthy individuals (control). RESULTS: MIP-1α mRNA levels were highest in AP (280 ± 23% higher than the control) also in comparison with CP. The difference in protein level was less pronounced. MMP-2 mRNA expression values were similar (300 ± 12% higher in comparison with control), but protein levels were lower, also when compared to CP. Only in CP MMP-9 mRNA levels were significantly higher than the control (30 ± 8%), while protein levels were again higher in AP. Both AP and CP showed a positive correlation between the level of MIP-1a and MMP-2 (0.879, and 0.954 respectively). However, a strong positive correlation was only found between the levels of MMP-2 and MMP-9 in CP (0.812). CONCLUSIONS: MIP-1α, MMP-2 and MMP-9 mRNA expression, along with the concentration of proteins in saliva in patients with periodontal disease, is higher than in healthy individuals and correlates with the severity of the disease.

CER 1 gene polymorphism in postmenopausal Roma and non-Roma Slovak women in connection with osteoporosis.

OBJECTIVE: The aim of our study was to investigate the relationship between the rs74434454 polymorphism of the CER1 gene and selected biochemical, densitometric and anthropometric markers in Slovak postmenopausal women of two ethnic groups: Roma and non-Roma. SUBJECTS AND METHODS: The scientific study included 303 postmenopausal women of the non-Roma and Roma populations who were divided into two groups based on densitometric measurements: control group (CG) and osteoporotic group (OG). Genomic DNA was isolated from peripheral blood using a commercial NucleoSpin® Blood kit following a standard protocol. The TaqMan Real-Time PCR method was used for genotyping. Biochemical markers were measured with Cobas e411 and Cobas Integra400 plus analysers. RESULTS: In the control group of postmenopausal Roma women, the occurrence of the risk genotype GG was not observed. In the group of Roma women with osteopenia and osteoporosis, the GG genotype occurred at a frequency of 3.03%. In the group of non-Roma women (between CG and OG) statistically significant differences were found in all monitored biochemical markers except CTx-I (p<0.66). In contrast, in the group of Roma women, statistical significance was only found in the osteoresorption marker CTx-I (p<0.007). In the population of Roma women, we did not find a statistically significant difference between the AA, AG and GG genotypes in any of the monitored markers. CONCLUSIONS: The results provide the first and unique insight on the distribution of genotypes and alleles of the rs74434454 CER1 gene polymorphism and its relationship to markers of bone metabolism in two ethnically distinct groups.

Possible prognostic biomarkers of periodontitis in saliva.

OBJECTIVE: While both first-line antioxidant enzymes and oxidation products have been considered as markers of periodontal disease, their assessment in the diagnosis of periodontal disease is more complicated. Some, such as superoxide dismutase (SOD, glutathione peroxidase (GPx) and reduced glutathione (GSH), have indicated significant differences between patients with chronic and aggressive periodontitis. PATIENTS AND METHODS: Participants (101) were divided into a control group of healthy individuals and, following diagnosis, patients with gingivitis, chronic periodontitis, and aggressive periodontitis. Compounds reflecting tissue destruction, inflammatory processes or antioxidant responses, such as sirtuins (SIRT-1, SIRT-2), metalloproteinases (MMP), SOD, GPx, GSH, and glutathione reductase (GR) were measured in saliva. RESULTS: SIRT-2 levels were significantly increased in all patients. In patients with gingivitis, MMP (p<0.05) and GPx (p<0.01) were significantly increased. In patients with chronic and aggressive periodontitis, SOD activities were increased (p<0.001) while GPx and GR were decreased (p<0.001). Relative activities of MMP were higher in patients with aggressive periodontitis. CONCLUSIONS: Measurements of SIRT-2 and SOD clearly showed increased levels of oxidative stress in cases of periodontitis with a subsequent inhibition of other antioxidant enzymes. Levels of GSH suggest reversibility of the conditions with appropriate intervention. With the assessment of the trends of these selected antioxidant markers, it is possible to determine the prognosis of the disease.

Preliminary results of ethnic divergence of G1181C (rs2073618) and C290T (rs9525641) OPG gene polymorphisms in groups of postmenopausal Slovak women.

OBJECTIVE: In this study, we focused on observation of the genetic polymorphisms of the OPG genes G1181C (rs2073618) and C290T (rs9525641), their interactions with biochemical markers and anthropometric parameters in groups of postmenopausal Slovak women (Roma and non-Roma, n = 311). PATIENTS AND METHODS: Genomic DNA was extracted and purified from peripheral blood leukocytes by the kit Ultraclean® Blood non-spin® (Carlsbad, CA, USA) using a standard protocol. Genotyping was performed by the TaqMan SNP genotyping assay. Biochemical markers were measured by the Cobas e411 (Roche Diagnostic, Tokyo, Japan) and Cobas Integra400 plus (Roche Diagnostic, Rotkreuz, Switzerland) analysers. RESULTS: We recorded a higher frequency of the T allele in the C290T polymorphism of the non-Roma control group (53.846%), in Roma groups: control (T - 56.618%) osteoporotic (T - 51.471%). In the G1181C polymorphism, the CC genotype occurred more in the osteoporotic group (34.286%) compared to the control group (27.885%). In the group of postmenopausal Roma women, a statistically significant difference (p<0.05) was found between osteoporotic and control in the biochemical parameters' osteocalcin, C-terminal telopeptide I, and age. Statistically significant differences (p<0.0001) were also found in bone mineral density and T-score. The high odds ratio suggests the association of G1181C with osteoporosis. A close relationship was found between haplotypes, BMD, T-score, and IL-6 in control; and BMI, WHR, T-score, and osteocalcin in osteoporotic groups of Roma and non-Roma women. CONCLUSIONS: The results point to differences in the occurrence of genotypes and associations of haplotypes with the manifestation of osteoporosis in Roma and non-Roma women. However, a larger number of samples is needed to determine whether or not there are differences between the Roma and non-Roma populations.

Epiduroscopic intervention in patients with a failed back surgery syndrome.

BACKGROUND: Epiduroscopy is a percutaneous endoscopic technique with an interesting diagnostic and therapeutic potential. AIM: The study was aimed to evaluate the effectiveness of this modern minimally invasive method in treating pain together with the accompanying neurological symptoms of patients with a failed back surgery syndrome (FBSS). RESULTS: In the retrospective multicentre clinical study, we monitored the effect of epiduroscopic performance in 40 patients with FBSS before epiduroscopy and after 6 and 12 months. The results of the study indicated a significant improvement in back pain after 6 months, at p<0.05. At the same time, a significant improvement was observed in pain radiating to the lower limbs after 6 and 12 months at p<0.05, as well as a significant improvement in the Oswestry Disability Index of the limbs after 6 months and after 12 months (p<0.05). CONCLUSION: The current summarised results indicate a significant positive effect of epiduroscopy in the treatment of chronic back pain with a reduction in radiating pain to the lower extremities and an increase in patients' quality of life after the procedure (Tab. 2, Fig. 1, Ref. 23).

Endoscopic discectomy as an effective treatment of a herniated intervertebral disc.

OBJECTIVE: Prospective observational multicentre two-arm parallel study describing clinical outcome after endoscopic discectomy provided via transforaminal and interlaminar approach. BACKGROUND: Endoscopic lumbar discectomy (ELD) is a percutaneous minimally invasive procedure for the treatment of herniated lumbar discs. Herniations at lumbar intervertebral disc levels of L1/2, L2/3, L3/4 and L4/5 are mostly accessed by the transforaminal (TF) approach. However, due to the anatomic position of the iliac crest, the L5/S1 level might not be reachable by the transforaminal approach, while the interlaminar (IL) percutaneous approach should be a suitable alternative. METHODS: In a prospective observational multicentre clinical trial NCT0274311, we compared the clinical outcomes of two groups of patients who underwent ELD via IL (83) and TF (103) approach. The subjects were followed for 12 months via planned examinations by pain physicians. The levels of leg pain and back pain intensity were assessed by an 11-point numerical ratings scale (NRS). Patient's functional disability was assessed by the Oswestry Disability Index (ODI). RESULTS: Study subjects showed a significant decrease in ODI scores in both groups (p<0.001) The values of mean preoperative ODI in TF and IL groups were 39.1±15.7 and 43.4±16, respectively. Postoperative values in the latter groups were 14.8±14.9 and 17.5±14.3, respectively. Significantly lower pain scores for leg pain (p<0.001) and back pain (p<0.001) were also recorded at 12-month follow-ups. CONCLUSION: Because both procedures are strictly percutaneous; they are now more commonly performed by interventional pain physicians as a safe and effective alternative to open surgical spine procedures (Tab. 3, Fig. 7, Ref. 19).

Incidence and Prognostic Value of Known Genetic Aberrations in Patients with Acute Myeloid Leukemia--a Two Year Study.

BACKGROUND: In this work, we evaluated the incidence and prognostic value of several genetic aberrations in patients with a diagnosis of acute myeloid leukemia (AML). PATIENTS AND METHODS: We analysed 90 patients: 42 males (mean age 54.5 years) and 48 females (mean age 59 years), with AML. The genetics of all leukemia samples was studied using conventional cytogenetics, the interphase fluorescence in situ hybridisation as well as the standardized RTPCR protocol. RESULTS: In 34.4% of patients, we detected at least one of the analysed genetic aberrations, except the CBFB MYH11, which we did not detect. Translocation t(8;21)/ AML1 ETO was found in 4.4% of patients with a mean age of 45.4 years, while none of these patients was older than 55 years. Translocation t(15;17)/ PMLRARA was found in 5.5% of patients with a mean age of 52.6 years and an almost equal distribution between younger and older patients. The MLL gene rearrangements were found in 6.6% of patients, the -5/ 5q- and/ or -7/ 7q- aberrations in 7.7% of patients, while the most frequent genetic abnormality in our study was trisomy 8 (10%). Moreover, we found a favorable clinical outcome in patients expressing fusion genes AML1-ETO or PMLRARA in contrast to an adverse clinical outcome with few remissions and death in AML patients with MLL, -5q/ -5 and -7q/ 7-. Finally, an intermediate prognosis was found in patients with trisomy 8. CONCLUSION: In this study, we found a good congruence with published literature on the incidence and prognostic value of several well established AML-associated genetic aberrations. This simple genetic-based classification system helps us to identify patients with a favorable, intermediate or unfavorable prognosis and to treat them with the best currently available therapy. However, analysis of new genetically defined abnormalities in AML is necessary for development of better therapeutic strategies and/or diagnostics.

Antioxidant potential of Aesculus hippocastanum extract and escin against reactive oxygen and nitrogen species.

OBJECTIVE: Antioxidant, anti-inflammatory and venoconstrictor properties have been attributed to extracts from Aesculus hippocastanum. These unusual and diverse properties may be possibly basically linked with ability to scavenge free radicals. MATERIALS AND METHODS: The scavenging capacity of dry horse chestnut extract of and escin have been investigated in vitro against superoxide anion radicals, hydroxyl radicals, nitrites and peroxynitrite. RESULTS: In general, the activity of the whole extract against superoxide radicals did not exceed 15% at pH 7.4, but the highest inhibition (46.11%) was recorded against hydroxyl radicals at a concentration of 100 µg.ml-1; however, the activity against other radicals was lower. Escin demonstrated a better ability to counteract nitric oxide oxidation products, nitrites. However, the efficiency of the whole extract completely disappeared as the concentration increased. Both extracts showed very low activity towards peroxynitrite. Escin was even able to induce peroxynitrite formation at the lower concentrations used. CONCLUSIONS: Whole extract showed better antiradical properties compared to its main active ingredient, escin, probably due to potential synergistic interaction with a mixture of compounds present in the plant extract. These findings can be the basis of both the presentation of side-effects and the persistence of disease in spite of ongoing treatment.

Effect of dietary microbially produced gamma-linolenic acid and plant extracts on enzymatic and non-enzymatic antioxidants in various broiler chicken organs.

Plant extracts and fungal fermented feed with gamma-linolenic acid-rich microbial oils are perspective additives for use in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for the prevention and treatment for certain pathological conditions, and as antioxidants. The activity of antioxidant enzymes and the level of reduced glutathione were measured in the plasma and in liver, heart and kidney mitochondria after 42 days of feeding broiler chickens both regular and combination diets. These were selected based on our previous experience. The administration of agrimony and gamma-linolenic acid resulted in a significant decrease in superoxide dismutase activity in all four bodies in contrast to plant extracts. We conclude that the decrease in activity is due to decreased production, and hence dismutation, of superoxide radicals to peroxides followed by lower activity of glutathione peroxidase, which was not seen in the case of only plant extract administration. Generally, higher glutathione reductase activity would be in response to increased demands on reduced glutathione as a cofactor for the reaction catalysed by glutathione peroxidase and the utilization of glutathione itself. However, measured levels of reduced glutathione showed no change. The results argue against any oxidative stress conditions. The application of agrimony extract appears to be suitable for the antioxidant effect against peroxidation of gamma-linolenic acid. As the efficacy of measuring the effects of diets on the oxidative stability of meat caused by selected antioxidant enzymes is rather low, additional data from the experiment will be processed to clearly assess the influence of this combination of diets.

Cytogenetic abnormalities predict treatment-free interval and response to therapy in previously untreated chronic lymphocytic leukemia patients.

We evaluated the prognostic impact of chromosomal abnormalities as detected by interphase fluorescence in situ hybridization (iFISH) in 86 chronic lymphocytic leukemia (CLL) patients. Overall, 39 of 86 (45%) patients displayed one (35%) or more (10%) chromosomal abnormalities, del13q (31%) being more frequently detected than trisomy 12 (19%) followed by del11q (17%), del17p (6%) and del6q (5%). Significant differences in the treatment free intervals (TFIs) were observed among individual cytogenetic subgroups (p=0.027) with the shortest mean TFIs in subgroups with del17p, del11q and trisomy 12 (10, 12 and 14 months, respectively) as compared to subgroups with normal cytogenetics (38 months) and del13q (68 months). Poor response to therapy was observed in subgroups with del11q (p=0.044) and trisomy 12 (p=0.047) while patients with normal cytogenetics had good response (p=0.003). Furthermore, del17p and del11q were associated with highest tumor burden and disease activity as reflected by corresponding laboratory data.

[Selenium supplementation in patients with severe acute pancreatitis]. / Suplementácia selénu u pacientov s t'azkou akútnou pankreatitídou.

UNLABELLED: Selenium supplementation improves antioxidant status in critically ill patients with severe acute pancreatitis. It depends on quantum of dosage and supplementation time. The aim of this analysis is point out on benefit of antioxidant therapy by supplementing selenium. METHODS: Patient with severe acute pancreatitis and developing septic shock was admitted on anesthesiology and intensive care department. Adjuvant supplementation therapy with selenium was started in continual infusion 750 mg/24 h during next six days. Activity of antioxidant enzyme glutathione peroxidase and others inflammatory markers were decrease. A case report presents the possibility to affect on systemic inflammatory response syndrome pathogenesis in initial phase. It has to improve therapeutic progress in patients with severe acute pancreatitis.

Anomalous maxillary lateral incisor--the only product of dental lamina, difficult classifiable in the time. Symptom of hypodontia syndrome non-described until present time.

The authors described anomalous development of maxillary lateral incisor non-yet published in stomatologic literature. Diagnostics and clinical findings of the anomaly and the prognosis of the affected tooth they based on the description of six longitudinal observed patients. The principal symptom of the anomaly is the disturbance of time-plan of the tooth development. Terms of the tooth mineralization, the time of eruption and root completion cannot be included either in deciduous or in permanent dentition. The tooth is the only product of the dental lamina; it has no predecessor or successor. Frequent disturbances of the eruption pathway result often in anomalous position of the tooth. Taking into consideration the fact that the anomaly appeared always in connection with hypodontia syndrome, it is possible to consider it to be the symptom of hypodontia syndrome. This fact is more significant as it enables to establish the diagnosis of hypodontia syndrome already in early deciduous dentition.

[Analysis of environmental indicators in relation to structural disorders of dental tissues]. / Rozbor nekterých ukazatelu zivotního prostredí ve vztahu k poruchám strukturyzubních tkání.

An epidemiological investigation revealed a high incidence of structural disorders of dental tissues in the child population of Prague. While hypoplastic disorders are steadily diminishing, there is an ever rising trend of hypomineralization. The observed disorders of hypomineralization are becoming a serious clinical problem. The aim of the present investigation was to assess to what extent exogenous harmful substances participate in the development of hypomineralization: infectious diseases, tetracycline antibiotics, excess fluorine intake and type of drinking water--from the water-mains or wells. The data were assembled by means of questionnaires. Two clinical investigations comprising 300 children, mean age 13 years, suggest that among the investigated indicators only the type of drinking water can be associated with the development of the observed changes. Children drinking from birth water from wells had significantly fewer disorders of mineralization than children who drank water from the water-mains. The submitted paper presents partial results of the state research plan of technical development.

[Comparative study of the prevalence of disorders of impaired structure of dental tissues in school children and university students]. / Srovnávací studie prevalence poruch struktury zubních tkánki u skolnich detí a vysokoskolských studentu.

The authors examined a group of 573 Prague children aged 12-15 years. In addition to the incidence of disorders of the structure of dental tissues they investigated the coriosity (DPE), oral hygiene (OHI) and condition of the gingiva (PI). Their main interest was focused on impaired mineralization of dental tissues the incidence of which is indemic in Prague children. The group was compared with 62 university students. It is of interest that in all indicator except impaired mineralization, the children had significantly better results. The rising curve of impaired mineralization signalises the ever deteriorating living environment in the capital.