Heart failure in patients is associated with downregulation of mitochondrial quality control genes.

BACKGROUND: Mitochondrial dysfunction is one of key factors causing heart failure. We performed a comprehensive analysis of expression of mitochondrial quality control (MQC) genes in heart failure. METHODS: Myocardial samples were obtained from patients with ischemic and dilated cardiomyopathy in a terminal stage of heart failure and donors without heart disease. Using quantitative real-time PCR, we analysed a total of 45 MQC genes belonging to mitochondrial biogenesis, fusion-fission balance, mitochondrial unfolded protein response (UPRmt), translocase of the inner membrane (TIM) and mitophagy. Protein expression was analysed by ELISA and immunohistochemistry. RESULTS: The following genes were downregulated in ischemic and dilated cardiomyopathy: COX1, NRF1, TFAM, SIRT1, MTOR, MFF, DNM1L, DDIT3, UBL5, HSPA9, HSPE1, YME1L, LONP1, SPG7, HTRA2, OMA1, TIMM23, TIMM17A, TIMM17B, TIMM44, PAM16, TIMM22, TIMM9, TIMM10, PINK1, PARK2, ROTH1, PARL, FUNDC1, BNIP3, BNIP3L, TPCN2, LAMP2, MAP1LC3A and BECN1. Moreover, MT-ATP8, MFN2, EIF2AK4 and ULK1 were downregulated in heart failure from dilated, but not ischemic cardiomyopathy. VDAC1 and JUN were only genes that exhibited significantly different expression between ischemic and dilated cardiomyopathy. Expression of PPARGC1, OPA1, JUN, CEBPB, EIF2A, HSPD1, TIMM50 and TPCN1 was not significantly different between control and any form of heart failure. TOMM20 and COX proteins were downregulated in ICM and DCM. CONCLUSIONS: Heart failure in patients with ischemic and dilated cardiomyopathy is associated with downregulation of large number of UPRmt, mitophagy, TIM and fusion-fission balance genes. This indicates multiple defects in MQC and represents one of potential mechanisms underlying mitochondrial dysfunction in patients with heart failure.

Drug-Eluting Balloons-A New Tool in the Treatment of Cardiac Allograft Vasculopathy: A Case Series.

Percutaneous coronary intervention in patients with cardiac allograft vasculopathy is burdened with a lot of difficulties. Although they have allowed significant progress in comparison with plain balloon angioplasty and bare metal stents, drug-eluting stents have not fully overcome problems of diffuse lesions and small-vessel disease that are so common in transplant coronary artery disease. There is growing evidence that drug-eluting balloons might be a better choice for patients with small vessel atherosclerotic coronary disease and yet there is no experience with this technology in patients with cardiac allograft vasculopathy. Herein we report a case series of successful percutaneous coronary interventions in patients with cardiac allograft vasculopathy.

PPARγ and IL-6 - 174G>C gene variants in Croatian patients with ischemic stroke.

AIM: Etiology of ischemic stroke (IS) is multifactorial and includes interaction of genetic and environmental factors. Different genes, their polymorphisms, host susceptibility, and inflammation processes play a role in IS development. The aim of this study was to evaluate the effect of PPAR-γ and IL-6 gene variants on IS onset. MATERIAL AND METHODS: A total of 301 subjects (144 males, 157 females) participated in the study, 114 patients with IS and 187 healthy controls. RESULTS: Statistically significant predictors of IS were male gender (OR 7.13, 95% CI 2.92-17.39, p<0.001), hypertension (OR 7.82. 95% CI 2.53-24.19, p<0.001), lowered HDL cholesterol (OR 8.20, 95% CI 2.41-27.94, p=0.001), elevated C-reactive protein (OR 5.26, 95% CI 1.92-14.41) and IL-6 -174 GC (OR 2.44 95% CI 1.01-5.91, p=0.0048) genotype. Males, compared to females, had 7 times higher odds for stroke. IL6 -174G/C genotype increased the odds for IS for 2.4 times. PPARγ was not statistically significantly associated with stroke. CONCLUSION: We can point to the IL-6 -174G>C polymorphisms as candidate gene marker and risk factor for the prediction of ischemic stroke.

Incidental extraction of a temporary epicardial pacemaker wire with right ventricular perforation during endomyocardial biopsy: a case report.

INTRODUCTION: Right ventricular perforation during endomyocardial biopsy is an unusual, although potentially life-threatening, complication caused with the tip of the bioptome. The majority of perforations in heart transplant patients can be managed without surgery owing to adhesions nearly obliterating pericardial space. CASE REPORT: We report a case of heart transplant patient who suffered right ventricular perforation as a consequence of incidental extraction of a temporary epicardial pacemaker wire during a routine endomyocardial biopsy sampling. CONCLUSIONS: The patient suffered no clinical consequences.

Dipeptidyl peptidase III in malignant and non-malignant gynaecological tissue.

Exopeptidases, in contrast to endopeptidases (proteinases) have been much less studied in relation to cancer. The aim of this study was to investigate one such enzyme, dipeptidyl peptidase III (DPP III), in gynaecological tissues, by measuring both the enzyme activity and enzyme content. DPP III activity was assessed in normal (n = 65), benign (n = 9) and malignant (n = 51) gynaecological tissues. A statistically significant higher DPP III activity was observed in endometrial (n = 40, P = 4.6 x 10(-7)) and ovarian (n = 11, P = 8.1 x 10(-4)) malignant tumours, whereas no significant difference was detected for leiomyomas (n = 8), if compared to the activity in normal tissue. A matched pair analysis of normal and cancerous endometrial tissue confirmed the significance of the DPP III activity increase in the transformed tissue (n = 7, P = 0.022). Western blot analysis revealed a significantly (P = 0.014) increased level of DPP III in endometrial cancer. Further, regression analysis showed a positive correlation between the activity and the content of DPP III in normal tissue (r = 0.637, P = 0.047) and in endometrial cancer (r = 0.574, P < 0.007). The increase of the DPP III activity was observed in the endometrial carcinomas of various histological types, grade or the depth of myometrial invasion. The easy-to-perform determination of this exopeptidase activity may serve as a potential indicator of endometrial and ovarian malignancies.

Epidemiology of Crohn's disease in Zagreb, Yugoslavia: a ten-year prospective study.

A ten-year prospective study of Crohn's disease was carried out in Zagreb, Yugoslavia. It included both inpatients and outpatients regardless of the extent and severity of the disease. The mean annual incidence rate was 0.7 per 100,000 between 1 January 1980 and 31 December 1989. There was no increase in the incidence of Crohn's disease during the study period. The prevalence of Crohn's disease was 8.3 per 100,000 on 31 December 1989. The results confirm the low frequency of Crohn's disease in central and southern Europe.

Ulcerative colitis in Zagreb, Yugoslavia: incidence and prevalence 1980-1989.

We present a ten-year incidence of ulcerative colitis in Zagreb, Yugoslavia. The study included both outpatients and inpatients regardless of the extent and severity of the disease. The mean annual incidence rate was 1.5 per 100,000 inhabitants for the period of 1 January 1980 through 31 December 1989. There was no increase in the incidence of ulcerative colitis during the study period. A prevalence rate estimate of 21.4 per 100,000 inhabitants was based on July 1985 official estimated population. The results confirm the low frequency of ulcerative colitis in central Europe.

Nutritional influences on growth and prevalence of anaemia in children during the war in Sarajevo.

OBJECTIVE: To assess growth and prevalence of anaemia in hospitalised children during the war in Sarajevo. DESIGN: A random sample of children who were patients in the paediatric clinic over a period of six months (1 June to 31 December 1993) were anthropometrically measured and blood samples taken to determine prevalence of anaemia. SETTING: The study was conducted at the paediatric clinic, Kosevo Hospital, Sarajevo. SUBJECTS: The sample comprised 542 children aged from 0 to 14 years of whom 50.6% were boys and 49.4% were girls. INTERVENTIONS: The children were weighed and had their heights measured. Nutritional status was assessed through calculating the weight for height percentile and comparing these with international standards. Blood samples were taken and low haematocrit and haemoglobin values were used to indicate nutritional anaemia. The cut-off points used were: haematocrit < 0.34 (6 months-5 years) and < 0.35 (6-14 years); haemoglobin < 105 g/l (6 months-5 years) and < 110 g/l (6-14 years). RESULTS: On average, 69% of boys and 71% of girls fell within the 10th to 90th percentile range (weight for height). 7% of boys and 5% of girls were below the 5th percentile. A total of 24.7% children aged from six months to five years and 16% of children aged from six to 14 years had haematocrit values below the cut-off points. The highest percentage of children with low haematocrit values came from the six months to one year age group. Anaemia as defined by haemoglobin value was diagnosed in 10.6% of children aged from six months to five years and 9.6% of six to 14 year olds. CONCLUSIONS: War and disruption of the food supply have not had the expected negative impact of child growth and prevalence of anaemia. Reasons for this may include: increased levels of breast feeding, distribution of supplementary food commodities enriched with micronutrients, and good parental care.

Glutathione S-transferase activity as an early marker for malignant tumors of corpus uteri.

Glutathione (GSH) concentrations and glutathione S-transferase (GST) activities were determined in 30 paired malignant human corpus uteri tumor samples and in samples of adjacent normal tissues. For GSH concentrations no difference was found between normal (126.0 +/- 52.4 nmol/mg protein) and tumor tissues (110.1 +/- 46.4 nmol/min/mg protein; p = 0.219). The GST activities were significantly higher in tumor tissues (322.4 +/- 135.54 nmol/min/mg protein) than in corresponding normal tissues (224.6 +/- 95.64 nmol/min/mg protein; p = 0.005). This activities were independent on the pathohistological and clinical factors, except for positive lymphovascular invasion and myometrial invasion (over 50%), where significantly lower GST activities were found. For normal tissues the positive correlation between GSH concentrations and GST activities was found (correlation coefficient = 0.50, p = 0.005), but not for tumor tissue (correlation coefficient = 0.20, p = 0.281). The prognosis of patients (according to the well established prognostic factors, such as tumor type, myometrial invasion and grades) who had lower GSH concentration and GST activity in normal tissue was similar to those with higher GSH concentration and GST activity. In conclusion, higher GST activities found in tumor of corpus uteri suggest, that GST activity could be used as a tumor marker for the early stages of these malignant tumors.

Plasminogen activator inhibitor type 2: potential prognostic factor for endometrial carcinomas.

The clinical determination of proteases which are involved in carcinogenesis, invasion and metastasis may contribute to the detection of the early stage of disease, and to the prognostic assessment of patients with the cancer. The aim of the present study was to determine the level of urokinase plasminogen activator (uPA), plasminogen activator inhibitor type 1 (PAI-1) and plasminogen activator inhibitor type 2 (PAI-2) in normal and malignant tissues of corpus uteri and to evaluate the possible correlation with clinical and histopathological prognostic factors. UPA, PA-I and PAI-2 were determined by the ELISA assay in tissue cytosol of matched pair samples from 27 patients with endometrial carcinoma. Results show that significantly higher levels of these proteins were found in malignant than in normal tissue samples (uPA: 1.266 versus 0.633 ng/mg protein, PAI-1:4.468 versus 1.958 ng/mg protein, and PAI-2:3.428 versus 0.483 ng/ml protein). The levels of uPA and PAI-1 did not correlate with clinical staging or pathohistological grading. However, in tumor tissues with clinical stages II and III, myometrial invasion > 50%, and lymphovascular invasion, increased levels of PAI-2 were determined. Our results indicate that components of the plasminogen activation cascade are up-regulated in endometrial cancer and suggest the role of PAI-2 in determining invasive potential of endometrial carcinomas.

Cardiac myxoma: diagnostic approach, surgical treatment and follow-up. A twenty years experience.

At the University Department of Cardiovascular Surgery in Zagreb, Croatia, we treated 81 patients with primary intracardiac myxoma, in a period from January 1975 to December 1994. There were 55 female and 26 male pts, in age from 1 month to 80 years, mean 46+/-15 years. Clinical manifestations varied from no symptoms and very poor or no clinical signs to various manifestations of chronic or acute congestive heart failure, syncope and arrhythmias with or without systemic findings such as high erythrocyte sedimentation rate, anaemia, leucocytosis, elevated gamma globulin, thrombocytopenia or low grade fever, as well as cerebrovascular accidents due to tumour embolization. Cardiac symptoms were predominant in 54 pts (66.6%) and cerebrovascular in 20 pts (24.7%). Seven pts (8.6%) were symptomless and discovered accidentally, mostly regarding on an unexplained heart murmur. In almost all the patients preoperative diagnosis of intracardiac myxoma was sufficiently established by echocardiography. The tumour was located in the left atrium in 62 pts (76.5%) and in the right atrium in 19 pts (23.5%). Delay from the onset of symptoms to the diagnosis was 6 months in average (range 10 days to 25 months). The average waiting for the operation was 9 days (range from 1 to 60 days). The echocardiographic diagnosis was confirmed during intraoperative examination followed by histological analysis. All pts underwent excision of myxoma using cardiopulmonary bypass with core and topical hypothermia and cold crystaloid cardioplegia. According to the additional preoperative and intraoperative findings, in 6 pts sinchronous mitral valve reconstruction, in 3 pts artificial mitral valve implantation and in 2 pts atrial wall reconstruction was performed. There was no perioperative mortality. After the operation, we could not evaluate all the patients long enough, mostly because of some paramedical circumstancies, such as war, migrations, etc. Twenty two pts undevent evaluation for at least 5 years after the operation. Among them there was no evidence of the tumour recurrence, 15 pts were asymptomatic and 7 had NYHA II class symptoms. For 17 pts with a left atrial myxoma preoperative and postoperative echocardiographic data were available for comparison, showing a significant reduction of the left atrial diameter (p<0.001) during the postoperative follow-up. Our data, presenting one of the biggest reports concerning cardiac myxomas, showed a broad spectrum of their clinical presentation, importance of echocardiography in diagnosing and postoperative follow-up and efficacy of a proper surgical intervention as a definite, curative therapy since there were no deaths and no significant cardiac dysfunction neither tumour reccurrence as well.

A method for the repair of vesicovaginal fistulas.

OBJECTIVE: To determine the efficacy of the sliding layers method for repairing vesicovaginal fistulas caused by surgical trauma. METHOD: Twenty one women suffering from vesicovaginal fistulas after surgical trauma, operated on between 1990 and 1995 using the sliding layers method, were included in an uncontrolled prospective study and followed 24 months after the procedure. Patient outcome was assessed according to subjective estimation, gynecological examination and objectively by instillation of diluted methylene blue to the urinary bladder. RESULTS: Sixteen out of 21 patients suffered vesicovaginal fistulas after being subjected to abdominal hysterectomy and five patients after vaginal surgery. Fourteen fistulas were located 5 to 10 mm over the interureteric edge, five fistulas were in the trigone and two fistulas were in the bladder neck region. Fistula size ranged from 5 to 35 mm in diameter but 72.1% of patient fistulas were less than 10 mm in diameter. Only one patient had a large bladder defect of 35 mm in diameter. According to subjective estimation and objective assessment the overall success rate of the procedure after 2 years was 95.2%. Only one patient had to be subjected to a second successful attempt for fistula closure. CONCLUSION: The sliding layers method is a safe and reliable vaginal surgical approach for the repair of vesicovaginal fistulas after surgical trauma.

A study of auditory afferent organization in children with dyslalia.

The auditory afferent (AA) control is an important feedback mechanism in the speech generation. A different organization of AA pathways in children with speech alterations is suggested. In order to investigate this possibility we recorded the auditory brainstem responses (ABR) and middle latency responses (MLR) on monoaural and binaural click stimulation in a group of 17 normal children with no alteration of the speech (N) and in 16 children with dyslalia (eight with systematic (S) and eight with non-systematic errors of the speech (NS)). All of children were normal hearing, with normal ORL and neurological status, right-handed and with the age approximately 7 years old. A lateralization effect was found in the S group. Normally, it was only found for wave I. The efficiency of both AA pathways was the same in NS group, indicating a more effective right pathway in more rostral areas. A prolonged latency (X = 0.25 ms) of wave III was registered on the right side in the NS group compared to normals, as well for wave V (X = 0.175 ms) with increased sweep rate (21 vs. 51 and 71). The effect of sweep rate augmentation was also studied (21-51-71) on latency values and inter-wave differences in these groups. A successive latency prolongation (X = 2.97 ms) of MLR wave Na was registered between the N-S-NS groups. In the S group a latency binaural interaction (BI) of MLR left wave Na was prolonged for 3.52 ms and in the NS group for a further 1.32 ms compared to normals. Only in the NS group was a prolongation of the BI of the right wave Pa detected (6.76 ms) compared to normals. Results suggest a different AA organization in children with dyslalia. Possible locations of alterations in functioning could be pons, and thalamocortical projections. ABR and MLR could evaluate the auditory-speech capability of children.

Acute laryngeal trauma: a comparison between peace time and war injuries.

Thirty-six patients with acute laryngeal trauma are presented. Sixteen patients had peace-time and 20 had war injuries. Peace-time injuries were mostly due to car accidents, violence and suicide using a knife, whereas the war injuries were caused by shell and mine fragments. Nine patients were treated conservatively (eight peace-time and one war-time) and 27 by surgery (eight peace-time and 19 war-time). Surgical experience showed that acute laryngeal trauma must be treated within the first 24 hours after injury. Primary treatment must be a definite one. In that way complications and uncontrolled healing can be successfully avoided.

Prognostic factors in carcinoma of the larynx: relevance of DNA ploidy, S-fractions and localization of the tumour.

The influence of the cell cycle profile and the site of the primary tumour on the overall survival were examined in 36 patients with squamous cell carcinoma of the larynx. DNA ploidy (p = 0.0091), the overall proliferative activity (p = 0.0001), the overall proliferative activity of diploid tumour cells (p = 0.0017) and primary tumour site (p = 0.0008) were found to be significant single prognostic factors of the overall survival. Multivariate analysis showed that only the overall proliferative activity was prognostically significant (p = 0.013). The results of the study show that the supraglottic site of the tumours correlates significantly with DNA ploidy (p = 0.0334) and the overall proliferative activity of tumour cells (p = 0.0159), whereas the correlation with proliferative activity of diploid tumour cells (p = 0.1416) has not been confirmed by this study. Glottic tumours showed a prognostically significant correlation with the overall proliferative activity (p = 0.0037) and proliferative activity of diploid tumour (p = 0.0054). Such a prognostic correlation was not found for DNA ploidy (p = 0.6542).

War injuries to the head and neck.

During the 1991-1992 war in Croatia, 7,043 wounded persons were treated at Dr Josip Bencevic General Hospital in Slavonski Brod. Among them, 728 (580 soldiers and 148 civilians) had war injuries to the head and neck and were admitted to the Department of Otorhinolaryngology and Cervicofacial Surgery. There were 541 (74.3%) patients with head injuries, 117 (16.0%) with neck injuries, and 70 (9.6%) with a combination of head and neck injuries. The wounds were mostly inflicted by shell and bomb fragments (542 wounds, or 74.5%). War injuries of the facial bones were preliminarily or definitively treated. Preliminary treatment was used for the wounds with multifragmentary fractures and extensive soft-tissue defects. Definitive treatment was used in lesions of cervical structures. Immediate exploration of the neck was performed on 84 patients with penetrating neck wounds. Exploration was positive in 49 patients. Concerning long-term complications, two cases of partial paralysis of the cranial plexus and one case each of quadriplegia, hemiplegia, and glottic paralysis were recorded. To our knowledge, primary closure of war wounds to the head and neck (supported by antibiotic therapy) and reconstruction of extensive laryngotracheal injuries with the medial layer of the cervical deep fascia were used for the first time as war surgery procedures.

Mortality caused by war wounds to the head and neck encountered at the Slavonski Brod Hospital during the 1991-1992 war in Croatia.

From July 1, 1991, until January 1, 1993, a total of 7,720 patients (soldiers and civilians) with war wounds were treated at Dr. Josip Bencevic General Hospital in Slavonski Brod, Croatia. Treatment was provided for 7,043 patients, whereas 677 individuals (8.8%) killed in action (KIA) were examined at the Forensic Department. There were 1,456 patients (18.9%) with head and neck wounds: 1,176 soldiers and 280 civilians. The mortality rate was significantly greater in patients with head and neck wounds (N = 271, 40.0%) than in those with injuries to the thorax (N = 163, 24.1%) and abdomen (N = 62, 9.2%; p < 0.01 for both). During treatment, 188 patients (2.4%) died of wounds (DOW). The DOW mortality was 5.2% (61 of 1,185), 4.0% (41 of 1,026), and 2.9% (25 of 867) for wounds of the head and neck, thorax, and abdomen, respectively. There was no significant difference in the mortality rate between head and neck and thorax wounds: however, the former exceeded the mortality rate recorded for abdominal wounds (p < 0.05). No significant differences were observed between soldiers and civilians with head and neck injuries either in the KIA (205 of 1,176, 17.4% vs. 66 of 280, 23.5%, respectively) or the DOW group (51 of 971, 5.3% vs. 10 of 214, 4.7%, respectively. According to the mechanism of head and neck wounding, there were 1,046 explosive (71.9%), 226 gunshot (15.5%), and 184 other (12.6%) wounds. Lethal outcome was significantly more common in gushot than in explosive wounds (79 of 226, 35% vs. 243 of 1,046, 23.2%; p < 0.01). The proportion of head and neck injuries did not differ significantly from literature reports on recent conventional wars. The site of wounding, i.e., at the battlefield or elsewhere, had no effect on the prognosis of wounds to the head and neck. Gunshot head and neck injuries showed a significantly higher mortality rate.

A tinnitus objectivization: how we do it.

The medical therapy of tinnitus should be oriented by objective measurement of the disorder. Preferably, it should be qualitative, indicating the exact neural mechanism to be neuromodulated by neuroprotective medication. The neurophysiological approach in objectivization of tinnitus is presented by means of auditory brainstem response and middle latency response. These tests could be applied in functional follow-up of medical therapy, as these are more sensitive and harmless methods as compared to standard morphological methods.

Reconstruction of laryngotracheal war injuries with the median layer of the deep cervical fascia.

Surgical exploration and immediate reconstruction with the median layer of the deep cervical fascia (MLDCF) was performed in 8 of 22 patients with exogenous war injuries of larynx and cervical trachea. A surgical technique of reconstruction with the median layer of the deep cervical fascia is described. The 7 surviving patients had good respiration without signs of stenosis of the larynx and/or the trachea. Four had good and 3 satisfactory, phonation, and none had swallowing difficulties. Owing to the simplicity of the surgical approach, its size and biological properties, the median layer of deep cervical fascia proved itself to be a suitable material in the immediate reconstruction of exogenerous war injuries of the larynx and cervical trachea.

Laryngeal war injuries.

During the war in Croatia in the region of Brodska Posavina at the Medical Center in Slavonski Brod, 7,043 wounded were treated. The Otolaryngology and Oral Surgery Service treated 728 wounded, of whom 20 had laryngeal injuries. Most of the injuries were caused by shrapnel from shells, mines, and hand grenades. Nineteen wounded had associated injuries of the neck, head, and the neck and/or other parts of the body. The authors performed in 8 surgical explorations and immediate reconstruction with the median layer of the deep cervical fascia (MLDCF). Upon termination of the treatment, the nineteen wounded had good respiration without signs of stenosis of the larynx. Twelve had good phonation, five satisfactory phonation, and two bad. None had swallowing difficulties. Among the eight wounded on whom the reconstruction of the larynx was done with cervical fascia, four had very good phonation, three satisfactory, and the eight died on the sixth postoperative day because of associated injuries to the cervical spine. The cervical fascia proved itself to be a suitable material in the immediate reconstruction of exogenous war injuries of the larynx.