Autochthonous starter cultures and indigenous grape variety for regional wine production.

AIMS: To characterize Oenococcus oeni strains isolated from North-Apulian wines where malic acid degradation is usually achieved by spontaneous fermentations, and to determine the influence of bacterial inoculation time on the malolactic performances in 'Nero di Troia' wine using a complete autochthonous microbial regime. METHODS AND RESULTS: Oenococcus oeni strains from wines produced with the autochthonous (Apulia Region, southern Italy) grape variety 'Uva di Troia' were isolated, selected and characterized. Multilocus sequence typing and variable number tandem repeat analysis were used to investigate intraspecific diversity. Oenococcus oeni strains were tested in co-inoculation and in sequential inoculation, with two autochthonous yeast strains previously isolated from 'Nero di Troia' wine. After a preliminary screening using co-inoculation regime, the O. oeni strains were grouped in reason of the different behaviour in malic acid performances. Results suggested that the efficient degradation of malic acid in co-inoculation is a strain-dependent characteristic. CONCLUSIONS: Autochthonous yeast/bacterium combinations were identified as starter culture, and used in a co-inoculation approach, for vinification of regional wines. SIGNIFICANCE AND IMPACT OF THE STUDY: The 'microbial terroir' of typical fermented food and beverage production represents a dynamic sector of applied research in food microbiology. In this work, we propose the use of autochthonous bacteria and yeast for wine production from an indigenous grape variety.

An innovative tool reveals interaction mechanisms among yeast populations under oenological conditions.

Alcoholic fermentation of grape must is a complex process, involving several yeast genera and species. The early stages in fermentation are dominated by non-Saccharomyces yeasts that are gradually replaced by the Saccharomyces cerevisiae species, which takes over the fermentation. Quantitative studies have reported the influence of non-Saccharomyces yeast species on wine quality and evaluated their biotechnological interest. The industrial yeast market, which, until recently, exclusively focused on S. cerevisiae, now offers S. cerevisiae/non-Saccharomyces (including Torulaspora delbrueckii) multi-starters. The development of these new mixed industrial starters requires a better understanding of the interaction mechanisms between yeast populations in order to optimize the aromatic impact of the non-Saccharomyces yeast while ensuring complete alcoholic fermentation thanks to S. cerevisiae. For this purpose, a new double-compartment fermentor was designed with the following characteristics: (1) physical separation of two yeast populations, (2) homogeneity of the culture medium in both compartments, (3) fermentation kinetics monitored by weight loss due to CO2 release, and (4) independent monitoring of growth kinetics in the two compartments. This tool was used to compare mixed inoculations of S. cerevisiae/T. delbrueckii with and without physical separation. Our results revealed that physical contact/proximity between S. cerevisiae and T. delbrueckii induced rapid death of T. delbrueckii, a phenomenon previously described and attributed to a cell-cell contact mechanism. In contrast, when physically separated from S. cerevisiae, T. delbrueckii maintained its viability and its metabolic activity had a marked impact on S. cerevisiae growth and viability. The double fermentor is thus a powerful tool for studying yeast interactions. Our findings shed new light on interaction mechanisms described in microorganism populations.

Evidence for autotetraploidy associated with reproductive isolation in Saccharomyces cerevisiae: towards a new domesticated species.

Partial or whole-genome duplications have played a major role in the evolution of new species. We have investigated the variation of ploidy level in a panel of domesticated strains of Saccharomyces cerevisiae coming from different geographical origins. Segregation studies and crosses with tester strains of different ploidy levels showed that part of the strains were well-balanced autotetraploids displaying tetrasomic inheritance. The presence of up to four different alleles for various loci is consistent with a polyploidization mechanism relying on the fusion of two nonreduced meiospores coming from two S. cerevisiae strains. Autotetraploidy was also in accordance with karyotype and flow cytometry analyses. Interestingly, most bakery strains were tetraploids, suggesting a link between ploidy level and human use. The null or drastically reduced fertility of the hybrids between tetraploid and diploid strains indicated that domesticated S. cerevisiae strains are composed of two groups isolated by post-zygotic reproductive barriers.

Histochemical differential diagnosis and polarization optical analysis of amyloid and amyloidosis.

Amyloidosis is characterized by extracellular deposition of protein fibrils of chemically heterogeneous composition. Early recognition and identification of amyloid deposits allows an early start of therapy, which may entail a better prognosis. Congo red staining according to Romhányi (1971) is a highly specific and sensitive method for early microscopic recognition of amyloidosis. The main and most important types of amyloidosis may be distinguished by classic histochemical methods of performate pretreatment according to Romhányi (1979), or by KMnO4 oxidation according to Wright (1977) followed by Congo red staining and viewed under polarized light. Differences in the speed of breakdown (disintegration) of amyloid deposits according to Bély and Apáthy allow a more precise distinction of various types of amyloid.

Histochemical and immunohistochemical differential diagnosis of amyloidosis--a brief illustrated essay and personal experience with Romhányi's method.

The histochemical and immunohistochemical differential diagnosis of amyloidosis in surgical pathology in a referral center is presented. Different forms of amyloidosis are considered e.g. systemic generalized amyloidosis: secondary (AA), primary (AL), senile, hemodialysis-associated, hereditary and organ (tissue)-limited (localized) amyloidosis: cerebral, dystrophic (age-related, so-called "senile"), endocrine-related, localized to tumours, focal (concentrated secretion), and isolated plasma cell (solitary plasmacytoma, B-cell) dyscrasia related amyloidosis. The amyloid deposits were identified and characterized histochemically by Congo red staining after performate pre-treatment at 20 degrees C for 1, 3, 5, 10, 15, 20 or 25 sec, and with oxidation induced proteolysis by trypsin digestion at 20 degrees C for 5, 10, or 30 sec, 1, 2, 3, 4, 5, 6 or 10 min and covered with gum-arabic according to Romhányi, and confirmed by streptavidin-biotin-complex/horseradish peroxidase immunohistochemical reactions. The "sensitivity" or "resistance" to pre-treatment of amyloid deposits depends on the type of amyloid, and the length of pre-treatment. Secondary (AA) amyloid is sensitive to KMnO4 oxidation, followed by trypsin digestion (for 1 min), and its green birefringence under polarized light disappears, while primary (AL) (for 1-5 min), senile (for 1-10 min), and most forms of organ (tissue)-limited (localized) amyloid (for 1-10 min) are resistant. Performate pre-treatment is followed by pronounced congophilia. Secondary (AA) is sensitive to performate pre-treatment (for 1 sec), while primary (AL) amyloid (for 1-20 sec), senile (for 1-25 sec), and most forms of organ (tissue)-limited (localized, isolated) amyloid deposits (for 1-25 sec) are resistant, and are constantly positively birefringent. Early identification and differentiation of amyloid deposits is important for the prognosis and for the choice of therapy. The authors conclude that the presented classical histochemical methods are useful as first line screens for the histological identification of amyloidosis.

A case of bilateral testicular lymphoma.

Authors report on a 75-year-old man with bilateral testicular lymphoma. He complained of painless right testicular enlargement. Orchidectomy was indicated by ultrasound examination and the diagnosis (large cell, non-Hodgkin lymphoma B-cell origin) was established by histology and immunohistochemistry. Two months later, the left testis enlarged, orchidectomy was performed, and a lymphoma with identical histology was found. PET revealed retroperitoneal spread of the tumor. Irradiation (18 Gy) was applied. Three months later, because of gastric metastases of the lymphoma the patient underwent CVP and CAVP (Cyclophosphamide, Adriablastin, Vincristin, Prednisolone) chemotherapy. Despite of the repeated courses, eleven months after the primary diagnosis the patient died due to of multiple metastases.

Mosaicplasty for the treatment of articular cartilage defects: application in clinical practice.

This article describes a one-step operative technique for the treatment of circumscript cartilage defects of weight-bearing surfaces of the knee. Since 1992, a total of 227 patients were treated using this technique for full-thickness lesions resulting from chondropathy, traumatic chondral defects, and osteochondritis dissecans; the procedure was evaluated in 57 patients who had > 3 years of follow-up. Magnetic resonance imaging, computed tomography arthrographies, ultrasound, and arthroscopy were used to evaluate the technique. Using the modified Hospital for Special Surgery (HSS) knee scoring system, 91% of the patients achieved a good or excellent result. The operative technique, clinical results, and complications are detailed.

[Generalized secondary amyloidosis in rheumatoid arthritis]. / Generalizált secunder amyloidosis rheumatoid arthritisben.

Generalized (systemic) secondary amyloidosis was a post mortem finding recorded in 28 patients (25.2%) by amyloid specific Congored staining according to Romhányi, from 111 randomized autopsy cases with rheumatoid arthritis. Amyloidosis was present most frequently in the gastro-intestinal tract, the heart, kidney, thyroid gland, spleen and in the suprarenal glands. The quantity of amyloid deposits is usually the highest in the organs that are most frequently have deposits. The amyloidosis is a progressive cumulative process. The conclusion is that the organs where there are frequently high quantities of amyloid is where the deposits begin and where there are infrequently any deposits or low quantities is where the deposits develop later. Biopsy of the most frequently involved organs are suggested for early diagnosis of amyloidosis. Comparing the laboratory parameters (at the last hospitalisation), the amyloidotic patients were anaemic, with low haemoglobin level (p < 0.05), the renal function significantly decreased, the carbamid (p < 0.001), and kreatinin level high (p < 0.05), with frequent proteinuria (p < 0.0001).

[Tumors in rheumatoid arthritis]. / Daganatos megbetegedések rheumatoid arthritisben.

Benign (bTu) and malignant tumours (mTu) were studied in a randomised autopsy material of 161 patients with rheumatoid arthritis (RA). The tissue specimens were fixed in 8% formaldehyde solution at pH 7.6 and embedded in paraffin. The tumours were diagnosed histologically and confirmed by immunohistochemical methods. Five benign (3.1%), and thirteen malignant tumours (8.1%) found observed in 18 (11.2%) of 161 cases. There was no significant difference between laboratory parameters of patients with malignant tumours and without tumours. One benign (0.62%) and seven malignant (4.35%) tumours led to death in 8 (4.97%) of 18 cases altogether. Neoplasms were detected clinically in 8 of 18 cases (44.4 rel%). None of tumorous patients received immunosuppressive treatment and only five had gold (Tauredon) therapy. Paraneoplastic syndromes with rheumatoid complaints may be excluded by the onset and duration of RA and tumours. Benign neurogenic tumours and malignant bronchioloalveolar carcinoma were frequently associated with RA not treated by immunosuppressive drugs. Our data do not support the assumption of a high risk of malignant lymphomas associated with RA treated with immunosuppressive therapy.

[Vasculitis in rheumatoid arthritis]. / Vasculitis rheumatoid arthritisben.

The frequency and histopathological characteristics of systemic vasculitis were studied in the autopsy material of 161 patients with rheumatoid arthritis. Systemic vasculitis was observed in 36 case (22.4%). In percentage of all cases with systemic vasculitis, the most frequently involved organs were the heart (66.7%), skeletal muscles (54.8%), and peripheral nerves (52%). The skin was involved only in about one third of 36 cases (36%). In most cases the arterioles and the small arteries were affected by vasculitis. Three types of vasculitis (non specific, fibrinoid necrotic, granulomatous) could be observed simultaneously in different vessels or combined in the same vessel. Different stages of inflammation could be found simultaneously, reflecting the relapsing nature of vasculitis. The frequency, the severity, and the recurrence of vasculitis are different aspects of the same phenomenon running usually parallel to each other in different organs and on different vessels. Vasculitis lead to local ischaemia and regressive changes depend on the number and size of the involved vessels. Systemic vasculitis led to death in 19 of 36 cases. Vasculitis was detected clinically in 7 of 36 cases. Exitus lethalis depend on the localization of the involved vessels. Vasculitis in the heart and brain are more life threatening, than that of localized to the skin.

[Causes of death as well as complications in rheumatoid arthritis]. / Halálokok, szövödmények rheumatoid arthritisben.

The frequency of basic diseases, complications, causes of death and associated illnesses were studied on non selected autopsy material of 161 patients with rheumatoid arthritis died at the National Institute of Rheumatology in 1970-1992. The authors determined the incidence of severe complications such as systemic vasculitis, generalized secondary amyloidosis, generalized septic infection and miliary epithelioid granulomatosis (probably miliary tuberculosis). In 122 cases (75.8%) the RA, while in the remaining 39 (24.2%) cases other basic disease was the underlying cause of death. Vasculitis was observed in 36 cases (22.4%), amyloidosis in 34 (21.1%), sepsis in 22 (13.7%), and miliary tuberculosis in 6 (3.7%) out of 161 RA patients. Vasculitis led to death in 19 (11.8%), amyloidosis in 17 (10.6%), sepsis in 22 (13.7%) cases. However, none of the miliary tuberculosis was direct cause of death. In 76 cases (47.2%) rheumatoid arthritis was not complicated by vasculitis, amyloidosis, sepsis or military tuberculosis and in 85 cases (52.8%) one or more of these complications existed in the necropsy material of 161 RA patients. In 74 patients (46%) only one complication and in 11 (6.8%) more than one from the mentioned complication existed simultaneously in the same patients. Vasculitis was detected clinically in 7 patients (relative frequency: 19.4%), amyloidosis in 8 (relative frequency: 23.5%), sepsis in 10 (relative frequency: 45.4%), however none of the 6 mT was detected clinically. Out of 98 complications in 85 Rheumatoid patients only 25 were recognized clinically (25.5%).

[Clinico-pathological and orthopedic-surgical aspects of essential osteolysis (Gorham disease)]. / Az osteolysis idiopathica (Gorham-kór) klinikopatológiai és ortopédsebészi vonatkozásai.

Authors present two cases. One of their patient was followed for 30 years. They discuss the pathological characters and diagnostic difficulties of the disease. They have observed that with radical operations of--according to necessity--serial surgical interventions the outcome of patients can be properly controlled against recent pessimistic opinions.

[Isolated metastasis in the gallbladder from malignant cutaneous melanoma discovered by ultrasound]. / Bórmelanoma ultrahanggal felfedezett izolált epehólyag metasztázisa.

The authors describe the metastasis in gallbladder of the malignant melanoma removed from dorsal skin. Due to the increasing incidence of dermal melanomas, they emphasize the occurrence of unexpected and unpredictable metastases and the need for a close, life-long follow-up of patients. The prognosis of visceral metastases is affected to a great extent by their early detection.

[The role of perisinusoidal (Ito) cells in the course of hepatocellular carcinoma: light and electron microscopic data to the pathogenesis of hepatocellular carcinoma]. / A perisinusoidalis (Ito-) sejtek szerepe a hepatocellularis carcinoma kórlefolyásában: fény- és elektronmikroszkópos adatok a hepatocellularis carcinoma patogeneziséhez.

The clinical history of a 49 year old female patient suggested a multifocal, rapidly progressive liver disease of one month duration, apparently due to metastatic tumour. An open needle biopsy of the liver revealed a primary hepatocellular carcinoma of low grade malignancy; the diagnosis was confirmed by histological, immunohistochemical and electron microscopic studies. Besides the ultrastructural examination of the liver biopsy disclosed an unusually marked proliferation of perisinusoidal (Ito)-cells. The authors assume that the myofibroblast proliferation and transformation of Ito-cells in the noncirrhotic liver led to the formation of multifocal areas. The perisinusoidal cell proliferation was presumably due to vitamin A intoxication caused by an extreme vegetarian diet (daily consumption of large amounts of carrot juice for years, as disclosed by a retrospectively obtained history). It is assumed that the vitamin A abuse, and perisinusoidal cell proliferation may have promoted the unusually rapid progression of the multifocal, but histologically low grade hepatocellular tumour. Spectacular clinical improvement could be observed after chemotherapy, combined with local hyperthermic treatment. Presumably, the change in diet (cessation of excessive retinol and carotene intake) also may have had a beneficial effect. After one year the clinical course suggests a slower progression of tumour growth which would be more in keeping with the prognosis based on the histologic appearance of the low grade hepatocellular carcinoma. This patient's case illustrates the importance of electron microscopy supplementing diagnostic histological and immunohistochemical examinations.

[Osteoblastoma in the iliac crest]. / Osteoblastoma a csípölapátban.

An 18 year old male patient is reported with a five year history of mild pain in his right hip. Radiological investigation showed a typical feature for osteoblastoma of iliac bone, with two niduses. En bloc excision was performed. The histological examination's result was also osteoblastoma.

[Fatal myocardial lesion in systemic lupus erythematosus]. / Halálos myocardialis elváltozás systemás lupus erythematodesben.

The authors present the case of a 47-year old female patient. She suffered from a moderate activity SLE for two years. The outcome of SLE was changed by an ulcer because of leg injury, which was repeatedly infected. Severe polyarthritis, leucopenia, thrombocytopenia, clinical signs of vasculitis and in the last two months fever, high ESR, and pericarditis appeared. The angina pectoris, the cardiac decompensation, the electrocardiogram, the echocardiogram was suspect to diffuse myocardial lesion. Cardiac decompensation caused the death of the patient. Besides the activation of her autoimmune disease, infection was suspect in the patient taken immunosuppressive and steroid drugs, though it could not be verified. Autopsy verified besides the recent necrosis of heart without reaction, and multifocal myocardial abscess, which appeared possible in the terminal phase.

[Molecular genetic, laboratory and clinical study of serum amyloid P]. / Szérum amiloid P molekuláris genetikai, laboratóriumi és klinikai vizsgálata.

Studying the gene polymorphism of serum amyloid P a common constituent of amyloid deposits two restriction fragment length polymorphism patterns were found among Hungarian rheumatoid arthritis (RA), juvenile rheumatoid arthritis (JRA), systemic lupus erythematosus (SLE) patients and healthy controls. Although no direct connection between DNA polymorphism and the frequency of amyloidosis in three patient groups was found, a marked predominance of the heterozygosity in all studied groups has been described. Authors found close correlation between the serum levels of SAP and CRP in juvenile rheumatoid arthritis, but failed to find the same correlation in SLE and healthy controls. Moreover, the serum level of CRP of heterozygous patients were significantly higher than that of homozygous patients within the juvenile rheumatoid arthritis patient groups.

[Simultaneous occurrence of hereditary angioneurotic edema and Crohn disease]. / Hereditaer angioneuroticus oedema és Crohn-betegség együttes elöfordulása.

Hereditary C1 esterase inhibitor deficiency is often associated with immunpathologic disorders. The authors present a case of the rare coincidence of hereditary angioedema (HAE) and Crohn's disease. The history of the patient is analysed along with the familial occurrence of the disease. Characteristic abdominal manifestations of C1 esterase inhibitor deficiency are compared to the clinical signs of Crohn's disease. Differential diagnostic pitfalls are described along with efficatious therapeutic options.

[Acute abdominal signs of B-cell non-Hodgkin lymphoma at a young age]. / Akut hasi tünetekkel járó fiatalkori B-sejtes non-Hodgkin lymphoma.

In this report the authors present the case of a highly malignant non-Hodgkin lymphoma in a young operated patient. They call attention to the gastrointestinal form which is difficult to diagnose. In this case the lymphoma was detected only by laparotomy. The prognosis of secondary generalized lymphoma is poor. In most cases just like in the on described above--it ends with sepsis and death.