Impact of Differential Rates of Disease Progression in Amyloid-Positive Early Alzheimer's Disease: Findings from a Longitudinal Cohort Analysis.

BACKGROUND: There is limited literature regarding the impact of differential rates of disease progression on longitudinal outcomes in individuals with early Alzheimer's disease (AD) and confirmed brain amyloid pathology. OBJECTIVES: To describe the underlying characteristics and long-term outcomes associated with different rates of disease progression among amyloid-positive individuals with early symptomatic AD. DESIGN: Retrospective observational study. SETTING: Data from the National Alzheimer's Coordinating Center (NACC) Uniform Data Set (UDS) in the United States (06/2005-11/2021). PARTICIPANTS: Individuals with a clinical assessment of mild cognitive impairment or dementia and Clinical Dementia Rating® Dementia Staging Instrument Sum of Boxes (CDR-SB) score 0.5-9.0 (inclusive; first visit defined as the index date) and confirmed amyloid positivity. Participants were stratified into No Progression (change ≤0), Slower Progression (0< change <2.0 points), Median Progression (2.0-point change), and Faster Progression (change >2.0 points) cohorts based on the observed distribution of changes in CDR-SB score between the index and first subsequent visit. MEASUREMENTS: For each cohort, the functional and neuropsychiatric outcomes were described at index and each subsequent visit for up to five years, and least-square (LS) mean changes from baseline were estimated using linear mixed-effects models adjusting for baseline demographic and clinical characteristics. RESULTS: Among 1,263 participants included in the analysis, the mean±standard deviation (SD) age at index was 72.7±9.7 years and 55.3% were males. Demographic characteristics and comorbidity profiles at index were similar across cohorts. However, at index, the Faster Progression (N=279) cohort had higher CDR-SB and Functional Assessment Questionnaire (FAQ) scores compared with the No Progression (N=474), Slower Progression (N=297), and Median Progression (N=213) cohorts. Adjusting for baseline characteristics, at year 5 after index the FAQ score increased by 23.6 points for Faster Progression cohort and 10.4, 15.8, and 19.2 points for the No, Slower, and Median Progression cohorts, respectively. The corresponding increases in Neuropsychiatric Inventory Questionnaire (NPI-Q) scores were 6.7 points for the Faster Progression cohort, and by 1.3, 3.1, and 8.3 points, for the No, Slower, and Median Progression cohorts, respectively. CONCLUSIONS: Despite similar demographic and clinical profiles at baseline, amyloid-positive individuals with greater deterioration based on CDR-SB early in the AD trajectory continue to experience worse functional and behavioral outcomes over time than those with more gradual deterioration in this metric.

Potential Implications of Slowing Disease Progression in Amyloid-Positive Early Alzheimer's Disease: Estimates from Real-World Data.

BACKGROUND: Emerging therapies have shown promising results for slowing the progression of Alzheimer's disease (AD). However, the potential impact of these therapies on real-world outcomes remains to be explored. OBJECTIVE: To examine the impact of slowing AD progression on functional abilities and behavioral symptoms. DESIGN: Retrospective observational study. SETTING: Data from the National Alzheimer's Coordinating Center (NACC) Uniform Data Set (UDS) in the United States (06/2005-11/2021, primary analysis) and the Alzheimer's Disease Neuroimaging Initiative (ADNI) database (09/2005-03/2022, sensitivity analysis) were used. PARTICIPANTS: Individuals with mild cognitive impairment (MCI) or mild dementia, Clinical Dementia Rating Scale Sum of Boxes (CDR-SB) score 0.5-9.0 (inclusive; first visit defined as the index date), and confirmed amyloid positivity were identified in NACC. In ADNI, individuals with at least one clinical center visit with a clinical assessment of MCI or mild dementia and confirmed amyloid positivity were identified. MEASUREMENTS: Hypothetical effects of slowing disease progression as assessed by CDR-SB on functional and behavioral outcomes including the Functional Activities Questionnaire (FAQ) score, Neuropsychiatric Inventory Questionnaire (NPI-Q) score, and the probability of complete dependence over five years were evaluated using multivariable regression among NACC participants, separately for the subgroups with MCI and mild dementia at baseline, respectively. For the ADNI sensitivity analysis, the hypothetical effects of slowing disease progression were evaluated for FAQ score using multivariable regression among the MCI participants only. RESULTS: Compared with natural disease progression, slowing progression by 20% over five years for NACC participants with MCI and mild dementia, respectively, would result in 1.7-point (10.8%) and 1.6-point (12.9%) less deterioration based on FAQ; 0.5-point (20.3%) and 0.5-point (19.3%) less deterioration based on NPI-Q; 4.7 percentage-point (22.2%) and 10.1 percentage-point (21.6%) lower probability of complete dependence. Among ADNI participants, delaying disease progression by 20% or 30% over 4 years would avert deterioration based on FAQ of 1.1 points (20.4%) and 1.6 points (29.6%), respectively, compared to natural disease progression. CONCLUSIONS: Slowing early AD progression could result in preservation of functional and behavioral attributes and functional autonomy for longer.

The increasing threat to European forests from the invasive foliar pine pathogen, Lecanosticta acicola.

European forests are threatened by increasing numbers of invasive pests and pathogens. Over the past century, Lecanosticta acicola, a foliar pathogen predominantly of Pinus spp., has expanded its range globally, and is increasing in impact. Lecanosticta acicola causes brown spot needle blight, resulting in premature defoliation, reduced growth, and mortality in some hosts. Originating from southern regions of North American, it devastated forests in the USA's southern states in the early twentieth century, and in 1942 was discovered in Spain. Derived from Euphresco project 'Brownspotrisk,' this study aimed to establish the current distribution of Lecanosticta species, and assess the risks of L. acicola to European forests. Pathogen reports from the literature, and new/ unpublished survey data were combined into an open-access geo-database (http://www.portalofforestpathology.com), and used to visualise the pathogen's range, infer its climatic tolerance, and update its host range. Lecanosticta species have now been recorded in 44 countries, mostly in the northern hemisphere. The type species, L. acicola, has increased its range in recent years, and is present in 24 out of the 26 European countries where data were available. Other species of Lecanosticta are largely restricted to Mexico and Central America, and recently Colombia. The geo-database records demonstrate that L. acicola tolerates a wide range of climates across the northern hemisphere, and indicate its potential to colonise Pinus spp. forests across large swathes of the Europe. Preliminary analyses suggest L. acicola could affect 62% of global Pinus species area by the end of this century, under climate change predictions. Although its host range appears slightly narrower than the similar Dothistroma species, Lecanosticta species were recorded on 70 host taxa, mostly Pinus spp., but including, Cedrus and Picea spp. Twenty-three, including species of critical ecological, environmental and economic significance in Europe, are highly susceptible to L. acicola, suffering heavy defoliation and sometimes mortality. Variation in apparent susceptibility between reports could reflect variation between regions in the hosts' genetic make-up, but could also reflect the significant variation in L. acicola populations and lineages found across Europe. This study served to highlight significant gaps in our understanding of the pathogen's behaviour. Lecanosticta acicola has recently been downgraded from an A1 quarantine pest to a regulated non quarantine pathogen, and is now widely distributed across Europe. With a need to consider disease management, this study also explored global BSNB strategies, and used Case Studies to summarise the tactics employed to date in Europe.

Comparison of analog and digitally evaluated volume of the female breast in reconstructive breast surgery. Validation of a noninvasive measurement method with 3D camera1.

BACKGROUND: Reconstructive surgery is established as a standard treatment option after mastectomy due to cancer. It is crucial to patients to achieve a natural and symmetric looking breast through reconstruction. Anthropometric measurements are used to assess the objective symmetry of the breast, which are prone to errors and difficult to reproduce. OBJECTIVE: The aim of this work is to validate breast volumetry using three-dimensional surface imaging. METHODS: We compared preoperatively analog and digitally evaluated volume of the breast with our gold standard, direct water displacement measurement of the mastectomy specimen. We examined 34 breast specimens in total. RESULTS: Each measurement method (Breast Sculptor, VAM, Breast-V) for breast volume/mass determination demonstrates acceptable agreement ranges when compared with resected volumes and masses. The strongest volumetry instrument is Breast Sculptor (digital), the weakest is Breast-V (analog). CONCLUSIONS: 3D surface imaging is a quick, effective, and convenient method to evaluate breast shape and volume. The accuracy, reproducibility, and reliability of 3D surface imaging were comparable with MRI in our study.This takes us a step closer to the long-term goal of establishing robust instruments to plan breast reconstructive surgery, achieve better surgical results, and contribute to quality assurance in breast surgery.

Role of clinical and CT findings in the identification of adult small-bowel intussusception requiring surgical intervention.

BACKGROUND: In adults, intussusception has been considered traditionally to have an underlying aetiology. The aim of this study was to determine CT and clinical features of small-bowel intussusceptions that required surgical intervention. METHODS: Adult patients were identified in whom small-bowel intussusceptions were noted on CT images. The appearance, number, type (enteroenteric versus enterocolic), length and maximum short-axis diameter of intussusceptions, and presence of bowel obstruction (short-axis diameter of proximal small bowel greater than 3 cm) were analysed. The outcome was defined as surgical (complicated) or self-limiting (uncomplicated). Associations between complicated and uncomplicated intussusceptions and patient characteristics were investigated. RESULTS: Among 75 patients (56 male) with a mean age of 45 years, 103 intussusceptions were identified, of which 98 (95 per cent) were enteroenteric and 5 (5 per cent) enterocolic. Only 12 of 103 intussusceptions (12 per cent) in 12 of 75 (16 per cent) patients required surgical therapy and were considered to be complicated, with half of these having a neoplastic lead point. Length (P < 0.001), diameter (P < 0.001) and type (P = 0.002) of intussusception as well as presence of vessels (P = 0.023) within an intussusception on a CT scan, clinical symptoms (P = 0.007) and signs of bowel obstruction (P < 0.001) were associated with a surgical outcome. CONCLUSION: Clinical symptoms, signs of bowel obstruction, type and length of intussusception, and a visible tumour within an intussusception on CT scan were critical signs of complicated intussusception, requiring surgical intervention.

Imaging of breast implant and implant-associated complications: Capsular contracture and intra- or extracapsular rupture.

BACKGROUND: In recent years, follow-up after breast reconstruction with silicone implants and the detection of complications have been relieved by the possibility of improved diagnostic methods. METHODS: Between January 2015 and December 2019 a total of 40 patients (29-84 years) with silicone implants were included in this retrospective study. The implants were examined clinically and with modern imaging: general ultrasound imaging (US), magnetic resonance imaging (MRI), high resolution computed tomography (CT) and positron emission tomography -computed tomography (PET-CT). If necessary, a histological/cytological sample was taken. The breast implants were assessed by three radiologists specialized in breast imaging. The grade of capsular contracture was classified according to the Baker classification. RESULTS: All 40 women obtained a clinical examination and an US diagnostic to identify early and more common complications such as implant folding and capsular fibrosis. Depending on the clinical examination and ultrasound findings additional MRI (n = 10), CT (n = 9) and/or PET-CT (n = 2) were performed. 16 patients had implants folding proven with US (n = 16), MRI (n = 6) and CT (n = 1). The grade of capsular fibrosis was determined according to the Baker classification. The following results were obtained in our study: 25 breast implants with Baker grade I and eleven breast implants with Baker grade II, both proven with US; one breast implants with Baker grade III and one breast implant with Baker grade IV, proven with US (n = 2), MRI (n = 1) and CT (n = 1). One patient had intracapsular rupture and one patient had extracapsular rupture, both detected on CT and surgically proven. No patient had a silicone accumulation in the lymph nodes. One patient had pathologically enlarged axillary lymph nodes, which were evaluated as inflammatory changes in PET-CT. Long-term complications such as the development of malignant breast tumors could not be observed. CONCLUSION: To detect early complications after breast implant surgery, a regular clinical examination is indispensable. Imaging methods complement each other and if they are used multimodal, it is easier to identify early complications. Modern diagnostic modalities like ultrasound and magnetic resonance imaging expand the spectrum and improve diagnostic safety.

Diagnostic performance of ultrasound strain elastography for differentiation of malignant breast lesions.

OBJECTIVE: Objective of the study was to assess the performance of ultrasound strain elastography combined with conventional B-mode ultrasound to diagnose suspicious breast lesions (BI-RADS 4 and 5). METHODS: Between January 2015 and December 2017, a total of 86 patients (55±17.9 years) with solid or unclear, non-cystic breast lesions were enrolled in the study. The breast lesions were evaluated by B-mode ultrasound and subsequent ultrasound strain elastography. Ultrasound features including echo pattern, lesion shape, marginal characteristics, calcifications, orientation and posterior features are useful for differentiating benign breast lesions from malignant ones. Malignant ultrasound features are irregular shape, hypoechoic pattern, spiculated margin, nonparallel orientation and posterior shadowing. Lesions with benign features such a circumscribed margin were classified as low conspicuous. In patients with highly conspicuous malignant masses ultrasound-guided core needle biopsy or surgical excision was performed. RESULTS: Among the 86 patients, 60 masses were histologically proven and 36 of these were malignant. Of the malignant lesions 26 invasive carcinomas were of no special type (NST), 1 invasive lobular carcinoma, 3 ductal carcinomas in situ and 6 metastases. 20 of these patients had breast density classified as ACR 3 or 4. The mean size of the masses was 1.2 cm (range: 0.5-5 cm). Based on ultrasound B-mode imaging, the lesions were classified as BI-RADS category 4 (n = 20; three 4a, nine 4b and eight 4c) and BI-RADS category 5 (n = 16). One patient had to be reclassified after strain elastographxy to BI-RADS 3.The sensitivity of ultrasound with strain elastography and additional B-Mode ultrasound for the characterization of solid breast lesions was 97%, for conventional B-mode alone 92%; the respective specificities were 82% and 73%, respectively. CONCLUSION: Ultrasound elastography can be a supplementary approach to conventional ultrasound to improve the diagnostic accuracy of malignant breast lesions.

Antigenic Variation in Streptococcus pneumoniae PspC Promotes Immune Escape in the Presence of Variant-Specific Immunity.

Genomic analysis reveals extensive sequence variation and hot spots of recombination in surface proteins of Streptococcus pneumoniae While this phenomenon is commonly attributed to diversifying selection by host immune responses, there is little mechanistic evidence for the hypothesis that diversification of surface protein antigens produces an immune escape benefit during infection with S. pneumoniae Here, we investigate the biological significance of sequence variation within the S. pneumoniae cell wall-associated pneumococcal surface protein C (PspC) protein antigen. Using pspC allelic diversity observed in a large pneumococcal collection, we produced variant-specific protein constructs that span the sequence variability within the pspC locus. We show that antibodies raised against these PspC constructs are variant specific and prevent association between PspC and the complement pathway mediator, human factor H. We found that PspC variants differ in their capacity to bind factor H, suggesting that sequence variation within pspC reflects differences in biological function. Finally, in an antibody-dependent opsonophagocytic assay, S. pneumoniae expressing a PspC variant matching the antibody specificity was killed efficiently. In contrast, killing efficacy was not evident against S. pneumoniae expressing mismatched PspC variants. Our data suggest that antigenic variation within the PspC antigen promotes immune evasion and could confer a fitness benefit during infection.IMPORTANCE Loci encoding surface protein antigens in Streptococcus pneumoniae are highly polymorphic. It has become a truism that these polymorphisms are the outcome of selective pressure on S. pneumoniae to escape host immunity. However, there is little mechanistic evidence to support the hypothesis that diversifying protein antigens produces a benefit for the bacteria. Using the highly diverse pspC locus, we have now characterized the functional and immune implications of sequence diversity within the PspC protein. We have characterized the spectrum of biological function among diverse PspC variants and show that pspC sequence diversity reflects functional differences. Further, we show that sequence variation in PspC confers an immune escape benefit in the presence of anti-PspC variant-specific immunity. Overall, the results of our studies provide insights into the functional implications of protein sequence diversity and the role of variant-specific immunity in its maintenance.

Contrast-enhanced ultrasound (CEUS) in an interdisciplinary intensive care unit (ICU): Diagnostic efficacy in the assessment of post-operative complications compared to contrast-enhanced computed tomography (CECT): First results.

To evaluate the effectiveness of high-resolution contrast-enhanced ultrasound (CEUS) to diagnose early post-operative complications in an interdisciplinary intensive care unit.In 50 patients (male 32; female 18) 64 CEUS examinations were performed in an intensive care unit (ICU) setting to detect post-operative complications. Multi-frequency transducer (1-5 and/or 6-9 MHz) were used. All 64 examinations were performed by one experienced examiner. CEUS findings were compared with contrast-enhanced computed tomography (CECT) findings. CECT images were acquired within 1-24 hours after CEUS examination in arterial and portal-venous phases of the abdomen using either a 16-slice computed tomography scanner or a 128-dual slice computed tomography scanner.In 56 out of 64 cases (88%) the CEUS-based diagnosis corresponded with the CECT diagnosis. Vascular stenosis in hepatic arteries and portal veins were recognised in both imaging modalities but were evaluated differently [5 cases]. In 3 cases (5%) small peripheral splenic infarction, retroperitoneal hematoma and fluid collection around the liver were not diagnosed by CEUS. CONCLUSION: CEUS in an ICU setting enables a reliable detection of postoperative abdominal complications as compared to CECT.

Mineralization of Alvinella polychaete tubes at hydrothermal vents.

Alvinellid polychaete worms form multilayered organic tubes in the hottest and most rapidly growing areas of deep-sea hydrothermal vent chimneys. Over short periods of time, these tubes can become entirely mineralized within this environment. Documenting the nature of this process in terms of the stages of mineralization, as well as the mineral textures and end products that result, is essential for our understanding of the fossilization of polychaetes at hydrothermal vents. Here, we report in detail the full mineralization of Alvinella spp. tubes collected from the East Pacific Rise, determined through the use of a wide range of imaging and analytical techniques. We propose a new model for tube mineralization, whereby mineralization begins as templating of tube layer and sublayer surfaces and results in fully mineralized tubes comprised of multiple concentric, colloform, pyrite bands. Silica appeared to preserve organic tube layers in some samples. Fine-scale features such as protein fibres, extracellular polymeric substances and two types of filamentous microbial colonies were also found to be well preserved within a subset of the tubes. The fully mineralized Alvinella spp. tubes do not closely resemble known ancient hydrothermal vent tube fossils, corroborating molecular evidence suggesting that the alvinellids are a relatively recent polychaete lineage. We also compare pyrite and silica preservation of organic tissues within hydrothermal vents to soft tissue preservation in sediments and hot springs.

Preoperative diagnosis of thyroid adenomas using high resolution contrast-enhanced ultrasound (CEUS).

To evaluate the impact of high-resolution contrast-enhanced ultrasound (CEUS), Color Coded and Power Doppler Sonography (CCDS/PD), in the preoperative recognition of thyroid adenomas. A total of 64 patients (36 female, median age 54 years) underwent surgery for thyroid adenoma. CEUS with linear multifrequency probes (6-9 or 6-15 MHz) and CCDS/PD was performed in all patients preoperatively by an experienced examiner and images were stored in PACS. Reading of the ultrasound images was performed in consensus by two experienced radiologists. Histopathology was achieved as standard of reference in all patients. Median diameter of the tumor lesions was 27 mm. 31 nodules were found on the left side. In B-Scan mode 13 (20%) adenomas showed complex cystic echo pattern and 44 (69%) had a complete and sharp hypoechoic edge. 43 (67%) thyroid adenomas had strong hypervascularization in CCDS/PD. In 61 (95%) adenomas early contrast enhancement was present in CEUS and 50 (78%) adenomas proved no wash-out or central wash-out with persisting edge enhancement. In 47 (73%) cases early contrast enhancement was combined with no washout or with central wash-out with persisting edge enhancement. 45 (70%) thyroid lesions had preoperatively been classified as benign lesions correctly. Dynamic evaluation of microcirculation by CEUS and CCDS/PD enables a more reliable preoperative characterization of thyroid adenomas.

Sodiomyces alkalinus, a new holomorphic alkaliphilic ascomycete within the Plectosphaerellaceae.

In this study we reassess the taxonomic reference of the previously described holomorphic alkaliphilic fungus Heleococcum alkalinum isolated from soda soils in Russia, Mongolia and Tanzania. We show that it is not an actual member of the genus Heleococcum (order Hypocreales) as stated before and should, therefore, be excluded from it and renamed. Multi-locus gene phylogeny analyses (based on nuclear ITS, 5.8S rDNA, 28S rDNA, 18S rDNA, RPB2 and TEF1-alpha) have displayed this fungus as a new taxon at the genus level within the family Plectosphaerellaceae, Hypocreomycetidae, Ascomycota. The reference species of actual Heleococcum members showed clear divergence from the strongly supported Heleococcum alkalinum position within the Plectosphaerellaceae, sister to the family Glomerellaceae. Eighteen strains isolated from soda lakes around the world show remarkable genetic similarity promoting speculations on their possible evolution in harsh alkaline environments. We established the pH growth optimum of this alkaliphilic fungus at c. pH 10 and tested growth on 30 carbon sources at pH 7 and 10. The new genus and species, Sodiomyces alkalinus gen. nov. comb. nov., is the second holomorphic fungus known within the family, the first one being Plectosphaerella - some members of this genus are known to be alkalitolerant. We propose the Plectosphaerellaceae family to be the source of alkaliphilic filamentous fungi as also the species known as Acremonium alcalophilum belongs to this group.

Galantamine-based hybrid molecules with acetylcholinesterase, butyrylcholinesterase and γ-secretase inhibition activities.

We previously designed novel peptides-containing galantamine analogues. These compounds we analyzed for their putative inhibitory effect towards acetylcholinesterase, butyrylcholinesterase and γ-secretase, three activities of which could be central to various neurodegenerative pathologies including Alzheimer's disease. These pharmacological agents were virtually equipotent on acetylcholinesterase activity but display drastically higher inhibitory activities towards butyrylcholinesterase with several compounds displaying an about 100-fold higher activity than that harboured by galantamine. Strikingly, two of the galantamine amides that displayed low activity towards acetylcholinesterase exhibited the highest inhibitory potency towards butyrylcholinesterase (106 to 133 times more active than galantamine). Interestingly, five compounds show a rather good γ-secretase inhibitory potency while they retain their ability to inhibit AChE and/or BuChE activity. Thus, we have been able to design novel compounds with significant inhibitory activity against several of the enzymes responsible for key dysfunctions taking place in several neurodegenerative diseases. These mixed inhibitors could therefore be envisioned as potential pharmacological tools aimed at circumventing the degenerative processes taking place in these major pathologies.

Image fusion of contrast enhanced ultrasound (CEUS) with computed tomography (CT) or magnetic resonance imaging (MRI) using volume navigation for detection, characterization and planning of therapeutic interventions of liver tumors.

AIM: To evaluate, whether image fusion of contrast enhanced ultrasound (CEUS) with CT or MRI affects the diagnosis and characterization of liver lesions or the therapeutic strategy of surgical or interventional procedures compared to the preliminary diagnosis. MATERIAL AND METHODS: In a retrospective study the image fusion scans of CEUS with contrast enhanced CT or MRI of 100 patients (71 male, mean age 59 years, 0.3-85 years) with benign or malignant liver lesions were evaluated. Fundamental B-scan, color Doppler imaging and CEUS were performed in all patients by an experienced examiner using a multifrequency convex transducer (1-5 MHz, LOGIQ 9/GE) and volume navigation (Vnav). After a bolus injections of up to 2.4 ml SonoVue® (BRACCO, Italy) digital raw data was stored as cine-loops up to 5 min. In 74 patients, CEUS was fused with a pre-existing ceCT, in 26 patients a ceMRI was used. RESULTS: In all 100 patients (100%) the image quality in all modalities (ceCT, ceMRI and CEUS) was excellent or with only minor diagnostic limitations. Regarding the number of lesions revealed in image fusion of CEUS/ceCT/ceMRI and the preceding diagnostic method, concordant results were found in 84 patients. In 12 patients, additional lesions were found using fusion imaging causing subsequently a change of the therapeutical strategy. In 15 out of 21 patients with either concordant or discordant results regarding the number of lesions, image fusion allowed a definite diagnosis due to a continuous documentation of the microcirculation of the tumor and its contrast enhancement. A significant coherency (p < 0.05) among image fusion with either ceCT or ceMRI and CEUS and a subsequent change of therapeutic strategy was found. CONCLUSION: Image fusion with volume navigation (VNav) of CEUS with ceCT or ceMRI frequently allows a definite localization and diagnosis of hepatic lesions in patients with primary hepatic carcinoma or metastatic diseases. This might cause a change of the therapeutic strategy in many patients with hepatic lesions.

Investigation of the spread of brucellosis among human and animal populations in southeastern Bulgaria, 2007.

Three human cases of brucellosis were reported in summer 2007 in the region of Haskovo in southeastern Bulgaria. Subsequently, the regional veterinary and public health authorities carried out investigations to determine the spread of infection in domestic animals and in the human population. As a result, over 90,000 animals were tested, and 410 were found infected with brucellosis. The screening of 561 people believed to have been at risk of infection yielded 47 positive results. The majority of these persons had direct contact with domestic animals or had consumed unpasteurised dairy products. The investigations revealed evidence of disease among animals in the region and a considerable risk to humans, thus emphasising the need for effective prevention and control programmes.

Induction of apoptosis by electrotransfer of positively charged proteins as Cytochrome C and Histone H1 into cells.

Cytochrome C (Cyt. C) is a mitochondrial protein inducing apoptosis when it is accumulated in the cytosol by a currently unknown mechanism, but regulated by the bcl-2 family of proteins. The linker Histone H1 is another basic protein with highly conservative structure, composition, and equal molecular weight, not changed during the evolution. An attempt was made to understand better the apoptotic processes by electroloading of leukemic cells, such as K562, HL-60, and SKW3, and human lymphocytes with positively charged proteins, such as Cyt. C, Histone H1, and methylated BSA albumin (mBSA). The triggering apoptotic processes followed by MTT test, FACS analysis, and DNA fragmentation after the electrotransfer of these proteins into the cells were observed. Histone H1 and mBSA induce the release of Cyt. C from rat liver mitochondria. Cytochrome C release was higher when mitochondria were in "high-energy" state. It is supposed that release of Cyt. C from mitochondria is due to the mechanical rupture of the outer mitochondrial membrane, rich in negatively charged groups, predominately due to cardiolipin. The reason for the morphological rupture of the outer mitochondial membrane could be the rigidification and segregation of the membrane and the destroyed membrane asymmetries of both monolayers in the presence of positively charged proteins at higher linear charges such as Histone H1. We suggested that Histone H1, at a given moment of activated signaling for apoptosis, could be not transported to the nucleus and could lead to the release of Cyt. C from the mitochondria in the cytoplasm. It is temping to speculate that Histone H1 has other physiological extranuclear functions involved in apoptosis.

[Urinary ascites and perirenal urinoma - a renoprotective "Complication" of posterior urethral valves]. / Urinaszites und perirenales Urinom - eine renoprotektive "Komplikation" posteriorer Harnröhrenklappen.

We report on a male newborn with urinary ascites and perirenal urinoma due to calyceal rupture caused by bladder outlet obstruction and posterior urethral valves. Despite the presence of left-sided high-grade reflux, the "pop-off" mechanism occurred in the non-refluxing right kidney. Impressive normalization of the upper urinary tract anomalies and renal function occurred after surgical treatment of the valves and removal of obstruction.

Determination of platinum(IV) by UV spectrophotometry.

A simple, rapid and sensitive spectrophotometric procedure for the determination of platinum has been elaborated. Pt traces were determined in the form of the PtCl(6)(2-) complex in hydrochloric acid solution whose concentration varies from 0.01 to 2 mol L(-1) by measuring the absorbance at 260 nm. The detection limit is 4.7 x 10(-7) mol L(-1), the linearity range from 2 x 10(-6) mol L(-1) to 7 x 10(-6) mol L(-1), and the correlation coefficient is r=0.9990. No significant interferences were observed from a majority of the investigated ions, such as Zn(II), Pb(II), Mn(II), Cd(II), Co(II) and Ni(II) with the exception of Cu(II), Sb(III), Fe(III), Pd(II), Sn(II) and I(-) ions. The method was successfully applied for the determination of Pt traces in different solid samples and the recovery from inorganic materials was studied.

Improvement of posttranslational bottlenecks in the production of penicillin amidase in recombinant Escherichia coli strains.

Using periplasmic penicillin amidase (PA) from Escherichia coli ATCC 11105 as a model recombinant protein, we reviewed the posttranslational bottlenecks in its overexpression and undertook attempts to enhance its production in different recombinant E. coli expression hosts. Intracellular proteolytic degradation of the newly synthesized PA precursor and translocation through the plasma membrane were determined to be the main posttranslational processes limiting enzyme production. Rate constants for both intracellular proteolytic breakdown (k(d)) and transport (k(t)) were used as quantitative tools for selection of the appropriate host system and cultivation medium. The production of mature active PA was increased up to 10-fold when the protease-deficient strain E. coli BL21(DE3) was cultivated in medium without a proteinaceous substrate, as confirmed by a decrease in the sum of the constants k(d) and k(t). The original signal sequence of pre-pro-PA was exchanged with the OmpT signal peptide sequence in order to increase translocation efficiency; the effects of this change varied in the different E. coli host strains. Furthermore, we established that simultaneous coexpression of the OmpT pac gene with some proteins of the Sec export machinery of the cell resulted in up to threefold-enhanced PA production. In parallel, we made efforts to increase PA flux via coexpression with the kil gene (killing protein). The primary effects of the kil gene were the release of PA into the extracellular medium and an approximately threefold increase in the total amount of PA produced per liter of bacterial culture.

Genotoxic effect of substituted phenoxyacetic acids.

The potential toxic and mutagenic action of 2,4-dichlorophenoxyacetic acid has been studied in different test systems, and the obtained results range from increased chromosomal damage to no effect at all. We reexamined the effect of this herbicide by simultaneous using three tests based on yeast, transformed hematopoietic, and mouse bone marrow cells. The results obtained demonstrated that 2,4-dichlorophenoxyacetic acid has cytotoxic and mutagenic effects. The positive response of yeast and transformed hematopoietic cells was verified in kinetics and dose-response experiments. The analysis of metaphase chromosomes indicated a statistically proved induction of breaks, deletions, and exchanges after the intraperitoneal administration of 2,4-dichlorophenoxyacetic acid in mice. The study of phenoxyacetic acid and its differently chlorinated derivatives showed that cytotoxicity and mutagenicity are induced by chlorine atoms at position 2 and/or 4 in the benzene ring. The mutagenic effect was abolished by introduction of a third chlorine atom at position 5. Thus 2,4,5-trichlorophenoxyacetic acid was found to have very weak, if any mutagenic effect; however, the herbicide preserved its toxic effect.