Localization of spots in FISH images of breast cancer using 3-D shape analysis.

The fluorescence in situ (FISH) belongs to the most often used molecular cytogenetic techniques, applied in many areas of diagnosis and research. The analysis of FISH images relies on localization and counting the red and green spots in order to determine HER2 status of the breast cancer samples. The algorithm of spot localization presented in the paper is based on 3-D shape analysis of the image objects. The subsequent regions of the image are matched to the reference pattern and the results of this matching influence localization of spots. The paper compares different shapes of the reference pattern and their efficiency in spot localization. The numerical experiments have been performed on the basis of 12 cases (patients), each represented by three images. Few thousands of cells have been analysed. The quantitative analyses comparing different versions of algorithm are presented and compared to the expert results. The best version of the procedure provides the absolute relative difference to the expert results smaller than 3%. These results confirm high efficiency of the proposed approach to the spot identification. The proposed method of FISH image analysis improves the efficiency of detecting fluorescent signals in FISH images. The evaluation results are encouraging for further testing of the developed automatic system directed to application in medical practice.

Clinical usefulness of PI3K/Akt/mTOR genotyping in companion with other clinical variables in metastatic renal cell carcinoma patients treated with everolimus in the second and subsequent lines.

BACKGROUND: The aim of this study was to search for predictive and prognostic factors in patients with metastatic renal cell carcinoma (mRCC) treated with everolimus among the components of PI3K/AKT/mTOR pathway. PATIENTS AND METHODS: In a prospective, one-arm, phase II study, patients with mRCC received everolimus (10 mg/day) using a 30-day cycle. A prospectively planned evaluation of potential biomarkers of PI3K/AKT/mTOR pathway. RESULTS: The median age of the 58 patients enrolled into the study was 60 years (range 41-78 years). In multivariate analysis, it was found that the adverse independent predictors for everolimus therapy were histological grade G1/2 {hazard ratio (HR): 2.68 [95% confidence interval (CI) 1.29-5.58, P = 0.0082]}, increased lactate dehydrogenase (LDH) level before treatment [HR: 2.55 (95% CI 1.30-4.99, P = 0.0064)] and the PIK3CA gene variant rs6443624 (HR: AC + AA versus CC = 2.08, 95% CI 1 11-3.89, P = 0.0254). In multivariate analysis, it was observed that the adverse independent prognostic factors were: elevated corrected calcium level [HR: 4.17 (95% CI 1.66-10.51; P = 0.0024)] and the PIK3CA gene variant rs6443624 [HR: AC + AA versus CC = 1.97 (95% CI 1.02-3.79; P = 0.0421)]. CONCLUSIONS: The PI3KCA gene polymorphism, LDH, and histologic grade can predict the effects of everolimus treatment. The corrected calcium level and the PIK3CA gene variant rs6443624 may be independent prognostic factors. Further investigation is needed to confirm and validate these findings prospectively in other RCC trials.

Multipartite entangled spatial modes of ultracold atoms generated and controlled by quantum measurement.

We show that the effect of measurement backaction results in the generation of multiple many-body spatial modes of ultracold atoms trapped in an optical lattice, when scattered light is detected. The multipartite mode entanglement properties and their nontrivial spatial overlap can be varied by tuning the optical geometry in a single setup. This can be used to engineer quantum states and dynamics of matter fields. We provide examples of multimode generalizations of parametric down-conversion, Dicke, and other states; investigate the entanglement properties of such states; and show how they can be transformed into a class of generalized squeezed states. Furthermore, we propose how these modes can be used to detect and measure entanglement in quantum gases.

Clinical and morphological evaluation of gold micro shunt after unsuccessful surgical treatment of patients with primary open-angle glaucoma.

PURPOSE: To report the clinicopathologic features in a series of patients after ineffective glaucoma surgery with gold micro shunts (GMS) 2 years after the procedure. METHODS: This was an interventional case series study including two cases of GMS and two of GMS+ removal. Each specimen was sectioned into three portions: proximal, middle, and distal, and embedded into paraffin blocks, cut into 3 µm sections and stained with hematoxylin and eosin and Masson's trichrome. In the case of inflammatory infiltrations a reaction with an LCA (CD45) monoclonal antibody was performed. RESULTS: Mean IOP before GMS removal was 28.8 ± 4.3 mm Hg, and the patients were administered 2.3 ± 0.5 anti-glaucoma drugs. The progression of changes in the visual field was observed in all cases. In three cases different grade intensification of corneal decompression was observed. Colonization of the connective tissue was found in the channels and around the microimplant in all cases. In two cases infiltration was detected from giant polynuclear and mononuclear cells. CONCLUSIONS: Connective tissue colonization was the cause of GMS obstruction. This can be a non-inflammatory process, but it may also result from chronic inflammation occurring in the suprachoroidal space.

Suberythemal ultraviolet B radiation alters the expression of cell cycle-related proteins in the epidermis of human subjects without leading to photoprotection.

BACKGROUND: Deregulation of the cell cycle proteins is one of the critical factors leading to cutaneous carcinogenesis. OBJECTIVES: To monitor the expression of cell cycle proteins in the epidermis of subjects after repeated exposure to ultraviolet (UV) B radiation, and to test for the development of photoprotection by subsequent irradiation with a single erythemal UVB dose. METHODS: A total of 26 healthy volunteers were divided into four groups: group 1 (n = 9) were given whole-body UVB irradiation for 10 consecutive days with 0.7 minimal erythema dose (MED), group 2 (n = 9) were irradiated as in group 1 followed 24 h later by a single UVB dose of 3 MED on buttock skin, group 3 (n = 4) were irradiated with a UVB dose of 3 MED on buttock skin, and group 4 (n = 4) were not irradiated. Skin biopsies were collected 24 h after the final irradiation and stained for cyclins A, B1, D1, and p16, p18, p21, p27, p53, pRB, Bax and Bcl-2. RESULTS: The expression of cyclin D1, p18 and p21 was significantly higher in groups 1 and 2 compared with the nonirradiated group 4 controls and, in group 2, the expression of pRB, p53 and Bax was also increased. In group 3, only p53 and Bax proteins were significantly elevated compared with group 4. The expression of cyclin D1, p16, p18, p27, pRB and Bcl-2 was higher in group 2 compared with group 3. CONCLUSIONS: Suberythemal UVB radiation was sufficient to cause changes in the expression of several epidermal cell cycle proteins. When tested by irradiation with a single erythemal UVB dose following the repeated exposures, no photoprotection against the UV-induced alteration in cell cycle protein expression was apparent.

Repeated low-dose ultraviolet (UV) B exposures of humans induce limited photoprotection against the immune effects of erythemal UVB radiation.

BACKGROUND: Exposure of human subjects to ultraviolet (UV) B radiation causes immunosuppression. Most experiments to date have not tested the effects of low daily doses of UVB radiation. OBJECTIVES: To ascertain whether photoprotection against several UV-induced immune effects might develop following repeated exposure. METHODS: Groups of approximately 30 healthy individuals were given whole-body UVB irradiation on each of 10 consecutive days with 0.7 minimal erythema dose, or whole-body irradiation as before followed by a single erythemal UVB dose on a small body area, or irradiated only with a single erythemal UVB dose on a small body area, or were not irradiated. They were sensitized with diphenylcyclopropenone (DPCP) 24 h after the final dose, and skin biopsies collected to assess cytokine mRNA expression and the number of cells with thymine dimers and expression cyclooxygenase (COX)-1 and COX-2. RESULTS: The contact hypersensitivity (CHS) response to DPCP was significantly lower in the three irradiated groups compared with the unirradiated controls, while cutaneous interleukin (IL)-1beta, IL-6, IL-10 and tumour necrosis factor-alpha mRNAs, COX-1 and COX-2 and thymine dimers were all significantly higher. When the single erythemal UVB dose was given following the repeated low exposures, a slight downregulation in cytokine expression and thymine dimer formation was indicated. CONCLUSIONS: The repeated low doses of UVB protected to a limited extent against the effects of an erythemal UVB dose on cytokine expression and thymine dimer formation, but not on CHS or COX enzymes.

BRCA1-positive breast cancers in young women from Poland.

We identified 4316 unselected incident cases of early-onset breast cancers (<51 ears of age at diagnosis) in 18 Polish hospitals between 1996 and 2003. We were able to obtain a blood sample for DNA analysis from 3472 of these (80.4%). All cases were tested for the presence of three founder mutations in BRCA1. The proportion of cases with a BRCA1 mutation was 5.7%. The hereditary proportions were higher than this for women with breast cancer diagnosed before age 40 (9%), for women with cancer of medullary or atypical medullary histology (28%), for those with bilateral cancer (29%) or with a family history of breast or ovarian cancer (13%). It is reasonable to offer genetic testing to women with early-onset breast cancer in Poland.

[Morphological, microstereological and immunohistoenzymological studies of the jejuni of infants with recurrent constructive bronchitis and chronic diarrhea due to cow's milk protein allergy]. / Badania morfologiczne, mikrostereologiczne i immunohistoenzymatyczne w blonie sluzowej jelita czczego u niemowlat z nawracajacymi obturacyjnymi zapaleniami oskrzeli i towarzyszaca biegunka przewlekla na tle uczulenia na bialka mleka krowiego.

Morphological, microstereological and immunohistoenzymatic technique were used to assess jejunal biopsy samples obtained from 17 infants suffering from chronic diarrhoea and bronchitis spastica (the study group), and 5 infants with chronic diarrhoea and weight deficiency (group G). In the first group, histopathological studies revealed a small degree of villous atrophy (II degree) in 12/17 of cases, and in 5/17, a moderate atrophy (III degree). The microstereological technique disclosed infiltration of the submucosa which was significantly more marked in the study group as compared with group G (p < 0.05). Positive immunohistoenzymatic reactions in mononuclear cells of the submucosa infiltration were observed in all cases, but was more distinct in the first group. The results of the study show jejunal biopsy to be helpful in confirming the allergic etiology of the disease in the study group.

Helicobacter pylori infection in the etiopathogenesis of duodenal ulcer in children.

The study evaluates the frequency of Helicobacter pylori (H. pylori) infection, as well as systemic cellular immune response to H. pylori in children with duodenal ulcer (DU). The study group comprised 47 children with DU, aged 6-17 (mean 13, 1 +/- 4, 2). H. pylori detection was based on urease test, histology, culture and serologic tests. Endoscopic and morphologic findings were analysed according to Sydney System criteria. In 12 children from the overmentioned group subsets of blood lymphocytes B and T (CD3, CD4, CD8, CD3/DR, CD19) and NK cells, some neutrophils functions (phagocytosis, chemiluminescence) and phagocytes receptors before and one month after H. pylori triple treatment were investigated. H. pylori infection was detected in 44 of the investigated children. In addition, pathologic examination revealed chronic gastritis in 44 children and chronic duodenitis in 42 of them. In immunosystemic examination decreased percentage of CD8 lymphocytes and NK cells, increased CD4/CD8 ratio, decreased mitogen-induced response and changes of function and receptor expression of neutrophils were found. After H. pylori treatment in follow-up endoscopy no ulcers were found and histologic examination did not reveal chronic active gastroduodenitis, while the rate of nonactive gastritis was increased. Eradication of H. pylori infection in 41 children and normalisation of immune parameters in 11 children were obtained. The results of our investigation indicate, that H. pylori infection plays an important role in the pathogenesis of DU in children.

[Chronic gastritis and ulcer disease in young men]. / Przewlekle zapalenie blony sluzowej zoladka, a choroba wrzodowa u mlodych mezczyzn.

Examination has been carried out on 34 young men (average age: 22 years) with chronic erosive gastritis. Chronic erosions were situated most frequently in the prepyloric part of the stomach (88.2%). Duodenal ulcer-was found in 14 patients (41.1%), gastric ulcer was found in 1 patient (2.9%). In 19 examined patients (56.0%) chronic erosive gastritis existed without any other disease of the gastrointestinal tract. Clinical picture, gastric secretion and histopatological examination were compared in two group: I-chronic erosive gastritis without any other changes within the gastrointestinal tract and group II-chronic erosive gastritis coexisting with duodenal ulcer in the clinical picture of the group I, pain usually occurred after a meal and was felt as a heaviness sensation in epigastrum, while in the group II patients felt suction, hunger pain night pain and heartburn. Gastric secretion in the two group did not differ much. Microscopic examination of the mucosa usually confirmed chronic active gastritis. A typical feature for both group was regenerative glandular hyperplasia of the gastric mucosa. Despite many common features, it is impossible to identify explicitly and univocally the relationship between chronic erosive gastritis and ulceration.

Diamine oxidase plasma activity and jejunal mucosa integrity in children with protracted diarrhoea.

The aim of this study was to answer the question whether diamine oxidase (DAO) plasma activity reflects jejunal mucosa integrity in children with protracted diarrhoea. DAO activity was estimated with the radioisotopic method described by Fogel [14]. Postheparin DAO plasma activity was significantly lower in children with jejunal mucosa atrophy compared to control subjects.

T lymphocytes and immunoglobulins in small and large intestinal mucosa in children with ulcerative colitis.

In 52 children, aged 4-16 years, with ulcerative colitis, behaviour of A,M,G,E globulins and T lymphocytes in the small and large intestinal mucosa was evaluated. The investigations were performed both in the acute phase and remission of the disease. In the final stage of the disease, numerous plasma cells with intensive reaction to IgA and IgM were found in the stroma, while on the epithelial surface they were less marked. An intensive reaction to IgA was observed in remission. Reaction to T lymphocytes was found in single mononuclear stroma cells both in the acute phase and in remission.

Morphological and immunohistochemical examinations of the dynamic changes of gastric mucosa associated with the treatment of helicobacter pylori infection in children.

The aim of this study was to investigate the consequence of Helicobacter pylori eradication on gastric mucosa and antral G and D-cells. Forty children, aged 5-17 years with Helicobacter pylori infection were assessed. Helicobacter pylori was detected by a urease test and identified by serological and microbiological methods. Twenty children were again assessed after the therapy (the combination of colloid bismuth subcitrate, amoxycillin and metronidazole). Gastroscopic examination was performed and at least six bioptic specimens were taken from the antrum, body and fundus. Tissue samples, processed with the paraffin method and stained with hematoxyllin and eosin, were assessed. Monoclonal antiserum Gastrin PAP kit 516 and somatostatin PAP kit 512 (DAKO) in the peroxidase-antiperoxidase technique (PAP) have been used to detect G and D-cells. Helicobacter pylori in the gastric mucosa was demonstrated with the Giemsa method. The results show the coincidence of Helicobacter pylori infection and the count of antral G and D-cells and active chronic gastritis in children. After the treatment Helicobacter pylori was eradicated in 70% of children. In 34% of these cases the eradication was followed by a diminution of activity of gastric antral mucosa inflammation and in 20% of these children the resolution of the inflammatory infiltration in the gastric mucosa was seen. A decrease of the antral G and D-cells count and also a diminution of G/D index in these cases were observed.

Chronic erosive gastritis on morphological analysis.

The present study was carried out to systematize the histopathological markers of chronic erosive gastritis referred to also as gastritis varioliformis (GV). The study population consisted of 74 patients aged 20 to 69 years with gastroscopic GV. The samples were obtained from the erosion site and representative areas of the prepyloric region and gastric body. In routine microscopic examinations we evaluated the severity, type and location of changes in the gastric mucosa. Helicobacter pylori in tissue samples was identified with Giemsa's staining. Gastroscopic GV is accompanied in 76% by microscopic gastritis. In the vicinity of GV the gastric mucosa shows inflammation signs but less severe. Regenerative glandular hyperplasia of the gastric pit layer is the most frequent microscopic marker of GV.

The level of immunoglobulins in the small and large intestinal mucosa from children with ulcerative colitis.

In 42 children 4 to 16 years old with ulcerative colitis by using peroxidase method and monoclonal antibodies we evaluated the presence of immunoglobulin A, M, G, E in the mucosa of the large and small intestines. It was found that the distribution of immunoglobulins in the acute phase was similar in the intestinal segments studied, which supports the opinion that the whole intestine has a common immune system playing a major role in the pathogenesis of the disease.

[Evaluation of the development of the respiratory system in big-city adolescents, e.g., students of the Technical School of Mechanics and the Grammar School in Lódz (II)]. / Ocena rozwoju ukladu oddechowego mlodziezy ze srodowiska wielkomiejskiego na przykladzie uczniów technikum Mechanicznego i Liceum Ogolnoksztalcacego w Lodzi (Czesc II).

In 104 students from Secondary Technical School of Mechanics (TM) and 37 students from Grammar School (LO, 15-19 year-old, a longitudinal study of respiratory system was carried out. During the whole period of studies such spirographic parameters as VC, FEV1, FIV1, IMBC and Ziemssen and Amar indices were assessed. Spirographic parameters were assessed on the Pulmotest Godard. The subjects from each school were divided into two sub-groups: those practising in a sporting club (sport group--SG) and those who do not practise any sport in an organized way (non-sport group--NSG). In the students from TM and LO spirographic parameters were increased during the whole period of investigation and they reached the highest level in the first and second form. No differences were observed between students of TM and LO as regards the test parameters. The highest values were observed in the sport subgroups--the differences in the first and second forms being statistically significant.

[Evaluation of motor skills of adolescents from a large city environment attending the Technical School of Mechanical Engineering and the Grammar School in Lódz]. / Ocena rozwoju sprawnosci motorycznej mlodziezy ze srodowiska wielkomiejskiego na przykladzie uczniów Technikum Mechanicznego i Liceum Ogólnoksztalcacego w Lodzi.

A long-term study on the motor skills of juveniles was carried out. The subjects were 104 students from the Secondary School of Mechanical Engineering (SSME) and 37 from Grammar School (GS). The subjects from each of the schools were divided into two subgroups: those undergoing training in a sporting club (sport group--SG) and those who did not practise any sports in an organised way (non-sport group--NSG). Motor skills were examined with the use of the test battery worked out by Denisiuk (60-metre run, 30-metre run with overturn, high force jump, standing long jump test, 300-metre run, 1000-metre run, medical ball throw). General motor skills were expressed in terms of a synthetic coefficient. The motor skills were found to be at high and intermediate levels. Those evaluated by the Denisiuk battery of tests in the SSME students were higher, as compared to those in GS students. Parameters tested in sport groups were higher than respective parameters in NSG groups.

[An hourglass-shaped enteric cyst of the cervico-thoracic region with extra- and intramedullary location]. / Klepsydrowata torbiel jelitowa (cystis enterica) pogranicza szyjno-piersiowego o zewnatrz-i wewnatrzrdzeniowym umiejscowieniu.

The authors report a case of a hourglass dysontogenetic tumour in a 7-year-old girl (enteric cyst) of the spine at the cervicothoracic region coexistent with a developmental anomaly of the spine and posture anomaly. Atypical symptoms (pains) suggesting a disease of the gastrointestinal tract or heart made the diagnosis very difficult. Contrast examination of the vertebral canal made possible a correct diagnosis and appropriate surgical intervention. Microsurgical technique permitted the tumour to be removed radically.

[Evaluation of somatic development and physical fitness of adolescents from a large-city environment attending the Secondary School of Mechanical Engineering and the Grammal School in Lódz II]. / Ocena rozwoju somatycznego i wydolnosci fizycznej mlodziezy ze srodowiska wielkomiejskiego na przykladzie uczniów Technikum Mechanicznego w Lodzi i Liceum Ogólnoksztalcacego w Lodzi (Czesc I).

A long-term study on the somatic development was carried out. The subjects were: students from the Secondary School of Mechanical Engineering (SSME) and Grammar School (GS). The subjects from each school were divided into two sub-groups: those undergoing training in a sporting club (sport group-SG) and those who do not practise any sport in an organized way (non-sport group-NSG). Throughout the period of studies such somatic parameters as height, body weight, physical fitness and strength of hand flexors were assessed. The energy cost of work was tested during the practice. The studies demonstrated better physical constitution and fitness in the SSME group as compared to GS group. Within similar studies in sub-groups, the parameters tested in SG-group were higher as compared to those in the NSG-group.