Protection of Personal Information Act No. 4 of 2013: Implications for biobanks.

The Protection of Personal Information Act (POPIA) No. 4 of 2013 is the first comprehensive data-protection regulation to be passed in South Africa (SA). Its objectives include giving effect to the constitutional right to privacy by regulating the way in which personal information must be processed, balancing the right to privacy against other rights, and establishing an Information Regulator to ensure that the rights protected by POPIA are respected. POPIA will have an impact on health research, including biobanks. As sharing of samples and data is a central feature of biobanks, POPIA could change the way in which data are obtained, shared and exported. In particular, the provisions regarding data minimisation, requirements pertaining to the transfer of data abroad, consent provisions and identification of the 'responsible person' will impact the operation of biobanks in SA. With POPIA soon to come into force, it is now time to consider its implications for biobanks in SA.

Medical care and treatment of allergic rhinitis: a population-based cohort study based on routine healthcare utilization data.

BACKGROUND: Health services research on medical care and treatment of allergic rhinitis (AR) is scarce. OBJECTIVES: To investigate the prevalence, incidence, comorbidities, and treatment of AR in a realistic setting. METHODS: A cohort of 1 811 094 German National Health Insurance beneficiaries in 2005 was followed until 2011. To avoid misclassification, the ICD-10 code for AR (J30) had to be documented at least twice to classify patients as having AR. Descriptive statistics and logistic regression models were used to describe the burden, comorbidities, and treatment of AR. RESULTS: A total of 111 394 patients (6.2%) had prevalent AR in 2005/2006. In another 60 145 individuals (3.3%), AR was newly diagnosed in 2007 to 2011 (incident cases). Patients with prevalent AR were three times more likely to develop asthma compared to patients without AR (age and sex-adjusted risk ratio (RR) 3.04; 95% confidence interval (95%CI) 2.98-3.10). Newly diagnosed recurrent depressive disorder (RR 1.61; 95%CI 1.55-1.68), anxiety disorder (RR 1.52; 95%CI 1.48-1.56), and ADHD (RR 1.21; 95%CI 1.13-1.29) were also related to prevalent AR. Approximately 20% of children and 36% of adults with AR were exclusively treated by general practitioners. Allergy immunotherapy (AIT) was prescribed for 16.4% of patients with AR. Subcutaneous immunotherapy was most frequently used (80% of AIT). CONCLUSIONS: This study highlights the significant burden of AR. Despite the established benefits of AIT to treat AR and prevent asthma, this study suggests significant undertreatment. Future research is necessary to develop and implement adequate measures to increase guideline adherence.

[Diseases of the hepatobiliary system as a cause of acute abdomen]. / Erkrankungen des hepatobiliären Systems als Ursache des akuten Abdomens.

Diseases of the liver and biliary system are common causes of acute abdominal pain and gallstone disease predisposes to cholecystitis and cholangiolithiasis. Sonography is the method of choice for the assessment of cholecystitis, whereas magnetic resonance cholangiopancreaticography (MRCP) is the standard technique to detect stones in the common bile duct. Multi-detector computed tomography (MDCT) is ideal for detection of associated complications, including abscess formation and gall stone ileus. Pyogenic, amebic and fungal liver abscesses are reliably diagnosed with MDCT which can also be used for interventional radiologic therapy of liver abscesses by percutaneous aspiration or drainage procedures. The second most common cause of liver rupture after blunt trauma is spontaneous rupture of hypervascular liver tumors (i.e., HCC, adenoma, angiosarcoma) and due to medical procedures. Multi-phase contrast-enhanced MDCT can reliably detect active bleeding to guide further therapy in these cases.

Identification of glucosinolates on the leaf surface of plants from the Cruciferae and other closely related species.

Leaf-surface extracts prepared from 18 non-cultivated (wild) plant species, derived from the Capparidaceae, Cruciferae, Resedaceae and Tropaeolaceae were ranked for their ability to stimulate oviposition by the cabbage root fly, and analysed for glucosinolates. A total of 28 different glucosinolates were identified. A clear relationship was detected between the indolyl-, benzyl- and the total glucosinolate composition on the leaf surface and oviposition preference by cabbage root fly females. However, as the results are not fully explained by differences in leaf surface glucosinolates, other important oviposition deterrents and stimuli on the leaf surface of these wild crucifers must also be present.

Fibrous tumors in children - a morphologic and interphase cytogenetic analysis of problematic cases.

We describe and discuss the findings by fluorescent in situ hybridization (FISH) for detection of non-random chromosomal gains, in a group of unusual fibrous lesions in children. Nuclear disaggregation was used to prepare slides from eight cases which were hybridized using alpha-satellite enumeration probes for chromosomes 8, 11 and 17. Trisomy 8 and 11 were detected in a high percentage of nuclei in cases of congenital/infantile fibrosarcomas (ranging from 45 to 80%), and in a low grade fibrosarcoma in an older child (23%). Only gains of chromosome 17 were detected in a case of infantile fibromatosis (22%). In this study we have found that given the unconventional histopathologic features, the detection of more than one non-random chromosomal gains by FISH, may aid in further defining fibrous tumors in children, and may be useful as an ancillary diagnostic test in the future.

Indeterminate-cell histiocytosis: immunophenotypic and cytogenetic findings in an infant.

BACKGROUND: The authors report the immunohistochemical, ultrastructural, and cytogenetic findings in a case of malignant histiocytic proliferation in an infant. PROCEDURE: The patient presented initially with bone lesions without skin or systemic involvement. Multiple biopsies were studied extensively by immunohistochemistry and electron microscopy. Cytogenetic studies of cell cultures supplemented with granulocyte-monocyte colony stimulating factor (GM-CSF) were also performed. RESULTS: Morphologically, the cells resembled Langerhans cells, although with greater pleomorphism, as evinced by cells with usual polylobated nuclei. These cells expressed markers for macrophages and antigen presenting cells and were CD1a- and S-100-positive, but lacked Birbeck granules. The cells grown in culture supplemented with GM-CSF showed a unique combination of numerical and structural abnormalities affecting chromosomes 1, 6, 8, and 10. The disease followed a malignant course leading to the patient's demise despite aggressive chemotherapy and bone marrow transplant. CONCLUSIONS: The findings suggest a malignant hematopoietic stem-cell neoplasm with a capacity for macrophage or dendritic-cell differentiation. Morphology and immunophenotypic features place this neoplasm within the group recently conceptualized as indeterminate-cell histiocytosis.

Rapid growth of cutaneous metastases after surgical resection of thrombospondin-secreting small blue round cell tumor of childhood.

In animal models, the importance of tumor-derived antiangiogenic factors in controlling metastases has been demonstrated by the growth acceleration of distant metastases after surgical excision of a primary tumor mass. We report the case of an infant who developed rapidly growing cutaneous metastases after surgical resection of a neoplasm of an upper extremity. The tumor was undifferentiated, with some morphological features of primitive neuroectodermal tumor. To test the possibility that the primary tumor was secreting an angiogenic inhibitor, cells from the primary tumor were grown in culture, and the culture medium was tested with an in vitro endothelial cell migration assay and Western blot. The cultured cells secreted sufficiently high levels of an angiogenic inhibitor to overcome the inducing ability of vascular endothelial growth factor and basic fibroblast growth factor. One of the secreted proteins was thrombospondin-1, a potent antiangiogenic glycoprotein. The rapid dissemination of distant metastases after resection of the primary tumor in this case suggests that tumor-derived angiogenic inhibitors are important in maintaining the local net balance of angiogenic mediators controlling the growth of micrometastasis.

Hemangiopericytoma of the liver: immunohistochemical observations, expression of angiogenic factors, and review of the literature.

PURPOSE: We describe an unusual case of a hemangiopericytoma in the liver of a child, review the literature, and characterize the tumor by immunohistochemistry and electron microscopy. We study the expression of basic fibroblast growth factor (bFGF) and of vascular endothelial growth factor (VEGF) in the tumor. MATERIALS AND METHODS: Clinical history and pathology were reviewed; sections of the tumor were studied by histology, electron microscopy, and immunohistochemistry using antibodies directed towards factor-XIIIa, HAM-56, bFGF and VEGF, among others. RESULTS: The expression of VEGF resembled that of "proliferating" hemangiomas; however, despite being markedly elevated in the urine, bFGF could not be unequivocally detected in the tumor. A subpopulation of factor XIIIa positive cells was identified, similar to the "interstitial" cells of the cellular hemangiomas of infancy. The nature and function of these cells remains speculative. CONCLUSIONS: Hemangiopericytomas are rare in the liver. When arising in this location in a child, they may clinically resemble a hemangioma, may express angiogenic factors in a similar fashion, and should be considered in the differential diagnosis.

Electrochemical Properties of

[Ru(edta)(H2O)]- is strongly adsorbed on a zirconium(IV) oxide-coated silica gel surface. The immobilized complex showed an electrochemical response due to the Ru(II)/Ru(III) redox couple. By substituting the coordinated water molecule in the adsorbed complex, the midpoint potentials shifted in the order (in mV) water, -290; thiocyanate, -200; pyridine, -180; 4-cyanopyridine, -80; and pyrazine, -50 vs SCE.

Extracellular matrix penetration by epithelial cells is influenced by quantitative changes in basement membrane components and growth factors.

We have previously shown that isolated mouse fetal choroid plexus epithelial (CPE) cells penetrate a basement membrane matrix (Matrigel) substrate in vitro to form single-layered epithelial vesicles embedded within the matrix. To determine which properties of the matrix are important for inducing or permitting cells to penetrate the substrate and organize into multicellular vesicles we have made quantitative changes to the basement membrane components and growth factors in cell cultures. Matrigel diluted to 33 or 10% with a collagen I gel was not permissive to cell invasion, and CPE cells formed a polarized epithelial monolayer on the substrate surface which had ultrastructural characteristics similar to those of CPE vesicles. Cells in these monolayers proliferated more rapidly than cells in epithelial vesicles. When deliberately embedded within a 33 or 10% Matrigel matrix, CPE cells were able to form vesicles, indicating that a dilute matrix is nonpermissive to cell invasion but promotes epithelial polarization and organization into vesicles. Cells embedded within a 100% collagen I matrix did not proliferate or form epithelial vesicles and the majority of cells did not remain viable. Addition of laminin to the collagen I gel promoted cell adhesion and cell survival, but did not promote the formation of extensive monolayers on the substrate nor the formation of epithelial vesicles within the matrix. Cell invasion into the 33% Matrigel matrix was induced by addition of laminin, nidogen, or a laminin-nidogen complex to the substrate or by addition of TGFbeta2 to the culture medium, but not TGFbeta1 or PDGF. These studies show that CPE cells are sensitive to quantitative changes in matrix composition, which influences their survival and proliferation and also their ability to penetrate the matrix and organize into multicellular epithelial vesicles.

Oviposition stimulants for the black swallowtail butterfly: Identification of electrophysiologically active compounds in carrot volatiles.

Headspace volatiles were collected from undamaged foliage of carrot,Daucus carota, a host-plant species of the black swallowtail butterfly,Papilio polyxenes. The volatiles were fractionated over silica on an open column, and the fractions were tested in behavioral assays withP. polyxenes females in laboratory experiments. The polar fractions, as well as the total mixture of volatiles, increased the landing frequency and the number of eggs laid on model plants with leaves bearing contact-oviposition stimulants. The nonpolar fraction, containing the most abundant compounds in carrot odor, was not stimulatory. Gas Chromatographic (GC) separation of the fractions was coupled with electroantennogram (EAG) recordings to identify the compounds perceived byP. polyxenes females. The EAG activity corresponded to the behavioral activity of the fractions. None of the nonpolar compounds, identified as various monoterpenes, evoked a major EAG response, but several constituents of the polar fractions elicited high EAG responses. Sabinene hydrate (both stereoisomers), 4-terpineol, bomyl acetate, and (Z)-3-hexenyl acetate were identified by GC-MS as active compounds.

Effects of the extracellular matrix on fetal choroid plexus epithelial cells: changes in morphology and multicellular organization do not affect gene expression.

We have developed a primary culture system for fetal mouse choroid plexus epithelial cells which maintains their differentiated phenotype. When grown on a reconstituted basement membrane substrate (Matrigel) epithelial cells formed aggregates which became embedded in the matrix and developed into characteristic and highly reproducible multicellular vesicular structures. These vesicles consisted of a squamous layer of epithelial cells with extensive attachment to the matrix substrate, surrounding a fluid-filled lumen. Electron microscopy showed that cells comprising these vesicles had a high degree of membrane specialization and polarized morphology which in many respects mimicked the in vivo morphology. Biochemical analyses demonstrated that under these culture conditions the tissue-specific pattern of gene expression of fetal choroid plexus epithelium was maintained. After 6 days in culture these cells contained approximately the same amount of transthyretin mRNA as the 12.5-day choroid plexus in vivo, and the level of total RNA per cell, which is proportional to the protein synthetic capability of the cells, was also maintained. The pattern of protein secretion was also very similar to that generated by fetal mouse choroid plexus cells in vivo. In contrast choroid plexus epithelial cells attached poorly to collagen I gels. Heterogeneous aggregates were formed in which cell-cell interactions were more extensive than cell-substrate interactions, and in no cases was a central lumen observed. Cells on the surface of large aggregates showed some evidence of membrane polarization, while the majority of cells in the cultures exhibited little evidence of polarized morphology. Despite the striking difference in morphology and multicellular organization these cells still expressed high levels of transthyretin mRNA and maintained the same pattern of protein synthesis as cells cultured on Matrigel. These results indicate that the basement membrane is important for the organization of choroid plexus epithelial cells into a functional epithelium in vitro and thus presumably the maintenance of the integrity of the blood-brain barrier in vivo. In contrast to several other epithelial systems which have been studied, the type of extracellular matrix does not appear to directly influence tissue-specific gene expression by choroid plexus epithelial cells. Thus the level of gene expression is not dependent on the cytoarchitecture and multicellular organization of this cell type.

Three-dimensional structure of the Fab fragment of a neutralizing antibody to human rhinovirus serotype 2.

The crystal structure of the antigen-binding fragment of a monoclonal antibody (8F5) that neutralizes human rhinovirus serotype 2 has been determined by X-ray diffraction studies. Antibody 8F5, obtained by immunization with native HRV2 virions, cross-reacts with peptides of the viral capsid protein VP2, which contribute to the neutralizing immunogenic site B in this serotype. The structure was solved by the molecular replacement method and has been refined to an R-factor of 18.9% at 2.8 A resolution. The elbow angle, relating the variable and constant modules of the molecule is 127 degrees, representing the smallest elbow angle observed so far in an Fab fragment. Furthermore, the charged residues of the epitope can be well accommodated in the antigen-binding site. This is the first crystal structure reported for an antibody directed against an icosahedral virus.

Do cultured vascular smooth muscle cells resemble those of the artery wall? If not, why not?

Ultrastructural stereological techniques were used to quantitate the phenotype of vascular smooth muscle cells grown in primary culture on a plastic substrate (control), on and within matrices of collagen type I, and on collagen type III, collagen type IV, or basement membrane Matrigel. The volume fraction of myofilaments (Vv myo) of freshly isolated cells at day 0 was 54.2 +/- 2.0%. By day 5 in culture, the Vv myo of cells grown on plastic, on collagen type I, and collagen type III had decreased significantly to 14.2 +/- 2.1%, 16.5 +/- 2.4%, and 16.3 +/- 2.9%, respectively. In contrast, smooth muscle cells grown within matrices of collagen type I, on collagen type IV, or on basement membrane Matrigel had a Vv myo of 30.7 +/- 1.5%, 30.8 +/- 4.2%, and 32.1 +/- 3.1%, respectively. These findings demonstrate that the presence of extracellular matrix components prevents modulation of smooth muscle phenotype early in culture and suggests that the extracellular matrix synthesized and secreted by smooth muscle in the normal vessel wall contributes to the maintenance of their contractile functional state.

Identification of host plant attractants for the carrot fly,Psila rosae.

Cold-trapped carrot leaf volatiles were analyzed by gas chro-matography with an outlet splitter to a flame ionization detector and to a carrot fly antennogram preparation as the second detector (GC-EAD). Strongest EAD responses were elicited by products whose elution temperatures corresponded to the propenylbenzenes,trans-methylisoeugenol (3,4-dimethoxy-1-propenylbenzene) andtrans-asarone (2,4,5-trimethoxy-1-propenylbenzene) and, to a lesser extent, by-products matching the elution temperatures of the leaf aldehydes hexanal, (E)-2-hexenal, and heptanal, and of the terpenes linalool and caryophyllene. The identity of the propenylbenzenes was confirmed by gas chromatography-mass spectrom-etry. GC-EAD permitted accurate estimation of the olfactory thresholds; it was lowest fortrans-asarone at 500 attogram (5 × 10(-16)g)/ml of air passing over the antenna. Both the leaf aldehydes and propenylbenzenes were attractive when tested individually in the field with yellow sticky traps; fly captures were linearly related to the quantity of propenylbenzenes applied per trap. A combination oftrans-asarone and hexanal was more attractive than either compound singly, suggesting that the fly is adaptively equipped to respond to a mixture of compounds emanating from carrot foliage. In laboratory choice tests, flies were more attracted by vapors from intact carrot foliage than by that from a nonhost; leaf odor alone also mediated oviposition. We conclude that through the selectivity and sensitivity of its response to foliar volatiles, the carrot fly may achieve host-plant orientation and also at close range, in union with its response to less volatile leaf surface components, selection of an oviposition site.

Purification and characterization of animal porphobilinogen synthases. I. Bovine liver porphobilinogen synthase.

Porphobilinogen synthase was purified from ox liver by ammonium sulfate fractionation, heat denaturation and column chromatography (purification: 400-fold; specific activity 4.72 nkat). The molecular weight of the native enzyme obtained by thin-layer gel filtration is about 280 000. Using 8M urea in the presence of dithiothreitol as reducing agent, the molecule breaks down into 8 subunits of molecular weight 36 000 (dodecylsulfate gel electrophoresis); the preparation of aminoethylated subunit is described. According to the above-mentioned molecular weight and to the above-mentioned molecular weight and to the quantitative amino acid analysis after total hydrolysis, the following compositon of the enzymes subunit was calculated ASX23-25 Thr7 Ser23-24 Glx29-31 Pro22-23 Gly22-24 Ala36-37 Val23-26 Met7 Ile9 Leu34-35 Tyr10 Phe11-12 Lys11-12 Cys6-7 His6-8 Arg22 Trp1-2. The subunits, having two free sulfhydryl groups, therefore consists of a chain of about 306 amino acids. The Dansyl-Edman procedure did not enable identification of any free N-terminal amino acid. The acyl group blocking the N-terminus is an acetyl group. It was identified, after hydrazinolysis of the enzyme, by means of chromatographic comparison with 1-formyl-2-dansyl-hydrazine and 1-acetyl-2-dansylhydrazine, whose syntheses and UV spectra are described.