First Report of Stemphylium globuliferum Causing Stemphylium Leaf Spot on Alfalfa (Medicago sativa) in the United States.

Stemphylium leaf spot occurs in most areas where alfalfa (Medicago sativa) is grown. In the United States, Stemphylium botryosum is reported to be the predominant pathogen (1), although S. vesicarium and S. herbarum are also observed. S. alfalfae was isolated on alfalfa in Australia (4) and S. globuliferum was reported in Egypt and Korea. In April and May 2012, alfalfa plants with leaf spot symptoms were observed in Rosemount and Waseca, MN, and in Arlington, Tomah, and Waupaca, WI. Initial symptoms consisted of white to tan spots with a brown border, 2 to 3 mm in diameter, circular to oval, enlarging to 5 to 8 mm in diameter. Large lesions often coalesced. Small, narrow, brown lesions occurred on petioles. Lower killed leaves remained attached to the primary stem. Spots were larger than those caused by the cool temperature biotype of S. botryosum. Conidia formed on lesions after 48 h in a moist chamber. Conidia were removed with a fine glass rod, germinated on 1% water agar, and single hyphae transferred to V8 agar (V8A). After 2 weeks under room light, plates were placed under UV light to stimulate spore production. Conidia on host material were borne singly on straight, unbranched, smooth conidiophores, medium brown at the apex. Conidia were medium to dark brown with small papillae, subspherical with 3 to 4 transverse and 3 to 4 complete or near complete longitudinal septa, with a distinct constriction at the median transverse septum. Conidia were 27.5 to 32.5 µm long × 20 to 22.5 µm wide with a length/width (L/W) ratio of 1.2 to 1.5. Conidia on V8A were smaller, 25 to 30 µm long × 12.5 to 19 µm wide with a L/W of 1.6 to 1.8. Ascostromata 300 µm in diameter formed on leaves held at 4°C for 2 months as well as on culture plates after 1 month. Ascospores from leaves were golden brown to reddish, 40 to 42.5 × 20 µm, slightly broader in the upper half of the spore, with 7 to 8 transverse septa and one complete longitudinal septum with several incomplete septa. Ascospores from culture were smaller, 27.5 to 30 × 12.5 to 15 µm wide. These morphological features are consistent with the description for S. globuliferum (3). DNA was extracted from pure cultures of SAr301 and SWp202, isolated from plants grown in Arlington and Waupaca, respectively, and used to amplify ITS1-5.8S-ITS2 rDNA using primers ITS1 and ITS4, GPD with primers GPD1 and GPD2, EF-1α with EF446f and EF1473R, and the intergenic spacer between vmaA and vpsA with primers ATPF2 and GTP604R (2). In sequence comparisons made by BLASTn searches of GenBank, the ITS (KF479193), GPD (KF479194), and EF-1α (KF479195) sequences from S. globuliferum were different from the gene sequences of S. botryosum but identical to those from S. vesicarium, S. herbarum, and S. alfalfae. The vmaA-vpsA spacer sequence (KF479196) of S. globuliferum had 3 nucleotide differences from S. vesicarium and S. herbarum and 4 nucleotide differences from S. alfalfae, demonstrating that this sequence is useful for species discrimination. Conidia from strains SAr301 and SWp 202 were suspended at 104/ml in sterile water with 0.01% Tween 20 and used to inoculate 12 alfalfa plants using a handheld sprayer. Plants were kept at 100% RH for 48 h, then grown at 20°C with a 16-h photoperiod. After 2 weeks, lesions similar to those seen in the field were observed on leaves of all plants. Symptomatic leaves placed in moist chambers produced conidia with the size and morphology of S. globuliferum within 48 h. This is the first report to our knowledge of S. globuliferum causing disease on alfalfa in the United States. Cultures were deposited in the University of Minnesota Mycological Culture Collection. References: (1) W. A. Cowling et al. Phytopathology 71:679, 1981. (2) P. Inderbitzin et al. Mycologia 101:320, 2009. (3) E. G. Simmons. Mycologia 61:1, 1969. (4) E. G. Simmons. Sydowia 38:284, 1985.

Severity of mastalgia in relation to milk duct dilatation.

OBJECTIVE: The etiology of mastalgia is poorly understood. Histology cannot detect any distinct characteristics. This investigation therefore applied ultrasonography to mastalgia patients to investigate morphological structures and to obtain further insights into the pathophysiology of mastalgia. The aim of the study was to analyze the significance of milk duct dilatation in patients with mastalgia. METHODS: A total of 335 premenopausal women participated in a genital and breast cancer screening program. Of these, 123 women were asymptomatic and 212 complained of breast pain (136 had cyclical mastalgia; 76 reported the noncyclical type). The width of milk ducts was measured by ultrasonography and correlated with the intensity of breast pain. The site of the pain was correlated with duct dilatation. Statistical analysis was performed using the t test, the chi(2) test, analysis of variance, and Pearson correlation. RESULTS: In asymptomatic women, the maximum mean width of the milk ducts was 1.8 +/- 0.84 mm, in cyclical mastalgia 2.34 +/- 1.10 mm, and 3.89 +/- 1.26 mm in noncyclical mastalgia (P <.001). The intensity of pain showed a significant, positive correlation with the width of the milk ducts (r =.5008, P <.001). Noncyclical mastalgia patients located the pain at the site where dilated ducts were detected ultrasonographically (P <.001). CONCLUSION: The results of this study demonstrate that duct ectasia is a major factor in mastalgia. The degree of duct dilatation correlates positively with the intensity of breast pain. In the noncyclical type of pain, there is a positive correlation between the site of duct dilatation and the site of pain.

[Changes in T-cell subpopulations in normal pregnancy and in patients with spontaneous abortion]. / Veränderungen von T-Zell-Subpopulationen während der normalen Schwangerschaft und bei Patientinnen mit Spontanaborten.

From the immunological point of view, pregnancy is a privileged allograft, with complex mechanisms of adaptation within the maternal immune system preventing rejection. The importance of lymphocytes and their specific subsets for this mechanism is a controversial issue. This study was designed to examine changes within T-cell subsets during uneventful pregnancy and to evaluate their significance by comparison with findings in patients undergoing spontaneous pregnancy loss. When compared to non-pregnant control subjects, normal pregnant women showed significantly decreased numbers for peripheral T-lymphocytes, for helper/inducer-T-cells and for the helper/suppressor-T-cell ratio (T4/T8). In contrast, patients undergoing spontaneous abortion showed a significantly elevated T4/T8-ratio, if compared to normal pregnant women. The clinical relevance of these findings is uncertain. Nevertheless, in individual cases of spontaneous miscarriage, the analysis of T-cell subsets might indicate a possible immunological aetiology.

Automated determination of serum (plasma) and urine iron: a comparative investigation of chromogens improved tripyridyltriazine micromethod.

Four different, well known colour reagents for iron determination were tested in a citrate buffer at pH 2.0 under the same automated standard conditions and compared with a manual reference method on human serum and plasma samples. Specific results were obtained only with the chromogen tripyridyl-s-triazine. A micromethod was developed, which is generally well suited for automated, direct serum iron determinations, with respect to good flow properties, simple reagent composition, high reagent stability, rapid colour development, stable colour complex and high specificity. This method was run on either a Gilford System 203 S, Gilford Impact 400 or a Greiner G-400 analyser and adapted to the Technicon SMAC II, Technicon RA 1000, Eppendorf Epos and Abbott Spectrum automated systems. The tripyridyl-s-triazine method permits the determination of ferrioxamine iron in urine after a simple sample pretreatment.

Comparative histological, histochemical, immunohistochemical and biochemical studies on oestrogen receptors, lectin receptors, and Barr bodies in human breast cancer.

The present study performed on a total of 567 cases of human female breast cancer compares the results of the biochemical assay (dextran-coated charcoal assay = DCC) for oestrogen receptor (ER) with those of several morphological methods developed for the detection of the ER or for the prediction of prognosis by use of other systems (FSA = fluorescent ligand binding assay, ER-ICA = monoclonal antibody assay for ER, LRA = lectin receptor assay using peanut agglutinin, and Barr body estimation). Whereas no correlation at all was observed among the results of the DCC and those of the FSA and Barr body estimation, the ER-ICA and the LRA showed an unanimous tendency towards higher values of ER with increasing intensity of the staining product. The results of the ER-ICA may be expressed by an immuno-reactive score (IRS) calculated from the staining intensity (SI) and the percentage of positive cells (PP). The morphological methods are evaluated with special regard to their correlation with the DCC, their theoretical basis, and their practical application. In summary, the ER-ICA appears to be the sole method directly visualizing the ER protein and--in contrast to the DCC--is therefore completely independent of the content of endogenous or exogenous oestrogens in the tumor tissue. The LRA provides valuable additional information concerning tumour differentiation and possible response to endocrine therapy, whereas the FSA and Barr body estimation should be considered as obsolete and should therefore be abandoned.

[Detection of lymphocyte subpopulations in EDTA whole blood samples using flow cytometry. Stability and normal values]. / Bestimmung von Lymphozyten-Subpopulationen in EDTA-Vollblutproben im Durchflusszytometer. Stabilitäts- und Normalwertstudien.

Series of T-cell-specific monoclonal antibodies (OKT 11 for peripheral T-lymphocytes, OKT 4 for helper/inducer T-cells, OKT 8 for suppressor/cytotoxic T-cells), one B-cell-specific antibody (anti-LEU-10) and the antibody anti-LEU-7 for LGL/NK-cells were used to detect the distribution of different lymphocyte subpopulations in the peripheral blood of 60 healthy, clinically examined test persons. The flow cytometry was performed by means of direct immunofluorescence method. The statistical evaluation showed that the results in all three age groups studied did not essentially differ with the exception of the so-called natural killer cells which increase significantly with advancing years. The described method allows the dispatch of lymphocytes samples even from places far away and the analysis of the specimen within 36 to 48 h.

[Frequency of hemoglobin abnormalities and hemoglobinopathies in Mozambique peoples (comparison between leprosy and non-leprosy subjects]. / Uber die Häufigkeit von Hämoglobinanomalien und Hämoglobinopathien bei moçambiquanischen Völkerschaften. (Mit Vergleich zwischen Leprösen und Nicht-Leprösen)

In 588 bloodsamples of negride natives from Moçambique, preferably Chuabo and Macua, haemoglobin analyses were performed. In 21 cases an increase of Hb A2 was found, indicating the presence of heterozygous beta-thalassaemia, in one case the changes in Hb-analysis were typical for beta-delta-thalassaemia, 18 samples could be shown to contain Hb S, typical for the heterozygous sickle cell trait. Futhermore in 7 cases Hb A2' was found. In two bloodsamples haemoglobin variants were observed, which according to their electrophoretical mobility were assumed to represent Hb D in one case, and Hb G in the other. In the Chuabo population the frequency of the thalassaemia gene was found to be more than twice as high as in the Macua population. In non-lepers Hb S was observed with a remarkable higher incidence than in lepers.

[Isolation and amino acid composition of two pathologically augmented proteins from psoriatic scales (author's transl)]. / Isolierung und Aminosäurenzusammensetzung zweier pathologisch erhöhter Proteine aus Psoriasisschuppen

Two protein bands of water soluble extracts of psoriatic scales, which appear intensified in disc-electrophoretical separation, were isolated and their amino acid compositions were determined. It could be demonstrated that band 9 was rich of glycine, alanine, aspartic acid, leucine, and serine and that band 10 was rich of glycine, threonine, and serine. Both proteins contained an amino-sugar (galactosamine).