First report of a Hypoderma diana infestation in alpaca (Vicugna pacos) in Germany.

A Hypoderma larva was removed from a painful swelling in the lumbar region of a 17-month-old male alpaca kept on a farm in the Brandenburg district, eastern Germany. Morphological analysis and sequencing of the 18S rRNA gene demonstrated it was a second instar larvae of Hypoderma diana. The main host of H. diana is the roe deer (Capreolus capreolus). This is the first description of hypodermosis caused by H. diana in a camelid species.

Comparison of MammaPrint and TargetPrint results with clinical parameters in German patients with early stage breast cancer.

The 70-gene expression profile MammaPrint is a powerful prognostic indicator for disease outcome in breast cancer patients with improved prediction of recurrence risk compared to currently used guidelines. The microarray-based test TargetPrint further provides reliable, quantitative assessment of mRNA expression levels of estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2). This study was performed as a validation of MammaPrint and TargetPrint in an unselected German breast cancer population and was designed to determine the degree of concordance with currently applied clinical parameters. One hundred and forty cases of breast cancer stage I and II were classified as being low or high risk for distant metastasis using MammaPrint. Results were compared to current clinical risk classifications and adjuvant treatment management. Immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH)/chromogenic in situ hybridization (CISH) assessments of ER, PR and HER2 were further compared with gene expression read-outs using TargetPrint. Thirty-two percent of patients (19/59) with a poor prognosis-signature identified via MammaPrint did not receive adjuvant systemic treatment apart from endocrine therapy and were potentially undertreated; whereas 42% (35/77) of patients with a good prognosis-signature received chemotherapy and were potentially overtreated. Comparison of microarray receptor results with IHC and FISH/CISH were concordant in 97% for ER; 86% for PR; and 94% for HER2. In this German study population, MammaPrint would have resulted in altered treatment advice for adjuvant systemic therapy in 40% of patients. Furthermore, TargetPrint presented high concordance for ER, PR and Her2 with IHC and FISH/CISH analysis.

The pattern of invasion of early carcinomas in Barrett's esophagus is dependent on the depth of infiltration.

The differential diagnosis "high-grade intraepithelial neoplasia" or "well-differentiated Barrett's adenocarcinoma limited to the mucosa" is controversial. We investigated 277 endoscopically resected specimens of early Barrett's carcinoma. Depth of infiltration was classified as follows: m 1=carcinoma limited to Barrett's mucosa; m 2=carcinoma infiltrating the neo-muscularis mucosae; m 3=infiltration of the original lamina propria of the esophageal mucosa; m 4=infiltration of the original muscularis mucosae; sm 1, sm 2, and sm 3=infiltration into the upper third, middle third, and lower third of the submucosa. The pattern of invasion was classified and graded as follows: tubular (D 0)=only neoplastic tubuli showing cytologic criteria of malignancy - no tumor cell dissociation; dissociation grade 1 (D 1)=few dissociated tumor cells; D 2=moderate amount of dissociated tumor cells; D 3=pronounced tumor cell dissociation. 74-96% of m 1-m 4 Barrett's carcinomas limited to the mucosa have a D 0-pattern. Tubular invasion decreases only when the submucosa has been infiltrated (sm 1: 70.4%, sm 2: 30.0%, sm 3: 24.0%). Our study shows that the pattern of invasion in early cancer in Barrett's esophagus statistically significantly depends on depth of infiltration.

Glucocorticoids increase alpha5 integrin expression and adhesion of synovial fibroblasts but inhibit ERK signaling, migration, and cartilage invasion.

OBJECTIVE: In rheumatoid arthritis (RA), integrins mediate cell adhesion, migration, and invasion, and their expression is regulated by cytokines and growth factors. The aim of this study was to investigate whether hormones such as cortisol or other steroids can influence integrin expression and function in the synovial cells of patients with RA. METHODS: We performed immunofluorescence and fluorescence-activated cell sorting analyses to quantify surface integrin levels. Adhesion and migration assays were performed to study the function of synovial fibroblasts (SFs). ERK activation was measured by cellular activation of a signaling enzyme-linked immunosorbent assay. Invasion of SFs into cartilage was determined in the SCID mouse coimplantation model of RA in vivo. RESULTS: In RA, expression of integrin subunits alpha5, alphav, and beta1 was higher at the site of invasion compared with the sublining zone. Testosterone and 17beta-estradiol had no influence on integrin levels, but cortisol up-regulated expression of the alpha5 subunit in a time-dependent and dose-dependent manner. In addition, cortisol increased the adhesion of SFs to fibronectin and inhibited ERK signaling upon integrin activation or upon stimulation with tumor necrosis factor. Small interfering RNA or a neutralizing antibody to alpha5 integrin increased SF migration, indicating that up-regulated alpha5 integrin is responsible for an immobile phenotype. In addition, in the SCID mouse model, SF invasion into cartilage was attenuated by glucocorticoid treatment in vivo. CONCLUSION: Glucocorticoids increase integrin expression and the adhesion of cells to fibronectin, inhibit ERK signaling, and down-regulate the invasiveness of SFs in vivo. This study demonstrates that an important antiinflammatory aspect of glucocorticoids is regulating the expression and function of alpha5 integrin.

Mandibular prognathism caused by acromegaly - a surgical orthodontic case.

A 22-year-old man presented for orthodontic surgery because of mandibular prognathism. Clinical symptoms suggested acromegaly, and diagnosis was verified by an endocrinologist as well as by radiograph. Bilateral mandibular prognathism often represents the first and most striking physical characteristic of acromegaly; usually, it is also the main reason why patients seek help from orthodontists or maxillo-facial surgeons. This case report recapitulates the clinical and histopathological findings in pituitary growth hormone (GH) adenomas and emphasises their importance in surgical orthodontic planning. Mandibular prognatism, macroglossia and abnormal growth of hands and feet represent strong indicators for the diagnosis of acromegaly. This disease and its complications not only affect the entire body but increase mortality if the pituitary gland tumour remains untreated.

MethyQESD, a robust and fast method for quantitative methylation analyses in HNPCC diagnostics using formalin-fixed and paraffin-embedded tissue samples.

Promoter hypermethylation occurs in various tumors and leads to silencing of tumor-relevant genes. Thus, promoter methylation analysis (MA) has been established as an important tool in cancer research and diagnostics. Here we present MethyQESD (methylation-quantification of endonuclease-resistant DNA) as a fast, easy, precise and reliable method for quantitative MA without the need of bisulfite-treatment or fluorescent probes. Though MethyQESD principally works with any gene promoter we established MethyQESD for the mismatch repair gene MLH1 and tested its utility to differentiate between sporadic microsatellite unstable (MSI-H) colorectal cancer and hereditary nonpolyposis colorectal cancer (HNPCC) by quantitative MLH1 MA. We investigated formalin-fixed and paraffin-embedded tissue samples from a previously published, well-characterized tumor collective comprising 25 HNPCC, 14 sporadic MSI-H CRC and 16 sporadic microsatellite stable (MSS) CRC. We found a high accuracy of MethyQESD by spiking experiments with dilution series of methylated (SW48 cancer cell line) and unmethylated (blood) DNA (Pearson's r=0.9997 (proximal MLH1 promoter region), r=0.9976 (distal MLH1 promoter region)). MethyQESD and conventional quantitative MA using of 96 formalin-fixed and paraffin-embedded CRC showed a high degree of concordance of both methods (Pearson's r=0.885). HNPCC tumors showed either null MLH1 methylation or a significantly lower degree of MLH1 methylation than sporadic MSI-H CRC (P<0.001). MLH1 methylation was negative in all MSS tumors. Receiver operating characteristic (ROC) curve analyses defined a cutoff value of 16.5% MLH1 methylation for specific and sensitive identification of sporadic MSI-H CRC (area under ROC curve: 1.000; asymptotic significance: P<0.001). Thus, quantitative MLH1 MA by MethyQESD provides a simple, fast and valuable tool to identify HNPCC candidates. Furthermore, MethyQESD works reliably with formalin-fixed paraffin-embedded tissue and simplifies DNA MA both for research and diagnostic purposes.

Lymph node preparation in resected colorectal carcinoma specimens employing the acetone clearing method.

For the histopathological evaluation of resected colorectal carcinoma specimens, the currently required minimum number of lymph nodes that need to be examined to ensure accurate staging is 12. In some cases, however, this number of lymph nodes cannot be obtained by the conventional preparation method, based on formalin-fixed fatty tissue. Since prognostic accuracy correlates with the number of lymph nodes examined, a repeat work-up of the fatty tissue after soaking in acetone for 24h is a means of achieving the required minimum of 12 lymph nodes in many cases. Our department, which deals with an average of 307 colorectal cancer resection specimens per year, received between May 2004 and November 2006 80 cases (10.4%) in which the number of detected lymph nodes was less than 12. Owing to the use of conventional preparation methods, the required number of 12 lymph nodes could not be found, and subsequent acetone clearance of fatty tissue for a repeat examination was carried out. On average, 4.4 additional lymph nodes having an average size of 2mm were discovered. Accessory lymph node metastases were found in 9 cases. In 2 of these, the UICC classification had to be revised. In 1 case, a lymph node metastasis was detected after acetone clearance in an initially nodal negative carcinoma. Acetone clearance is an appropriate option to identify accessory lymph nodes in colorectal carcinoma specimens for daily routine use, and also constitutes an excellent instrument for internal quality control.

Use of Ovsynch in dairy herds--differences between primiparous and multiparous cows.

Ovsynch protocols are used to increase service rate and decrease days open and cullings for infertility. Recent reports have indicated better results after Ovsynch in primiparous than in older cows. However, this was not observed in all investigations on the subject. The objective of the study was to evaluate differences between primiparous and multiparous cows after synchronization of ovulation with an Ovsynch protocol in six trials. A total of 1584 cows (583 primiparous and 1001 multiparous cows, respectively) on three dairy farms were synchronized with an Ovsynch protocol consisting of a GnRH-analogue at Days 0 and 9, and a prostaglandin F(2alpha) analogue on Day 7. AI was carried out in all cows 16-20 h after the last treatment. Cows were categorized into primiparous and multiparous cows for analysis. Conception rate (CR) to timed AI, to further AI, overall conception rate and proportion of cows pregnant by 200 days in milk were compared between the age groups. Finally, two logistic regression models were calculated with conception to first service and conception by 200 DIM as the outcome variables. Independent variables were trial (categorical) and age group (primiparous versus multiparous). Conception rates to TAI were higher in primiparous than in older cows (37.9% versus 31.6%, P=0.015). Likewise pregnancy rates by 200 DIM were higher in primiparous cows (81.8% versus 75.4%, P=0.003). However, the extent of the difference varied between trials. Results indicate that Ovsynch protocols are more effective in primiparous than in older cows.

Influence of stage of lactation and milk production on conception rates after timed artificial insemination following Ovsynch.

Conception rates after timed artificial insemination (TAI) are of paramount importance for the success of protocols based on synchronization of ovulation. Stage of lactation and milk production level are known factors that influence dairy cow fertility. It was the objective of this study to analyse the effect of stage of lactation and milk production level on conception rates and pregnancy rates by 200 days in milk (DIM) in dairy cows synchronized with the Ovsynch protocol (Day -10, Day -1: 0.1 mg of D-Phe6-gonadorelin, Day -3: 0.5 mg of cloprostenol, Day 0: AI). A total of 1,288 dairy cows were assigned to two groups and classified in three production levels (high, average, low). Cows of all milk production levels in Group 1 (Simultaneous Ovsynch, SO) were synchronized with the Ovsynch protocol simultaneously for TAI between 73 and 81 DIM. In Group 2 cows with average milk production were synchronized at the same time as Group 1, while low producing cows were synchronized 3 weeks earlier and high producing cows were synchronized 3 weeks later than Group 1, respectively. First service conception rates (FSCRs) were lower (P<0.05) in cows synchronized earlier than in cows of the same production level synchronized later (low production: 14.4% (22/153) versus 34.5% (51/148); high production: 28.2% (40/142) versus 41.4% (53/128)). Milk production level had no significant impact on conception rates after TAI in cows synchronized at the same stage of lactation. At 200 DIM fewer cows with high production level were pregnant than cows with average or low production (P<0.05). This effect was independent of the stage of lactation at the initiation of Ovsynch. Endometritis at a postpartum examination did not influence conception rates after TAI. In conclusion, stage of lactation, but not milk production level, has a major influence on conception rates after TAI. Early AI after Ovsynch is less efficient and therefore its return on investment should be evaluated carefully.