Whole blood miRNA expression analysis reveals miR-3613-3p as a potential biomarker for dedifferentiated liposarcoma.

BACKGROUND: Liposarcoma constitute about 13% of all soft tissue sarcoma and are associated with a high risk of metastases. As the preoperative differentiation between benign and malign lipomatous tumors is restricted to magnetic resonance imaging, computed tomography and biopsy, we performed a miRNA array to distinguish dedifferentiated liposarcoma patients from healthy controls and lipoma patients. METHODS: Blood samples of patients with dedifferentiated liposarcoma, healthy controls and lipoma patients were collected. Whole blood RNA was extracted and samples of patients with dedifferentiated liposarcoma (n= 6) and of healthy donors (n= 4) were analyzed using an Affymetrix GeneChip miRNA Array v. 4.0. qRT-PCR was carried out to confirm the most differentially expressed miRNA; being further analyzed in an independent cohort of healthy controls as well as in lipoma patients. RESULTS: As shown by the microarray, two miRNAs (miR-3613-3p, miR-4668-5p) were shown to be significantly upregulated (fold change: > 2.5; p< 0.05) in patients with dedifferentiated liposarcoma (n= 6) as compared to healthy controls (n= 4). miR-3613-3p was further validated by qRT-PCR to be significantly upregulated in dedifferentiated liposarcoma patients compared to an independent cohort of healthy controls (n= 3) and lipoma patients (n= 5). CONCLUSION: We identified a specific whole blood miRNA (miR-3613-3p) that may serve to distinguish between dedifferentiated liposarcoma patients and healthy controls, thus potentially serving as a specific biomarker for dedifferentiated liposarcoma.

Levels of activated platelet-derived microvesicles in patients with soft tissue sarcoma correlate with an increased risk of venous thromboembolism.

BACKGROUND: Microvesicles are small vesicles expressing specific antigens from their cells of origin. Elevated levels of microvesicles have been shown to be associated with coagulation disorders as well as with different types of malignancies. This study aims to evaluate a possible correlation of different microvesicle subpopulations with a positive history of venous thromboembolism (VTE) in patients with soft tissue sarcoma. METHODS: Annexin V - positive microvesicles, leukocyte (CD45-positive), platelet (CD61-positive), activated platelet (CD62P-, CD63-positive), endothelium-derived (CD62E-positive) and tissue-factor (CD142-positive) microvesicles were identified in the peripheral blood of patients with soft tissue sarcoma (n = 39) and healthy controls (n = 17) using fluorescence-activated cell sorting (FACS). RESULTS: Both the total amount of Annexin V-positive microvesicles and levels of endothelium-derived (CD62E-positive) microvesicles were shown to decrease significantly after tumor resection (n = 18, p = 0.0395 and p = 0.0109, respectively). Furthermore, the total amount of Annexin V - positive microvesicles as well as leukocyte (CD45-positive) and endothelium-derived (CD62E-positive) microvesicles were significantly higher in patients with grade 3 (G3) soft tissue sarcoma (n = 9) compared to healthy controls (n = 17) (p = 0.0304, p = 0.0254 and p = 0.0357, respectively). Moreover, patients with G3 soft tissue sarcoma (n = 9) presented higher levels of Annexin V-positive and endothelium-derived (CD62E-positive) microvesicles compared to patients with grade 2 (G2) soft tissue sarcoma (n = 8) (p = 0.0483 and p = 0.0045). Patients with grade 1 (G1) soft tissue sarcoma (n = 3) presented with significantly lower levels of platelet (CD61-positive) microvesicles than patients with G3 soft tissue sarcoma (n = 9) (p = 0.0150). In patients with a positive history of VTE (n = 11), significantly higher levels of activated platelet (CD62P- and CD63-positive) microvesicles (p = 0.0078 and p = 0.0450, respectively) were found compared to patients without a history of VTE (n = 28). CONCLUSION: We found significantly higher levels of Annexin V-positive and endothelium-derived (CD62E-positive) microvesicles to be circulating in the peripheral blood of patients with G3 soft tissue sarcoma compared to patients with G2 soft tissue sarcoma. Furthermore, we showed that high counts of activated platelet-derived microvesicles correlate with the occurrence of VTE. Thus, the detection of these microvesicles might be an interesting new tool for early diagnosis of soft tissue sarcoma patients with increased risk for VTE, possibly facilitating VTE prevention by earlier use of thromboprophylaxis.

Distribution of histamine in the CNS of different spiders.

Immunohistochemistry is used to demonstrate histamine-immunoreactivity in the CNS of spiders. We found histamine-immunoreactivity in the photoreceptors of different spiders. Therefore, we suggest that histamine is a neurotransmitter of photoreceptors in all arthropods, since it is also known to occur in the photoreceptors of the other main arthropod taxa (Merostomata, Crustacea, and Insecta). We also describe a system of only six omnisegmental histamine-immunoreactive neurons within the central nervous system. These histamine-immunoreactive neurons can be divided into two subgroups: a dorsal system with two cells per hemisphere and a ventral system with only one cell per hemisphere. All six cells have extended arborizations in both the motor and the sensory areas of all neuromeres in the suboesophageal ganglionic mass. In contrast to araneomorph spiders, two additional sets of histamine-immunoreactive neurons were detected in mygalomorph spiders. The first set consists of seventeen cells with their cell bodies located in the cheliceral ganglion and projecting to central areas of the protocerebrum. The second set contains many if not all sensory projections from the tarsal organs on all eight legs and the pedipalps to the Blumenthal neuropil.

Leucokinin-like immunoreactive neurones in the central nervous system of the spider Cupiennius salei.

Leucokinin is a member of the myokinin peptide family. These myotropic peptides are widely distributed in arthropods. A specific antiserum immunoreactive to the neuroactive octapeptide leucokinin I (LKI) has been used to map neurones within the central nervous system of the central American wandering spider Cupiennius salei. The antiserum labels nine pairs of cell bodies and their axons. The somata are grouped near the joint of the optic lobes in the dorsal part of the supraoesophageal ganglion. The axons descend in a bundle, pass the oesophagus and divide to innervate the suboesophageal ganglion at three levels. In the dorsal-most layer, five small parallel fibres travel within the medio-central tract towards the opisthosomal neuromeres and give off small varicose projections into all leg neuromeres. In the middle layer, two neurones in the sensory-longitudinal tract 3 project into each leg neuromere. In the ventral layer, the two largest fibres (about 8 microm) run in the medio-ventral tract, each giving off a branch into all leg neuromeres. These first order arborizations have small second order arborizations in the ventral part of the leg neuromere and ascend to the five dorsal fibres in the medio-ventral tract where they form varicosities. The second order arborizations form extensive varicosities in the ventral part of the leg neuromere. These neurones may play a role in the intercellular communication between the sensory input and the motor output system, because they are represented in the dorsal sensory and the ventral motor neuropile. The ventral fibres resemble intersegmental interneurones and may function as modulators for leg motor neurones.

[Closed or open periodontitis treatment. A review of the literature]. / Geschlossene oder offene Parodontitisbehandlung. Eine Literaturübersicht.

Clinical data characterizing the results of non-surgical, conservative versus surgical modalities of periodontal treatment are presented and summarized, as they appeared in the international literature. Primarily, the clinical disease status prior to treatment is discussed. Thereafter, the therapeutic success of either modality was measured on the basis of the following parameters: reduction of probing depth, changes in attachment levels, cleanliness of root surfaces, elimination of inflammation, and longlasting tooth survival. The literature review revealed that the choice of either treatment modality influences the therapeutic success only indirectly. More important is that the diseased root surface is meticulously cleaned from all bacterial debris. In the presence of shallow (1 to 3 mm) and medium-sized (4 to 6 mm) pockets, surgical and non-surgical treatment provides equally good results. Deep pockets (7 mm or deeper) and, in particular, crater-like bony pockets as well as furcation involvement respond with better results, if surgical treatment is assigned. In the long run, however, therapeutic success can be secured only by means of a consequent periodical recall and with support of the patient's willingness to perform optimal oral hygiene.

[Comparative probing with an electronic and a manual periodontal probe]. / Vergleichende Sondierung mit einer elektronischen und einer manuellen Parodontalsonde.

The goal of this study was to compare a new electronic probe (Peri-Probe) which uses a defined probing force and allows measurement of probing depth in 0.1 mm increments, with a standard manual probe. 24 sites were probed in each of 12 subjects (6 treated for periodontal disease and 6 untreated). Duplicate measurements with the electronic probe were made with the probe tip remaining in contact with gingival tissue. Similarly executed measurements using the manual probe followed during the same appointment. One week later, measurements were repeated in reversed sequence. Duplicate measurements (within appointments) resulted in measurement errors of +/- 0.18 mm for the manual probe. 98.6% of the electronic measurements and 99.3% of the manual measurements differed by < 1 mm. Replicated duplicated measurements (between appointments) resulted in measurements errors of +/- 0.86 mm for the electronic probe and +/- 0.65 mm for the manual probe. A difference of < 1 mm was found in 89.2% and 95.5% of the measurement repetitions with the electronic probe and the manual probe, respectively. The electronic probe exhibited greater variation of measurements than the manual probe. In addition, probing using the electronic instrument was more arduous than using the manual instrument, resulting in less reliable measurements.

Melphalan-resistant multiple myeloma: results of treatment according to the M-2 protocol.

A group of 46 patients with melphalan-resistant multiple myeloma was treated according to the M-2 protocol with melphalan, prednisolone, BCNU, cyclophosphamide, and vincristine. According to the Salmon and Durie classification, four patients had stage II A; 36, stage III A; and six, stage III B disease. Treatment resulted in five patients (11%) entering remission, while 21 (46%) had stable and 20 (43%) had progressive disease. The median survival for all patients was 12.5 months, patients in remission surviving longer (median 46 months) than those with stable disease (median 15.4 months) or progressive disease (median 6.9 months). Compared with other treatment regimens used in melphalan-resistant myeloma, the remission rate is low but the median survival exceeds that reported by most other authors.