Miracle-dz/UniFAR-FTUs huggingface dataset:
Induction of secreted matrix metalloproteinase-9 activity in human melanoma cells by extracellular matrix proteins and cytokines

Degradation of matrix proteins that constitute the dermal–epidermal junction and dermis by proteolytic enzymes is an essential step of melanoma invasion and metastasis, and this is primarily achieved by the matrix metalloproteinases. In this report, using zymography, we compared the basal secretion levels of active matrix metalloproteinase-2 and matrix metalloproteinase-9 to levels in response to various extracellular matrix proteins, cytokines, and growth factors in normal human melanocyte cells and melanoma cell lines from different stages of neoplastic progression. Basal matrix metalloproteinase-9 activity was only detected in vertical growth phase and metastatic melanoma cell lines, suggesting that matrix metalloproteinase-9 is a candidate biomarker for identifying vertical growth phase and metastatic melanomas. Most melanoma cell lines and cultured normal melanocytes produced high levels of matrix metalloproteinase-2. In addition, both tumor necrosis factor-&agr; and interleukin-1&bgr; are strong inducers of active matrix metalloproteinase-9 in vertical growth phase melanoma cell lines, indicating a possible role of these cytokines in the switch from radial growth phase to vertical growth phase. We propose that these proinflammatory cytokines promote melanoma invasion in part through upregulating matrix metalloproteinase-9. Both these cytokines are released from keratinocytes in the epidermis by ultraviolet radiation. Thus, our study suggests that the microenvironment of melanoma cells is an important feature in melanoma progression, and ultraviolet-radiation-induced cytokines might promote the progression of melanoma through the release or activation of matrix metalloproteinases.
Human CD4+ T Cells Displaying Viral Epitopes Elicit a Functional Virus-Specific Memory CD8+ T Cell Response1

The Ag-specific cellular recall response to herpes virus infections is characterized by a swift recruitment of virus-specific memory T cells. Rapid activation is achieved through formation of the immunological synapse and supramolecular clustering of signal molecules at the site of contact. During the formation of the immunological synapse, epitope-loaded MHC molecules are transferred via trogocytosis from APCs to T cells, enabling the latter to function as Ag-presenting T cells (T-APCs). The contribution of viral epitope expressing T-APCs in the regulation of the herpes virus-specific CD8+ T cell memory response remains unclear. Comparison of CD4+ T-APCs with professional APCs such as Ag-presenting CD40L-activated B cells (CD40B-APCs) demonstrated reduced levels of costimulatory ligands. Despite the observed differences, CD4+ T-APCs are as potent as CD40B-APCs in stimulating herpes virus-specific CD8+ T cells resulting in a greater than 35-fold expansion of CD8+ T cells specific for dominant and subdominant viral epitopes. Virus-specific CD8+ T cells generated by CD4+ T-APCs or CD40B-APCs showed both comparable effector function such as specific lysis of targets and cytokine production and also did not differ in their phenotype after expansion. These results indicate that viral epitope presentation by Ag-specific CD4+ T cells may contribute to the rapid recruitment of virus-specific memory CD8+ T cells during a viral recall response.
Cross-modal Interactions during Perception of Audiovisual Speech and Nonspeech Signals: An fMRI Study

During speech communication, visual information may interact with the auditory system at various processing stages. Most noteworthy, recent magnetoencephalography (MEG) data provided first evidence for early and preattentive phonetic/phonological encoding of the visual data stream—prior to its fusion with auditory phonological features [Hertrich, I. , Mathiak, K. , Lutzenberger, W. , & Ackermann, H. Time course of early audiovisual interactions during speech and non-speech central-auditory processing: An MEG study. Journal of Cognitive Neuroscience, 21, 259–274, 2009]. Using functional magnetic resonance imaging, the present follow-up study aims to further elucidate the topographic distribution of visual–phonological operations and audiovisual (AV) interactions during speech perception. Ambiguous acoustic syllables—disambiguated to /pa/ or /ta/ by the visual channel (speaking face)—served as test materials, concomitant with various control conditions (nonspeech AV signals, visual-only and acoustic-only speech, and nonspeech stimuli). (i) Visual speech yielded an AV-subadditive activation of primary auditory cortex and the anterior superior temporal gyrus (STG), whereas the posterior STG responded both to speech and nonspeech motion. (ii) The inferior frontal and the fusiform gyrus of the right hemisphere showed a strong phonetic/phonological impact (differential effects of visual /pa/ vs. /ta/) upon hemodynamic activation during presentation of speaking faces. Taken together with the previous MEG data, these results point at a dual-pathway model of visual speech information processing: On the one hand, access to the auditory system via the anterior supratemporal “what” path may give rise to direct activation of “auditory objects. ” On the other hand, visual speech information seems to be represented in a right-hemisphere visual working memory, providing a potential basis for later interactions with auditory information such as the McGurk effect.
Various regions within the alpha‐helical domain of the COL1A1 gene are fused to the second exon of the PDGFB gene in dermatofibrosarcomas and giant‐cell fibroblastomas

Dermatofibrosarcoma protuberans (DFSP) and its juvenile form, giant‐cell fibroblastoma (GCF), are uncommon infiltrative tumors of the dermis, which present unique cytogenetic features, such as the reciprocal translocation t(17;22) or, more commonly, supernumerary ring chromosomes containing sequences from chromosomes 17 and 22. We have recently shown that these aberrations are cytogenetic manifestations of gene fusions between the platelet‐derived growth factor B‐chain gene (PDGFB), the cellular equivalent of the v‐sis oncogene, and the collagen type I alpha 1 gene (COL1A1), the major protein constituent of the extracellular matrix in connective tissue of skin. We now report characterization of COL1A1/PDGFB chimeric genes at the RNA and DNA sequence levels in a series of DFSPs and GCFs. All 16 tumors studied contained the COL1A1/PDGFB gene. The location of breakpoints within COL1A1 varied greatly, but was always limited to the region encoding the alpha‐helical domain. The PDGFB segment of the chimeric transcript always starts with exon 2, placing PDGFB under the control of the COL1A1 promoter and removing all known elements negatively controlling PDGFB transcription and translation. Production of these aberrant transcripts in fibroblasts, the suspected cell of origin of DFSP/GCF, likely causes autocrine stimulation and cell proliferation. No specific function has yet been assigned to exon 2 of PDGFB, and this exon does not encode for the mature growth factor. Its retention in all chimeric COL1A1/PDGFB genes suggests that it is important for the normal processing of the PDGFB polypeptide. Genes Chromosomes Cancer 23:187–193, 1998. © 1998 Wiley‐Liss, Inc.
Probing the Kinesin-Microtubule Interaction*

Kinesin is a mechanoenzyme that couples adenosine triphosphate hydrolysis to the generation of force and movement along microtubules. To gain insight into the interactions of kinesin and microtubules, cross-linking, mapping, and proteolysis experiments were executed. The motor domain of kinesin was consistently cross-linked to both α- and β-tubulin subunits. Initial mapping of the cross-linked kinesin suggested that amino acids within the N- and C-terminal cyanogen bromide fragments of the motor domain formed cross-links to both α- and β-tubulin subunits. Mapping of the cross-linked tubulin suggested that cross-linking to kinesin motors occurred within the negatively charged, C-terminal cyanogen bromide fragments of α- and β-tubulin subunits. Treatment of microtubules with subtilisin, a protease that cleaves C-terminal fragments from α- and β-tubulin, reduced their ability to be cross-linked to kinesin motors supporting the idea that C-terminal sequences of α- and β-tubulin may interact with kinesin motors. Finally, of three synthetic peptides, a peptide consisting of the last 12 C-terminal amino acids of β-tubulin competitively interfered with the microtubule-stimulated adenosine triphosphatase activity of the kinesin motor, further suggesting that C-terminal sequences of β-tubulin may be involved in kinesin binding.
The application potential of sintered beta-dicalcium pyrophosphate in total joint arthroplasty.

An in vitro bone cell culture model was used to evaluate the potential application of sintered beta-dicalcium pyrophosphate (SDCP) in arthroplasty surgery. Primary osteoclasts and osteoblasts were cocultured with different sizes of SDCP particles. The changes in cell counts and the synthesis and secretion of alkaline phosphatase, acid phosphatase, and prostaglandin E(2) in response to the SDCP particles were monitored. When bone cells were cultured with SDCP particles smaller than 53 microm, both the osteoblast and osteoclast cell counts decreased significantly. When the SDCP particles were larger than 177 microm, although the osteoblast population increased significantly, the osteoclast population decreased significantly. Simultaneously, the titer of prostaglandin E(2) in the medium and the cytoplasmic prostaglandin E(2) increased significantly. We concluded that SDCP is a potentially useful bioceramic for the prevention of osteoclast-mediated periprosthetic osteolysis.
Synergistic Effects of Long-Term Antioxidant Diet and Behavioral Enrichment on β-Amyloid Load and Non-Amyloidogenic Processing in Aged Canines

A long-term intervention (2.69 years) with an antioxidant diet, behavioral enrichment, or the combined treatment preserved and improved cognitive function in aged canines. Although each intervention alone provided cognitive benefits, the combination treatment was additive. We evaluate the hypothesis that antioxidants, enrichment, or the combination intervention reduces age-related β-amyloid (Aβ) neuropathology, as one mechanism mediating observed functional improvements. Measures assessed were Aβ neuropathology in plaques, biochemically extractable Aβ40 and Aβ42 species, soluble oligomeric forms of Aβ, and various proteins in the β-amyloid precursor protein (APP) processing pathway. The strongest and most consistent effects on Aβ pathology were observed in animals receiving the combined antioxidant and enrichment treatment. Specifically, Aβ plaque load was significantly decreased in several brain regions, soluble Aβ42 was decreased selectively in the frontal cortex, and a trend for lower Aβ oligomer levels was found in the parietal cortex. Reductions in Aβ may be related to shifted APP processing toward the non-amyloidogenic pathway, because α-secretase enzymatic activity was increased in the absence of changes in β-secretase activity. Although enrichment alone had no significant effects on Aβ, reduced Aβ load and plaque maturation occurred in animals receiving antioxidants as a component of treatment. Aβ measures did not correlate with cognitive performance on any of the six tasks assessed, suggesting that modulation of Aβ alone may be a relatively minor mechanism mediating cognitive benefits of the interventions. Overall, the data indicate that multidomain treatments may be a valuable intervention strategy to reduce neuropathology and improve cognitive function in humans.
Trapped Platelets Activated in Ischemia Initiate Ventricular Fibrillation

Background— We tested the hypothesis that ischemia-induced ventricular fibrillation (VF) is facilitated by platelets, trapped regionally in the ischemic zone and activated to release arrhythmogenic secretome. Methods and Results— In a randomized study in blood-free, buffer-perfused isolated rat hearts, ischemic zone territory (34±1% of left ventricle) was selected so that ischemia evoked VF in only 42% of controls. VF incidence was increased to 91% (P<0.05) by coronary ligation–induced trapping of freshly prepared autologous platelets (infused before and during coronary ligation, with trapping confirmed by 111In-labeled platelet autoradiographic imaging). Trapping of platelet secretome prepared ex vivo, or platelet-sized fluorospheres, did not increase ischemia-induced VF incidence. Secretome alone did, however, evoke VF in 2 sham coronary-ligated hearts. Perfusion did not activate infused platelets in sham coronary-ligated hearts, whereas ligation activated trapped platelets (assessed by thromboxane release). In a separate study, trapping whole-heparinized blood mimicked the ability of trapped platelets to increase VF incidence. This effect was not prevented by >5 days oral pretreatment in vivo with clopidogrel (10 mg/kg per day) or indomethacin (2.4 mg/kg per day). Conclusions— Platelets facilitate VF during acute ischemia independently of their ability to participate in occlusive thrombosis. Moreover, the effect is unresponsive to antiplatelet drugs commonly used. Labile secretome constituents appear to be responsible. This opens a novel avenue for antiarrhythmic drug research.
Tangential and Radial Epileptic Spike Activity: Different Sensitivity in EEG and MEG

Objective: Observations in epileptic patients show that interictal spikes are sometimes only visible in electroencephalography (EEG) and sometimes only in magnetoencephalography (MEG). This observation cannot readily be explained by the theoretical sensitivities of EEG and MEG based on analytical models. In this context, we aimed to study the directional sensitivity of radial and tangential spike activity in numerical simulations using realistic head models. Methods: We calculated the signal-to-noise ratio (SNR) of simulated spikes at varying orientations and with varying background activity in 12 brain regions in 4 volunteers. Different levels of background activity were modeled by adjusting the amplitudes of several thousand dipoles distributed in the cortex. Results: For a fixed realistic background activity, we found a higher SNR for MEG spikes for spike orientations that deviated not >30° from the tangential direction. In contrast, we found a higher SNR for EEG spikes that deviated not >45° from the radial direction. When the radial background activity was selectively increased, the sensitivity of EEG for radially oriented spikes decreased; when the tangential background activity selectively increased, the sensitivity of MEG for tangentially oriented spikes was decreased. Conclusions: Our simulations provide a possible explanation for the clinically observed differences in epileptic spike detection between EEG and MEG. Epileptic spike detection can be improved by analyzing a combination of EEG and MEG data.
Family study and segregation analysis of Tourette syndrome: evidence for a mixed model of inheritance.

To investigate the transmission of Tourette syndrome (TS) and associated disorders within families, complex segregation analysis was performed on family study data obtained from 53 independently ascertained children and adolescents with TS and their 154 first-degree relatives. The results suggest that the susceptibility for TS is conveyed by a major locus in combination with a multifactorial background. Other models of inheritance were definitively rejected, including strictly polygenic models, all single major locus models, and mixed models with dominant and recessive major loci. The frequency of the TS susceptibility allele was estimated to be . 01. The major locus accounts for over half of the phenotypic variance for TS, whereas the multifactorial background accounts for approximately 40% of phenotypic variance. Penetrance estimates suggest that all individuals homozygous for the susceptibility allele at the major locus are affected, whereas only 2.2% of males and 0.3% of females heterozygous at the major locus are affected. Of individuals affected with TS, approximately 62% are heterozygous and approximately 38% are homozygous at the major locus. While none of the families had two parents affected with TS, 19% of families had two parents affected with the broader, phenotype, which includes TS, chronic tic disorder, or obsessive-compulsive disorder.
Early-stage retinal melatonin synthesis impairment in streptozotocin-induced diabetic wistar rats.

PURPOSE Retinal melatonin synthesis occurs in the photoreceptor layer in a circadian manner, controlling several physiologic rhythmic phenomena, besides being the most powerful natural free radical scavenger. The purpose of the present work was to evaluate the diurnal profile of retinal melatonin content and the regulation of its synthesis in the retina of streptozotocin-induced diabetic rats. METHODS Diabetes was induced in male Wistar rats (12 hour-12 hour light/dark cycle) with streptozotocin. Control, diabetic, and insulin-treated diabetic animals were killed every 3 hours throughout the light-dark cycle. Retinal melatonin content was measured by high-performance liquid chromatography, arylalkylamine N-acetyltransferase (AANAT) activity was analyzed by radiometric assay, Bmal1 gene expression was determined by qPCR, and cyclic adenosine monophosphate (cAMP) content was assessed by ELISA. RESULTS Control animals showed a clear retinal melatonin and AANAT activity daily rhythm, with high levels in the dark. Diabetic rats had both parameters reduced, and the impairment was prevented by immediate insulin treatment. In addition, the Bmal1 expression profile was lost in the diabetic group, and the retinal cAMP level was reduced 6 hours after lights on and 3 hours after lights off. CONCLUSIONS The present work shows a melatonin synthesis reduction in diabetic rats retinas associated with a reduction in AANAT activity that was prevented by insulin treatment. The Bmal1-flattened gene expression and the cAMP reduction seem to be responsible for the AANAT activity decrease in diabetic animals. The melatonin synthesis reduction observed in the pineal gland of diabetic rats is also observed in a local melatonin tissue synthesizer, the retina.
General Strategy to Humanize a Camelid Single-domain Antibody and Identification of a Universal Humanized Nanobody Scaffold*

Nanobodies, single-domain antigen-binding fragments of camelid-specific heavy-chain only antibodies offer special advantages in therapy over classic antibody fragments because of their smaller size, robustness, and preference to target unique epitopes. A Nanobody differs from a human heavy chain variable domain in about ten amino acids spread all over its surface, four hallmark Nanobody-specific amino acids in the framework-2 region (positions 42, 49, 50, and 52), and a longer third antigen-binding loop (H3) folding over this area. For therapeutic applications the camelid-specific amino acid sequences in the framework have to be mutated to their human heavy chain variable domain equivalent, i.e. humanized. We performed this humanization exercise with Nanobodies of the subfamily that represents close to 80% of all dromedary-derived Nanobodies and investigated the effects on antigen affinity, solubility, expression yield, and stability. It is demonstrated that the humanization of Nanobody-specific residues outside framework-2 are neutral to the Nanobody properties. Surprisingly, the Glu-49 → Gly and Arg-50 → Leu humanization of hallmark amino acids generates a single domain that is more stable though probably less soluble. The other framework-2 substitutions, Phe-42 → Val and Gly/Ala-52 → Trp, are detrimental for antigen affinity, due to a repositioning of the H3 loop as shown by their crystal structures. These insights were used to identify a soluble, stable, well expressed universal humanized Nanobody scaffold that allows grafts of antigen-binding loops from other Nanobodies with transfer of the antigen specificity and affinity.
Luminescence based enzyme-labeled phage (Phazyme) assays for rapid detection of Shiga toxin producing Escherichia coli serogroups

Most diagnostic approaches for Shiga toxin producing Escherichia coli (STEC) have been designed to detect only serogroup O157 that causes a majority, but not all STEC related outbreaks in the United States. Therefore, there is a need to develop methodology that would enable the detection of other STEC serogroups that cause disease. Three bacteriophages (phages) that infect STEC serogroups O26, O103, O111, O145 and O157 were chemically labeled with horseradish peroxidase (HRP). The enzyme-labeled phages (Phazymes) were individually combined with a sampling device (a swab), STEC serogroup-specific immunomagnetic separation (IMS) beads, bacterial enrichment broth and luminescent HRP substrate, in a self-contained test device, while luminescence was measured in a hand-held luminometer. The O26 and O157 Phazyme assays correctly identified more than 93% of the bacteria tested during this study, the O123 Phazyme assay identified 89.6%, while the O111 and O145 Phazyme assays correctly detected 82.4% and 75.9%, respectively. The decreased specificity of the O111 and O145 assays was related to the broad host ranges of the phages used in both assays. The Phazyme assays were capable of directly detecting between 105 and 106 CFU/ml in pure culture, depending on the serogroup. In food trials, the O157 Phazyme assay was able to detect E. coli O157:H7 in spinach consistently at levels of 1 CFU/g and occasionally at levels of 0.1 CFU/g. The assay detected 100 CFU/100 cm2 on swabbed meat samples and 102 CFU/100 ml in water samples. The Phazyme assay effectively detects most STEC in a simple and rapid manner, with minimal need for instrumentation to interpret the test result.
How Followers Differing in Career Motivation Gain Career Profits from Transformational Leaders: A Longitudinal Moderated Mediation Model

Although, transformational leadership is among the most thoroughly examined leadership theories, knowledge regarding its association with followers' career outcomes is still limited. Furthermore, the underlying mechanisms explaining how transformational leaders affect their employees' career success are yet not well-understood. Based on theoretical assumptions about the processes involved in setting the goal of “making a career,” we propose an indirect effect of transformational leadership on subjective and objective career success via development opportunities that depends on the level of career motivation of employees. We conducted a longitudinal study with two measurement occasions separated by 13 months with 320 employees of a large IT company. Respondents provided ratings online on their direct supervisor's transformational leadership, their own development opportunities, and career motivation at T1; subjective career success was rated at both time points, whereas objective indicators of career transitions were rated at T2 retrospectively. Using structural equation modeling, we tested the proposed moderated mediation model. The results indicated that transformational leadership increased subordinates' subjective career success via development opportunities. In addition, and contrary to theoretical reasoning, the indirect effect was not significant for employees with high career motivation. Thus, employees high in career motivation appeared not to benefit from the development opportunities offered by transformational leaders. The results are discussed in light of tailored leadership that takes the aspirations, and needs of followers into account.