Additively manufactured biomedical Ti-Nb-Ta-Zr lattices with tunable Young's modulus: Mechanical property, biocompatibility, and proteomics analysis.

Some ß-Ti alloys, such as Ti-Nb-Ta-Zr (TNTZ) alloys, exhibit a low Young's modulus and excellent biocompatibility. These alloys are promising new generation biomedical implant materials. Selective laser melting (SLM) can further enable customer-specific manufacturing of ß-Ti alloys to satisfy the ever-increasing need for enhanced biomedical products. In this study, we quantitatively determined the relationships between porosity, yield strength, and Young's modulus of SLM-prepared TNTZ lattices. The study constitutes a critical step toward understanding the behavior of the lattice and eventually enables tuning the Young's modulus to match that of human bones. Fatigue properties were also investigated on as-printed lattices in terms of the stress limit. The biocompatibility study included a routine evaluation of the relative cell growth rate and a proteomics analysis using a common mouse fibroblast cell line, L929. The results indicated that the as-printed TNTZ samples exhibited evidence of protein proliferation of the L929 cells, particularly P06733, and that those proteins are responsible for biological processes and molecular functions. They in turn may have promoted cell regeneration, cell motility, and protein binding, which at least partially explains the good biocompatibility of the as-printed TNTZ at the protein level. The study highlights the promising applications of additively manufactured TNTZ as a bone-replacing material from mechanical and biocompatibility perspectives.

[Intravenous leiomyomatosis]. / Die intravenöse Leiomyomatose.

Intravenous leiomyomatosis (IVLM) is an unusual neoplasm derived from uterine smooth muscle cells seen in patients with uterine leiomyomas. The typical histological features of IVLM consist of benign smooth muscle cells present within venous vascular spaces of the uterine wall. Increasing intravascular and intracardial spread of IVLM may lead to life-threatening clinical complications. Thorough pathological study of routine hysterectomy specimens may lead to the diagnosis of IVLM. Most affected patients will be cardiologically asymptomatic at the time of diagnosis. Herein, the relatively unknown clinical and morphological aspects of IVLM are presented and discussed.

In vitro biodegradation testing of Mg-alloy EZK400 and manufacturing of implant prototypes using PM (powder metallurgy) methods.

The study is focussing towards Metal Injection Moulding (MIM) of Mg-alloys for biomedical implant applications. Especially the influence of the sintering processing necessary for the consolidation of the finished part is in focus of this study. In doing so, the chosen high strength EZK400 Mg-alloy powder material was sintered using different sintering support bottom plate materials to evaluate the possibility of iron impurity pick up during sintering. It can be shown that iron pick up took place from the steel bottom plate into the specimen. Despite the fact that a separating boron nitrite (BN) barrier layer was used and the Mg-Fe phase diagram is not predicting any significant solubility to each other. As a result of this study a new bottom plate material not harming the sintering and the biodegradation performance of the as sintered material, namely a carbon plate material, was found.

Production, characterisation, and cytocompatibility of porous titanium-based particulate scaffolds.

Despite its non-matching mechanical properties titanium remains the preferred metal implant material in orthopaedics. As a consequence in some cases stress shielding effect occurs, leading to implant loosening, osteopenia, and finally revision surgery. Porous metal scaffolds to allow easier specialised cells ingrowth with mechanical properties closer to the ones of bone can overcome this problem. This should improve healing processes, implant integration, and dynamic strength of implants retaining. Three Ti-6Al-4V materials were metal injection moulded and tailored porosities were effectively achieved. After microstructural and mechanical characterisation, two different primary cells of mesenchymal origin (human umbilical cord perivascular cells and human bone derived cells which revealed to be two pertinent models) as well as one cell line originated from primary osteogenic sarcoma, Saos-2, were bestowed to investigate cell-material interaction on genomic and proteome levels. Biological examinations disclosed that no material has negative impact on early adhesion, proliferation or cell viability. An efficient cell ingrowth into material with an average porosity of 25-50 µm was proved.

Theileria parva 104 kDa microneme--rhoptry protein is membrane-anchored by a non-cleaved amino-terminal signal sequence for entry into the endoplasmic reticulum.

The 104 kDa microneme-rhoptry protein (p104) is the only known apical complex organelle-specific protein of Theileria parva. p104 exhibits striking structural similarities to circumsporozoite protein and sporozoite surface protein 2 of Plasmodium yoelii. Their primary sequences contain two hydrophobic segments, located at the amino-and the carboxy-terminus. The p104 amino-terminal hydrophobic region was suggested to be a signal peptide for entry into the endoplasmic reticulum and the extreme carboxy-terminal region to function as a membrane anchor. We have studied the biogenesis of p104 in a cell-free expression system and found that p104 is co-translationally transported into membranes derived from endoplasmic reticulum. The amino-terminal signal peptide is not cleaved off and anchors the protein in the membrane with the carboxy-terminal portion translocated into the lumen. We suggest that in vivo p104 is co-translationally integrated into the membrane of the endoplasmic reticulum, from where it is further transported to the microneme-rhoptry complex. Thus, p104 appears to be a suitable marker to study the development of micronemes and rhoptries in T. parva.

Characterization of a secretory type Theileria parva glutaredoxin homologue identified by novel screening procedure.

The schizont stage of the protozoan parasite Theileria parva induces features characteristic of tumor cells in infected bovine T-cell lines. Most strikingly T. parva-infected cell lines acquire unlimited growth potential in vitro. Their proliferative state is entirely dependent on the presence of a viable parasite within the host cell cytoplasm. It has been postulated that parasite proteins either secreted into the host cell or expressed on the parasite surface membrane are involved in the parasite-host cell interaction. We used an in vitro transcription-translation-membrane translocation system to identify T. parva-derived cDNA clones encoding secretory or membrane proteins. Within 600 clones we found one encoding a 17-kDa protein which is processed by microsomal membranes to a 14-kDa protein (11E), presumably by signal peptidase. The processed form is expressed in the T-cell line TpM803 harboring viable parasites. By immunolocalization we show that the 11E protein mostly resides within the parasite, often in close vicinity to membranous structures, but in addition it appears at the surface membrane. Amino acid sequence comparison suggests that 11E belongs to the glutaredoxin family, but is unique so far in containing a signal sequence for endoplasmic reticulum membrane translocation.

Breast neoplasms: T2* susceptibility-contrast, first-pass perfusion MR imaging.

PURPOSE: To evaluate the differentiation of benign from malignant breast tumors with T2*-weighted perfusion magnetic resonance (MR) imaging (blood volume imaging) versus that with dynamic T1-weighted contrast agent-enhanced MR imaging. MATERIALS AND METHODS: Ten healthy adult volunteers and 18 adult patients with benign or malignant lesions underwent both conventional T1-weighted dynamic contrast-enhanced breast MR imaging and repetitive first-pass, single-section, dynamic T2*-weighted perfusion MR imaging. Images were obtained before, during, and after injection of 20 mL of gadopentetate dimeglumine; peak gadopentetate dimeglumine concentrations were calculated from the maximal signal intensity loss on T2*-weighted images. RESULTS: No perfusion effect was detectable in healthy breast parenchyma. A strong susceptibility-mediated signal intensity loss occurred in malignant breast tumors. No or only minor perfusion effects were seen in fibroadenomas, in spite of their rapid enhancement at T1-weighted dynamic imaging. Perfusion imaging was possible after conventional dynamic contrast-enhanced breast MR imaging. CONCLUSION: T2*-weighted perfusion imaging exploits the susceptibility-mediated signal intensity loss of a first-pass bolus of gadopentetate dimeglumine within the capillary bed. First-pass perfusion imaging of breast lesions is feasible. It is promising in the differentiation of benign from malignant, rapidly enhancing lesions.

Trisomy 8 and 18 as frequent clonal and single-cell aberrations in 185 primary breast carcinomas.

For cytogenetic investigations short-term cultures of 185 breast carcinomas (135 invasive ductal, 21 invasive lobular, 12 invasive ductal with intraductal components, seven heterogeneous, six intraductal, four invasive ductal and lobular) were prepared. Cytogenetic examinations revealed clonal abnormalities in 39 cases with a predominance of simple numerical chromosome changes, i.e., trisomies of chromosomes 7, 8, and 18. One hundred forty-six tumors did not show clonal abnormalities, but single-cell aberrations other than monosomies occurred in 79 of these tumors. Compared to cells of epithelial hyperplasia of the breast, amniotic fluid cells, and cells from pleomorphic adenomas cultivated using the same medium, the frequency of single-cell trisomies was significantly higher. Trisomy 8 was not only found as a clonal aberration in 10 cases but was also the most frequent non-clonal aberration. Trisomy 7 and 18 were also frequent clonal as well as non-clonal cytogenetic deviations.

Sequence and expression of a 90-kilodalton heat-shock protein family member of Theileria parva.

A Theileria parva specific full-length cDNA clone, T7, which encodes a protein with more than 60% homology to heat shock protein 90 (hsp90) of other organisms, has been identified. T7 appears to be a single copy gene. The gene is expressed as a protein of 87 kDa in both the sporozoite and schizont stages of T. parva. The protein was not found in the piroplasm stage, although the corresponding transcript was detected, suggesting post-transcriptional regulation of the gene. In the schizont stage the T7 protein is upregulated upon heat shock and localized in the cytoplasm.

Intracellular transport of rubella virus structural proteins expressed from cloned cDNA.

The structural proteins of rubella virus consist of a nucleocapsid protein (C) and two membrane-embedded spike glycoproteins (E1 and E2). Since many reports have suggested that rubella virus buds intracellularly, we have examined the intracellular transport of the structural proteins in the absence of virion formation, particularly whether the membrane glycoproteins are retained inside the cell or are transported to the cell surface. We have expressed the structural proteins from cloned cDNA either alone or in different combinations, have examined the intracellular location of the proteins by immunofluorescence and using biochemical methods, and have looked for plasma membrane-localized E1 or E2 using a cell surface biotinylation assay. The C protein was found in the Golgi complex when expressed with E2 and E1; without the membrane glycoproteins, C appeared to remain in the endoplasmic reticulum (ER). When expressed alone, E1 was retained in a pre-Golgi compartment, and was not detected at the cell surface in any cell line. When E2 was expressed alone a small fraction could be detected at the cell surface, but the majority was retained intracellularly, apparently in the ER and the Golgi. Both proteins were transported to the surface when they were expressed together, albeit with low efficiencies in all cell lines. These data suggest that, although neither glycoprotein carries a dominant intracellular retention signal, E2 and E1 are largely retained in the Golgi even when present as a transport-competent heterodimer.

Reduced adenosine 3',5'-cyclic monophosphate levels in patients with reversible obstructive airways disease.

1. Patients were grouped into categories of 'no airways disease', 'obstructive airways disease without response to bronchodilator' and 'obstructive airways disease with bronchodilator responsiveness'. 2. Cyclic nucleotides were assayed in specimens of lung tissue that were excised during surgery. 3. Reduced levels of adenosine 3',5'-cyclic monophosphate (cyclic AMP) were found in pulmonary tissue obtained from patients with reversible obstructive airways disease, lending support to the beta-adrenergic theory of asthma.