Virtual planning, simultaneous dental implantation and CAD/CAM plate fixation: a paradigm change in maxillofacial reconstruction.

Prosthetic rehabilitation in patients undergoing reconstructive surgery using vascularized free flaps is challenging, and functional rehabilitation of the patient with a fixed prosthesis is rare. Virtually planned maxillofacial reconstruction including simultaneous dental implantation according to the prosthodontic ideal position of the implants could further enhance dental rehabilitation. The data of 21 patients undergoing fibula free flap reconstructive surgery with CAD/CAM patient-specific reconstruction plates during the years 2015-2018 were analysed, including the applicability of the virtual plan, flap survival, duration of surgery, ischemia time, simultaneous dental implantation, implant exposure, and postoperative complications. The virtual plan could be translated to surgery in all cases. In total, 76 dental implants were simultaneously placed during primary reconstruction in the 21 patients. For 38.1% of these patients, the implants could be uncovered in secondary surgery; the mean duration until exposure was 7.6 months. The implant survival rate was 97.4% (74/76). Wound infection requiring a secondary intervention occurred in 23.8% of patients during follow-up. Virtually planned reconstruction with a fibula free flap, simultaneous dental implantation, and CAD/CAM plates allows early and functional dental rehabilitation. A dental workflow should be integrated into the virtual planning, and prosthetically favourable implant positions should determine the position of the fibula segments.

The influence of lymph node ratio on survival and disease recurrence in squamous cell carcinoma of the tongue.

This study was performed to report the outcomes of patients with oral squamous cell carcinoma (OSCC) of the tongue over a 10-year period with the aim of testing the hypothesis that the lymph node ratio (LNR) has a significant influence on loco-regional recurrence. The charts of 227 patients with OSCC of the mobile tongue treated at the University Hospital of Zurich from 2003 to 2012 were screened. Following the application of the exclusion criteria (prior chemotherapy, radiotherapy, or surgery, perioperative death, N3 disease, unresectable disease, synchronous second primary, no signed informed consent, and follow-up <3years), prospective data were collected and a retrospective analysis performed for 88 of these patients who were treated with selective neck dissection. During a mean follow-up period of 78 months (standard deviation 37 months), loco-regional recurrence was diagnosed in 25 patients (28%). The overall and disease-specific survival rates for the study population were 72% and 80%, respectively. Perineural invasion was identified as an independent risk factor for decreased disease-specific survival, whereas LNR was not. LNR did not show an influence on disease recurrence. Thus, its prognostic value in patients with tongue cancer remains uncertain and the decision regarding adjuvant therapy should not be made solely on the basis of LNR.

Sentinel lymph node biopsy for early stage tongue cancer-a 14-year single-centre experience.

This study was performed to report the usage of sentinel lymph node biopsy (SLNB) in clinical stage I or II tongue cancer patients with cN0 necks seen over a 14-year period. Data were collected prospectively, and a retrospective analysis was performed of 41 patients with early stage oral squamous cell carcinoma of the tongue and a cN0 neck. Sentinel lymph node (SLN)-positive patients underwent elective neck dissection, whereas SLN-negative patients were kept under careful observation. Seven of the 41 (17%) patients enrolled in the study were found to have occult metastases. The patients were followed up for a mean duration of 92 months (range 60-144 months). The neck recurrence rate for SLN-positive patients was 0% and for SLN-negative patients was 3%. The authors recommend the routine use of SLNB in patients with early stage oral squamous cell carcinoma of the tongue and a cN0 neck. Furthermore, special focus should be placed on isolated tumour cells, as their presence is of high clinical relevance.

Total virtual workflow in CAD-CAM bony reconstruction with a single step free fibular graft and immediate dental implants.

The Surgical reconstruction of defects of the face is challenging. Local and regional flaps have an important part to play, but large defects of bone and soft tissue are a greater problem. Microvascular tissue transfer has become the standard for such patients, and preoperative planning of bony reconstructions is now common. To use these preplanning tools best the implants should be placed in the prosthetically ideal place, and the bone positioned to surround the implants - that is, truly backward planning of the position of the bone. The buccolingual angulation and the actual position of the implants during operation can be difficult to verify. Using commonly available software and 3-dimensional printing solutions, therefore, we have constructed an algorithm to optimise the position of these implants during the operation, and to get their position as close to the planned outcome as possible. This algorithm is adaptable to any implant system and is potentially possible in any implant or preplanning software unit.

Complication rate in mandibular angle fractures-one vs. two plates: a 12-year retrospective analysis.

PURPOSE: Treatment of mandibular angle fractures using one or two osteosynthesis plates is still a controversial topic. Fracture, treatment, and patient-dependent influencing factors could affect the overall outcome. In the present retrospective study, complication rates of mandibular angle fractures treated by open reduction were assessed according to type of treatment. MATERIALS AND METHODS: We analyzed retrospective medical records using the search terms "mandibular angle fracture." We included all patients presenting with a mandibular angle fracture treated by open reduction and internal fixation at our department between 2002 and 2012. RESULTS: We included 186 patients treated with open reduction and miniplate fixation (84 one plate; 102 two plates). The early complication rate was significantly higher for the double-plate group (72.5% vs. 47.6%, respectively; p = 0.001). Most common findings in the postoperative period were transient hypoesthesia and tissue swelling. In the two-plate group, a significantly increased operation time of 183 min versus 150 min in the one-plate group was found (p < 0.001). Late complications did not differ significantly between both groups (21.4% single-plate group; 30.4% two-plate fixation group; p = 0.32). CONCLUSION: We found a significantly increased early complication rate in the two-plate group. Long-term complications did not differ between both groups.

Evaluation of the immunochromatographic CORIS Giardia-Strip test for rapid diagnosis of Giardia lamblia.

The aim of this study was to evaluate the performance of the CORIS Giardia-Strip test (CORIS Bioconcept, Gembloux, Belgium) as a rapid initial method for the routine diagnosis of giardiasis. Compared to a commercial ELISA-coproantigen test (ProSpect Giardia-ELISA-microplate assay; Remel, Lenexa, KS, USA), the commercial strip test had a sensitivity of 58%, a specificity of 99%, a positive predictive value of 93% and a negative predictive value of 93% (n=158). These results are comparable to those obtained using microscopy of direct wet-mounted stool. Since the CORIS Giardia-Strip test is simpler to perform, it can replace direct wet-mounted stool microscopy for the rapid diagnosis of giardiasis; however, its sensitivity is inferior to that of other immunochromatographic antigen detection tests and fresh stool samples are required for its use. Nevertheless, the results suggest that a positive CORIS Giardia-Strip test outcome does not need confirmation, while samples with negative results should be re-examined using another, more sensitive, test.

Comparison of different PCR protocols for the detection and diagnosis of Plasmodium falciparum.

An assessment of differing PCR protocols for the diagnosis of Plasmodium falciparum infection was performed on samples from an area of holoendemic malaria transmission in western Burkina Faso. The PCR protocols had generally high sensitivities (>92%) and specificities (>69%), but the negative predictive values (NPV) were moderate and differed widely among the PCR protocols tested. These PCR protocols that amplified either the P. falciparum pfcrt gene or the small subunit ribosomal DNA were the most reliable diagnostic tools. However, the moderate NPV imply that more than one PCR protocol should be used for diagnosis in holoendemic areas.

Adherence of Plasmodium falciparum infected erythrocytes to CHO-745 cells and inhibition of binding by protein A in the presence of human serum.

Adhesion of erythrocytes infected with the malaria parasite Plasmodium falciparum to human host receptors is a process associated with severe malarial pathology. A number of in vitro cell lines are available as models for these adhesive processes, including Chinese hamster ovary (CHO) cells which express the placental adhesion receptor chondroitin-4-sulphate (CSA) on their surface. CHO-745 cells, a glycosaminoglycan-negative mutant CHO cell line lacking CSA and other reported P. falciparum adhesion receptors, are often used for recombinant expression of host receptors and for receptor binding studies. In this study we show that P. falciparum-infected erythrocytes can be easily selected for adhesion to an endogenous receptor on the surface of CHO-745 cells, bringing into question the validity of using these cells as a tool for P. falciparum adhesin expression studies. The adhesive interaction between CHO-745 cells and parasitized erythrocytes described here is not mediated by the known P. falciparum adhesion receptors CSA, CD36, or ICAM-1. However, we found that CHO-745-selected parasitized erythrocytes bind normal human IgM and that adhesion to CHO-745 cells is inhibited by protein A in the presence of serum, but not in its absence, indicating a non-specific inhibitory effect. Thus, protein A, which has been used as an inhibitor for a recently described interaction between infected erythrocytes and the placenta, may not be an appropriate in vitro inhibitor for understanding in vivo adhesive interactions.

Protein transport and trafficking in Plasmodium falciparum-infected erythrocytes.

The human malarial parasite Plasmodium falciparum extensively modifies its host erythrocyte, and to this end, is faced with an interesting challenge. It must not only sort proteins to common organelles such as endoplasmic reticulum, Golgi and mitochondria, but also target proteins across the 'extracellular' cytosol of its host cell. Furthermore, as a member of the phylum Apicomplexa, the parasite has to sort proteins to novel organelles such as the apicoplast, micronemes and rhoptries. In order to overcome these difficulties, the parasite has created a novel secretory system, which has been characterized in ever-increasing detail in the past decade. Along with the 'hardware' for a secretory system, the parasite also needs to 'program' proteins to enable high fidelity sorting to their correct subcellular location. The nature of these sorting signals has remained until relatively recently, enigmatic. Experimental work has now begun to dissect the sorting signals responsible for correct subcellular targeting of parasite-encoded proteins. In this review we summarize the current understanding of such signals, and comment on their role in protein sorting in this organism, which may become a model for the study of novel protein trafficking mechanisms.

Antimalarial activity of extracts of Abutilon grandiflorum G. Don - a traditional Tanzanian medicinal plant.

The Tanzanian medicinal plant Abutilon grandiflorum G. Don was studied for its in vivo and in vitro antiplasmodial effects. The ethyl acetate extract showed prominent in vivo activity against P. vinckei vinckei in mice and in vitro against P. falciparum strains HB3 and FCB. The extract was only moderately cytotoxic if tested in vitro against the colon cell line HT29. In the in vivo study, the results were significantly influenced by the treatment schedule used, i.e. early treatment with higher doses was more successful than applying the same overall amount over a longer period. Phytochemical analysis of the extract provided no conclusive evidence for the observed parasitological effects.

A human schwannoma cell line supports the in vitro adhesion of Plasmodium falciparum infected erythrocytes to chondroitin-4-sulfate.

The paucity of human cell lines expressing defined receptors for the cytoadhesion of erythrocytes infected with the human malarial parasite Plasmodium falciparumhas hampered the investigation of this important virulence property. Here, we investigate a permanent cell line derived from a human, malignant schwannoma, termed HMS-97, and show that this cell line expresses chondroitin-4-sulfate as the only surface receptor to which P. falciparum-infected erythrocytes can cytoadhere. Other common receptors for parasite adhesion, including CD36, vascular cellular adhesion molecule-1 (VCAM), intercellular adhesion molecule-1 (ICAM-1), and E-selectin are absent. Thus, HMS-97 cells are a useful tool for the study of P. falciparum adhesion to chondoitin-4-sulfate, the main receptor for parasite sequestration in the placenta. As chondoitin-4-sulfate can be readily cleaved from the cells, HMS-97 cells are also an ideal system for expressing recombinant adhesion receptors and studying their function in binding assays.

Subcellular localization and characterization of chorismate synthase in the apicomplexan Plasmodium falciparum.

The resurgence of drug-resistant apicomplexa, in particular Plasmodium falciparum, the most fatal human malarial parasite, has focused attention on the recent discovery of the shikimate pathway in these organisms, as it may provide the urgently required, novel drug targets resulting from the absence of this pathway in mammals. The direction of a parasiticidal drug design programme obviously requires knowledge of the subcellular localization and indeed full characterization of the possible enzyme targets. Here, we report the cloning and characterization of chorismate synthase from P. falciparum and present the first biochemical and immunological studies of an enzyme of the shikimate pathway from an apicomplexan parasite. We show that this chorismate synthase does not possess an intrinsic flavin reductase activity and is therefore monofunctional like the plant and bacterial chorismate synthases. Highest immunological cross-reactivity was found with a plant chorismate synthase. However, in contrast to the plant enzyme, which is located to the plastid, P. falciparum chorismate synthase is found in the parasite cytosol, akin to the fungal enzymes that possess an intrinsic flavin reductase activity (i.e. are bifunctional). Thus, P. falciparum chorismate synthase has a combination of properties that distinguishes it from other described chorismate synthases.

A superfamily of variant genes encoded in the subtelomeric region of Plasmodium vivax.

The malarial parasite Plasmodium vivax causes disease in humans, including chronic infections and recurrent relapses, but the course of infection is rarely fatal, unlike that caused by Plasmodium falciparum. To investigate differences in pathogenicity between P. vivax and P. falciparum, we have compared the subtelomeric domains in the DNA of these parasites. In P. falciparum, subtelomeric domains are conserved and contain ordered arrays of members of multigene families, such as var, rif and stevor, encoding virulence determinants of cytoadhesion and antigenic variation. Here we identify, through the analysis of a continuous 155,711-base-pair sequence of a P. vivax chromosome end, a multigene family called vir, which is specific to P. vivax. The vir genes are present at about 600-1,000 copies per haploid genome and encode proteins that are immunovariant in natural infections, indicating that they may have a functional role in establishing chronic infection through antigenic variation.

Single-cell measurements of ion concentrations within the intracellular parasite.

Rapid progress has been made in the study of intracellular ion activities of eukaryotic cells through the recent combination of high-resolution microscopy with fluorimetric ion-specific probes. This technique allows a specific ion concentration within a single living cell to be monitored on-line with high temporal and spatial resolution. In this report, Stefan Wünsch, Paul Horrocks, Michael Gekle and Michael Lanzer evaluate the application of single-cell fluorimetry to the study of transport processes in Plasmodium falciparum.

Mutational analysis identifies a five base pair cis-acting sequence essential for GBP130 promoter activity in Plasmodium falciparum.

Here we describe the functional characterization of a Plasmodium falciparum promoter region, identifying a discrete five base pair sequence element that is responsible for efficient promoter activity. This sequence element binds nuclear factors in a sequence-specific manner. It shares no homology with any known eukaryotic transcription factor binding site, supporting the notion that the protozoan parasite P. falciparum has evolved a transcriptional machinery distinct from that of its human and mosquito hosts. This report represents the first description of a minimal and necessary cis-acting sequence element for efficient promoter activity in P. falciparum.

Differences in nucleosome organization over episomally located plasmids coincides with aberrant promoter activity in P. falciparum.

Here we investigated whether the Plasmodium falciparum GBP130 promoter maintains its developmental activity during the intraerythrocytic cycle when located on an episomal plasmid introduced using transient transfection. Comparing its activity with that of the endogenous chromosomally located GBP130 promoter indicates that the episomally located GBP130 promoter looses its developmental restriction, being rendered constitutively active. Loss of developmental restriction coincides with the absence of phased nucleosomal arrays over the episome. These data suggest that epigenetic factors may play a role in developmentally regulated gene expression in P. falciparum.

Control of gene expression in Plasmodium falciparum.

Transfection has facilitated a functional analysis of transcriptional processes in the human malarial parasite Plasmodium falciparum, providing the first fascinating glimpses into the mechanisms regulating parasite development and pathogenicity. Here we review our rapidly evolving knowledge of what constitutes a promoter, what factors regulate promoter activity and how this activity affects the manifestation of the disease.

Antigenic variation in malaria: in situ switching, relaxed and mutually exclusive transcription of var genes during intra-erythrocytic development in Plasmodium falciparum.

Members of the Plasmodium falciparum var gene family encode clonally variant adhesins, which play an important role in the pathogenicity of tropical malaria. Here we employ a selective panning protocol to generate isogenic P.falciparum populations with defined adhesive phenotypes for CD36, ICAM-1 and CSA, expressing single and distinct var gene variants. This technique has established the framework for examining var gene expression, its regulation and switching. It was found that var gene switching occurs in situ. Ubiquitous transcription of all var gene variants appears to occur in early ring stages. However, var gene expression is tightly regulated in trophozoites and is exerted through a silencing mechanism. Transcriptional control is mutually exclusive in parasites that express defined adhesive phenotypes. In situ var gene switching is apparently mediated at the level of transcriptional initiation, as demonstrated by nuclear run-on analyses. Our results suggest that an epigenetic mechanism(s) is involved in var gene regulation.

Characterization and cloning of the gene encoding the vacuolar membrane protein EXP-2 from Plasmodium falciparum.

As a contribution to the characterization of the parasitophorous vacuolar membrane from Plasmodium falciparum we have begun the identification of vacuolar membrane proteins. Exported protein-2 (EXP-2) is a vacuolar membrane protein exposed into the vacuolar space. To further characterize EXP-2, it was purified, and the 45 N-terminal amino acids were determined by micro-sequencing. Based on this information, partial cDNA and genomic fragments were amplified by PCR and used as probes for the isolation of complete cDNA and genomic DNA clones. The single copy gene is located on chromosome 14, and is transcribed during the ring stage of parasite development. The open reading frame encodes an N-terminal signal sequence which is cleaved from the mature protein. The amino acid composition of EXP-2 is characterized by charged amino acids, with a high abundance of aspartate residues in the C-terminal portion of the protein. In contrast to EXP-1, an integral protein of the vacuolar membrane, EXP-2 lacks a typical hydrophobic transmembrane domain. We suggest that EXP-2 may associate with the vacuolar membrane via an amphipathic helix located in the N-terminal half of the protein.