Bordetella pertussis-infected innate immune cells drive the anti-pertussis response of human airway epithelium.

Pertussis is a severe respiratory tract infection caused by Bordetella pertussis. This bacterium infects the ciliated epithelium of the human airways. We investigated the epithelial cell response to B. pertussis infection in primary human airway epithelium (HAE) differentiated at air-liquid interface. Infection of the HAE cells mimicked several hallmarks of B. pertussis infection such as reduced epithelial barrier integrity and abrogation of mucociliary transport. Our data suggests mild immunological activation of HAE by B. pertussis indicated by secretion of IL-6 and CXCL8 and the enrichment of genes involved in bacterial recognition and innate immune processes. We identified IL-1ß and IFNγ, present in conditioned media derived from B. pertussis-infected macrophage and NK cells, as essential immunological factors for inducing robust chemokine secretion by HAE in response to B. pertussis. In transwell migration assays, the chemokine-containing supernatants derived from this HAE induced monocyte migration. Our data suggests that the airway epithelium on its own has a limited immunological response to B. pertussis and that for a broad immune response communication with local innate immune cells is necessary. This highlights the importance of intercellular communication in the defense against B. pertussis infection and may assist in the rational design of improved pertussis vaccines.

Synthetic Long Peptide Influenza Vaccine Containing Conserved T and B Cell Epitopes Reduces Viral Load in Lungs of Mice and Ferrets.

Currently licensed influenza vaccines mainly induce antibodies against highly variable epitopes. Due to antigenic drift, protection is subtype or strain-specific and regular vaccine updates are required. In case of antigenic shifts, which have caused several pandemics in the past, completely new vaccines need to be developed. We set out to develop a vaccine that provides protection against a broad range of influenza viruses. Therefore, highly conserved parts of the influenza A virus (IAV) were selected of which we constructed antibody and T cell inducing peptide-based vaccines. The B epitope vaccine consists of the highly conserved HA2 fusion peptide and M2e peptide coupled to a CD4 helper epitope. The T epitope vaccine comprises 25 overlapping synthetic long peptides of 26-34 amino acids, thereby avoiding restriction for a certain MHC haplotype. These peptides are derived from nucleoprotein (NP), polymerase basic protein 1 (PB1) and matrix protein 1 (M1). C57BL/6 mice, BALB/c mice, and ferrets were vaccinated with the B epitopes, 25 SLP or a combination of both. Vaccine-specific antibodies were detected in sera of mice and ferrets and vaccine-specific cellular responses were measured in mice. Following challenge, both mice and ferrets showed a reduction of virus titers in the lungs in response to vaccination. Summarizing, a peptide-based vaccine directed against conserved parts of influenza virus containing B and T cell epitopes shows promising results for further development. Such a vaccine may reduce disease burden and virus transmission during pandemic outbreaks.

Increased proportion of perforin-expressing CD8+T-cells indicates control of herpesvirus reactivation in children after stem cell transplantation.

Hematopoietic stem cell transplantation (HSCT) is frequently complicated by viral reactivations. Early diagnosis of viral reactivations and preemptive therapy relies on frequent viralload monitoring. An easy marker of effective cytotoxicity in lymphopenia is lacking and therefore we studied perforin-expression in CD8+T-cells in children following HSCT. Prospectively, we weekly monitored viral loads and perforin-expression of CD8+T-cells in whole blood by FACS, until 4months after HSCT in children. 27 patients were included (median age 4,3, range 0.3-20,1years) of whom 19 developed viral reactivations. These patients showed higher percentages of perforin-expressing CD8+T-cells (17,2%, range 0-63%) than those without (6,8%; range 0-16%) (p=0.001). The increased percentage of perforin-expressing CD8+T-cells coincided with a decrease in viral load with a median interval between maximum viral load and maximum level of perforin-expression of 0,4weeks (range 0.1-7.1). We conclude that perforin-expression in CD8+T-cells may be a marker for effective antiviral T-cell reconstitution early after HSCT in children.

High level of perforin expression in T cells: An early prognostic marker of the severity of herpesvirus reactivation after allogeneic stem cell transplantation in adults.

BACKGROUND: Epstein-Barr virus (EBV) and cytomegalovirus reactivations are frequent complications of hematopoeitic allogeneic stem cell transplantation (SCT) because of a lack of T cell control after immunosuppression. Early diagnosis of reactivation and subsequent preemptive therapy relies on frequent viral load measurement. Additional virus-specific T cell reconstitution data could improve the predictive value of viral load detection for viral complications after transplantation. Here, we studied perforin expression in CD8(+) T cells as a measure of cytotoxic T cell capacity in relation to the occurrence of viral reactivation. METHODS: In a prospective study, we monitored 40 patients during the first 3 months after transplantation and measured viral loads in combination with intracellular perforin expression in CD8(+) T cells. RESULTS: Median perforin expression in CD8(+) T cells throughout follow-up was higher in patients with viral reactivations than in patients without viral reactivations (4.9% vs 2.3%; P = .001). The median percentage of perforin-expressing CD8(+) T cells in patients with high viral reactivations exceeding 1000 copies/mL (10.7%) was statistically significantly higher than that in patients with minor reactivations of 50-1000 copies (4.0%), that in patients with detectable EBV loads that did not exceed the detection limit of 50 copies/mL (2.9%), and that in patients without reactivations (0.8%). Patients with high viral reactivations reached a high percentage of perforin-expressing CD8(+) T cells (>10.2%) more often and faster than did patients with low viral loads (1000 copies/mL) or without viral reactivations. High perforin expression preceded high viral loads. CONCLUSION: Perforin-expressing CD8(+) T cells may be useful as an easy-to-measure prognostic marker for identifying patients at risk for severe viral reactivation very soon after SCT.

The "Red Lady" ages gracefully: new ultrafiltration AMS determinations from Paviland.

The "Red Lady" partial human skeleton found at Goat's Hole, Paviland, in south Wales by William Buckland in 1823 is one of the iconic relics of the British Paleolithic. Originally thought to be Roman, a Paleolithic age has been suspected from the middle of the 19th century. Several attempts have been made at directly radiocarbon dating the "Red Lady," and here we report new determinations that suggest that it is, by a significant margin, the oldest of a group of 'rich,' Mid-Upper Paleolithic burials. We list similar Gravettian-aged burials from Europe, which have been dated recently for comparison. In this paper, we also reconsider the chronology of human use of the cave, apart from as a burial location.

Isotope evidence for the intensive use of marine foods by Late Upper Palaeolithic humans.

We report here on direct evidence for the intensive consumption of marine foods by anatomically modern humans at approximately 12,000 years ago. We undertook isotopic analysis of bone collagen from three humans, dating to the late Palaeolithic, from the site of Kendrick's Cave in North Wales, UK. The isotopic measurements of their bone collagen indicated that ca. 30% of their dietary protein was from marine sources, which we interpret as likely being high trophic level marine organisms such as marine mammals. This indicates that towards the end of the Pleistocene modern humans were pursuing a hunting strategy that incorporated both marine and terrestrial mammals. This is the first occurrence of the intensive use of marine resources, specifically marine mammals, that becomes even more pronounced in the subsequent Mesolithic period.

Reconfigurable systems for sequence alignment and for general dynamic programming

Reconfigurable systolic arrays can be adapted to efficiently resolve a wide spectrum of computational problems; parallelism is naturally explored in systolic arrays and reconfigurability allows for redefinition of the interconnections and operations even during run time (dynamically). We present a reconfigurable systolic architecture that can be applied for the efficient treatment of several dynamic programming methods for resolving well-known problems, such as global and local sequence alignment, approximate string matching and longest common subsequence. The dynamicity of the reconfigurability was found to be useful for practical applications in the construction of sequence alignments. A VHDL (VHSIC hardware description language) version of this new architecture was implemented on an APEX FPGA (Field programmable gate array). It would be several magnitudes faster than the software algorithm alternatives.

Sonographic detection of echogenic fluid and correlation with culdocentesis in the evaluation of ectopic pregnancy.

OBJECTIVE: Because the presence of echogenic fluid on transvaginal sonography has been shown to correlate well with hemoperitoneum in patients with possible ectopic pregnancy, the aim of this study was to compare echogenic fluid on sonography with the results of culdocentesis in predicting hemoperitoneum. MATERIALS AND METHODS: Free fluid on transvaginal sonography and the results of culdocentesis were correlated with the presence or absence of hemoperitoneum in 46 patients at surgery. Forty ectopic pregnancies and six nonectopic pregnancies were found. Echogenic fluid was the criterion used to establish hemoperitoneum on sonography. For statistical analysis, negative and nondiagnostic culdocentesis results were combined. The sensitivity, specificity, and positive and negative predictive values of each diagnostic technique were compared. RESULTS: In 40 of 46 patients with ectopic pregnancy, the sensitivity and specificity of echogenic fluid for establishing hemoperitoneum were 100% and 100%, respectively, compared with 66% and 80%, respectively, for culdocentesis. More important, the negative predictive value of a nondiagnostic culdocentesis was 25% compared with 100% for echogenic fluid in the ectopic subgroup of patients. In two patients with incomplete abortions, sonography failed to detect small amounts of hemoperitoneum at surgery performed 4 hr and 7 days after sonography. CONCLUSION: Sonography is more sensitive than culdocentesis in the detection of hemoperitoneum. Culdocentesis is invasive, and nondiagnostic results cannot be used to exclude hemoperitoneum. Culdocentesis should play no role in the evaluation of ectopic pregnancy except in the unusual circumstance in which high-resolution sonography cannot be readily performed.

Pins, dowels, and other retentive devices in posterior teeth.

Devices used to increase retention of restorations to severely damaged posterior teeth are described, with an emphasis on pins and dowels. Advantages and disadvantages of different techniques and measures to help prevent and correct problems are presented.

A comparison of the abrasiveness of six ceramic surfaces and gold.

A type III gold alloy and six different ceramic surfaces were secured in an abrasion machine opposing extracted teeth to determine their relative abrasiveness and resistance to wear. The rankings of restorative materials from least abrasive to most abrasive were: gold alloy, polished; cast ceramic, polished; porcelain, polished; cast ceramic, polished and shaded; porcelain, polished and glazed; cast ceramic, cerammed skin shaded; and cast ceramic, cerammed skin unshaded. The ranking of materials from most wear-resistant to least wear-resistant was: gold alloy, cast ceramic cerammed, cast ceramic cerammed and shaded, porcelain polished, porcelain glazed, cast ceramic polished and shaded, and cast ceramic polished.

The ceramic reverse three-quarter crown for anterior teeth: laboratory procedures.

The laboratory procedures for making a Hi-ceram porcelain anterior reserve three-quarter crown are described. A core of high-fusing aluminum oxide is used as a coping on which tooth contours are restored with a compatible porcelain. Standard dental laboratory equipment is used and the technique can be quickly learned by any experienced ceramics technician.

The ceramic reverse three-quarter crown for anterior teeth: preparation design.

A new preparation design of the ceramic reverse three-quarter crown for anterior teeth offers a way to improve the appearance of anterior teeth with slight to moderate structural damage. Advantages and indications of the new design are discussed.

An intraoral lift technique for refining porcelain margins.

A method has been described by which a paste of shoulder porcelain and light-curing resin is used to correct deficient porcelain margins. The resin-bonded porcelain is fired in an oven for final hardening.