The Impact of Delirium on Recovery in Geriatric Rehabilitation after Acute Infection.

OBJECTIVES: Delirium is common during acute infection in older patients and is associated with functional decline. Geriatric rehabilitation (GR) can help older patients to return to their premorbid functional level. It is unknown whether delirium affects GR outcomes in patients with acute infection. We evaluated whether delirium affects trajectories of activities of daily living (ADL) and quality of life (QoL) recovery in GR after COVID-19 infection. DESIGN: This study was part of the EU-COGER study, a multicenter cohort study conducted between October 2020 and October 2021. SETTING AND PARTICIPANTS: Participants were recruited after COVID-19 infection from 59 GR centers in 10 European countries. METHODS: Data were collected at GR admission, discharge, and at the 6-week and 6-month follow-ups. Trajectories of ADL [using the Barthel index (BI)] and QoL [using the EuroQol-5 Dimensions-5 Level (EQ-5D-5L)] recovery were examined using linear mixed models. RESULTS: Of the 723 patients included (mean age 75.5 ± 9.9 years; 52.4% male), 28.9% had delirium before or during GR admission. Participants with delirium recovered in ADL at approximately the same rate as those without (linear slope effect = -0.13, SE 0.16, P = .427) up to an estimated BI score of 16.1 at 6 months. Similarly, participants with delirium recovered in QoL at approximately the same rate as those without (linear slope effect = -0.017, SE 0.015, P = .248), up to an estimated EQ-5D-5L score of 0.8 at 6 months. CONCLUSIONS AND IMPLICATIONS: Presence of delirium during the acute phase of infection or subsequent GR did not influence the recovery trajectory of ADL functioning and QoL.

Post-acute COVID-19 geriatric rehabilitation : A European perspective.

Coronavirus disease 2019 (COVID-19) poses a threat to the health and independence of older people in particular. In this article we elaborate on the content and importance of post-acute COVID-19 geriatric rehabilitation from a European perspective. We explain the geriatric rehabilitation paradox and how this can and should be solved. We also present what post-acute COVID-19 geriatric rehabilitation should entail. This might not only help us to develop better geriatric rehabilitation services, but it should also inform pandemic preparedness in the future.

Pan-European Study on Functional and Medical Recovery and Geriatric Rehabilitation Services of Post-COVID-19 Patients: Protocol of the EU-COGER Study.

OBJECTIVES: There is insufficient knowledge about the functional and medical recovery of older people infected with SARS-CoV-2. This study aims to gain insight into the course of functional and medical recovery of persons who receive geriatric rehabilitation (GR) following SARS-CoV-2 infection across Europe. Special attention will be paid to the recovery of activities of daily living (ADL) and to the GR services offered to these patients. DESIGN: A multi-center observational cohort study. SETTING AND PARTICIPANTS: This study will include several European countries (EuGMS member states) each providing at least 52 comparable routine datasets (core dataset) of persons recovering from a SARS-CoV-2 infection and receiving geriatric rehabilitation. The routine data will be anonymously collected in an online CASTOR database. The ethical regulations of each participating country will be followed. PRIMARY OUTCOME: ADL functioning. SECONDARY OUTCOMES: length of stay, discharge destination, hospital readmission and mortality. Other variables that will be collected are quality of life, treatment modalities, complications, cognition, frailty, mood/anxiety, BMI, nutrition and pain. All variables will be reported at admission and compared with follow-up scores (discharge, 6 weeks and 6 months follow-up). CONCLUSION: This study will explore the effect of geriatric rehabilitation on post-COVID-19 patients, especially on ADL recovery, and the variety of geriatric rehabilitation services across Europe. Information from this study may help improve recovery of older persons infected with SARS-CoV-2 and improve geriatric rehabilitation services in the ongoing COVID-19 pandemic.

Kinetic effect of different ground conditions on the sole of the claws of standing and walking dairy cows.

For the first time, we analyzed the direct kinetic effects of concrete and rubber flooring on the soles of live dairy cows' claws while standing and walking. Ten adult dairy cows were equipped with foil-based pressure sensors (HoofSystem, Tekscan Inc., Boston, MA) under their left hind leg using a leather claw shoe. These sensors captured parameters of pressure distribution and vertical ground reaction forces while the cows walked on the 2 tested flooring types. The mean pressure was 15.1 to 21.1% lower on rubber flooring compared with concrete; mean pressure values (± standard deviation) were 36.32 ± 7.77 N/cm2 for static measures and 57.33 ± 11.77 N/cm2 for dynamic measures. We observed an even more obvious relief on rubber flooring in the maximum pressure loads, which were 30.1 to 32.7% lower on rubber flooring compared with concrete; mean pressure values (± standard deviation) were 98.79 ± 14.49 N/cm2 for static measures and 150.45 ± 20.87 N/cm2 for dynamic measures). The force-time curves of the dynamic measures essentially showed biphasic curve progression, with local peaks at 29 and 79% of the stance phase. However, we found considerable differences in curve progression between individuals and between the lateral and medial claws, which may be verified in further investigations with more animals. The study showed a noticeable reduction in mechanical load during standing and walking on rubber flooring compared with concrete.

Does the range of motion in the bovine interphalangeal joints change with flooring condition? A pilot study using biplane high-speed fluoroscopic kinematography.

This study had 2 objectives. The first objective was to investigate motion patterns and the range of motion of the bovine proximal and distal interphalangeal joints on concrete during the stance phase. The second objective was to determine whether the range of motion and the locomotive stability of the interphalangeal joints differ on concrete and 3 different commercially available rubber mats (Karera, Kura, and Pedikura; Kraiburg Elastik GmbH & Co. KG, Tittmoning, Germany). Biplane high-speed fluoroscopic kinematography (72 ± 2.5 kV and 112.5 ± 12.5 mA, refresh rate 500 frames per second, shutter 0.5 ms) was applied to record 1 stance phase of the right forelimb of 2 Holstein Friesian heifers (15 mo old, 440 ± 10 kg; ± standard deviation) on each flooring. Three-dimensional digital animations were generated with a marker-supported manual animation technique based on the recordings and computer tomographic bone models. The mean maximum range of motion of each of the 4 interphalangeal joints in terms of flexion/extension, abduction/adduction, and internal/external rotation were calculated as well as the mean number of local extrema as a measure of stability during the stance phase. The main degree of freedom in all interphalangeal joints was flexion and extension with a range of motion of 17.7 to 25.9°. The second largest degree of freedom differed between abduction/adduction (7.7-10.0°) and internal/external rotation (6.5-9.6°) depending on the joint. Remarkably, although smaller, these extrasagittal directions still contribute to the overall motion to a considerable degree. In addition, the interphalangeal joints of the lateral digit showed a tendency to move less during the stance phase than their medial counterparts. Comparing concrete to the rubber mats, the interphalangeal joints tend to have to cover a larger range of motion on concrete with the exception of the distal interphalangeal joint in terms of flexion/extension. The unyielding surface of concrete seems to force the flexible parts of the animal-ground-interaction into extended motion. Furthermore, there tends to be more instability in all 3 degrees of freedom in all 4 joints on concrete, implying a greater effort of the soft tissues to achieve a balanced motion. Detailed biomechanical research contributes to the development of adequate flooring systems by evaluating the mechanical strain on claws and joints and working toward lameness prevention and thus animal welfare.

Effect of different flooring types on pressure distribution under the bovine claw - an ex vivo study.

BACKGROUND: Mechanical interactions between hard floorings and the sole of bovine claws can be reasonable to cause traumatic claw lesions. In this ex vivo study, the direct kinetic impact of concrete and three types of rubber mats on the sole of dairy cattle claws was analyzed. In order to apply uniform loads, isolated distal hind limbs of adult Holstein Friesian dairy cows were functionally trimmed according to the Dutch method and attached to a load applicator. Kinetic data were recorded using a thin, foil-based pressure measurement system (Hoof™ System, Tekscan®). RESULTS: On concrete, the load distribution between the lateral and medial claw was less balanced than on the rubber floorings. The loaded area was significantly smaller on concrete (32.2 cm2) compared to all rubber mats (48.3-58.0 cm2). Average pressures (Pav) and maximum pressures (Pmax) were significantly higher on concrete (Pav 44.7 N/cm2; Pmax 130.3 N/cm2) compared to the rubber floorings (Pav 24.9-29.7 N/cm2; Pmax 71.9-87.2 N/cm2). Pressure peaks occurred mainly in plantar and abaxial parts of the lateral claw and in apical and plantar regions of the medial claw. Load distribution displayed a widely unloaded slope region, but considering the pressure distribution under the claw, none of the zones showed a generally lower pressure exposure. CONCLUSIONS: Altogether, rubber floorings lead to a significant mechanical relief of the sole compared to concrete. Furthermore, relevant differences between the tested rubber mats could be determined. Therefore the used system may be applied to compare further flooring types.

Validation of biplane high-speed fluoroscopy combined with two different noninvasive tracking methodologies for measuring in vivo distal limb kinematics of the horse.

REASON FOR PERFORMING STUDY: Biplane high-speed fluoroscopy is a new method for gait analysis of the equine distal extremity. This is the first study validating the noninvasive tracking possibilities (Autoscoping and Scientific Rotoscoping) taking equine anatomy into account. OBJECTIVES: To determine the resolution with which Autoscoping and Scientific Rotoscoping depict motion of the equine phalanges in comparison to the invasive gold standard marker-based registration. STUDY DESIGN: Comparative ex vivo study. METHODS: In 5 distal extremities of slaughtered ponies, 3 or 4 tantalum beads with 1 mm diameter were implanted in each of the proximal, middle and distal phalangeal bones. Three-dimensional models of the bones were reconstructed using computed tomographic data (120 kV, 50 mA, slice thickness 1 mm, increment 0.5). The beads were digitally removed from the bone models. Biplane fluoroscopic videos were taken at 69.5 ± 3.5 kV, 102.5 ± 22.5 mA, 500 frames/s and 0.5 ms shutter speed. The 5 specimens were moved in the trial field of the biplane fluoroscopic setup in a step-like motion (simulation of landing, main stance phase, lift-off). Marker-based registration, Autoscoping and Scientific Rotoscoping were carried out. For statistical analysis agreement was computed as percentiles, mean and s.d. RESULTS: The medians of Scientific Rotoscoping ranged from 0.16 to 0.66 mm in translations and 0.43 to 2.78° in rotations, while values for Autoscoping were 0.13-0.70 mm and 0.28-2.39° respectively. With 2 exceptions, all differences between methods were statistically significant. Scientific Rotoscoping is more time efficient than Autoscoping and results in smaller maximum errors. MAIN LIMITATIONS: The experimental set-up was specifically designed to accommodate in vivo requirements. Autoscoping was not manually corrected but rather expected to work automatically. CONCLUSIONS: It is possible to noninvasively apply both Autoscoping and Scientific Rotoscoping for gait analysis of the equine phalanges with high precision. The summary is available in Chinese - see supporting information.

Dormancy and activation of human oocytes from primordial and primary follicles: molecular clues to oocyte regulation.

STUDY QUESTION: Do specific transcriptome dynamics in human oocytes from primordial and primary follicles identify novel pathways in oocyte activation? SUMMARY ANSWER: The transcriptomic profiles in oocytes from primordial and primary follicles, respectively, revealed several new canonical pathways as putative mediators of oocyte dormancy and activation. WHAT IS KNOWN ALREADY: Cellular signaling pathways including PI3K/AKT and AKT/mTOR as well as TGF-ß and IGF signaling are known to regulate the primordial-to-primary transition in mammalian follicle development. STUDY DESIGN, SIZE, DURATION: We performed a class comparison study on human oocytes from primordial (n = 436) and primary (n = 182) follicles donated by three women having ovarian tissue cryopreserved before chemotherapy. PARTICIPANTS/MATERIALS, SETTING, METHODS: RNA was extracted from oocytes from primordial and primary follicles isolated by Laser Capture Microdissection, and submitted to the HiSeq Illumina platform. Data mapping, quality control, filtering and expression analysis were performed using Tophat (2.0.4), Cufflinks (2.0.2), BWA (0.6.2) and software R. Modeling of complex biological systems was performed using the IPA® software. Finally, qPCR and immunohistochemistry were employed to explore expression and localization of selected genes and products in human ovarian tissue. MAIN RESULTS AND THE ROLE OF CHANCE: We found 223 and 268 genes down-regulated and up-regulated, respectively, in the oocytes during the human primordial-to-primary follicle transition (P < 0.05 and/or FPKM fold-change >2). IPA® enrichment analysis revealed known pathways ('mTOR Signaling', 'PI3K/AKT Signaling' and 'PTEN Signaling') as well as enriched canonical pathways not previously associated with human ovarian follicle development such as 'ErB Signaling' and 'NGF Signaling' in the down-regulated category and 'Regulation of eIF4 and P70S6K Signaling' and 'HER-2 Signaling in Breast Cancer' in the up-regulated group. Additionally, immunohistochemistry on human ovarian tissue explored the intraovarian localization of VASA, FOXO1 and eIF4E. LARGE SCALE DATA: http://users-birc.au.dk/biopv/published_data/ernst_2017/. LIMITATIONS, REASONS FOR CAUTION: This is a descriptive analysis and no functional studies were performed. The study was based on a limited number of patients and the experimental design could not take into account the natural biological variance in human samples. Therefore, qPCR was used to confirm selected genes alongside immunohistochemical stainings. WIDER IMPLICATIONS OF THE FINDINGS: This study shows, for the first time, a detailed molecular description of global gene transcription activities in oocytes from primordial and primary follicles, respectively. Knowing the global transcription profiles of human oocyte dormancy and activation are important in developing new clinical applications. STUDY FUNDING/COMPETING INTEREST(S): E.H.E. was supported by Health Faculty, Aarhus University and Kong Christian Den Tiendes Fond. K.H. and S.F. were supported by an MRC (UK) project grant MR/M012638/1. K.L.H. was supported by grants from Fonden til Lægevidenskabens Fremme, Kong Christian Den Tiendes Fond. K.L.H. and L.S. were supported by the IDEAS grant from Aarhus University Research Foundation (AUFF). There are no conflicts of interest.

Validation of 2 noninvasive, markerless reconstruction techniques in biplane high-speed fluoroscopy for 3-dimensional research of bovine distal limb kinematics.

Lameness severely impairs cattle's locomotion, and it is among the most important threats to animal welfare, performance, and productivity in the modern dairy industry. However, insight into the pathological alterations of claw biomechanics leading to lameness and an understanding of the biomechanics behind development of claw lesions causing lameness are limited. Biplane high-speed fluoroscopic kinematography is a new approach for the analysis of skeletal motion. Biplane high-speed videos in combination with bone scans can be used for 3-dimensional (3D) animations of bones moving in 3D space. The gold standard, marker-based animation, requires implantation of radio-opaque markers into bones, which impairs the practicability for lameness research in live animals. Therefore, the purpose of this study was to evaluate the comparative accuracy of 2 noninvasive, markerless animation techniques (semi-automatic and manual) in 3D animation of the bovine distal limb. Tantalum markers were implanted into each of the distal, middle, and proximal phalanges of 5 isolated bovine distal forelimbs, and biplane high-speed x-ray videos of each limb were recorded to capture the simulation of one step. The limbs were scanned by computed tomography to create bone models of the 6 digital bones, and 3D animation of the bones' movements were subsequently reconstructed using the marker-based, the semi-automatic, and the manual animation techniques. Manual animation translational bias and precision varied from 0.63 ± 0.26 mm to 0.80 ± 0.49 mm, and rotational bias and precision ranged from 2.41 ± 1.43° to 6.75 ± 4.67°. Semi-automatic translational values for bias and precision ranged from 1.26 ± 1.28 mm to 2.75 ± 2.17 mm, and rotational values varied from 3.81 ± 2.78° to 11.7 ± 8.11°. In our study, we demonstrated the successful application of biplane high-speed fluoroscopic kinematography to gait analysis of bovine distal limb. Using the manual animation technique, kinematics can be measured with sub-millimeter accuracy without the need for invasive marker implantation.

Serum antibodies against native and denaturated hemagglutinin glycoproteins detected by ELISA as correlates of protection after influenza vaccination in healthy vaccinees and in kidney transplant recipients.

The microneutralization assay is the standard method to investigate immune responses to influenza vaccination. However there remains some uncertainty as to whether ELISA results are a true measure of immunity in healthy or immuno-compromised vaccines. Furthermore it has been questioned if antibodies against native ("folded") and against denaturated ("unfolded") viral glycoproteins can equally be used as a marker of protection. In this study, two different quantitative IgG-ELISA assays detecting (i) antibodies against unfolded recombinant hemagglutinin (HA) (r-ELISA) and (ii) antibodies against the native HA on the influenza virus surface captured by fetuin-linkage (f-ELISA) were compared to microneutralization titers in sera from 29 healthy vaccinees (n=87 sera) and 39 kidney transplant recipients (n=117 sera) collected before, three weeks after and six months after vaccination against influenza A (H1N1) 2009. With both ELISAs a significant increase in antibody levels was detected after vaccination and linear regression analysis demonstrated that r-ELISA and f-ELISA correlated with microneutralization (R=0.622 for r-ELISA vs. R=0.56 for f-ELISA). For the healthy vaccinees both ELISAs were found to be adequate to distinguish protected from non-protected individuals (sensitivity and specificity: 87.5%/85.3% for r-ELISA and 87.5%/88.3% for f-ELISA). Results from the transplant recipients showed a slightly reduced sensitivity of 73.3% for r-ELISA while the f-ELISA demonstrated similar sensitivity and specificity as in the healthy vaccinees. However, in order to obtain these assay performances the cut-off-values for protection had to be adjusted for both assays and both investigation cohorts respectively limiting their application in routine laboratories.

[Concepts, effectiveness, and perspectives of pandemic and seasonal influenza vaccines]. / Konzepte, Wirksamkeit und Perspektiven pandemischer und nichtpandemischer Influenzaimpfungen.

For the first time in history, the conditions to influence the course of an influenza pandemic through vaccination were set during the influenza A H1N1 pandemic in 2009. The specific requirements for pandemic vaccines are to be highly immunogenic in immunologically naive individuals and to be producible quickly in large quantities. In contrast, seasonal influenza vaccines induce a booster response and a broadening of preexisting immunity. In this article the concepts of seasonal and pandemic influenza vaccines and data on their immunogenicity and clinical efficacy are reviewed and discussed. In the upcoming years, seasonal influenza vaccination will continue to be based on inactivated split-virion and subunit vaccines or the live attenuated cold-adapted vaccine. The pandemic vaccines used in 2009 proved to be more immunogenic than expected from prepandemic vaccine trials, while the adverse events observed with AS03-adjuvanted vaccines call their future use into question. However, neither seasonal nor pandemic influenza vaccines can be regarded to be an ideal solution, because they have to be frequently adapted to new virus strains and they lack effectiveness in particular risk groups. They can be regarded as interim approaches to highly immunogenic vaccines that hopefully become available in the future. The underlying principles of future vaccines are also presented in this article.

Epidemiology and clinical characteristics of pandemic (H1N1) 2009 influenza infection in pediatric hemato-oncology and hematopoietic stem cell transplantation patients.

BACKGROUND: For children with hemato-oncologic diseases, especially after hematopoietic stem cell transplantation (HSCT), the risk for developing complications related to pandemic influenza A (H1N1) 2009 (pH1N1) infection is largely unknown. METHODS: A retrospective chart study was performed of pH1N1 cases diagnosed between October 2009 to January 2010 in the hemato-oncologic unit of the University Children's Hospital of Düsseldorf, Germany. FINDINGS: In total, 21 children were diagnosed with laboratory-confirmed pH1N1; in 16 patients with malignancies (acute leukemia 7, lymphoma 4, solid tumors 2, others 3) and in 5 with benign hematologic disorders. Five patients had undergone prior HSCT, although 1 patient was diagnosed during conditioning therapy with high-dose chemotherapy in preparation for haploidentical HSCT. Most frequent symptoms were fever (>38.5°C) and cough (in 100%), and rhinorrhea (57%). The 2 patients acquiring pH1N1 infection under high-dose or intensive chemotherapy did not require intensive care or mechanical ventilation, and both recovered under antiviral therapy. Oseltamivir was administered to 11 patients; in 1 patient, therapy was switched, on a compassionate-use basis, to intravenous zanamivir because of lack of clinical improvement after oseltamivir therapy. Complications were hospitalization (19%), demand of oxygen supplementation, delay/interruption of antineoplastic therapy, and prolonged administration of antibiotics and antipyretics. CONCLUSION: In the investigated patient population, pH1N1 was mild in most cases, but was associated with substantial morbidity in a proportion of patients and led to interruption and delay in anticancer treatment.

[Typical intraoperative complications in laparoscopic surgery]. / Typische intraoperative Komplikationen in der laparoskopischen Chirurgie.

For all common laparoscopic procedures (e. g. cholecystectomy, appendectomy, inguinal hernia repair, fundoplication and colorectal resection) it has been possible to demonstrate in systematic reviews and meta-analyses that they produce better results in terms of perioperative outcome than open surgery. Accordingly, there are very few publications that report on intraoperative complications and their management. In this respect a distinction must be made between positioning complications, access complications and complications related to the pneumoperitoneum, which can manifest in all laparoscopic procedures, as well as the specific complications associated with individual procedures.The main focus of any consideration of intraoperative complications must of course be on strategies to prevent the occurrence. If intraoperative complications have occurred, the most important aspect is the diagnosis and control with prime importance accorded to which complications can still be controlled using a laparoscopic approach and when an open procedure must be used. In general a switch to open surgery should be made in the event of serious complications. Only a highly experienced laparoscopic surgeon will be able to safely manage complications once they have occurred without putting the patient at further risk. In doubtful situations the approach that poses least risk is open surgery for complications that have already occurred.

Comparison of hemagglutination inhibition assay, an ELISA-based micro-neutralization assay and colorimetric microneutralization assay to detect antibody responses to vaccination against influenza A H1N1 2009 virus.

The hemagglutination inhibition (HI) assay has been the main method used to investigate immune responses to vaccination against influenza H1N1 (2009) virus. However microneutralization tests (MNT) have been shown to be more sensitive and more specific. In this study, the three methods of choice: (i) the HI assay, (ii) an ELISA-based conventional MNT and (iii) a colorimetric MNT in terms of their ability to detect antibody responses in serum pairs collected from 43 healthy individuals before and 21 days after vaccination were compared. The colorimetric MNT was established yielding intra- and inter-run imprecisions of 7.5% and 12.4%, respectively. Testing of antisera to seasonal influenza viruses demonstrated the assay to be specific for antibodies to influenza H1N1 (2009) virus. A good correlation between the three methods was found, being highest for the ELISA-MNT and the colorimetric MNT (r=0.714 for geometric mean titers (GMT) and r=0.695 for titer increases). Similar rates of fourfold titer increases were detected: 95.3% in the ELISA-MNT vs. 93.0% in colorimetric MNT and 95.3% in HI assay. The ELISA-based MNT demonstrated the highest titer range leading to the highest postvaccination GMT and the highest titer increase (>50-fold). The lowest GMTs were measured with the HI assay, while the colorimetric MNT detected the highest GMT in prevaccination sera. Taken together, similar seroconversion rates were obtained with the three assays. The ELISA-MNT appeared to be the best method to compare absolute pre- and postvaccination GMTs. The colorimetric MNT, being less labour-intensive than the ELISA-MNT, seems to be a suitable tool in vaccination studies.

Outbreak of influenza virus A/H1N1 in a hospital ward for immunocompromised patients.

In February 2008, five patients were infected with the H1N1 subtype of influenza A virus in one hospital ward for immunocompromised patients at a hospital in North Rhine-Westphalia, Germany. All of these patients had an established haematologic disease and tested positive either for viral RNA or antigen shortly after the beginning of respiratory illness. In three of the patients, influenza virus was repeatedly detected, and four of the patients died in coincidence with the virus infection. Sequencing of the amplified (HA1) haemagglutinin yielded identical nucleotide sequences in isolates from three of the patients, whereas one nucleotide difference was found in the isolate of the fourth patient, resulting in an amino acid substitution (G153R). To investigate the source of infection, the medical staff (n = 104) of the hospital unit was tested and found negative for influenza virus RNA and antigen in pharyngeal lavages. Testing for influenza virus antibodies by immunofluorescence assay revealed that 12 staff members were positive for influenza virus A IgA antibodies. These findings suggest that wild-type influenza virus infections occurred within the medical staff at the same time the patients were infected and that the staff might have contributed to the circulation of virus in the hospital ward.

Expression plasmids are only useful for the investigation of co-receptor tropism and fusion capacity of short HIV-1 envelope domains.

Expression vectors have been used widely to identify functionally important domains in HIV-1 glycoproteins. Env domains such as the V3 loop were amplified by polymerase chain reaction (PCR) and inserted into plasmids carrying the backbone of an HIV-1 reference strain like NL4-3. The hypothesis of the present approach was that cloning large domains of wild type envelopes yields constructs that are non-functional in co-receptor-expressing HeLaCD4 cells, in contrast to laboratory-adapted HIV-1 strains. The background for this assumption was that primary HIV-1 virions are frequently less infectious and lack fusion capacity in HeLaCD4 cells compared to laboratory-adapted (LA) viruses. To address this hypothesis, env domains of different length were amplified from a panel of X4-tropic HIV-1 clinical isolates cultured in peripheral blood lymphocytes (PBLs) and cloned into the backbone of NL4-3 env. Constructs bearing either the V3 loops or 312 nucleotides of the intracellular trunk (ICT) of gp41 led to a similar fusion capacity as NL4-3. In contrast, none of the plasmids carrying the 2322 N-terminal nucleotides of primary isolates led to similar syncytium formation. These results have an effect on studies that investigate pathogenic effects of Env regions with chimeric constructs in the backbone of HIV reference envelopes.

[Laparoscopic appendectomy. The new standard?]. / Laparoskopische Appendektomie. Der neue Standard?

Since the introduction of laparoscopic appendectomy by Semm in 1983, the role of this minimally invasive surgical technique has been the focus of controversial discussion. Meta-analyses have identified its advantages as having significantly lower wound infection rates, less postoperative pain and earlier resumption of normal everyday activities. The disadvantages are higher rates of intra-abdominal abscesses, longer operating times and higher inpatient treatment costs. However, some of the advantages identified by meta-analyses have been called into question by the results obtained from research into aspects of care. These discrepancies are attributable to the different surgeons involved in the various studies. The results are greatly influenced by the qualifications and experience of the surgeons. Therefore conventional appendectomy using a right lower lateral McBurney incision should continue to be the gold standard. Surgeons who have extensive experience in the field of laparoscopic surgery can achieve better results with minimally invasive appendectomy than with open surgery. This also holds true for specific situations such as complicated appendicitis and for morbidly obese patients. However, the operating costs incurred for laparoscopic appendectomy are higher because it has been shown that removal of the appendix with a linear stapler is the most reliable method.

The microglial/macrophagic response at the tumour-brain border of invasive meningiomas.

AIMS: Little is known about the immune response of the brain to invasive meningiomas. The present study was based upon the hypothesis that the microglial/macrophagic response towards brain-invasive meningiomas is dependent on the intactness of the pial-glial basement membrane. METHODS: We immunostained sections from 40 brain-invasive meningiomas that were graded according to World Health Organization (WHO) 2007 criteria. Thirty-three tumours were histologically WHO grade II (18, 'otherwise benign', and 15, 'otherwise atypical'), and seven, grade III. Microglial/macrophagic cells were labelled with antibodies directed against major histocompatibility complex class II, CD68, CD14 and CD163. Anti-collagen IV was used to visualize basement membranes. RESULTS: Twenty-five per cent (10/40) meningiomas (1/18 WHO grade II 'otherwise benign', 3/15 grade II 'otherwise atypical' and 6/7 WHO grade III) contained microglial/macrophagic cells at the tumour-brain border. The presence of these cells correlated with the absence of the pial-glial basement membrane (BM) and with WHO grade III. The monocytic response was of two kinds: one consisted of a dense layer of mononuclear cells at the tumour-brain border in nine cases, the other of an elevated number of microglial cells expressing CD14 or CD163 (two cases). CONCLUSIONS: The immune response at the tumour-brain interface correlates with the absence of the pial-glial BM and with malignancy grade. It remains to be established whether the mononuclear cells at the tumour-brain border are native microglia or blood-derived macrophages.

Extrapolation to zero-flow pressure in cerebral arteries to estimate intracranial pressure.

BACKGROUND: Cerebral perfusion pressure (CPP) is commonly calculated from the difference between arterial blood pressure (AP) and intracranial pressure (ICP). ICP can be considered the effective downstream pressure of the cerebral circulation. Consequently, cerebral circulatory arrest would occur when AP equals ICP. Estimation of AP for zero-flow pressure (ZFP) may thus allow estimation of ICP. We estimated ZFP from cerebral pressure-flow velocity relationships so that ICP could be measured by transcranial Doppler sonography. METHODS: We studied 20 mechanically ventilated patients with severe head injury, in whom ICP was monitored by epidural pressure transducers. AP was measured with a radial artery cannula. Blood flow velocity in the middle cerebral artery (V(MCA)) ipsilateral to the site of ICP measurement was measured with a 2 MHz transcranial Doppler probe. All data were recorded by a microcomputer from analogue-digital converters. ZFP was extrapolated by regression analysis of AP-V(MCA) plots and compared with simultaneous measurements of ICP. RESULTS: ZFP estimated from AP-V(MCA) plots was linearly related to ICP over a wide range of values (r=0.93). There was no systematic difference between ZFP and ICP. Limit of agreement (2 SD) was 15.2 mm Hg. Short-term variations in ICP were closely followed by changes in ZFP. CONCLUSION: Extrapolation of cerebral ZFP from instantaneous AP-V(MCA) relationships enables detection of severely elevated ICP and may be a useful and less invasive method for CPP monitoring than other methods.