SARS CTL Vaccine Candidates — HLA Supertype, Genome-Wide Scanning and Biochemical Validation

An effective SARS vaccine is likely to include components that can induce specific cytotoxic T-cell (CTL) responses. The specificities of such responses are governed by HLA-restricted presentation of SARS-derived peptide epitopes. Exact knowledge of how the immune system handles protein antigens would allow for the identification of such linear sequences directly from genomic/proteomic sequence information. The latter was recently established when a causative coronavirus (SARS CoV) was isolated and full-length sequenced. Here, we have combined advanced bioinformatics and high-throughput immunology to perform an HLA supertype, genome-wide scan for SARS-specific cytotoxic T cell epitopes. The scan includes all nine human HLA supertypes in total covering >99% of all major human populations. For each HLA supertype, we have selected the 15 top candidates for test in biochemical-binding assays. At this time (approximately 6 months after the genome was established), we have tested the majority of the HLA supertypes and identified almost 100 potential vaccine candidates. These should be further validated in SARS survivors and used for vaccine formulation. We suggest that immunobioinformatics may become a fast and valuable tool in rational vaccine design.     

No correlation between number of MRI-evident lesions in cerebrum and the soleus stretch reflex in multiple sclerosis patients

The aim of the study was to investigate if the stretch reflex of the soleus muscle was useful in quantifying upper motor neuron lesions. The soleus stretch reflex was recorded in 10 healthy subjects and 20 patients with active relapsing-remitting multiple sclerosis and correlated to the number of MRI lesions in cerebrum and clinical scores (expanded disability status scale and regional functional scoring system). The short latency stretch reflex was elicited by rotating the left ankle joint 4 degrees with a rise time in the interval of 40-640 ms. The amplitude of the stretch was larger in multiple sclerosis patients being 88.5 microV in patients and 12.8 microV in controls, P = 0.007. The sensitivity of the stretch reflex expressed as the slope of the best linear fit was increased in MS patients to 2.6 microVs/degree compared with 0.6 microVs/degree (0.1-2.2) in controls, P = 0.009. There was no correlation between amplitude of the stretch reflex and number of MRI lesions (r = -0.03). In conclusion, the soleus stretch reflex might be useful to quantify spasticity but is not useful in detecting dysfunction of upper motor neurons in MS.     

Evidence for an association between hairy cell leukemia and renal cell and colorectal carcinoma.

BACKGROUND. Hairy cell leukemia (HCL) has been associated with several disease states. In this study, a possible association is reported between HCL and renal cell carcinoma (RCC) and colorectal carcinoma (CRC). METHODS. A retrospective study of the case records of 50 patients with HCL in a study of alpha-interferon (alpha-IFN) treatment of HCL. RESULTS. Three of 50 patients with HCL studied had RCC, and 2 of these also had CRC. In addition, two other patients had CRC. The other malignant lesions developed either before or after the diagnosis of HCL. In all patients, the HCL responded to alpha-interferon (alpha-IFN), but in four patients, the second lesion was diagnosed during IFN treatment. CONCLUSIONS. These findings could indicate that IFN does not correct a possible common basic etiologic defect and shows that even early CRC and RCC do not respond to the IFN doses administered. These findings should be considered in future trials of IFN treatment of these diseases. The authors also recommend a reevaluation of the frequency of second malignant lesions in HCL; this may be important particularly with the increased survival in patients with HCL who receive alpha-IFN treatment.     

Excitatory amino acid agonists. Enzymic resolution, X-ray structure, and enantioselective activities of (R)- and (S)-bromohomoibotenic acid

The enantiomers of alpha-amino-4-bromo-3-hydroxy-5-isoxazolepropionic acid (4-bromohomoibotenic acid, Br-HIBO, 1) a selective and potent agonist at one class of the central (S)-glutamic acid receptors, were prepared with an enantiomeric excess higher than 98.8% via stereoselective enzymic hydrolysis of (RS)-alpha-(acetylamino)-4-bromo-3-methoxy-5-isoxazolepropionic acid (4) using immobilized aminoacylase. The absolute configuration of the enantiomers of Br-HIBO was established by X-ray crystallographic analysis, which confirmed the expected preference of the enzyme for the S form of the substrate 4. (S)- and (RS)-Br-HIBO were potent neuroexcitants on cat spinal neurones in vivo, while (R)-Br-HIBO was a very weak excitant. Correspondingly, the S enantiomer of Br-HIBO (IC50 = 0.34 microM) was considerably more potent than the R form (IC50 = 32 microM) as an inhibitor of [3H]-(RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ([ 3H]AMPA) binding to rat brain synaptic membranes in vitro. In contrast, (S)- and (R)-Br-HIBO were approximately equipotent (IC50 values of 0.22 and 0.15 microM, respectively) as inhibitors of [3H]-(S)-glutamic acid binding in the presence of CaCl2. The enantiomers of Br-HIBO showed no significant affinity for those binding sites on rat brain membranes which are labeled by [3H]kainic acid or [3H]-(R)-aspartic acid.     

Lymphatic transport from normal and synovitic knees in rabbits

The resorption pattern of synovial fluid through the lymphatic system from normal and synovitic knee joints in rabbits was studied with ^99mHechnetium-rhenium-sulfur colloid injected intraarticularly and monitored for 14 hours with a gamma camera.

On the normal side the regional lymph nodes were visualized after I hour and after 14 hours still 75 percent activity remained in the knee. In the synovitic knees no lymphatic transport could be detected; and the radiotracer was unstable with rapid liberation of technetium, which was excreted in the urine. This radiolysis was not found in vitro in synovitic joint fluid.

The lymphatic transport from normal rabbit knees is low. We found a clear difference in lymphatic transport between normal and synovitic knee joints.     

Allergic airway disease caused by methyl tetrahydrophthalic anhydride in epoxy resin

Work-associated rhinitis and asthma occurred in a subject exposed to methyl tetrahydrophthalic anhydride (MTHPA) used as a curing agent in an epoxy resin system. He displayed bronchial hyperreactivity in provocation with methacholine, as well as skin prick test positivity and specific immunoglobulin E (IgE) serum antibodies, against a conjugate of MTHPA and human serum albumin. Thus, it seems likely that the disease was caused by an IgE-mediated allergy.     

Select types of supporting cell in the inner ear express aquaporin-4 water channel protein

Aquaporins (AQPs) confer a high water permeability on cell membranes and play important parts in secretory and absorptive epithelia in kidney and other organs. Here we investigate whether AQPs are expressed in the sensory epithelia of the inner ear, where a precise volume regulation is crucial. By use of specific antibodies it was found that the inner ear contains AQP1 and 4 while being devoid of detectable levels of AQP2, 3 or 5. Immunofluorescence and postembedding immunogold labelling revealed a strictly non-epithelial distribution of AQP1, confirming previous data. In contrast, AQP4 protein and mRNA (visualized by in situ hybridization) were concentrated in select types of supporting cell, including Hensen's cells and inner sulcus cells. Immunogold particles signalling AQP4 were confined to the basolateral plasma membrane of Hensen's cells and to the basal plasma membrane of Claudius cells and inner sulcus cells. AQP4 was also found in supporting cells of the vestibular end organs, but was absent from transitional epithelial cells and dark cells. Strong labelling for AQP4 and AQP4-mRNA was associated with the central part of the cochlear and vestibular nerves. Hair cells were consistently unlabelled. Our findings indicate that AQP4 may facilitate osmotically driven water fluxes in the sensory epithelia of the inner ear and thus contribute to the volume and ion homeostasis at these sites.     

Classification treesA possible method for iso-resource grouping in intensive care

Classification and grouping of clinical data into defined categories or hierarchies is difficult in intensive care practice. Diagnosis-related groups are used to categorise patients on the basis of diagnosis. However, this approach may not be applicable to intensive care where there is wide heterogeneity within diagnostic groups. Classification tree analysis uses selected independent variables to group patients according to a dependent variable in a way that reduces variation. In this study, the influence of three easily identified patient attributes on their length of intensive care unit stay was explored using classification analysis. Two thousand five hundred and forty-five critically ill patients from three hospitals were classified into groups so that the variation in length of stay within each group was minimised. In 23 out of 39 terminal groups, the interquartile range of the length of stay was ≤ 3 days.     

Abdominal ultrasound versus intravenous urography in the evaluation of infravesically obstructed males

In a prospective study intravenous urography (IVP) and abdominal ultrasonography (US) were compared in 100 consecutively admitted male patients with symptoms of infravesical obstruction. No pathology was found in 67 patients on IVP and in 61 patients on US. Both modalities disclosed 5 bilateral, 4 unilateral hydronephroses and one patient with contracted kidneys. IVP found 7 renal masses: one solid tumour, 2 "possibly solid" tumours and 4 "possibly cysts", whereas US found one solid tumour and 16 cysts. Both modalities detected 3 kidney stones, 5 bladder stones and 3 bladder tumours. It is concluded that "imaging" of the urinary tract is only indicated in cases of haematuria, elevated creatinine, history of renal calculous disease and other clinical suspicion of upper urinary tract disease. It is further concluded that US is preferable to IVP in this patient category due to (1) better characterization of renal masses, (2) the possibility of investigating the liver and the retroperitoneum in the same setting, (3) better evaluation of the prostate with respect to size, (4) better evaluation of the bladder, and (5) last but not least for economical reasons. However, bone metastases will be missed.     

Modulation of heteronymous reflexes from ankle dorsiflexors to hamstring muscles during human walking

In 16 human subjects, stimulation of the common peroneal nerve (CPN) was applied during walking and standing. The effect of the stimulation was evaluated from the rectified and averaged biceps femoris (BF) electromyographic (EMG) activity. In the swing phase of walking, the CPN stimulation evoked a suppression in the BF EMG in 12 of the subjects. In the early stance phase, the suppression was replaced by facilitation at a similar latency in 9 of the subjects. Of the other 3 subjects, in whom a suppression was observed during swing, a decrease in the suppression was observed in the stance phase in two of them. During a voluntary co-contraction of BF and tibialis anterior while standing, a suppression similar to that observed in the swing phase was observed. The thresholds of the suppression and facilitation were identical, suggesting that afferents of similar diameter were responsible. Cutaneous stimuli, which mimicked the sensation evoked by the CPN stimulation, but without activation of muscle afferents, did not produce similar effects in the BF EMG activity. It is suggested that the observed response and reflex reversal may reflect opening of an excitatory group I pathway in the early stance phase of walking with a concomitant shut-down of heteronymous group I inhibition.