Search

The HIV-1 quasispecies in peripheral blood mononuclear cells (PBMC) is considered to be a mix of actively replicating, latent, and archived viruses and may be genetically distinct from HIV-1 variants in plasma that are considered to be recently produced. Here we analyzed the genetic relationship between gp160 env sequences from replication competent clonal HIV-1 variants that were isolated from PBMC and from contemporaneous HIV-1 RNA in serum and HIV-1 proviral DNA in PBMC of four longitudinally studied therapy naïve HIV-1 infected individuals. Replication competent clonal HIV-1 variants, HIV-1 RNA from serum, and HIV-1 proviral DNA from PBMC formed a single virus population at most time points analyzed. However, an under-representation in serum of HIV-1 sequences with predicted CXCR4 usage was sometimes observed implying that the analysis of viral sequences from different sources may provide a more complete assessment of the viral quasispecies in peripheral blood in vivo., (Copyright 2010 Elsevier Inc. All rights reserved.)

Neutralizing antibodies (NAb) able to react to heterologous viruses are generated during natural HIV-1 infection in some individuals. Further knowledge is required in order to understand the factors contributing to induction of cross-reactive NAb responses. Here a well-established model of experimental pathogenic infection in cynomolgus macaques, which reproduces long-lasting HIV-1 infection, was used to study the NAb response as well as the viral evolution of the highly neutralization-resistant SIVmac239. Twelve animals were infected intravenously with SIVmac239. Antiretroviral therapy (ART) was initiated ten days post-inoculation and administered daily for four months. Viral load, CD4(+) T-cell counts, total IgG levels, and breadth as well as strength of NAb in plasma were compared simultaneously over 14 months. In addition, envs from plasma samples were sequenced at three time points in all animals in order to assess viral evolution. We report here that seven of the 12 animals controlled viremia to below 10(4) copies/ml of plasma after discontinuation of ART and that this control was associated with a low level of evolutionary divergence. Macaques that controlled viral load developed broader NAb responses early on. Furthermore, escape mutations, such as V67M and R751G, were identified in virus sequenced from all animals with uncontrolled viremia. Bayesian estimation of ancestral population genetic diversity (PGD) showed an increase in this value in non-controlling or transient-controlling animals during the first 5.5 months of infection, in contrast to virus-controlling animals. Similarly, non- or transient controllers displayed more positively-selected amino-acid substitutions. An early increase in PGD, resulting in the generation of positively-selected amino-acid substitutions, greater divergence and relative high viral load after ART withdrawal, may have contributed to the generation of potent NAb in several animals after SIVmac239 infection. However, early broad NAb responses correlated with relatively preserved CD4(+) T-cell numbers, low viral load and limited viral divergence.

Sinus lift elevation is an interesting method to restore bone mass at the maxilla in edentulated patients. We have investigated the histological effects of beta tricalcium phosphate (beta-TCP) combined with autograft bone for sinus lift elevation. A series of 14 patients who were candidate for dental implantation were grafted with beta-TCP granules and morcellized autograft bone harvested at the chin. beta-TCP was characterized by scanning electron microscopy and optical profilometry. Before implant placement, a small bone biopsy (2mm in diameter) was done. The amount of residual material and newly formed bone were determined by microcomputed tomography. Histological analysis was done on undecalcified sections stained by Goldner's trichrome and osteoclast identification (TRAcP). beta-TCP served as a template for bone apposition by osteoblasts onto the granules' surface. The material was simultaneously resorbed by TRAcP positive osteoclasts and macrophages. Fragments of the material remained buried in bone trabeculae as long as 12 months post-graft but the formed bone onto the granules surface had a lamellar texture. beta-TCP combined with autograft bone appears a suitable biomaterial for sinus lift augmentation before the placement of bone implants. The material favors the apposition of lamellar bone by osteoblasts and is simultaneous resorbed by two types of cells., (Copyright 2009 Elsevier Ltd. All rights reserved.)

This study describes the neuropsychological assessment of 34 patients with questionable Alzheimer's disease (QAD) followed up for 3 years. Several measures were selected from the California Verbal Learning Test (CVLT) and compared to other cognitive tasks to assess the best neuropsychological indices for (a) detecting early memory impairment in QAD and (b) predicting conversion to AD. Concerning detection, the results indicated that a recall measure depending on semantic categorization (short-delay cued recall) signaled a memory deficit in stable QAD patients, suggesting that episodic and semantic memory problems are involved in the early cognitive impairments of stable QAD patients. However, the conversion to AD was best predicted by the initial performance at the recency index (score reflecting high reliance on working memory), corroborating the idea that AD patients (even at the questionable stage) essentially rely on preserved phonological loop functioning in memory tasks. Finally, an additional impairment in visuospatial memory (Rey's figure) provided a good discriminant value to distinguish converters from stable QAD patients, showing that various cognitive disabilities deteriorate in AD.

Alzheimer's disease (AD) is characterized by a progressive loss of controlled cognitive processes, and neuroimaging studies at early stages of AD provide an opportunity to tease out the neural correlates of controlled processes. Accordingly, controlled and automatic memory performance was assessed with the Process Dissociation Procedure in 50 patients diagnosed with questionable Alzheimer's disease (QAD). The patients' brain glucose metabolism was measured using FDG-PET. After a follow-up period of 36 months, 27 patients had converted to AD, while 23 remained stable. Both groups showed a similar decrease in controlled memory processes but preserved automatic processes at entry into the study. Voxel-based cognitive and metabolic correlations showed that a decrease in controlled memory processes was preferentially correlated with lower activity in the dorsomedial prefrontal and posterior cingulate cortices in very early AD patients. In stable QAD patients, reduced controlled performance in verbal memory correlated with impaired activity in the left anterior hippocampal structure. The results demonstrate the central role of a medial frontal-posterior cingulate network for controlled processing of episodic memory in the early stages of AD.

Cadmium (Cd) is a highly toxic metal that enters the food chain. Following oral ingestion, the intestinal epithelium is the first biological barrier crossed by Cd and is also an important target tissue. In the present study, the human intestinal Caco-2 cell line was used to evaluate the impact of a low level of exposure on both undifferentiated and differentiated intestinal cells. As revealed by the LC(50) values estimated with the 3-[4,5-dimethyl-2-thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, mature Caco-2 cells were more resistant to Cd. However, following a 24-h exposure to non-cytotoxic levels of Cd (10 microM) or zinc (Zn, 100 microM), threefold increases were obtained in the LC(50) values of 7-day-old cells, whereas increased resistance in 21-day-old cells was observed exclusively with Zn. Induction of MT-IIa and HSP70 mRNAs was higher in undifferentiated cells and an increase in cellular glutathione (GSH) content was observed exclusively in these cell cultures. However, the results obtained with cycloheximide used for inhibiting protein synthesis and with L-buthionine sulfoximine (BSO), which inhibits GSH synthesis, revealed that protein synthesis is not a prerequisite to the development of resistance. The presence of 100 mM 3-amino-1,2,4-triazole (3AT), a catalase inhibitor, prevented Cd-induced but not Zn-induced resistance, as well as sensitized cells to Cd toxicity. These results show for the first time differences in constitutive and acquired resistance to Cd as a function of enterocytic differentiation status and suggest the involvement of different mechanisms for Cd- and Zn-induced adaptation in the intestinal cells. Redox signals may trigger Cd-induced adaptation mechanisms but pro-oxidant conditions would eliminate proliferative intestinal cells capability to develop resistance. This would be critical for Cd- but not Zn-induced mechanisms of resistance since Cd but not Zn may cause oxidative stress.

In order to better understand environmental disturbances in the French coastal Mediterranean lagoons, we used an ecotoxicological approach based on the measurement of the toxicity of the sediments using oyster embryo bioassay that provides a basis for assessing the effects on the fauna of contaminants adsorbed on the sedimentary particles. The study covers all of the main lagoons of the French Mediterranean coasts of Languedoc Roussillon, Camargue, and Provence (Berre and Bolmon lagoons), where 188 stations were sampled. The toxicity tests provide evidence of variable levels of toxicity in sediments. Contaminated lagoons such as La peyrade, Le canet, and Ingrill and locally affected lagoons such as Bages-Sigean, Vaccares, Bolmon, and Berre have sampling stations with 100% of larval abnormalities during 24-h development. In all of the lagoons, the toxicity was mainly located close to local harbors and rivers. Salses Leucate (Languedoc roussillon) lagoon was found very clean, with no important toxicity. The results are discussed in terms of environmental disturbances of the coastal lagoons and with regard to the long-term monitoring of the impact of contaminants on the coastal environment.

Bone autograft remains a very useful and popular way for filling bone defects. In maxillofacial surgery or implantology, it is used to increase the volume of the maxilla or mandible before placing dental implants. Because there is a noticeable delay between harvesting the graft and its insertion in the receiver site, we evaluated the morphologic changes at the light and transmission electron microscopy levels. Five patients having an autograft (bone harvested from the chin) were enrolled in the study. A small fragment of the graft was immediately fixed after harvesting and a second one was similarly processed at the end of the grafting period when bone has been stored at room temperature for a 20 min +/- 33 s period in saline. A net increase in the number of osteocyte lacunae filled with cellular debris was observed (+41.5%). However no cytologic alteration could be observed in the remaining osteocytes. The viability of these cells is known to contribute to the success of autograft in association with other less well-identified factors.

Thirty-four subjects with questionable Alzheimer's disease (QAD) were included in a 3-year prospective study and underwent neuropsychological testing and measurement of brain metabolism using FDG-PET at entry. Seventeen patients (50%) did not convert to AD during the follow-up period. Compared to elderly controls of similar age, the cerebral activity of non-converters was reduced in the dorsolateral prefrontal cortex. Moreover, the variability of metabolism in the posterior cingulate cortex was related to their visuospatial memory performance, while disparity in parietal activity was related to their verbal memory performance. These results demonstrate the cerebral metabolic heterogeneity of patients with QAD. Initial functional images of converters showed that activity was already impaired in the posterior cingulate, lateral temporal cortex, anterior cingulate and orbitofrontal cortex. This metabolic pattern is consistent with a pre-dementia stage of AD, and highlights the fact that significant frontal metabolic involvement may be associated with impaired activity in posterior associative cortices in very early AD.

Immunodominant and public T cell receptor (TCR) usage is relatively common in many viral diseases yet surprising in the context of the large naive TCR repertoire. We examined the highly conserved Vbeta17:Valpha10.2 JM22 T cell response to the influenza matrix peptide (58-66)-HLA-A*0201 (HLA-A2-flu) through extensive kinetic, thermodynamic, and structural analyses. We found several conformational adjustments that accompany JM22-HLA-A2-flu binding and identified a binding "hotspot" within the Vbeta domain of the TCR. Within this hotspot, key germline-encoded CDR1 and CDR2 loop residues and a crucial but commonly coded residue in the hypervariable region of CDR3 provide the basis for the substantial bias in the selection of the germline-encoded Vbeta17 domain. The chances of having a substantial number of T cells in the naive repertoire that have HLA-A2-flu-specific Vbeta17 receptors may consequently be relatively high, thus explaining the immunodominant usage of this clonotype.

T-cell recognition of the antigenic peptides presented by MHC class I molecules normally triggers protective immune responses, but can result in immune enhancement of disease. Cross-reactive T-cell responses may underlie immunopathology in dengue haemorrhagic fever. To analyze these effects at the molecular level, the functional MHC class I molecule HLA-A*1101 was crystallized bound to six naturally occurring peptide variants from the dengue virus NS3 protein. The crystals contained high levels of solvent and required optimization of the cryoprotectant and dehydration protocols for each complex to yield well ordered diffraction, a process that was facilitated by the use of a free-mounting system.

Microcomputed tomography is a new technique for the non-destructive study of porous biological materials. We examined 11 wisdom teeth that had been removed prophylactically by microcomputed tomography operating in the cone beam mode. The two-dimensional sections of the teeth were reconstructed with surface rendering software to provide three-dimensional models that were observed and handled in virtual reality. The tooth itself, or the pulp chamber and canals, can be reconstructed and observed separately or simultaneously. Many teeth looked dystrophic with abnormal distributions of roots and canals. Microcomputed tomography seems to be a promising way of studying dental anatomy.

The immunoproteasome (IP) is usually viewed as favoring the production of antigenic peptides presented by MHC class I molecules, mainly because of its higher cleavage activity after hydrophobic residues, referred to as the chymotrypsin-like activity. However, some peptides have been found to be better produced by the standard proteasome. The mechanism of this differential processing has not been described. By studying the processing of three tumor antigenic peptides of clinical interest, we demonstrate that their differential processing mainly results from differences in the efficiency of internal cleavages by the two proteasome types. Peptide gp100(209-217) (ITDQVPSFV) and peptide tyrosinase369-377 (YMDGTMSQV) are destroyed by the IP, which cleaves after an internal hydrophobic residue. Conversely, peptide MAGE-C2(336-344) (ALKDVEERV) is destroyed by the standard proteasome by internal cleavage after an acidic residue, in line with its higher postacidic activity. These results indicate that the IP may destroy some antigenic peptides due to its higher chymotrypsin-like activity, rather than favor their production. They also suggest that the sets of peptides produced by the two proteasome types differ more than expected. Considering that mature dendritic cells mainly contain IPs, our results have implications for the design of immunotherapy strategies.

Long-term nonprogression during acute HIV infection has been strongly associated with HLA-B*5701 or HLA-B*5703. In this study, we present the high resolution crystal structures of HLA-B*5703 complexes with three HIV-1 epitopes: ISPRTLNAW (ISP), KAFSPEVIPMF (KAF-11), and KAFSPEVI (KAF-8). These reveal peptide anchoring at position 2 and their C termini. The different peptide lengths and primary sequences are accommodated by variation in the specific contacts made to the HLA-B*5703, flexibility in water structure, and conformational adjustment of side chains within the peptide-binding groove. The peptides adopt markedly different conformations, and trap variable numbers of water molecules, near a cluster of tyrosine side chains located in the central region of the peptide-binding groove. The KAF-11 epitope completely encompasses the shorter KAF-8 epitope but the peptides are presented in different conformations; the KAF-11 peptide arches out of the peptide-binding groove, exposing a significant main chain surface area. Bioinformatic analysis of the MHC side chains observed to contribute to the peptide anchor specificity, and other specific peptide contacts, reveals HLA alleles associated with long-term nonprogression and a number of related HLA alleles that may share overlapping peptide repertoires with HLA-B*5703 and thus may display a similar capacity for efficient immune control of HIV-1 infection.

Crystallization trials at the Division of Structural Biology in Oxford are now almost exclusively carried out using a high-throughput workflow implemented in the Oxford Protein Production Facility. Initial crystallization screening is based on nanolitre-scale sitting-drop vapour-diffusion experiments (typically 100 nl of protein plus 100 nl of reservoir solution per droplet) which use standard crystallization screening kits and 96-well crystallization plates. For 294 K crystallization trials the barcoded crystallization plates are entered into an automated storage system with a fully integrated imaging system. These plates are imaged in accordance with a pre-programmed schedule and the resulting digital data for each droplet are harvested into a laboratory information-management system (LIMS), scored by crystal recognition software and displayed for user analysis via a web-based interface. Currently, storage for trials at 277 K is not automated and for imaging the crystallization plates are fed by hand into an imaging system from which the data enter the LIMS. The workflow includes two procedures for nanolitre-scale optimization of crystallization conditions: (i) a protocol for variation of pH, reservoir dilution and protein:reservoir ratio and (ii) an additive screen. Experience based on 592 crystallization projects is reported.

A new mixed barium zirconium oxalate, triaquatetra-mu-oxalato-dibarium(II)zirconium(IV), Ba(2)Zr(C(2)O(4))(4).3H(2)O or [Ba(2)Zr(C(2)O(4))(4)(H(2)O)(3)](n), has been synthesized. The complex is built from eightfold-coordinated Zr atoms and eleven- and sixfold-coordinated Ba atoms, linked by oxalate groups. The Zr atom, the two Ba atoms and one water O atom lie on crystallographic twofold axes, so that each coordination polyhedron has imposed C2 symmetry. Packing in the crystal is also assumed through hydrogen bonds.

Gastrointestinal symptoms, including diarrhoea and abdominal pain, are one of the earliest and most frequently reported signs of Fabry disease, a rare X-linked lipid storage disorder. As the disease progresses, renal, cardiac and cerebrovascular complications develop, resulting in more serious symptoms and early mortality. The present study evaluated the effects of enzyme replacement therapy (ERT) with agalsidase alfa on the gastrointestinal symptoms of Fabry disease. Following 6 months of treatment, both the severity ( p < 0.02) and frequency ( p < 0.02) of abdominal pain decreased. For those patients who had received agalsidase alfa for more than 6 months, the observed improvement was generally maintained. This is the first study indicating a significant beneficial effect of ERT on gastrointestinal symptoms in a group of patients treated for Fabry disease.

Objectives: Human Granulocytic Ehrlichiosis (HGE) is a recently discovered zoonosis and, in Europe, not always included in laboratory testing when a patient presents with a history of tick bite. The available serology results indicate that HGE should be included in the screening panel when a tick-borne disease is suspected., Methods: Serological methods were applied; i.e. indirect immunofluorescence and Western Blot analysis. Sixty-five serum samples from 47 patients were analysed, of six patients sequential samples were available., Results: 33.8% of the submitted samples were found positive in indirect immunofluorescence, Western Blot confirmed 46.1% of these positive samples., Conclusions: Although the causative agent and the vector for HGE, Ixodes ticks, are present in Belgium, serology for HGE is seldom solicited. Ehrlichiosis is apparently not always considered as a plausible or possible cause for illness, even when the patient presents with a history of tick bite. We present here a, true be it, incomplete picture of the present situation in Belgium, but nevertheless indicating that it is warranted to test patients with a history of tick bite not only for Lyme disease, but also for HGE.

The anti-influenza CD8+ T cell response in HLA-A2-positive adults is almost exclusively directed at residues 58-66 of the virus matrix protein (MP(58-66)). V(beta)17V(alpha)10.2 T cell receptors (TCRs) containing a conserved arginine-serine-serine sequence in complementarity determining region 3 (CDR3) of the V(beta) segment dominate this response. To investigate the molecular basis of immunodominant selection in an outbred population, we have determined the crystal structure of V(beta)17V(alpha)10.2 in complex with MP(58-66)-HLA-A2 at a resolution of 1.4 A. We show that, whereas the TCR typically fits over an exposed side chain of the peptide, in this structure MP(58-66) exposes only main chain atoms. This distinctive orientation of V(beta)17V(alpha)10.2, which is almost orthogonal to the peptide-binding groove of HLA-A2, facilitates insertion of the conserved arginine in V(beta) CDR3 into a notch in the surface of MP(58-66)-HLA-A2. This previously unknown binding mode underlies the immunodominant T cell response.

A purification procedure is described for the isolation of recombinant HIV-2 reverse transcriptase expressed in Escherichia coli. The p68 subunit is expressed, in the absence of induction, and use of a heparin-Sepharose column produces substantially pure protein. Concentration of the homodimeric p68 reverse transcriptase pool, followed by incubation at room temperature for several days, results in full conversion by E. coli proteases to the heterodimer (p68/p55). This extended incubation simplifies the purification process and improves the yield of heterodimeric reverse transcriptase, which shows a truncation of the smaller subunit to 427 residues. The protein is then purified further by hydroxyapatite and gel-filtration chromatography to homogeneity. The HIV-2 RT is active and has been used to produce crystals that diffract to beyond 3.0 A.

In order to determine the prevalence of human granulocytic ehrlichiosis (HGE) in Belgium, the sera of 216 patients previously diagnosed with Borrelia burgdorferi infection were analysed for possible coinfection with the agent of HGE. For this purpose, an indirect immunofluorescence assay was applied, and positive results were confirmed by Western blot using a 44-kilodalton recombinant protein (rP44) specific for the agent of HGE. Sixteen of the 216 (7.4%) sera tested were positive for the HGE agent using indirect immunofluorescence assay, and seven (3%) of them were confirmed positive by Western blot. These data suggest the agent for HGE is present in Belgium and may cause coinfection in patients infected with Borrelia burgdorferi, as has been reported in the USA and elsewhere in Europe. This is the first report documenting the identification of this agent in Belgium.

Hybrids between different species or genera of the single-stranded RNA coliphages have not been found in nature. Here, it has been shown that viable hybrids between different phage species can easily be generated in the laboratory by in vivo recombination. cDNA of species I phage MS2 located on a plasmid and lacking part of its 5' untranslated leader (5' UTR) was complemented with another plasmid carrying the 5' half of the genome of fr, a species I phage, or of KU1, a species II representative with low sequence similarity. When the two plasmids were present in the same cell there was spontaneous production of hybrid phages. Interestingly, these hybrids did not arise by a double or single crossover that would replace the missing MS2 sequences with those of fr or KU1. Rather, hybrids arose by attaching the complete 5' UTR of fr or KU1 to the 5' terminus of the defective MS2 phage. Several elements of the 5' UTR then occurred twice, one from KU1 (or fr) and the other from MS2. These redundant elements are in most cases deleted upon evolution of the hybrids. As a result, the 5' UTR of KU1 (or fr) then replaced that of MS2. It was earlier shown that this 5' UTR could assume two alternating structures that facilitated transient translation of the proximal maturation gene. Apparently, this timer function of the 5' UTR was exchangeable and could function independently of the rest of the genome. When hybrids were competed against wild-type, they were quickly outgrown, probably explaining their absence from natural isolates.

We report on three cases, two with myelodysplastic syndrome (MDS) and one with acute lymphoblastic leukemia (ALL), displaying trisomy 16 as the sole cytogenetic anomaly. In none of these cases was a concomitant inv(16)(p13q22) detected by fluorescence in situ hybridization (FISH) or reverse transcription polymerase chain reaction (RT-PCR). Summarizing the literature, only six other cases cytogenetically characterized by an isolated trisomy 16 have been reported in hematological malignancies. These patients had either MDS, acute myeloblastic leukemia (AML), myelofibrosis, or ALL. All but one of these cases were aged less than 50.

A case of chronic myeloid leukemia displaying an uncommon t(21;22)(q22;q11) is reported. For the first time, this translocation has been characterized by fluorescence in situ hybridization (FISH) and the reverse transcriptase polymerase chain reaction (RT-PCR). FISH, with the use of whole-chromosome painting probes and probes specific for the BCR and ABL genes, showed a three-way variant Philadelphia translocation (9;22;21)(q34;q11;q22) with a BCR/ABL fusion residing on the der(22). In addition, RT-PCR demonstrated a b2a3 BCR/ABL fusion transcript. Underlying mechanisms and prognostic implications are discussed.

We report on a case of biopsy-proven progressive multifocal leukoencephalopathy (PML) in a patient without obvious immunodeficiency. Analysis of the cerebrospinal fluid revealed the simultaneous presence of JC virus DNA and of locally produced, anti-JC virus antibodies. The intrathecal humoral immune response increased throughout the course of the disease, whereas the detection of the JC genome became ultimately negative in spite of the continuous extension of the lesions with fatal outcome.

Frontotemporal dementia (FTD) was diagnosed in 74 outpatients with a standardized assessment including neuropsychological tests, behavioural scale, structural and functional imaging. Clinical characteristics were consistent with the literature data. The cohort was followed for 2-6 years to determine the reliable variable for evaluating the progression of FTD. Every fourth patient died after a mean duration of 7 years. At first, FTD manifests itself in behavioural changes with relatively stable global cognition although language, verbal fluency and memory tests were reliable tools to follow the progression of the disease. Below 18 of Mini-Mental State Examination, mutism and apathy prevented from neuropsychological testing within the next 6 months. Behavioural disorders evolved with time but restlessness and hyperorality were long-lasting. Imaging showed the progression of a consistent pattern of anterior abnormalities with frequent leukoaraiosis.

Costimulatory molecules B7-1 (CD80) and B7-2 (CD86) are differently involved in T cell stimulation. In chronic experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis (MS), B7-1 was preferentially involved in pathophysiology of relapses. We used reverse transcription polymerase chain reaction (RT-PCR) to amplify the mRNA coding for these molecules in cerebrospinal fluid (CSF) cells and peripheral blood mononuclear cells (PBMC) from 18 MS patients and 21 other neurological patients. In CSF cells of MS cases, B7-1 mRNA was only detected in some patients who showed clinical signs of acute relapse at the time of the spinal tap, while B7-2 mRNA was widely detectable without difference between active or stable MS and controls. mRNA coding for transforming growth factor-beta (TGF-beta) was detectable in the majority of cases, with higher expression in CSF cells of MS and other inflammatory neurological diseases (OIND) than in noninflammatory controls, and higher expression in PBMC of MS patients than in all other cases. Finally, mRNA coding for interleukin (IL)-12p40 was only detected in a very few number of MS and inflammatory cases. These findings were related to previous detection of other cytokines in the same cases, showing relationships in CSF cells between high expression of B7-1, IL-12p40 and TNF-alpha.

AL amyloidosis is an infiltrative disorder characterized by the extracellular deposition of insoluble fibrillar immunoglobulin light chains whose production results from a plasma cell dyscrasia. Treatment with melphalan has resulted in an improvement in a few patients. Recently, intensive chemotherapy followed by autologous or syngeneic stem cell support has been shown to offer potential benefit. Allogeneic stem cell support after intensive therapy would retain the benefits of autologous transplantation, with the additional advantages of a tumor-free graft and of a possible graft-versus-tumor effect. We report a patient with AL amyloidosis and significant proteinuria. She improved after an allogeneic bone marrow transplantation.

We report a case of cluster-like headache in a patient with a trigeminal neurinoma. Symptomatic cluster headache was suspected because of the absence of typical periodicity and the persistence of background headache. Magnetic resonance imaging findings were consistent with a trigeminal neurinoma.

Reversible dementia and polyneuropathy in an elderly woman: a case of late onset systemic lupus erythematosus. We report the case of a 76-year-old woman with undiagnosed systemic lupus erythematosus. She presented with severe dementia. She was given prednisolone 20 mg daily with dramatic improvement occurring within one month. Electrophysiological study showed evidence of axonal neuropathy.

We report a study of astigmatism and postoperative complications in 170 eyes following phacoemulsification and implantation of a flexible silicone intraocular lens (SI-18NB) in the capsular bag. No significant difference existed between the preoperative mean astigmatism and the postoperative mean astigmatism after 7 days and after 1 month. Complications were infrequent and compared with those associated with PMMA intraocular lenses.

Contrast venography, the accepted gold standard for the diagnosis of venous thrombosis of the leg, is a painful and invasive procedure with late side effects. There is controversy in literature reports about the sensitivity of 99Tcm-macroaggregated albumin (MAA) phlebography: indeed, using 99Tcm-MAA, poor results are obtained when one has to detect calf vein thrombosis. As with other isotopic procedures requiring a pedal injection of the tracer, the use of an injectable solution of 81Krm is a nonspecific method, based upon the abnormality in flow in the deep venous system which results from a deep venous thrombosis. However, when compared with 99Tcm-MAA, 81Krm offers theoretical advantages for phlebographic studies of the lower limbs. In this work 24 patients were studied both with contrast phlebography and with 81Krm. Although 81Krm provided images of high quality, there was a lack of sensitivity below the knee, where false negative results were observed. This could be explained by the fact that the radionuclide venographic procedures usually visualize only one or, in some cases, two of the three deep veins of the calf. Moreover, accurate differentiation between superficial and deep veins in the calf often appears difficult, even using a tourniquet.

Multiparous Holstein cows at six universities were utilized to examine effects of ruminally protected methionine and lysine on lactational performance. Three hundred and four cows began the study; 259 cows were included in the production analysis. Following a 21-d standardization period, cows received a basal diet of corn silage and ground corn supplemented with one of five dietary treatments, which were supplements of soybean meal or corn gluten meal, the latter with zero and three combinations of protected methionine and lysine (methionine; methionine and lysine; methionine and double (2x) lysine). Treatment effects were evaluated during early, mid, late, and total lactation (22 to 112, 113 to 224, 225 to 280, and 22 to 280 d postpartum, respectively). On a DM basis, ratios of forage to concentrate (50:50, 60:40, and 70:30) increased, and dietary CP (16.0, 14.5, and 13.0%) decreased during the three periods of lactation. Amount of amino acid supplementation also decreased (15, 12, and 9 g/d methionine; 20, 16, and 12 g/d lysine; and 40, 32, and 24 g/d 2x lysine) with period of lactation. Actual and least squares means for milk, FCM, and milk protein yields were greater for soybean than for corn gluten meal during early, mid, and total lactation. In addition, these variables responded linearly to lysine in early lactation. Response to lysine was quadratic during mid and total lactation for these variables. Differences in nutrient intake explained production responses to protein sources but not to lysine. Serum amino acid responses primarily reflected differences in dietary protein source and rumen-protected amino acid.

Salivary inclusion is seen, on panoramic films of the maxilla and the mandible, in the form of a mandibular osseous lacuna. Such an indicative diagnostic image, the prognosis for which is often sobering, requires CT scan exploration. The latter permits the diagnosis of Stafne's lacuna, the nature of which is benign. Quick and sure.

Based on 27 rare forms of carcinoma of the endometrium (adenoacanthomas, clear cell adenocarcinomas, mulleroblastomas, leiomyosarcomas), the authors discuss their observations and compare their findings with literature data. The only forms with a more favorable prognosis than cancer of the endometrium is pure adenoacanthoma. Delays in diagnosis remain all too frequent. Post-surgery treatment including chemotherapy appears preferable to surgery alone for these severe lesions which result more often in local recurrences than in pulmonary metastases. New chemotherapy associations (ADR-DTIC) have given results which justify their use, while radiotherapy and curietherapy reduce the number of recurrences, except for leiomyosarcomas.

Data were from 45 Holstein cows (23 multiparous, 22 primiparous) assigned by calving date and parity within groups to one of two isonitrogenous (16% crude protein) diets. The diets were 50% forages (corn silage, alfalfa silage) and 50% concentrate, dry basis. In diet A, soybean meal supplied 34.2% of total crude protein; in diet B, ground sweet white lupin seeds provided 37.9% of total crude protein. Cows were fed once daily during the experimental period (d 4 to 116 postpartum). Cows fed lupins consumed significantly less dry matter, produced 1.8 kg/d less milk (but not significantly different), and had lower milk protein percent. Milk fat and total solids percents were similar. Reasons for reduced intake of cows fed lupins were not evident. Traces of alkaloids (.005% dry basis) were present in diet B. Combined results of in vitro continuous culture fermentation and in situ degradation measurements indicated that crude protein from lupins was more degradable than that of soybean meal. Poor performances of cows fed lupins could be partly due to a reduced true protein supply to the small intestine.

A particularly high hypercalcemia (141 mg/ml) was observed in a man with Graves' disease. An intense muscle asthenia, with lack of dynamism and vomiting which may cause dehydration, are the most suggestive signs of hypercalcemia. Bone biopsy and above all parathormone estimations permit one to eliminate associated hyperparathyroidism. The efficacy of mithramycin used alone, without any other hypocalcemic drug, was remarkable. The direct responsibility of thyrotoxicosis as a cause of the calcium disorder seems undoubted but the precise mechanism of the hypercalcemia remains unexplained.