Salivary Gland Proteins Research Articles

Salivary glands of Amblyomma cajennense (Acari: Ixodidae): a histological and an ultrastructural overview

The present study on the salivary glands of semi-engorged Amblyomma cajennense females has identified the various cell types present in this tissue and allowed its morphohistochemical characterization. Marking techniques were applied to detect polysaccharides (PAS), proteins (bromophenol blue), lipids (Nile blue) and calcium (von Kossa), as well as those of scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results obtained by TEM showed and confirmed that these individuals' glands are also formed by round acini that are connected to the common excretory duct through acinar and intermediate ducts. Histological data as well as ultrastructural ones showed that the glands are formed by types I, II and III acini. In this study with salivary glands polysaccharides, proteins, lipids and calcium were observed in the cytoplasm and/or cell secretion granules-both free or forming complexes, as the intensity of the marking varied according to the cell as well as the type of acini analyzed, showing the structural and functional complexity of the tick salivary glands, characteristics that give the multifunctional character to this organ.

Glycosylation of proteins in healthy and neoplastic human salivary glands – a preliminary study

In cancer tissue, altered glycosylation of proteins is observed. There are some typical changes; for example, sialyl-Lewis(a/x) glycoforms are more abundant in many types of cancers. The current study investigated the differences in glycosylation of proteins between neoplastic and healthy human salivary glands. Sugar structures on proteins with a molecular mass above 30 kDa were determined by enzyme-linked immunosorbent assay (ELISA) with biotinylated lectins. The expression of sialic acid in cancer tissues was higher in comparison with healthy ones. The same observations were revealed for Fuc α1-6, α1-2, T, Tn antigens and α1-6 mannose. Glycosylation of proteins in cancer salivary gland tissues is altered in comparison with healthy tissue.

Detection of AA-type amyloid protein in labial salivary glands

Among the diverse forms of amyloidosis, secondary type is the most frequent one. Diagnosis of amyloid deposition is based on the identification of the fibrillary protein amyloid by means of Congo Red (CR) or crystal violet (CV) stains, but these techniques do not differentiate between the different types of amyloid fibrils. The aim of this study was to identify by immunofluorescence (IF) AA amyloid a pathological fibrillar low-molecular-weight protein formed by cleavage of serum amyloid A (SAA) protein in labial salivary gland (LSG) biopsies from patients with secondary amyloidosis. 98 LSG were studied, 65 were from patients with secondary amyloidosis and 33 from subjects with chronic inflammatory diseases without evidence of this anomaly. All sections were stained with hematoxylin and eosin (H &E), CV, CR and IF using anti-AA antibodies. Positive and negative controls were used for all techniques. CV and CR demonstrated that the amyloid substance was found mainly distributed periductally (93.8%), followed by periacinar and perivascular locations (p <0.001); however, the IF demonstrated that amyloid AA substance predominates in the periacinar area (73.8%), followed by periductal and perivascular locations (p <0.001). IF has a sensitivity of 83%, 100% of specificity, 100% of predictive positive value and 75% of predictive negative value. The results of this study confirm the efficacy of the LSG biopsy as a highly reliable method for diagnosis of secondary amyloidosis.

Reduced statherin expression in labial glands of diabetic subjects

Many oral diseases are associated to diabetes and, correspondingly, the salivary composition is strongly altered in subjects affected by this metabolic disorder with respect to healthy ones. Periodontal disease, tooth demineralization, caries, and oral infections, which frequently trouble diabetics, are generally ascribed to changes in some secretory protein expression. We focused our attention on the expression of the phosphoprotein statherin, which seems to play important roles in maintaining the buccal health, in human salivary glands. Recently, we reported the ultrastructural localization of this protein in human major and minor salivary glands and a significant reduction of its expression in submandibular glands of diabetic patients versus normal ones. In this study, we compare the subcellular immunoreactivity of statherin in diabetic and non diabetic labial glands. Surgical samples were fixed with a glutaraldehyde and paraformaldehyde mixture, embedded in Epon, and treated for immunogold histochemistry using a polyclonal antibody specific for statherin. Specific immunoreactivity was revealed in serous cells, where gold particles were deposited onto a lot of secretory vesicles and tubules diffused throughout the cytoplasm, Golgi apparatus and RER. The gold particles were counted and the labelling density was expressed as number of gold particles /μm2. The results obtained with diabetic samples were then compared with those in the controls. Statistical analysis showed a significant reduction of statherin expression in the diabetic versus normal glands. These data suggest that statherin secretion is affected by diabetes not only in major but also in minor salivary glands, supporting the belief that scarce amounts of this protein could be partly responsible for oral disturbances. Since the best known statherin functions are related to the formation of the acquired enamel pellicle and inhibition of calcium salt precipitation, reduced amounts could determine defects in the pellicle and in calcium metabolism, thus supplying inadequate protection to the teeth.

Hsp70 expression in Chironomus ramosus exposed to gamma radiation

Purpose: A tropical species of midge, Chironomus ramosus has been recently reported to be one of the radio-tolerant groups of organisms. The present study was undertaken to examine the protein profile and expression of Heat shock protein-70 (Hsp70) in gamma radiation stress, which has also been reported as a common biomarker for different type of stressors.Materials and methods: Metabolic labelling of salivary gland (SG) proteins with [35S]-methionine showed over-expression of a 70 kDa protein band up to 4 hours (h) of observation in the post exposure recovery period. For confirmation of the expression of Hsp70 in SG cells after gamma radiation exposure, semi-quantitative real-time-polymerase chain reaction (RT-PCR), Western blotting and immuno-fluorescence detection of Hsp70 were carried out.Results: Results showed elevated levels of Hsp70 mRNA and protein in SG cells of larvae immediately after gamma radiation exposure. The levels dropped to basal values by 48 h in the recovery period.Conclusions: The present study confirmed that radio-tolerant midge, C. ramosus expressed Hsp70 upon gamma radiation exposure and Hsp70 might be one of the gamma radiation-induced stress proteins required during the early stages of radiation stress management in aquatic midge larvae. This is the first report of its kind from the juvenile stage of any aquatic insect group.

Differential Expression of Salivary Gland Genes in the Female Sand Fly Phlebotomus papatasi (Diptera: Psychodidae)

Saliva from blood-sucking arthropods modulates host homostasis and immunity, making salivary components potential candidates to be used against pathogens transmitted by these biting insects. Functional characterization of salivary molecules is fundamental to gain a better understanding into their roles during blood feeding and to determine under which conditions such molecules are expressed in the insect saliva. In the current study, we investigated the expression profile of 10 salivary genes from the sand fly Phlebotomus papatasi (Scopoli) (Diptera: Psychodidae), a principal vector of Leishmania major. Our analyses using quantitative polymerase chain reaction were aimed at defining whether diet or senescence influences the expression of P. papatasi salivary gland-expressed genes in laboratory-reared female sand flies. Our results demonstrate that at least one of the most abundant salivary transcripts, SP44, is consistently modulated by either senescence or diet. In contrast, another abundant transcript, SP32, was expressed without any influence from the diet received or the age of the sand fly. Differential expression of the other eight transcripts was not consistently regulated by either diet or age, suggesting that other factors may have a greater influence on differential expression of these salivary gland proteins.

Evolution of Vertebrate Hemostatic and Inflammatory Control Mechanisms in Blood-Feeding Arthropods

Arthropods evolved blood-feeding lifestyles on more than 20 independent occasions. A comparison of sialomes (secretory salivary gland protein family repertoires) corroborates this and indicates that unique protein families are found for different blood-feeding lineages. Closely related lineages considered to have evolved blood-feeding behavior independently might share similar sets of protein families in their sialomes. No orthologous relationships or conserved functional mechanisms have, however, been found for these apparently related protein family members. This implies that sialomes were already defined before the divergence of these related lineages, but that the functions involved in blood feeding evolved at a later stage. Protein families prone to lineage-specific expansion (gene duplication) tend to be abundant with specialized functions associated with various gene duplicates. A central theme that seems to emerge from comparative sialomic analysis is that of the convergent evolution of modulatory strategies that targets host defenses even if the molecular mechanisms differ.

Selective cloning, characterization, and production of the Culicoides nubeculosus salivary gland allergen repertoire associated with equine insect bite hypersensitivity

Salivary gland proteins of Culicoides spp. have been suggested to be among the main allergens inducing IgE-mediated insect bite hypersensitivity (IBH), an allergic dermatitis of the horse. The aim of our study was to identify, produce and characterize IgE-binding salivary gland proteins of Culicoides nubeculosus relevant for IBH by phage surface display technology. A cDNA library constructed with mRNA derived from C. nubeculosus salivary glands was displayed on the surface of filamentous phage M13 and enriched for clones binding serum IgE of IBH-affected horses. Ten cDNA inserts encoding putative salivary gland allergens were isolated and termed Cul n 2 to Cul n 11. However, nine cDNA sequences coded for truncated proteins as determined by database searches. The cDNA sequences were amplified by PCR, subcloned into high level expression vectors and expressed as hexahistidine-tagged fusion proteins in Escherichia coli. Preliminary ELISA results obtained with these fusions confirmed the specific binding to serum IgE of affected horses. Therefore, the putative complete open reading frames derived from BLAST analyses were isolated by RACE-PCR and subcloned into expression vectors. The full length proteins expressed in Escherichia coli showed molecular masses in the range of 15.5–68.7 kDa in SDS-PAGE in good agreement with the masses calculated from the predicted protein sequences. Western blot analyses of all recombinant allergens with a serum pool of IBH-affected horses showed their ability to specifically bind serum IgE of sensitized horses, and ELISA determinations yielded individual horse recognition patterns with a frequency of sensitization ranging from 13 to 57%, depending on the allergen tested. The in vivo relevance of eight of the recombinant allergens was demonstrated in intradermal skin testing. For the two characterized allergens Cul n 6 and Cul n 11, sensitized horses were not available for intradermal tests. Control horses without clinical signs of IBH did not develop any relevant immediate hypersensitivity reactions to the recombinant allergens. The major contribution of this study was to provide a repertoire of recombinant salivary gland allergens repertoire from C. nubeculosus potentially involved in the pathogenesis of IBH as a starting basis for the development of a component-resolved serologic diagnosis of IBH and, perhaps, for the development of single horse tailored specific immunotherapy depending on their component-resolved sensitization patterns.

Las glándulas salivales de dos flebotominos vectores de Leishmania: Lutzomyia migonei (França) y Lutzomyia ovallesi (Ortiz) (Diptera: Psychodidae)

Leishmaniasis is a vector-borne disease transmitted by the intradermal inoculation of Leishmania (Kinetoplastida: Trypanosomatidae) promastigotes together with saliva during the bite of an infected sand fly. The salivary glands were compared from two vector species, Lutzomyia ovallesi (Ortiz,1952) and Lutzomyia migonei (França,1920) (Diptera: Psychodidae). Protein profiles by SDS PAGE of salivary glands were compared among species as well as their development at several times post feeding. First, mice were immunized to salivary proteins by exposure to biting by L. ovallesi and of L. migonei. Antibodies in these mice against salivary gland-specific proteins were evaluated by immunoblotting. No apparent change was revealed in the kinetic expression of salivary proteins induced by the different physiological states post feeding. Qualitative and quantitative variations were detected in16-18 polypeptides with molecular weights ranging from 6 to 180 kDa. Species-specific proteins were demonstrated for L. migonei and L. ovallesi. In addition, antibodies against salivary gland specific proteins were found in mice immunized by the saliva of both species. Basic information was obtained concerning the nature of salivary gland proteins of L. migonei and L. ovallesi. This information helps to elucidate the role of salivary proteins and their potential as effective tools in screening risk factors in human and other vertebrate hosts.

PLUNC protein expression in major salivary glands of HIV-infected patients

To analyse and compare the expression of Palate, Lung, and Nasal Epithelium Clone (PLUNC) proteins in salivary glands from patients with and without AIDS (control group) using autopsy material. We analysed the expression of PLUNCs using immunohistochemistry in parotid (n = 45), submandibular (n = 47) and sublingual gland (n = 37) samples of AIDS patients [30 with normal histology, 21 with mycobacteriosis, 14 with cytomegalovirus (CMV) infection, 30 with chronic non-specific sialadenitis, and 30 HIV-negative controls. In situ hybridization (ISH) for SPLUNC 2 in the HIV-negative group was performed. SPLUNC 1 expression was detected in the mucous acini of submandibular and sublingual glands, and SPLUNC 2 were seen in the serous cells. LPLUNC 1 expression was only positive in the salivary ducts. There was a higher expression of SPLUNC 2 in AIDS patients with CMV infection and mycobacteriosis when compared with all other groups. The intensity of staining for SPLUNC 2 was greater around the lesions than the peripheral ones. ISH for SPLUNC 2 showed perinuclear positivity in the serous cells in all HIV-negative cases. SPLUNC 1 and LPLUNC 1 proteins were similarly expressed in the salivary glands of AIDS patients and non-HIV patients. CMV infection and mycobacteriosis increase SPLUNC 2 expression in serous cells in the salivary gland of AIDS patients.

Overexpression of sphingosine kinase 1 is associated with salivary gland carcinoma progression and might be a novel predictive marker for adjuvant therapy

BackgroundOverexpression of sphingosine kinase-1 (SPHK1) has been demonstrated to be associated with the development and progression in various types of human cancers. The current study was to characterize the expression of SPHK1 in salivary gland carcinomas (SGC) and to investigate the association between SPHK1 expression and progression of SGC.MethodsThe expression of SPHK1 was examined in 2 normal salivary gland tissues, 8 SGC tissues of various clinical stages, and 5 pairs of primary SGC and adjacent salivary gland tissues from the same patient, using real-time PCR and western blot analysis. Furthermore, the SPHK1 protein expression was analyzed in 159 clinicopathologically characterized SGC cases by immunohistochemistry. Statistical analyses were performed to determine the prognostic and diagnostic associations.ResultsSPHK1 expression was found to be markedly upregulated in SGC tissues than that in the normal salivary gland tissues and paired adjacent salivary gland tissues, at both mRNA and protein levels. Statistical analysis revealed a significant correlation of SPHK1 expression with the clinical stage (P = 0.005), T classification (P = 0.017), N classification (P = 0.009), M classification (P = 0.002), and pathological differentiation (P = 0.013). Patients with higher SPHK1 expression had shorter overall survival time, whereas patients with lower SPHK1 expression had better survival. Importantly, patients in the group without adjuvant therapy who exhibited high SPHK1 expression had significantly lower overall survival rates compared with those with low SPHK1 expression. Moreover, multivariate analysis suggested that SPHK1 expression might be an independent prognostic indicator for the survival of SGC patients.ConclusionsOur results suggest that SPHK1 expression is associated with SGC progression, and might represent as a novel and valuable predictor for adjuvant therapy to SGC patients.

Molecular cloning of a small heat shock protein (sHSPII) from the cattle tick Rhipicephalus ( Boophilus) annulatus salivary gland

Immunoscreening of a cDNA expression library of the Rhipicephalus ( Boophilus) annulatus tick with purified rabbit anti- R annulatus salivary glands antigens polyclonal antibodies led to the identification of a 661 bp sequence. The sequence includes an open reading frame of 543 bp encoding a protein of 180 amino acids with calculated molecular weight of 20.51 kDa, isoelectric point of 9.071 and with no signal sequence. Comparison of the deduced amino acids with protein data bank showed that the identified polypeptide belongs to the alpha crystallin small heat shock proteins superfamily and shows sequence similarity of 62% and 55% to Ixodes scapularis fed tick salivary gland protein and Ornithodoros parkeri alpha-crystallin protein, respectively. Accordingly, this protein was called Ra-sHSPII. The Ra-sHSPII protein was expressed in E. coli under T7 promotor of the pET-30b vector, purified under denaturation conditions and the immunogenicity and cross-reactivity of the recombinant Ra-sHSPII were evaluated. Direct ELISA showed that the Ra-sHSPII is a strong immunogen. In immunoblotting assay the anti-rRa-sHSPII antisera reacted specifically with purified rRa-sHSPII, with several proteins in R. annulatus whole tick, larval and gut protein extracts in addition to Hyalomma dromedarii and Ornithodoros moubata whole tick protein extracts, as examples of hard and soft tick species, respectively. The rRa-sHSPII protein confers thermal protection to other proteins in vitro as found in other sHSPs. E. coli cell extracts containing the protein were protected from heat-denatured precipitation when heated up to 100 °C, whereas extracts from cells not expressing the protein were heat-sensitive at 60 °C.

Infectious arthritis and immune dysregulation: lessons from Lyme disease

Borrelia burgdorferi colonization of the joints induces an inflammatory response, which in some individuals progresses to chronic arthritis. In this review, we discuss novel pathways that are implicated in disease development by modulating host defenses to B. burgdorferi infection. The use of transgenic mice and gene expression analyses has revealed novel pathways involved in pathogenesis of Lyme disease. It is now clear that B. burgdorferi exploits an array of salivary gland proteins of the tick to evade immune responses in the mammalian host. The spirochete also modulates its surface protein profile upon infection and induces anti-inflammatory cytokines, favoring survival of the pathogen. The host defense involves toll-like receptors (TLRs), such as TLR2 and others, in B. burgdorferi recognition. To further dissect the genetic predisposition to treatment-refractory Lyme arthritis, HLA-DR transgenic mice have been used. The cause and pathogenesis of Lyme arthritis are complex. Elucidating the mechanisms that govern this chronic inflammatory response will provide direct insights into other infectious arthritides and the development of novel therapeutic approaches against B. burgdorferi infection.

No Major Role for Binding by Salivary Proteins as a Defense Against Dietary Tannins in Mediterranean Goats

We investigated whether Mediterranean goats use salivary tannin-binding proteins to cope with tannin-rich forages by determining the affinity of salivary or parotid gland proteins for tannic acid or quebracho tannin. Mixed saliva, sampled from the oral cavity, or parotid gland contents were compared to the intermediate affinity protein bovine serum albumin with a competitive binding assay. Goats that consume tannin-rich browse (Damascus) and goats that tend to avoid tannins (Mamber) were sequentially fed high (Pistacia lentiscus L.), low (vetch hay), or zero (wheat hay) tannin forages. Affinity of salivary proteins for tannins did not differ between goat breeds and did not respond to presence or absence of tannins in the diet. Proteins in mixed saliva had slightly higher affinity for tannins than those in parotid saliva, but neither source contained proteins with higher affinity for tannins than bovine serum albumin. Similarly, 3 months of browsing in a tannin-rich environment had little effect on the affinity of salivary proteins for tannin in adult goats of either breed. We sampled mixed saliva from young kids before they consumed forage and after 3 months of foraging in a tannin-rich environment. Before foraging, the saliva of Mamber kids had higher affinity for tannic acid (but not quebracho tannin) than the saliva of Damascus kids, but there was no difference after 3 months of exposure to tannin-rich browse, and the affinity of the proteins was always similar to the affinity of bovine serum albumin. Our results suggest there is not a major role for salivary tannin-binding proteins in goats. Different tendencies of goat breeds to consume tannin-rich browse does not appear be related to differences in salivary tannin-binding proteins.

Salivary Gland Protein Repertoire fromAedes aegyptiMosquitoes

Diseases caused by arthropod-borne viruses are a significant threat to the health of human and animal populations throughout the world. Better knowledge of the molecules synthesized in the salivary gland and saliva of hematophagous arthropods could be of use for improving the control of pathogen transmission. Recently, a sialome analysis of three Aedes aegypti mosquito colonies (PAEA, Rockefeller, and Formosus) carried out in our laboratory allowed us to identify 44 saliva proteins. Of these secreted proteins, none was exclusively expressed in one colony, suggesting that expression of salivary proteins is highly conserved across populations. In another study, we reported that some of these salivary proteins could be used as the genus-specific markers for travelers' exposure to mosquito vectors. Here, comparison of salivary gland protein profiles between these same three Ae. aegypti colonies was performed using the one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) difference gel electrophoresis method. As observed at the saliva level, no significant differences were detected between these three colonies. The salivary gland protein repertoire from the Ae. aegypti mosquito was analyzed using a proteomic approach. One hundred and twenty proteins were identified in these salivary glands representing the largest description of the Ae. aegypti salivary gland protein catalog. We succeeded in identifying 15 secreted proteins, some of which have already been reported as being involved in blood feeding. A comparison of the proteins identified between the salivary glands and the sialome is discussed.

Blood‐feeding and immunogenic Aedes aegypti saliva proteins

Mosquito-transmitted pathogens pass through the insect's midgut (MG) and salivary gland (SG). What occurs in these organs in response to a blood meal is poorly understood, but identifying the physiological differences between sugar-fed and blood-fed (BF) mosquitoes could shed light on factors important in pathogens transmission. We compared differential protein expression in the MGs and SGs of female Aedes aegypti mosquitoes after a sugar- or blood-based diet. No difference was observed in the MG protein expression levels but certain SG proteins were highly expressed only in BF mosquitoes. In sugar-fed mosquitoes, housekeeping proteins were highly expressed (especially those related to energy metabolism) and actin was up-regulated. The immunofluorescence assay shows that there is no disruption of the SG cytoskeletal after the blood meal. We have generated for the first time the 2-DE profiles of immunogenic Ae. aegypti SG BF-related proteins. These new data could contribute to the understanding of the physiological processes that appear during the blood meal.

Transcriptome Changes in &lt;I&gt;Culex quinquefasciatus&lt;/I&gt; (Diptera: Culicidae) Salivary Glands During West Nile Virus Infection

Persistent West Nile virus (WNV) infection in the mosquito Culex quinquefasciatus Say (Diptera: Culicidae) is associated with pathological changes in the salivary glands, including apoptotic cell death and a corresponding reduction in virus transmission over time. The vector host response to WNV infection and the molecular basis of WNV pathogenesis in Cx. quinquefasciatus was investigated using oligonucleotide microarrays designed to detect differences in the salivary gland transcriptome between WNV-infected mosquitoes and uninfected controls. Transcripts with increased abundance in infected salivary glands included those related to immunity, transcription, protein transport and degradation, amino acid and nucleotide metabolism, signal transduction, and cellular detoxification. Microarray-based analysis detected a decrease in transcript levels of a Culex inhibitor of apoptosis gene (IAP-1) and a decrease in abundance of 11 transcripts encoding salivary gland proteins. Transcript levels for an endonuclease, a proline-rich mucin, and several D7 protein family members also decreased. Transcripts with the greatest change in abundance during infection had either no similarity to sequences found in GenBank, VectorBase, and FlyBase, or were similar to sequences with uncharacterized protein products. These transcripts represent exciting targets for future analysis. Results from this study suggest that WNV infection influences transcriptional changes in an invertebrate host target tissue that may confer an advantage to the replicating virus, induce a host defense response, and alter the composition of vector saliva. The ramifications of these changes are discussed in terms of mosquito vector competence and WNV pathogenesis.

Immunization with Adenoviral-expressed salivary gland proteins (SALPs) decreases spirochete load in a murine model of Lyme borreliosis (52.2)

Abstract It is known that tick saliva contains a vast array of proteins that function as anti-haemostatic, anti-inflammatory, and immunomodulatory factors when delivered to the vertebrate host. Previous studies have also demonstrated that prior exposure to tick saliva induced specific immunity to tick infestation, as well as affording protection against tick-transmitted Borrelia burgdorferi infection in the mammalian host. In our current studies we explored a vaccination technique to modulate tick feeding, using an adenovirus (Ad) expression system to deliver four salivary proteins (previously shown to modulate host immunity) derived from Ixodes scapularis; Salp15, Salp25A, Salp25D, and ISAC. C3H/HeJ mice were immunized individually with Ad-Salp15 and Ad-ISAC constructs, as well as all four constructs simultaneously. Immunized mice were then challenged with B. burgdorferi-infected I. scapularis ticks. Utilizing quantitative PCR, spirochete burden in target organs like heart and bladder was reduced by 55% compared to animals immunized with an empty Ad-cassette. The generation of anti-salivary protein antibodies in a murine model was also confirmed. Our results indicate that adenovirus expressed tick salivary proteins can stimulate dendritic cell expression of tick salivary proteins, as well as tick-specific humoral and cellular immune responses in mice, affording at least partial protection against subsequent tick-transmitted B. burgdorferi infection.

Spinal Cord Sarcoidosis

Sarcoidosis is a granulomatous disorder of unknown cause that affects the spinal cord in fewer than 1% of patients who suffer from it. We conducted a retrospective case-control study of 31 patients with spinal cord sarcoidosis and compared them to 30 patients with myelopathies of other causes to analyze their clinical, laboratory, and magnetic resonance imaging (MRI) profiles and to assess their long-term prognoses. Thirty-one patients presented with clinical signs of myelopathy and were diagnosed with sarcoidosis. Twenty-two of these patients had biopsy-proven noncaseating granulomas. In 9 patients, sarcoidosis involved only a neurologic localization. Patients in the control group were mainly diagnosed with multiple sclerosis or optic neuromyelitis. Patients with sarcoidosis were more likely to have elevated levels of C-reactive protein (CRP), elevated lactate dehydrogenase (LDH), and hypergammaglobulinemia in serum, as well as a higher protein content and white blood cell count in cerebrospinal fluid. Spinal cord MRIs performed in 26 patients with spinal cord sarcoidosis revealed T2-hyperintensities that were extensive and heterogeneous with a central distribution in axial slides. Twenty-six patients with spinal cord sarcoidosis presented neurologic sequelae after follow-up (mean, 64 +/- 8 mo), although 2 patients completely recovered. Neurologic sequelae correlated with cerebrospinal fluid white blood cell counts. One-third of the patients had a monophasic course of the disease, another third had a relapsing-remitting course, and the remaining third had a progressive course. Four patients had pulmonary embolism during follow-up. Spinal cord sarcoidosis remains a diagnostic dilemma since neurologic localization is frequently the only manifestation. Because treatment for spinal cord sarcoidosis is far different from treatment for other myelopathies, such as multiple sclerosis and optic neuromyelitis, diagnosis of sarcoidosis remains an important challenge. Here, we show that spinal cord MRI and blood and cerebrospinal markers may be useful tools in the diagnosis of spinal cord sarcoidosis. We suggest that accessory salivary gland biopsies, chest X-rays, protein electrophoresis, and blood levels of CRP and LDH should be obtained for each patient with subacute myelopathy. We also recommend paying careful attention to thromboembolism in patients with spinal cord sarcoidosis because of systemic disease and their decreased mobility.

Anticoagulant activity in salivary gland homogenates of Thyrsopelma guianense (Diptera: Simuliidae), the primary vector of onchocerciasis in the Brazilian Amazon

In this study, anticoagulant activity was detected in salivary gland homogenates (SGHs) of Thyrsopelma guianense (Diptera: Simuliidae). The SGH yielded 1.07 microg +/- 0.03 (n = 15) of total soluble protein per pair of glands. In addition, following SDS-PAGE (12.5% gel) and silver nitrate staining, 12 polypeptides with molecular weights ranging from 14-69 kDa were detected in all physiological ages analyzed (12 h, 24 h, 48 h and 72 h following emergence). Coagulation bioassays showed that the SGHs had activities that interacted at all levels of coagulation (the intrinsic, extrinsic and common pathways), by extending the plasma recalcification time, prothrombin time, thrombin time. This is the first report on the activity of salivary gland proteins from the main vector of onchocerciasis in Brazil. We also suggest detailed studies on the morphology and function of the salivary glands in order to understand the role of these proteins in host/vector interactions.