The D7 family of salivary proteins in blood sucking diptera

The D7 subfamily of salivary proteins is widespread in blood sucking Diptera and belongs to the superfamily of pheromone/odourant binding proteins. Although D7 proteins are among the most abundant salivary proteins in adult female mosquitoes and sand flies, their role in blood feeding remains elusive. In the present work we report the sequence of seventeen novel D7 proteins, and propose an evolutionary scenario for the appearance of the several forms of this protein, based on a total of twenty-one sequences from Culex quinquefasciatus, Aedes aegypti, Anopheles gambiae, An. arabiensis, An. stephensi, An. darlingi mosquitoes and Lutzomyia longipalpis and Phlebotomus papatasi sand flies.     

The diagnosis and management of cow milk protein intolerance in the primary care setting

Cow milk protein intolerance (CMPI) affects 3% of infants under the age of 12 months and is often misdiagnosed as GERD or colic, risking dangerous exposure to antigens. Most infants out grow CMPI by 12 months; however, those with IgE-mediated reactions usually continue to be intolerant to cow's milk proteins and also develop other allergens including environmental allergens that cause asthmatic symptoms. Clinical manifestations of CMPI include diarrhea, bloody stools, vomiting, feeding refusal, eczema, atopic dermatitis, urticaria, angioedema, allergic rhinitis, coughing, wheezing, failure to thrive, and anaphylaxis. The research and literature showed that CMPI is easily missed in the primary care setting and needs to be considered as a cause of infant distress and clinical symptoms. This article focuses on correctly diagnosing CMPI and managing it in the primary care setting.     

Partial characterization of abnormal salivary proteins associated with rheumatoid arthritis

Isoelectric focusing of parotid saliva from 33 patients with rheumatoid arthritis but without clinical evidence of salivary gland involvement revealed multiple abnormal proteins of pI 3.95-4.25 in all cases, whereas only two out of 16 samples from normal healthy individuals exhibited any similar bands. Although rheumatoid factors of abnormally low isoelectric point and restricted heterogeneity were detected in this pI region in five of the diseased samples, these could not account for all of the bands even in these cases. The abnormal proteins did not otherwise cross-react with antisera to immunoglobulin (Ig) G, IgA or albumin. Analysis of rheumatoid arthritis parotid saliva by a method involving adsorption of rheumatoid factors to immobilized rabbit immunoglobulin revealed the presence of salivary rheumatoid factors (in seven patients) of all the major immunoglobulin classes, including secretory-IgA in two patients. Although these abnormal proteins are as yet uncharacterized, their presence could form the basis of a non-invasive procedure in the diagnosis of salivary gland and connective tissue disorders.     

Imaging of the major salivary glands

The major salivary glands, submandibular, parotid and sublingual glands play an important role in preserving the oral cavity and dental health. Patients with problems of the major salivary glands may present with symptoms such as dry mouth, dysphagia and obstruction of duct, inflammation, severe dental caries or swelling. Imaging plays an important role in visualization of morphology and function, to establish a diagnosis, for treatment, and for surgical planning. There are several options for diagnostic imaging: plain radiography, sialography, ultrasound (US), magnetic resonance imaging (MRI), computed tomography (CT), salivary gland scintigraphy and ¹⁸F‐FDG positron emission tomography (PET). We present an overview of the modalities in relation to common salivary gland disease.     

The transcriptome of adult female Anopheles darlingi salivary glands

Anopheles (Nyssorhynchus) darlingi is an important malaria vector in South and Central America; however, little is known about molecular aspects of its biology. Genomic and proteomic analyses were performed on the salivary gland products of Anopheles darlingi. A total of 593 randomly selected, salivary gland-derived cDNAs were sequenced and assembled based on their similarities into 288 clusters. The putative translated proteins were classified into three categories: (S) secretory products, (H) housekeeping products and (U) products with unknown cell location and function. Ninety-three clusters encode putative secreted proteins and several of them, such as an anophelin, a thrombin inhibitor, apyrases and several new members of the D7 protein family, were identified as molecules involved in haematophagy. Sugar-feeding related enzymes (alpha-glucosidases and alpha-amylase) also were found among the secreted salivary products. Ninety-nine clusters encode housekeeping proteins associated with energy metabolism, protein synthesis, signal transduction and other cellular functions. Ninety-seven clusters encode proteins with no similarity with known proteins. Comparison of the sequence divergence of the S and H categories of proteins of An. darlingi and An. gambiae revealed that the salivary proteins are less conserved than the housekeeping proteins, and therefore are changing at a faster evolutionary rate. Tabular and supplementary material containing the cDNA sequences and annotations are available at http://www.ncbi.nlm.nih.gov/projects/Mosquito/A_darlingi_sialome/     

Zinc enhancement of nonviral salivary gland transfection

Gene transfer to exocrine glands, including the major salivary glands, presents an attractive method to deliver proteins for therapeutic applications. Previous efforts using nonviral gene delivery to these glands have resulted in limited success. In this report, zinc and other divalent transitions were coadministered with plasmid DNA in an effort to improve nonviral salivary gland transfection efficiency. The inclusion of zinc into plasmid DNA solutions resulted in a 20-fold enhancement in transgene expression without noticeable inflammation compared to a solution of plasmid DNA only. This observed enhancement in transgene expression was dependent upon the DNA dose and correlated with the accumulation of plasmid DNA by salivary gland tissues.     

Molecular cloning and sequencing of salivary gland-specific cDNAs of the blood-sucking bug Triatoma brasiliensis (Hemiptera: Reduviidae)

Haematophagous insects produce pharmacological substances in their saliva to counteract vertebrate host haemostasis events such as coagulation, vasoconstriction and platelet aggregation. To investigate the bioactive salivary molecules of the triatomine bug Triatoma brasiliensis, we produced subtraction-enriched cDNAs of salivary-gland specific genes using suppression subtractive hybridization. Six full-length differentially expressed cDNAs (Tb113, Tb125, Tb152, Tb169, Tb180 and Tb198) were selected, cloned and sequenced. Sequence similarity searches of the databases using the putative amino acid sequence of our clones gave the following results: Tb152 - Triabin, an antithrombin induced platelet aggregation factor found in salivary gland extracts of T. pallidipennis. Tb169 - Pallidipin, an anticollagen induced platelet aggregation factor also found in T. pallidipennis salivary homogenates. Tb180 - Procalin, the major allergen of T. protracta saliva. The other three salivary-gland specific cDNAs produced no obvious homologies. Comparison of these salivary gland-specific cDNAs of with those of other triatomines combined with functional studies using recombinant proteins will allow a better understanding of the co-evolutionary process occurring between these insects and their vertebrate hosts, and may also lead to the discovery of novel antihaemostatic agents.     

Gerbil bite anaphylaxis

Household pets are well known to cause allergic symptoms in susceptible individuals, most commonly conjunctivitis, rhinitis, bronchospasm or urticaria. The increasing prevalence of exotic pets, including rodents, may introduce novel allergens into the household setting. We describe the case of a 16-year-old female who presented to the emergency department (ED) with an immediate systemic reaction consistent with anaphylaxis following a bite injury from a pet Mongolian gerbil. Although rare, gerbil bite injury represents another possible allergen source for precipitating a severe allergic reaction.     

Capillary feeding of specific dsRNA induces silencing of the isac gene in nymphal Ixodes scapularis ticks

Ixodes scapularis transmits several pathogens including Borrelia burgdorferi. Bioactive compounds in tick saliva support tick feeding and influence pathogen transmission to the mammalian host. These studies utilized oral delivery of dsRNA to silence an anticomplement gene (isac) in I. scapularis nymphs. Silencing of isac significantly reduced fed-tick weight compared to delivery of control lacZ dsRNA, and immunoblots specific for FlaB protein indicated a reduction in spirochete load in isac-silenced infected nymphs. SDS-PAGE demonstrated that isac gene silencing affected expression of a number of salivary and non-salivary gland proteins in ticks. Finally, multiple isac cDNA homologues were cloned, and these may represent a new gene family coexpressed during tick feeding. This work presents a novel oral delivery approach for specific gene silencing in I. scapularis nymphs and characterizes the effect of isac on blood-feeding in an attempt to block transmission of B. burgdorferi.     

A role for intracellular immunization in chemosensitization of tumor cells?

Acquired drug resistance represents a major cause of chemotherapy failure in patients with cancer. The characterization of the molecular pathways involved in drug resistance has provided us with new targets to overcome this problem. Many of these target proteins are often overexpressed in human cancers. A number of gene therapy strategies, including antisense oligonucleotides, ribozymes and single-chain antibodies, have been developed to achieve the selective modulation and inhibition of various cellu- lar proteins. Thus, these approaches can be exploited to modulate the resistance phenotype of tumor cells. These gene therapy strategies represent a novel and unique way to enhance the sensitivity of tumor cells to chemotherapeutic drugs. This review will focus on the use of intracellular immunization as a means to modulate the expression of specific genetic determinants involved in the drug resistance phenotype.     

Prolonged transgene expression in murine salivary glands following non-primate lentiviral vector transduction.

Salivary glands are an accessible organ for gene therapy, enabling expression of recombinant proteins for both exocrine and endocrine secretion. Lentivirus-based vectors have many advantages for gene therapy, including their ability to infect nondividing cells and to stably integrate into the host genome, enabling long-term transgene expression without eliciting an inflammatory immune response. In the present study, murine salivary glands were inoculated with feline immunodeficiency virus (FIV)-based lentiviral vectors expressing various reporter genes. Luciferase expression was observed as early as 24 h posttransduction, peaked at 17-21 days, and remained stable for more than 80 days. Staining with X-gal suggested that mucous acinar cells were effectively transduced. FIV vector transduction with the secreted alkaline phosphatase gene increased serum levels in treated animals for up to 45 days, and the FIV vector harboring the interferon-gamma (IFN-gamma) expression cassette induced an increase in IFN-gamma serum levels as well as in the supernatant of salivary gland explant cultures. These results demonstrate that the transduction of salivary glands with nonprimate lentiviral vectors may provide a novel and highly effective vehicle for long-term endocrine transgene expression.     

RNA interference‐mediated depletion of N‐ethylmaleimide Sensitive Fusion Protein and Synaptosomal Associated Protein of 25 kDa results in the inhibition of blood feeding of the Gulf Coast tick,Amblyomma maculatum

The signalling pathways in tick salivary glands that control ‘sialo‐secretome’ secretion at the tick?host interface remain elusive; however, this complex process is essential for successful feeding and manipulation of the host haemostatic response. Exocytosis of the sialo‐secretome in the salivary glands requires a core of soluble N‐ethylmaleimide‐sensitive fusion (NSF) attachment proteins (SNAPs) and receptor proteins (SNAREs). SNAREs have been identified as the key components in regulating the sialo‐secretome in the salivary gland cells. In this study, we utilized RNA interference to investigate the functional role of two Amblyomma maculatum SNARE complex proteins, AmNSF and AmSNAP‐25, in the tick salivary glands during extended blood feeding on the vertebrate host. Knock‐down of AmNSF and AmSNAP‐25 resulted in death, impaired feeding on the host, lack of engorgement and inhibited oviposition in ticks. Depletion also led to important morphological changes in the collapse of the Golgi apparatus in the salivary gland cells. Our results imply a functional significance of AmNSF and AMSNAP‐25 in prolonged tick feeding, and survival on the host. Further characterization of the factors that regulate exocytosis may lead to novel approaches to prevent tick‐borne diseases.     

CT tomography of normal parotid and mandibular salivary glands

Computed tomography (CT) allows the differentiation of tissues by measurement of their roentgenologic densities. There are a few controversial literature data on the roentgenologic densities of the parotid and submandibular glands. These differences are, in our opinion, due to several factors: 1. The complex anatomic structure of the facial region of the skull, that demands an exact centering. In daily practice the respective imaging parameters are not always kept constant (including partial volume effects), thus introducing density errors. 2. Metallic prostheses and obturators as well as the high-density bones of the facial skull cause artefacts that impair image quality and density measurements. 3. The small density differences between the salivary glands and the adjacent soft tissues makes their differentiation difficult. 4. Different devices and parameters are used for scanning, resulting in different densities. Considering these facts, we have devised a complex method for the investigation of the parotid and mandibular salivary glands.     

House dust sensitivity and environmental control

Allergy to foreign proteins in house dust is extremely common. Despite the fact that immunotherapy with house dust extract has been utilized for over sixty years, environmental control is still infrequently employed as a therapeutic measure. In this article the major allergen content of house dust is defined and strategies for decreasing the levels of these allergens in homes are described.     

Hemangioma with phleboliths in the sublingual gland: as a cause of submental opacity

Hemangiomas are the most common lesions of the major salivary glands during infancy and early childhood. Changes in blood flow dynamic within hemangioma results in thrombus and phleboliths. There have been a number of reports of hemangiomas with phleboliths in parotid and submandibular glands. We present the first case of a hemangioma with multiple phleboliths in the subligual gland as a cause of submental opacity, and discussed the diagnosis of radiopaque masses in the sublingual and submental regions.     

Heat-shock protein-peptide complex-96 for the treatment of cancer

Heat-shock proteins (HSPs) are the most abundant and ubiquitous soluble intracellular proteins. Members of the HSP family bind peptides, including antigenic peptides generated within cells. HSPs also interact with antigen-presenting cells (APCs) through CD91 and other receptors, eliciting a cascade of events that includes representation of HSP-chaperoned peptides MHC, translocation of NF-kappaB into the nuclei, and maturation of dendritic cells. These consequences point to a key role of HSPs in fundamental immunologic phenomena such as activation of APCs, indirect presentation (or crosspriming) of antigenic peptides, and chaperoning of peptides during antigen presentation. The properties of HSPs also allow them to be used for immunotherapy of cancers and infections in novel ways. This paper reviews the development and clinical trial progress of vitespen, an HSP peptide complex vaccine based on tumor-derived glycoprotein 96.