A simple and inexpensive particle agglutination test to distinguish recent from established HIV-1 infection.

OBJECTIVES: We sought to modify the Serodia HIV-1/HIV-2 particle agglutination assay (PA), a simple and cost-effective HIV assay that is used globally for the detection of HIV antibodies, as a sensitive/less sensitive test (S/LS) to identify recently infected individuals and to estimate HIV incidence. METHODS: The Serodia PA test was modified as an S/LS test (PA-LS) by using HIV antigen-coated gelatin particles at a dilution of 1:68 and a specific diluent, and calibrated using 37 HIV clade B seroconversion panels (309 samples) from Trinidad and from a commercial source that were tested at dilution intervals from 1:10 to 1:80,000. The greatest sensitivity for correctly classifying samples from recent and established infections was determined by receiver operator curve (ROC) analysis. RESULTS: At a 1:40,000 sample dilution and a days post-seroconversion cutoff of 190 days, the PA-LS test yielded a 97% sensitivity for classifying recent and established infection samples. Furthermore, at a 1:20,000 dilution, the positive predictive value for correctly identifying recently infected individuals was 99%. The PA-LS test offers a 30-44-fold cost saving over currently available S/LS tests. CONCLUSION: A modified, low cost and simple-to-perform PA test is appropriate for use in resource-limited countries, and has exhibited excellence in distinguishing recent from established HIV infection.

Efficient gene transfer to airway epithelium using recombinant Sendai virus.

Clinical studies of gene therapy for cystic fibrosis (CF) suggest that the key problem is the efficiency of gene transfer to the airway epithelium. The availability of relevant vector receptors, the transient contact time between vector and epithelium, and the barrier function of airway mucus contribute significantly to this problem. We have recently developed recombinant Sendai virus (SeV) as a new gene transfer agent. Here we show that SeV produces efficient transfection throughout the respiratory tract of both mice and ferrets in vivo, as well as in freshly obtained human nasal epithelial cells in vitro. Gene transfer efficiency was several log orders greater than with cationic liposomes or adenovirus. Even very brief contact time was sufficient to produce this effect, and levels of expression were not significantly reduced by airway mucus. Our investigations suggest that SeV may provide a useful new vector for airway gene transfer.

Tenofovir and abacavir combination therapy: lessons learned from an urban clinic population.

Regimens containing abacavir (ABC), tenofovir (TDF), and lamivudine (3TC) have recently been demonstrated to have high failure rates. This poses a clinical dilemma of how to manage patients currently being treated with other regimens containing tenofovir/abacavir. We evaluated the outcomes of tenofovir/abacavir regimens in our clinical practice through a retrospective review of 2655 charts. Two hundred patients (7%) were on a tenofovir/abacavir-containing regimen. Fifty-nine patients met the criteria for analysis and were grouped into three groups: (1) antiretroviral naïve, (2) virally suppressed patients switched to TDF/ABC, and (3) patients with failure of their first antiretroviral regimen. Rates of viral suppression in the naïve, switch, and first-failure groups were 95%, 86%, and 46%, respectively. In the first-failure group, viral suppression was 66% without and 18% with a preexisting M184V. A composite analysis of the groups revealed a success rate of 86% when the regimen contained zidovudine (ZDV) and 62% when it did not. No K65R mutations were noted. These findings support continued caution in the use of TDF/ABC in combination. However, these data suggest that this combination may be successfully used in selected situations such as in combination with ZDV. In patients already virally suppressed on a TDF/ABC-containing regimen, considerations include continuing the regimen or adding zidovudine, in the attempt to protect against the development of a K65R mutation and/or virologic failure, versus changing a stable regimen.

All three potential N-glycosylation sites of the dog kidney (Na+ + K+)-ATPase beta-subunit contain oligosaccharide.

The beta-subunit of dog kidney (Na+ + K+)-ATPase is a sialoglycoprotein and contains three potential N-glycosylation sites. In this study, the oligosaccharide chains of purified dog kidney beta-subunit were labeled with tritium by oxidation with sodium periodate or galactose oxidase followed by NaB3H4 reduction. The beta-subunit was extensively digested by trypsin and the radioactive peptides were purified by HPLC. The enzyme, glycopeptidase A, which catalyzes the removal of N-linked oligosaccharide chains and the conversion of the glycosylated Asn residue to Asp, was used to demonstrate that a number of purified beta-subunit tryptic peptides were glycosylated. Amino-acid analysis of these beta-subunit peptides following glycopeptidase-A treatment revealed the expected Asn to Asp conversion for Asn-157, Asn-192 and Asn-264, demonstrating that all three potential N-glycosylation sites of the dog kidney beta-subunit are glycosylated. In addition, amino-acid sequence data suggest that a disulfide bond exists between Cys-158 and Cys-174.

Inhibition of ion pump ATPase activity by 3'-O-(4-benzoyl)benzoyl-ATP (BzATP): assessment of BzATP as an active site-directed probe.

The interaction of 3'-O-(4-benzoyl)benzoyl-ATP (BzATP) with the renal (Na+ + K+)-ATPase, the sarcoplasmic reticulum Ca-transport ATPase, and the gastric (H+ + K+)-ATPase has been investigated in order to determine whether BzATP is a suitable probe for the labeling and identification of a peptide from the ATP binding sites of these ion pumps. After ultraviolet irradiation BzATP inhibited the enzymatic hydrolysis of ATP by each of the ion pumps, and also was covalently incorporated into the 100 000 dalton polypeptides of each protein. The presence of excess ATP in the reaction solution did not prevent either the inactivation of ATPase activity or the labeling of the catalytic polypeptides by BzATP. Prior modification of the ATPases with fluorescein-5'-isothiocyanate (FITC), however, prevented much of the labeling of the 100 000 dalton polypeptides by BzATP. BzATP competitively inhibited the high-affinity binding of ATP to the ion pumps, but ATP did not block the high-affinity binding of BzATP by the enzymes. BzATP binds to the membrane-bound ATPases at a high-affinity site with a Kd of 0.8-1.2 microM and a Bmax of 2-3 nmol/mg, and also binds to at least one low-affinity, high-capacity site on the membranes. HPLC separation of the soluble peptides from a tryptic digest of BzATP-labeled (Na+ + K+)-ATPase revealed the presence of several labeled peptides, none of which was protected by either ATP or FITC. Although BzATP can displace ATP from a high-affinity binding site on the ion pumps, it appears, therefore, that inactivation of enzymatic activity is the result of reactions between BzATP and the proteins at locations outside this site. Thus, it is concluded from these experiments that BzATP is not likely to be a useful probe for the ATP binding sites on the ion transport ATPases.

Subunit requirements for expression of functional sodium pumps in yeast cells.

Na+/K(+)-ATPase from animal cell membranes is known to consist of an alpha-subunit and a beta-subunit. Amino acids within the alpha-subunit have been shown to participate in the catalytic functions of the enzyme and in the binding of cardioactive steroids. Although the function of the beta-subunit is not known, expression of both alpha- and beta-subunits is required for the functional enzyme. A putative third subunit, the gamma-subunit, has been suggested to be a part of the functional Na+/K(+)-ATPase complex, based on experiments showing that both the catalytic alpha-subunit and a small peptide of M(r) = 11,000 can be labeled by a photoreactive ouabain analog. Although the primary structure for the putative gamma-subunit from rat and sheep was recently deduced from cDNA clones, participation of this small protein in the catalytic activity of the Na+/K(+)-ATPase has not been demonstrated. In experiments described here, the heterologous expression of Na+/K(+)-ATPase in yeast cells was used to investigate whether the gamma-subunit is an essential component of the Na+/K(+)-ATPase. Yeast cells do not contain an endogenous Na+/K(+)-ATPase. The alpha- and beta-subunits or the alpha-, beta- and the putative gamma-subunits of Na+/K(+)-ATPase were expressed in the yeast Saccharomyces cerevisiae and ouabain-sensitive ATPase, p-nitrophenylphosphatase, and 86Rb uptake activities were measured either in membranes prepared from transformed yeast cells, or in intact yeast cells. Nontransformed yeast cells or yeast cells transformed with the gamma-subunit alone served as controls. Northern analysis and Western blots demonstrated that yeast cells do not contain an endogenous peptide with significant sequence homology to the putative gamma-subunit. Yeast samples containing only Na+/K(+)-ATPase alpha and beta subunits were capable of ouabain-inhibitable enzymatic activity and 86Rb transport. No gamma-subunit-dependent differences in the measured enzymatic activities or transport properties were detected in the different samples. These observations establish that the alpha beta-subunit complex is the minimum structural unit required for all the ouabain-sensitive reactions of Na+/K(+)-ATPase.

Orientation of the beta subunit polypeptide of (Na+ + K+)ATPase in the cell membrane.

Although the animal cell (Na+ + K+)-ATPase is composed of two polypeptide subunits, alpha and beta, very little is known about the beta subunit. In order to obtain information about the structure of this polypeptide, the beta subunit has been investigated using proteolytic fragmentation, chemical modification of carbohydrate residues, and immunoblot analysis. The sialic acid moieties on the oligosaccharide groups on the beta subunit of (Na+ + K+)-ATPase were labeled with NaB3H4 after oxidation by sodium periodate, or the penultimate galactose residues on the oligosaccharides were similarly labeled after removal of sialic acid with neuraminidase and oxidation by galactose oxidase. All of the carbohydrate residues of the protein are located on regions of the beta subunit that are found on the non-cytoplasmic surface of the membrane. Cleavage of the galactose oxidase-treated, NaB3H4-labeled beta subunit by chymotrypsin at an extracellular site produced labeled fragments of 40 and 18 kDa, indicating multiple glycosylation sites along the polypeptide. Neither the 40 kDa fragment nor the 18 kDa fragment was released from the membrane by chymotrypsin digestion alone, but after cleavage the 40 kDa fragment could be removed from the membrane by treatment with 0.1 M NaOH. This indicates that the 40 kDa fragment does not span the lipid bilayer. The 40 kDa fragment and the 18 kDa fragment are also linked by at least one disulfide bond. The 18 kDa fragment also contains all of the binding sites found on the (Na+ + K+)-ATPase for anti-beta subunit antibodies. Both the 40 kDa fragment and the 18 kDa fragment were also generated using papain or trypsin to cleave the beta subunit. These data indicate that the beta subunit of (Na+ + K+)-ATPase contains multiple sites of glycosylation, that it inserts into the cell membrane near only one end of the polypeptide, and that one region of the polypeptide is particularly sensitive to proteolytic cleavage relative to the rest of the polypeptide.

Molecular cloning and sequence analysis of the (Na+ + K+)-ATPase beta subunit from dog kidney.

cDNA complementary to mRNA coding for the beta subunit of dog renal (Na+ + K+)-ATPase has been cloned into lambda gt11 and the nucleotide sequence of the DNA has been determined. The amino acid sequence of the beta subunit polypeptide has also been deduced from the DNA. The mature form of the dog kidney beta subunit contains 302 amino acids with three potential asparagine-linked attachment sites for carbohydrate. The initiation methionine is removed during processing of the polypeptide to its mature form. Although the beta subunit is an integral membrane protein there is no signal sequence for the polypeptide, and hydropathy analysis predicts that the beta subunit polypeptide spans the cell membrane only once. Secondary structure predictions and a model for the structure of the beta subunit are proposed. DNA sequencing of the 5' non-coding region of the mRNA revealed a 200 bp inverted repeat from the coding region. Blot hybridization of a fragment of the beta subunit cDNA identified a single mRNA species of 2.7 kb in dog kidney and several rat tissues. RNA from rat liver was deficient in mRNA that hybridized to the dog kidney beta subunit cDNA, although mRNA that hybridized to an alpha subunit cDNA was detected. RNA from a human hepatoma cell line, HepG2, however, contained comparable levels of mRNA for both the alpha and the beta subunits.

An intermediate state of the gamma-aminobutyric acid transporter GAT1 revealed by simultaneous voltage clamp and fluorescence.

The rat gamma-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering GAT1 function. Voltage-jump relaxation analysis showed that fluorescence increased slightly and monotonically with hyperpolarization; the fluorescence at -140 mV was approximately 0. 8% greater than at +60 mV. The time course of the fluorescence relaxations was mostly described by a single exponential with voltage-dependent but history-independent time constants ranging from approximately 20 ms at +60 mV to approximately 150 ms at -140 mV. The fluorescence did not saturate at the most negative potentials tested, and the midpoint of the fluorescence-voltage relation was at least 50 mV more negative than the midpoint of the charge-voltage relation previously identified with Na(+) binding to GAT1. The presence of gamma-aminobutyric acid did not noticeably affect the fluorescence waveforms. The fluorescence signal depended on Na(+) concentration with a Hill coefficient approaching 2. Increasing Cl(-) concentration modestly increased and accelerated the fluorescence relaxations for hyperpolarizing jumps. The fluorescence change was blocked by the GAT1 inhibitor, NO-711. For the W68L mutant of GAT1, the fluorescence relaxations occurred only during jumps to high positive potentials, in agreement with previous suggestions that this mutant is trapped in one conformational state except at these potentials. These observations suggest that the fluorescence signals monitor a novel state of GAT1, intermediate between the E*(out) and E(out) states of Hilgemann, D.W., and C.-C. Lu (1999. J. Gen. Physiol. 114:459-476). Therefore, the study provides verification that conformational changes occur during GAT1 function.

Effects of a calcium supplement on bone mineral density in male cyclists.

AIM: The purpose of this study was to maintain or improve bone density in male road cyclists through provision of calcium and vitamin D3 supplementation ingested before cycling. METHODS: Participants were male cyclists (N=17), with a mean (±SD) age of 42.7 (9.4) years. Measurements of lumbar spine and hip areal bone mineral density (aBMD) were performed at the start and end of a cycling season. Cyclists were randomized into the calcium supplement (CAL) or the control group (CON) group based on lumbar spine T-scores. The CAL group was instructed to consume 1600 mg calcium and 1000 IU vitamin D3 prior to cycling for the 5-month period. RESULTS: Femoral trochanter aBMD significantly decreased during the 5 month cycling season. There was no difference in aBMD between CAL and CON groups. CONCLUSION: Negative effects of competitive cycling on aBMD in hip structures can be observed within 5 months. Calcium and vitamin D3 ingested prior to cycling does not ameliorate this effect. This proof of concept paper provides evidence that more work is needed to find mechanisms to protect cyclists from destructive bone loss in hip structures.

Mumps disease and its health impact: an outbreak-based report.

An outbreak of mumps in a middle school (grades 6 to 8) in Ohio during 1981 was investigated to describe the clinical findings, health impact, and costs. Individuals with clinical mumps in the middle school and in family members were questioned concerning symptoms, complications, hospitalizations, school days absent, and parental work days missed. There were 62 cases of clinical mumps in the middle school and 13 cases among family members. Parotitis lasted an average of 7.4 days and fever (if present) lasted an average of 2.5 days with a mean temperature of 38.6 degrees C (101.4 degrees F). The duration of parotitis and fever increased with age. Complications included encephalitis, aseptic meningitis, orchitis, oophoritis, mastitis, and pancreatitis. Visits to physicians were made by 62.7% (47/75) of the individuals with mumps for a total of 63 visits, and two patients were hospitalized for a total of six days. Persons who attended middle school missed an average of 4.9 days of school. The estimated direct and indirect costs associated with this outbreak were $2,460 and $1,353, respectively, or $51 per case. States lacking mumps immunization requirements experienced a three-times greater incidence of mumps in 1982 than States that required all school pupils to be immunized. We recommend that all States institute compulsory mumps school immunization laws for all school children.

Site-directed mutagenesis of amino acids in the cytoplasmic loop 6/7 of Na,K-ATPase.

The loop between transmembrane helices 6 and 7 (L6/7) of P-type ATPases has been suggested to be important for the functional linkage of ion binding and enzyme phosphorylation or to be a site of initial cation binding. To investigate the role of L6/7 in Na,K-ATPase, alanine substitutions were made for charged and conserved residues in L6/7 of the human alpha1 subunit and the proteins were expressed in yeast for analysis. All mutants except the triple mutant E825A/E828A/D830A bound ouabain. Although the equilibrium dissociation constant for ouabain binding by most mutants was similar to the wild-type value, the K(d) of R837A for ouabain binding was approximately 15-fold higher than the wild-type K(d). (18)O exchange measurements indicated that the apparent affinity of this mutant for Pi was reduced about 3-fold. The concentration dependence of KCl inhibition of ouabain binding or of NaCl inhibition of ouabain binding revealed 2-4-fold changes in the apparent affinity for cations in the E825A, E828A, and R837A mutants. The E825A and E828A mutants lost the ability to bind ouabain after extraction with 0.1% SDS or after brief heating, indicating that these mutations affected the stability of the enzyme. The ATPase activity of the other mutants was measured after extraction of crude yeast membranes with 0.1% SDS. For all mutants except R834A, R837A, and R848A, the activity was at least 50% of wild-type activity.

Ecology and behavior of the desert burrowing cockroach,Arenivaga sp. (Dictyoptera, Polyphagidae).

Measurements were made of environmental conditions in the microhabitat ofArenivaga sp., and results were correlated with their diurnal migratory behavior. The animals live in sand dunes with less than 1% moisture most of the year. During the day in spring, summer and fall they borrow in the sand at a depth of 20-60 cm, while at night, when surface temperatures have cooled, they borrow within 1-3 cm of the surface. In winter they are rarely found near the surface, remaining active at lower levels during both night and day. Adult males were collected on the surface or at a depth of 20-60 cm, but they were never observed burrowing near the surface at 1-3 cm.The larvae and adult females are photonegative and remain at lower levels during the day, even though daytime temperature and humidity are sometimes favorable near the surface. In the summer, they migrate to the surface about 2 h later after darkness than in spring and fall. Temperature measurements indicated this was probably due to delayed nighttime cooling of the surface in the summer months. By burrowing near the surface the animals can experience cooler nighttime temperatures and water-loss may be reduced during the summer. This behavior may also facilitate disperasal and mating.The cockroaches feed on decaying leaves and the roots of desert shrubs. Since the latter have 35-38% moisture, they are probably the main source of water for these animals.

Development of segments and appendages in embryos of the desert scorpion Paruroctonus mesaensis (Scorpiones: Vaejovidae).

The scanning electron microscope was used to study the changing features of scorpion embryos from the blastula through early stages in the development of appendages. The earliest scorpion fossils (Silurian period) have structures more advanced than the embryos herein, so the possibility is considered that these embryos still retain and display some features indicative of evolutionary patterns in adult pre-Silurian ancestors. The blastodisc stage is followed by a knob-like germinal center that gives rise to most of the embryo body. The germinal center elongates on the ventral surface of the spherical yolk mass. The broad cephalic lobe is first delineated from the following pedipalpal segment. The limbbuds for the pedipalps and anterior walking legs appear, as additional segments are added at a growth zone at the rear of the embryo body. Initially, in the cephalic lobe there are no limbbuds; then the cheliceral buds emerge from the posterior part of the lobe. The stomodeum appears first in the anterior half of the cephalic lobe, but an oral groove forms and the mouth is displaced posteriorly within the groove. This repositioning allows space anteriorly for invagination (semilunar grooves) of epithelium for the brain and medial eyes. The mouth is directed ventrally in all stages of this study. The widespread chelicerae are initially posterior to the mouth, but later move anterior and dorsal to it. Small limbbud bulges on mesosomal segments disappear later and never become protruding appendages. Metasomal segments are produced free from the yolk surface in a ventral flexure beneath the embryo body. The telson starts as two spherical lobes, but later elongates and tapers distally, not yet developing the sharp sting (aculeus) seen in Silurian and all subsequent scorpions. The walking legs are digitigrade, as in most fossil aquatic scorpions. Segments are delineated in the appendages; the chelicerae and pedipalps are divided distally for chela (claw) formation. Bilateral swellings (limbbuds) on the third abdominal segment become larger than the others, indicating the site of pectine formation. The early fin-like pectines are somewhat posterior in the mesosoma, suggesting ancestral swimming, maneuvering, and balancing for the elongate abdomen. The pectinal surface is initially smooth but later transverse striations increase the surface area as a possible respiratory adaptation. Pectinal teeth (present in Silurian and all subsequent scorpions) and forward movement and merging of anterior abdominal segments are not yet evident in embryos of this study.

Febrile spontaneous abortion and the IUD.

A case-control study was done to determine the risk and pathogenesis of febrile spontaneous abortion for intrauterine device (IUD) wearers compared to non-wearers. Four groups of women coming to a large city hospital between 1970 and 1975 were compared: 1) women presenting with febrile spontaneous abortion, 2) women presenting with afebrile spontaneous abortion, 3) women presenting with afebrile spontaneous abortion which subsequently became febrile, and 4) women delivering a live infant. Pregnant women with an IUD in situ had a 5-fold higher risk of both febrile and afebrile spontaneous abortion compared to pregnant women without an IUD in situ. Women who had a spontaneous abortion that shifted from afebrile to febrile were somewhat more likely to be IUD wearers than the other 2 spontaneous abortion groups. In our study population, the increased risk of febrile spontaneous abortion for IUD wearers appeared primarily due to the increased risk of the spontaneous abortion event itself, rather than a primary IUD-related infection causing febrile spontaneous abortion.

The ultrastructure of hemocytopoietic organs in the desert scorpion, Paruroctonus.

The light and electron microscopes were used to examine possible hemocytopoietic tissue in the desert scorpion, Paruroctonus mesaensis. Results agree with earlier light microscopic studies that cells are released into the blood from the two lateral lymphoid organs and the supraneural gland. The former are sacciform structures attached by their anterior ends to the diaphragm. The supraneural gland forms the thickened wall of the supraneural artery in the mesosoma from the first to the third abdominal ganglia. The lateral lymphoid glands have an acellular stroma in which are embedded granular and agranular cells. The stroma is apparently formed by specialized cells which release membranous cell fragments that become the matrix of the gland. Cells are released into the body cavity from the periphery of the two organs. The supraneural gland has a fibrous stroma in which are embedded a variety of cell types. The cells appear to be released in greatest abundance into the blood in the lumen of the gland. The gland has cells with opaque granules (0.9-1.4 micron diameter) and agranular cells of variable shape. The most abundant cell, possibly the stem-cell for the others, is about 10 micron diameter and often has processes of variable length. In addition, muscle cells at various stages of differentiation are found at the inner margin of the gland. These cells have thick and thin myofilaments (24-32 and 5-8 nm diameter) and dense bodies which sometimes become organized into sarcomeres with Z-bands before the cells are released into the gland lumen. The function of these muscle cells is unknown, but possibly they contribute to the maintenance of blood pressure and the release of cells into the blood from the inner margin of the gland.

Regulation of air and blood flow through the booklungs of the desert scorpion, Paruroctonus mesaensis.

Injections of dye, latex and India ink were used to reveal the path of hemolymph circulation through the scorpion booklungs. Fine, branched arteries carry blood directly to muscle and other organs. The blood returns through venous channels to the ventral mesosoma where it passes laterally through the booklungs and into the pneumocardial veins just beneath the pleural cuticle. Blood flows dorsally through these veins to the pericardial sinus and heart. The scorpion has four pairs of booklungs located in the anterior segments of the ventral mesosoma. Each booklung has a spiracle which opens into an atrium enclosed by cuticular membrane. Air passes from the atrium into the booklung lamellae. Agitation of the animal or application of CO(2) causes retraction of the anterior and posterior atrial membrane. This expands the atrial chamber and allows gas exchange in the booklung lamellae. The posterior atrial membrane has a specialized region which forms a springy valve. This normally closes the spiracle unless pulled open by contraction of the attached poststigmaticus muscle. The pectens and receptors within the atrium may mediate the responses to CO(2). Slender hypocardial ligaments containing muscle fibers extend from the heart (dorsal mesosoma) to the booklungs in the ventral mesosoma. Heart movements thus cause dorso-ventral movement of the booklungs. The significance of these movements is as yet unclear. They may increase ventilation, help force blood to the heart and/or agitate the blood and booklung lamellae and thereby aid gas exchange. Passage of blood through the booklungs is regulated by dorsal and ventral muscles attached to the atrium at the lateral edge of the booklung. Contraction of the ventral atrial muscle closes the excurrent channel for passage of blood from the booklung into the pneumocardial vein. Electrical stimulation of the segmentai nerves from the subesophageal and first three abdominal ganglia causes spiracle opening and contraction of muscles attached to the atrial membrane. A previous study showed that these same segmental nerves also modulate heart activity. They thus provide a major pathway for regulation of the respiratory and circulatory systems.

Comparison of the ultrastructure of stimulated and unstimulated mechanoreceptors in the taste hairs of the blowfly Phaenicia serricata.

The structure of mechanoreceptors at the base of labellar taste hairs of the blowfly Phaenicia serricata were examined in stimulated and unstimulated conditions (i.e. with the hair bent or unbent). Physiological recordings from the mechano-receptor showed that the receptors responded when the hair is bent dorsally or ventrally and when the hair is bent at extreme angles. These conditions are the same as those placed on hairs in the anatomical studies. Bending the hair toward the ventral labellar surface caused the hair base to compress and indent the tubular body and its surrounding membrane and sheath at the distal end of the mechanoreceptor dendrite. In compressed tubular bodies, microtubules oriented longitudinally were bent and separated a greater distance from each other. Separation as much as 70 nm was observed in compressed tubular bodies as compared with a maximum of 26 nm between micro-tubules in tubular bodies of unbent hairs. The dense amorphous material between microtubules of compressed tubular bodies formed prominent bridges 18 nm thick connecting the microtubules at intervals of 48-74 nm. Thin 10 nm filaments were also evident in the spaces between microtubules. When the hair was bent toward the proximal end of the proboscis, the tip of the tubular body was bent about 15 degrees. The tubular body appears to function as a firm but resilient structure over which the dendritic membrane can be stretched during mechanostimulation. Comparison of morphology of bent and unbent hairs suggests a means by which mechanical force from the movement of the hair is transferred to the receptors by structures in the hair socket region. No differences were found in ciliary structures of stimulated and unstimulated receptors.

Antennal chemoreceptors of the desert burrowing cockroach, Arenivaga sp.

The antennae of Arenivaga have six types of chemoreceptor sensilla. Some of these have unusual morphological features which may be adaptations for survival in a dry habitat. The sensory dendrites are well protected by cuticular structures, and in some receptors stimulatory molecules must pass through long channels or through pores filled with strands to reach the sensory cells. Large grooved pegs (possibly pheromone receptors) are numerous on antennae of adult males, and grooved sensilla are described here in detail for the first time. Thin-walled pegs, present in males and females, do not have pore tubules or hollow filaments as observed in many other insects. Rather, they contain structures designated here as pore strands, since they have a dense core rather than a light center as previously described for pore tubules and filaments. These strands do not appear to be evaginations of the dendritic membrane, but are probably formed in association with the cuticular structures of the sensilla.