Prolonged Attachment Research Articles

Tick extracellular vesicles undermine epidermal wound healing during hematophagy.

Wound healing has been extensively studied through the lens of inflammatory disorders and cancer, but limited attention has been given to hematophagy and arthropod-borne diseases. Hematophagous ectoparasites, including ticks, subvert the wound healing response to maintain prolonged attachment and facilitate blood-feeding. Here, we unveil a strategy by which extracellular vesicles (EVs) ensure blood-feeding and arthropod survival in three medically relevant tick species. We demonstrate through single cell RNA sequencing and murine genetics that wildtype animals infested with EV-deficient Ixodes scapularis display a unique population of keratinocytes with an overrepresentation of pathways connected to wound healing. Tick feeding affected keratinocyte proliferation in a density-dependent manner, which relied on EVs and dendritic epidermal T cells (DETCs). This occurrence was linked to phosphoinositide 3-kinase activity, keratinocyte growth factor (KGF) and transforming growth factor β (TGF-β) levels. Collectively, we uncovered a strategy employed by a blood-feeding arthropod that impairs the integrity of the epithelial barrier, contributing to ectoparasite fitness.

The Effectiveness of Using Communicative Language Teaching in ESL Classroom by EFL Teachers; An Exploratory Study

Success of any innovation in the field of education directly associated with learners’ perceptions about that. Communicative Language Teaching is an innovation and this article attempts to discover learners’ attitude and perceptions regarding this. This study will be very useful in Pakistan where implementation of CLT is in hot discussion and mostly researchers worked on the perceptions and attitude of teachers. This study is quantitative in nature. Data was collected from students through questionnaire that was adapted from the study of Sandra J.Savignon and Chaochang Wang (2003). The goal of the study is to explore: what reputation Grammar Translation Method and Communicative Language Teaching are enjoying, what new is introduced to fit CLT in Pakistani educational system, whether learners have positive attitude towards communication based activities or not, what are the problems that challenge the implementation of CLT in classrooms, what perceptions do students have towards communication based activities, which classroom activities are serious threat in meeting the goal of communicative ability. Findings show that teachers are aware about the importance of CLT and learners have positive attitude and perception towards communication based activities. Functional English and English Comprehension is taught that resulted learners can understand classic literature and English movies as well. There is a fair possibility of adopting CLT but their prolonged attachment with Grammar Translation Method, examination system, emphasis on accuracy, preference of mother tongue are the hindrances that blur the concept of implementation of CLT in Pakistan’ classrooms.

Design and Testing of a Superelastic Nitinol Tissue Attachment Mechanism for Long-Term Gastrointestinal Device Retention

AbstractIngestible devices have become a popular means for diagnosing and treating the gastrointestinal (GI) tract due to their noninvasive nature. However, their residency period in the GI tract is limited by the transit time through it. In previous work, we designed a tissue attachment mechanism (TAM) inspired by parasitic worms' attachment methods, which were tested for implanting biosensors or drug delivery payloads to the small intestine with a swallowable capsule robot. In that work, the attachment success rate was 91.7%, and the average attachment duration of the TAM was 32.2 h after factorial optimization of major design factors. This work develops a novel nitinol TAM (NTAM) for improving the attachment duration using the shape-changing properties of nitinol. The attachment strength of the NTAM to the intestinal tissue was assessed both ex vivo and in vivo. The attachment duration of the NTAMs in live porcine models was evaluated from radiographic images, and histological analysis of the attachment location of an NTAM was performed after euthanasia. The NTAM was 100% successful in an attachment strength study and achieved a maximum attachment duration of 13 days, while the average attachment duration was 85.63 ± 77.83 h. Histological analysis did not report any permanent damage to the tissue. This work shows a 2.7-fold improvement in attachment duration over the previous design. This work has demonstrated a method of prolonged attachment to the intestinal wall through a swallowable device, which can be used for long-term drug delivery or biosensing.

A sudden and widespread change in symptoms and incidence of vascular streak dieback of cocoa (Theobroma cacao) linked to environmental change in Sulawesi, Indonesia

Unstable new-encounter relationships between plants and pathogens form when organisms are transported between once-isolated biogeographic regions. Ceratobasidium theobromae is a new-encounter pathogen that has plagued the cocoa industry causing vascular streak dieback in Southeast Asia and western Melanesia. The disease underwent a sudden change in symptoms associated with an increase in disease incidence in Indonesia and Malaysia from about 2004. These changes have not been linked to changes in the pathogen or host and this shift could signal an effect of region-wide environmental changes in the responses of cocoa trees to infection. To study the effect of ambient temperature on symptom expression, this study assessed disease symptoms in cocoa trees along altitudinal gradients in Sulawesi, Indonesia. It recorded types of VSD symptoms (necrotic or chlorotic) in clones and local hybrids on managed farms, and in a reciprocal transplant experiment. The results found that consistently across landscapes, in trees of varying inherent susceptibilities to VSD, the incidence of new, necrotic symptoms decreased with altitude and was restricted to lowlands, while the chlorotic symptoms that originally characterised the disease were found throughout the altitudinal range. The new symptom pattern in cocoa appears to be an environmentally driven change in tree responses to the pathogen, possibly related to recent climate warming or elevated atmospheric concentration of CO2 due to widespread burning of forest and peat beds in Indonesia. The increased occurrence of a necrotic response of leaves to invasion by C. theobromae is typical of a hypersensitive response often associated with resistance to invasion by pathogens and could indicate enhanced resistance, although prolonged attachment of diseased leaves allows increased sporulation of the fungus and increased disease incidence.These are the first results to show an altered plant host–fungus pathogen response likely due to climate change, indicating that global warming may cause subtle but ecologically important changes in plant-parasite interactions. New-encounter relationships that are less stable than long-term co-evolved relationships might provide early-warning of ecological changes due to climate change. Determining whether the changed symptoms are a new threat is vital knowledge for farmers' livelihoods in this region.

Impact of hypnotic safety on disorders of gut-brain interaction: A pilot study

ABSTRACT Functional gastrointestinal disorders, i.e., abdominal conditions without identifiable structural etiologies, are seen frequently in primary care and specialty practices. As subtle physiological processes have been identified as potential contributing factors to these functional disorders, these disorders have been recently relabeled, Disorders of Gut-Brain Interaction (DGBI). Moreover, some of these processes, e.g., sympathetic nervous system activity and inflammation, are being increasingly related to psychosocial factors such as situational stress and histories of trauma, abuse, and neglect. As the activity of the autonomic nervous system (ANS) has been long considered to be a contributory factor for DGBI, the present study utilized a theory-driven model based on the Polyvagal Theory to optimize ANS activity for the promotion of healthy digestive activity. Specifically, a hypnotic intervention to increase neuroception of safety was employed with three female college students diagnosed with functional dyspepsia and irritable bowel syndrome in a single-subject design. This intervention was found to be associated with increases in the experience of safe/warm positive affect and decreases in symptoms of functional dyspepsia and irritable bowel syndrome as well as depression and anxiety. The hypnotic intervention for the promotion of a sense of safety is recommended for the treatment of other functional somatic disorders as well as trauma-related conditions. Potential complications related to individuals with prolonged trauma and attachment issues also are reviewed.

Synthesis and Characterization of Thiolated PVP–Iodine Complexes: Key to Highly Mucoadhesive Antimicrobial Gels

The aim of this study was to synthesize iodine containing polymeric excipients for mucosal treatment of microbial infection exhibiting a prolonged mucosal residence time by forming an adhesive gel on the mucosal surface. In order to achieve this aim, 2-(2 acryloylamino-ethyldisulfanyl)-nicotinic acid (ACENA) was copolymerized with N-vinylpyrrolidone (NVP) to obtain thiolated polyvinylpyrrolidone (PVP) for complexation with iodine. The average molecular mass of different thiolated PVP variants was determined by size exclusion chromatography. The structure of thiolated PVP was confirmed by 1H NMR. Thiolated PVP variants were characterized for thiol content, cytotoxicity, iodine loading capacity, rheological behavior, and adhesion time on mucosa. The highest achieved degree of thiolation was 610 ± 43 μmol/g, and the maximum recorded iodine loading was 949 ± 31 μmol/g of polymer. Thiolated PVP variants (0.5% m/v) showed no toxicity after incubation on Caco-2 cells for the period of 3 and 24 h, respectively. Thiolated PVP and thiolated PVP-iodine complexes exhibited a 5.4- and 4.4-fold increased dynamic viscosity in porcine mucus in comparison to PVP and PVP-iodine complex, respectively. Compared to PVP and PVP-iodine complex thiol-functionalized PVP and PVP-iodine complexes demonstrated significantly prolonged attachment to mucosal surface over a period of 3 h. Thiol functionalized PVP proved to be a promising novel excipient for complexation with iodine and to exhibit strongly improved mucoadhesive properties.

The numerical simulation of the wing kinematics effects on near wake topology and aerodynamic performance in hovering Drosophila flight

The parallel large-scale unstructured finite volume method based on an Arbitrary Lagrangian–Eulerian (ALE) formulation in Erzincanli and Sahin (2013) has been employed in order to investigate the effects of the wing kinematics on the near wake topology produced by a pair of flapping Drosophila wings in hover flight. The three-dimensional wing shape and the baseline wing kinematics are the same as those of the robotic fly by Dickinson et al. (1999). The simulations are used to quantify the important wing kinematic parameters for the wake topology and as well as their correlations with the force production. The angle-of-attack is shown to be very effective for producing lift during the wing translational motion. However, for larger values the angle-of-attack limits the angle during the stroke reversal and reduces the rotational lift. The angle-of-attack at which the maximum lift coefficient occurs is in excellent agreement with the earlier experiments in the literature. In addition, the numerical results confirm that the increase in the wing stroke amplitude leads to a prolonged attachment of the leading edge vortex (LEV) over a relatively large distance and increases force production. The variations in the heave angle include the figure-of-eight pattern, figure-of-O pattern and figure-of-U pattern. The constant heave angle and figure-of-eight pattern are found to have a more profound influence on the magnitude of force production. In addition, some asymmetric variations in the wing kinematics are introduced in order to assess the important parameters determining forward and backward flights. The paddling wing motion is shown to be very effective to initiate forward and backward acceleration.

Vulnerability and resilience after early institutional care: The Greek Metera study.

The aim of the study was to examine possible influences on individual differences in adolescence in response to early institutional care in infancy not involving either generalized privation or subnutrition. Fifty-two adopted adolescents aged 13 years who received institutional care in infancy at the Metera Babies Centre and 36 adolescents of the same age who were raised in their biological families participated in the study. Adolescents' attachment relationships, cognition, behavioral adjustment and use of psychological services were assessed. Marked heterogeneity in outcomes was found. No predictive effects were found for preinstitutional features or for adoptive circumstances. By contrast a large effect was found for institutional care extending beyond the age of 2 years and for quality of institutional care as experienced at an individual level. There was a close association between prolonged institutional care and disorganized attachment while in the institution, but the main institutional effect derived from the length of time in the institution.

Immunization with a recombinant subunit OspA vaccine markedly impacts the rate of newly acquired Borrelia burgdorferi infections in client-owned dogs living in a coastal community in Maine, USA.

BackgroundIn North America, Borrelia burgdorferi is the causative bacterial agent of canine Lyme borreliosis and is transmitted following prolonged attachment and feeding of vector ticks, Ixodes scapularis or Ixodes pacificus. Its prevention is predicated upon tick-avoidance, effective on-animal tick control and effective immunization strategies. The purpose of this study is to characterize dogs that are newly seropositive for Borrelia burgdorferi infection in relation to compliant use of a recombinant OspA canine Lyme borreliosis vaccine. Specifically, Preventive Fractions (PF) and Risk Ratios (RR) associated with the degree of vaccine compliancy (complete versus incomplete) are determined.Methods6,202 dogs were tested over a five year period in a single veterinary hospital utilizing a non-adjuvanted, recombinant OspA vaccine according to a 0, 1, 6 month (then yearly) protocol. Rates of newly acquired “Lyme-positive” antibody test results were compared between protocol compliant and poorly compliant (incompletely and/or non-vaccinated) dogs.ResultsOver the five-year span, one percent (range 0.39 - 1.3) of protocol compliant vaccinated, previously antibody negative dogs became seropositive for infection. Approximately twenty-one percent (range 16.8 – 33.3) of incompletely vaccinated dogs became positive for infection-specific antibodies. The Preventative Fraction for testing positive for antibodies specific for infection with Borrelia burgdorferi in any given year based on optimal vaccine compliance was, on average, 95.3% (range 93.29 - 98.08). The Risk Ratio for becoming infected with Borrelia burgdorferi antibodies in any given year if vaccine non-compliant was 21.41 (range 14.9 – 52.1). There was a high statistically significant relationship (p = <0.0001) in the observed data in terms of vaccination protocol compliance and the probability of Borrelia burgdorferi infection in each of the five years under study.ConclusionsThe recombinant outer surface protein A (rOspA) vaccine for dogs is highly effective in preventing new seropositive cases of Borrelia burgdoferi infection over a five-year period in dogs living in an endemic area. Dogs that were vaccine protocol-compliant were significantly less likely to become infected (as indirectly assessed by antibody) with the agent of canine Lyme borreliosis as measured by Preventive Fraction and Risk Ratio calculations.

PH sensitive N-succinyl chitosan grafted polyacrylamide hydrogel for oral insulin delivery

pH sensitive PAA/S-chitosan hydrogel was prepared using ammonium persulfate (APS) as an initiator and methylenebisacrylamide (MBA) as a crosslinker for oral insulin delivery. The synthesized copolymer was characterized by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) study; morphology was observed under scanning electron microscope (SEM). The PAA/S-chitosan with ∼38% of insulin loading efficiency (LE) and ∼76% of insulin encapsulation efficiency (EE), showed excellent pH sensitivity, retaining ∼26% of encapsulated insulin in acidic stomach pH 1.2 and releasing of ∼98% of insulin in the intestine (pH 7.4), providing a prolonged attachment with the intestinal tissue. The oral administration of insulin loaded PAA/S-chitosan hydrogel was successful in lowering the blood glucose level of diabetic mice. The bioavailability of insulin was ∼4.43%. Furthermore, no lethality or toxicity was documented after its peroral administration. Thus, PAA/S-chitosan hydrogel could serve as a promising oral insulin carrier in future.

The kinematics of cytotoxic lymphocytes influence their ability to kill target cells.

Cytotoxic lymphocytes (CTL) have been reported to show a range of motility patterns from rapid long-range tracking to complete arrest, but how and whether these kinematics affect their ability to kill target cells is not known. Many in vitro killing assays utilize cell lines and tumour-derived cells as targets, which may be of limited relevance to the kinetics of CTL-mediated killing of somatic cells. Here, live-cell microscopy is used to examine the interactions of CTL and primary murine skin cells presenting antigens. We developed a qualitative and quantitative killing assay using extended-duration fluorescence time-lapse microscopy coupled with large-volume objective software-based data analysis to obtain population data of cell-to-cell interactions, motility and apoptosis. In vivo and ex vivo activated antigen-specific cytotoxic lymphocytes were added to primary keratinocyte targets in culture with fluorometric detection of caspase-3 activation in targets as an objective determinant of apoptosis. We found that activated CTL achieved contact-dependent apoptosis of non-tumour targets after a period of prolonged attachment – on average 21 hours – which was determined by target cell type, amount of antigen, and activation status of CTL. Activation of CTL even without engagement of the T cell receptor was sufficient to mobilise cells significantly above baseline, while the addition of cognate antigen further enhanced their motility. Highly activated CTL showed markedly increased vector displacement, and velocity, and lead to increased antigen-specific target cell death. These data show that the inherent kinematics of CTL correlate directly with their ability to kill non-tumour cells presenting cognate antigen.

Network based Load-Aware Mobility Management in IEEE 802.11 Wireless Mesh Networks

One major concern for mobile networks consists of finding efficient way s of handling user mobility so that the handover process has minimum effect on users' ongoing sessions. Although COAP provides an efficient way for mobility management in IEEE 802.11 wireless mesh networks, it does not control load among multiple MAPs in the network. Thus, in many cases, the attached MAP is overloaded, and extensive delays are experienced during the routing process. To tackle this problem, this paper proposes a load-aware mobility management scheme. The overloaded MAP is detected according to the load estimation. Then the overloaded MAP initiates the search procedure to find the underutilized MAP and transfe r the MN's attachment request to it. The proposed scheme can efficiently balance the handover load among MAPs at the cost of sligh tly prolonged attachment delay and increased attachment messages compared with COAP.

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Nonmuscle myosin II exerts tension but does not translocate actin in vertebrate cytokinesis

During vertebrate cytokinesis it is thought that contractile ring constriction is driven by nonmuscle myosin II (NM II) translocation of antiparallel actin filaments. Here we report in situ, in vitro, and in vivo observations that challenge this hypothesis. Graded knockdown of NM II in cultured COS-7 cells reveals that the amount of NM II limits ring constriction. Restoration of the constriction rate with motor-impaired NM II mutants shows that the ability of NM II to translocate actin is not required for cytokinesis. Blebbistatin inhibition of cytokinesis indicates the importance of myosin strongly binding to actin and exerting tension during cytokinesis. This role is substantiated by transient kinetic experiments showing that the load-dependent mechanochemical properties of mutant NM II support efficient tension maintenance despite the inability to translocate actin. Under loaded conditions, mutant NM II exhibits a prolonged actin attachment in which a single mechanoenzymatic cycle spans most of the time of cytokinesis. This prolonged attachment promotes simultaneous binding of NM II heads to actin, thereby increasing tension and resisting expansion of the ring. The detachment of mutant NM II heads from actin is enhanced by assisting loads, which prevent mutant NM II from hampering furrow ingression during cytokinesis. In the 3D context of mouse hearts, mutant NM II-B R709C that cannot translocate actin filaments can rescue multinucleation in NM II-B ablated cardiomyocytes. We propose that the major roles of NM II in vertebrate cell cytokinesis are to bind and cross-link actin filaments and to exert tension on actin during contractile ring constriction.

The Effect of Adriamycin Exposure on the Notochord of Mouse Embryos

The notochord has important structural and signaling properties during vertebrate development with key roles in patterning surrounding tissues, including the foregut. The adriamycin mouse model is an established model of foregut anomalies where exposure of embryos in utero to the drug adriamycin leads to malformations including oesophageal atresia and tracheoesophageal fistula. In addition to foregut abnormalities, treatment also causes branching, displacement, and hypertrophy of the notochord. Here, we explore the hypothesis that the notochord may be a primary target of disruption leading to abnormal patterning of the foregut by examining notochord position and structure in early embryos following adriamycin exposure. Treated (n = 46) and control (n = 30) embryos were examined during the crucial period when the notochord normally delaminates away from the foregut endoderm (6-28 somite pairs). Transverse sections were derived from the anterior foregut and analyzed by confocal microscopy following immunodetection of extracellular matrix markers E-cadherin and Laminin. In adriamycin-treated embryos across all stages, the notochord was abnormally displaced ventrally with prolonged attachment to the foregut endoderm. While E-cadherin was normally detected in the foregut endoderm with no expression in the notochord of control embryos, treated embryos up to 24 somites showed ectopic notochordal expression indicating a change in characteristics of the tissue; specifically an increase in intracellular adhesiveness, which may be instrumental in structural changes, affecting mechanical and signaling properties. This is consistent with disruption of the notochord leading to altered signaling to the foregut causing abnormal patterning and congenital foregut malformations.

Effects of Smart Position Only (SPOT) Tag Deployment on White Sharks Carcharodon carcharias in South Africa

We present 15 individual cases of sub-adult white sharks that were SPOT tagged in South Africa from 2003–2004 and have been re-sighted as recently as 2011. Our observations suggest SPOT tags can cause permanent cosmetic and structural damage to white shark dorsal fins depending on the duration of tag attachment. SPOT tags that detached within 12–24 months did not cause long term damage to the dorsal fin other than pigmentation scarring. Within 12 months of deployment, tag fouling can occur. After 24 months of deployment permanent damage to the dorsal fin occurred. A shark survived this prolonged attachment and there seems little compromise on the animal's long term survival and resultant body growth. This is the first investigation detailing the long term effects of SPOT deployment on the dorsal fin of white sharks.

Adaptive timing of detachment in a tick parasitizing hole-nesting birds

In non-permanent parasites with low intrinsic mobility such as ticks, dispersal is highly dependent on host movements as well as the timing of separation from the hosts. Optimal detachment behaviour is all the more crucial in nidicolous ticks as the risk of detaching in non-suitable habitat is high. In this study, we experimentally investigated the detachment behaviour of Ixodes arboricola, a nidicolous tick that primarily infests birds roosting in tree-holes. We infested great tits with I. arboricola larvae or nymphs, and submitted the birds to 2 experimental treatments, a control treatment in which birds had normal access to nest boxes and an experimental treatment, in which the birds were prevented access to their nest boxes for varying lengths of time. In the control group, most ticks detached within 5 days, whereas in the experimental group, ticks remained on the bird for as long as the bird was prevented access (up to 14 days). This prolonged attachment caused a decrease in survival and engorgement weight in nymphs, but not in larvae. The capacity of I. arboricola larvae to extend the duration of attachment in non-suitable environments with no apparent costs, may be an adaptation to unpredictable use of cavities by roosting hosts during winter, and at the same time may facilitate dispersal of the larval instars.

ON THE PROLONG ATTACHMENT OF LEADING EDGE VORTEX ON A FLAPPING WING

In this paper, we take a fundamental approach to investigate the effect of spanwise flow on the prolonged attachment of leading edge vortex (LEV) on a flapping wing. By imposing a constant acceleration-constant velocity flow on elliptic wings of various sweep angles and angles of attack, our experimental and numerical results show that while spanwise flow per se has negligible influence on the prolong attachment of the LEV, vortex stretching can significantly delay detachment of the LEV, even for a small spanwise flow.

Rhipicephalus appendiculatus: Characterization of a testis-associated protein

A novel gene coding for Rhipicephalus appendiculatus Male-specific Protein (RAMP) was identified in a cDNA library constructed from the testis/vas deferens of R. appendiculatus ticks. This gene encodes a secreted protein exclusively expressed in the testis/vas deferens. The putative RAMP amino acid sequence contains a signal peptide and has 29% amino acid identity with male-specific Is5 gene of Ixodes scapularis. Gene expression studies revealed that RAMP mRNA was up-regulated in male ticks during blood feeding. RAMP was detected not only in the testis/vas deferens of males but also in postcoitum female ticks based on Western blotting, indicating that this protein is transferred to the female tick during copulation. Virgin female ticks, microinjected with recombinant RAMP, had significantly prolonged attachment duration during feeding, but there was no effect on fed weight. These results suggest that RAMP is a male-specific molecule in the spermatophore, and is related to female attachment behavior in R. appendiculatus ticks.