Natural, accidental, and intentional translocation of bats, both intra- and intercontinentally, has been documented. Some bats have been translocated while incubating infectious diseases, including rabies or related lyssavirus infections; others have escaped confinement en route to or at their destinations, while others have been released deliberately. Known events and potential consequences of bat translocation are reviewed, including a proposed solution to the attendant problems. Herpestes javanicus) in rabies transmission on Caribbean islands  and related lyssaviruses , Red Bats , Seminole Bats (L. seminolus), and Silver-haired Bats (Lasionycteris noctivagans), all species from which RABV has been isolated  landed on a ship over 322 km from the coast of Argentina (Vampyressa pusilla) evidently boarded a vessel passing through the Panama Canal and was later found aboard when the ship was between Australia and Tasmania  roosted aboard a ship that frequently traveled from Canada to Europe, flying ashore after arrival in the Netherlands and England , which is similar to the Big Brown Bat (E. fuscus) but with a slightly more massive skull. The Serotine has been reported in North Africa and England and across Europe and Asia to Korea. Hundreds of ill or dead Serotines have been found infected with the RABV-related European bat lyssavirus 1 (EBLV-1) in Europe, where one or two persons have died of the infection after bat bites  was discovered in Victoria, British Columbia, in a shipment of lettuce from California ((Pipistrellus javanicus) was discovered in a container transported by ship from Japan to New Zealand  was recovered from a plane that had just arrived in Texas from Mexico  was discovered flying aboard a U.S. Air Force cargo plane en route from California to Hawaii. This bat was also negative for RABV, although rabies has been diagnosed in the species in California. Evidently the bat was loaded into the aircraft within a shipment of fruit.An Asiatic Pipistrelle bat was captured May 25, 1993, aboard an airliner en route from Tokyo to San Francisco. This bat was negative for RABV. The next month a Yuma Myotis bat (Tadarida brasiliensis), but closer inspection indicated differences, although the bat belonged to a family with similar representatives in warm areas worldwide. After extensive study, I determined the specimen to be a Wrinkle-lipped bat (Chaerephon pumila), known throughout sub-Saharan Africa, Madagascar, and southern Arabia. Further research disclosed the transported bat’s African origin. This species supports experimental replication of Ebola virus without showing disease signs , an insectivorous species in which RABV has been reported  family have been reported throughout their geographic range, which extends from northern Mexico south to Chile and Uruguay  live on land masses, including islands, from Madagascar, India, Southeast Asia, the East Indies, the Philippines, and Australia to the Samoan and Cook Islands of the South Pacific Ocean. They have been popular zoo attractions for many years. RABV was reported in a Pteropus in India  ascribed to Pteropus origin have proven pathogenic or fatal to people and domestic animals. Four species of Australian Pteropus bats in Queensland carry Hendra virus without developing symptoms. These bats disseminate virus in urine or placental fluid during birthing, and the virus is later ingested by pregnant horses that amplify the virus, which then spreads to people and causes a fatal pneumonia   is a widespread Old World fruit bat that readily reproduces in captivity; thus colonies occur in some zoos. This species has been implicated in several viral infections in Africa (West Nile virus (WNV) had been reported in the Rousettus species in Uganda and Israel  from California during the late 1800s to establish bat populations for insect control purposes, but the attempts were evidently unsuccessful.Bats have been translocated and released in attempts to establish bat populations in new areas for reasons such as insect control and experimental study. Such efforts are sometimes supplemented by providing living quarters or shelters for bats ranging from elevated boxlike structures to tunnels. Before the knowledge that some insectivorous bats might be infected with rabies or other pathogens, bats were sometimes transported great distances over land or overseas and released in efforts to establish populations at the new location. Tomich  in the air, each transporting a small time-activated fire bomb. The objective was to start thousands of simultaneous fires in adversary target areas, achieved after each bat had sought out a hideaway in various available structures , the chances of successful introduction of the species are minimized. Populations would more likely develop should large numbers be freed in places favorable to survival. Although a single escaped bat might not survive long or reproduce, it would seek shelter in places frequented by local bats to which it might transmit pathogens. As has been observed, introduced pathogens include RABV, other lyssaviruses, or various other agents.(Trypanosoma evansi) to horses and cattle, Vampire Bats can also be temporary biologic as well as mechanical vectors of Venezuelan equine encephalomyelitis virus and foot-and-mouth disease. They are likely effective mechanical vectors if not biologic vectors of any bloodborne pathogen, including the AIDS virus (Vampire Bats can be especially problematic in view of their possible colonization in warm climates and their dependence on a diet of blood, thus necessitating their biting vertebrates, including man and domestic animals. As reported, in addition to their known role as biologic vectors of rabies to humans and domestic animals and surra Accidental or planned translocations of bats between land masses happens almost certainly with far greater frequency than is reported. Such events can be embarrassing, and although incidents that result in successful containment are more likely to be reported, failed efforts can remain unpublicized. Relevant reporting requirements do not exist. Personnel involved in the various described incidents generally have performed very well in efforts to resolve the problems, often with immediately contrived solutions. Inspectors at entry centers are usually exceptionally competent because they must cover a broad array of subject areas, but their competency must be taxed at times. For example, most bats are exceptionally adept at avoiding capture, and even bat scientists with special equipment frequently are outmaneuvered. Some inspectors contact specialists for help in emergencies, but help is not always available or is displaced by previous commitments and economic necessities. Previous contractual arrangements with institutions such as universities, natural history museums, zoos, or specialized commercial services could dispel most relevant problems, including funding, and maintain program continuity. Unaffiliated specialized personnel would be expected to maintain or acquire relevant competency, but incidents, such as those cited here, show some lapses. Ideally, the services of a bat expert are required. For example, if bats are to be excluded from any vehicle of conveyance, the usual procedures and equipment should be reviewed by responsible persons very familiar with bats, their capabilities, their capture, their confinement, and their exclusion in order to recognize flaws that permit bats to be transported. Thus, experts can help establish and maintain more effective programs.


To the Editor: West Nile virus  has been perpetuating in North America since 1999 , 2 Campylorhamphus trochilirostris, and 1 each of Elaenia flavogaster, Coereba flaveola, Thraupis palmarum, and Anisognathus flavinucha.Serum samples from birds and horses from 33 locations in Venezuela  were scrT. leucomelas [pale-breasted thrush], 1 C. flaveola [bananaquit], and 1 G. gallus [domestic chicken] with the earliest collection date in February 2006); virus neutralization titers ranged from 80 to 320. One serum sample cross-reacted with other flaviviruses tested, with equivalent titers to WNV, Saint Louis encephalitis virus (SLEV), and Ilheus virus (ILHV) and was thus considered infected with an undetermined flavivirus. Seven serum samples were negative (antibody titers <20), and 1 sample was not tested because of insufficient sample volume.WNV was confirmed as the etiologic agent of infection in 5 adult birds  were confirmed positive for WNV infection by PRNT; viral titers ≥640 occurred in half of these horses. The earliest collection date for a WNV-positive horse was February 2004 and the most recent was May 2006. Specific WNV-reactive equine serum samples were distributed in valley regions , savannah grasslands (2.4%), the western region of Zulia (0.4%) and the Central Lake Basin (0.3%). A total of 46 (5.8%) equine serum samples were positive for neutralizing antibody to SLEV, and 8 (1.0%) samples were positive for neutralizing antibodies to ILHV. Forty-nine samples neutralized at least 2 of the 3 viruses and were classified as undetermined flaviviruses. Serum samples from 2 horses were negative in neutralization assays; 2 others were not tested because of insufficient sample volume.Culex spp.), which are present in the area in consistently high numbers, acquired the virus through hematophagous feeding on recently infected, migrating birds. Once introduced to local mosquitoes, virus is amplified among susceptible resident birds fed upon by ornithophilic mosquitoes. This pattern allows perpetuation and subsequent establishment of virus in a continuous transmission cycle, as opposed to infection of dead-end hosts, e.g., horses. This is the first report of WNV infection in South American birds and definitive establishment of the virus in South America.WNV-infected resident birds, rather than an importation event, are the basis of establishment of WNV in South America. We hypothesize that ornithophilic mosquitoes (such as some 2 test) would be virus introduction by migrating birds by an eastern migration route.We observed varying WNV seroprevalence rates in birds and horses across regions in Venezuela . These dExistence of several closely related flaviviruses in the American tropics (We encourage those involved in the public and animal health systems in Venezuela to consider zoonotic flaviviruses in the differential diagnoses of human and equine cases of encephalitis and to consider ecologic surveillance for zoonotic flaviviruses in mosquito and vertebrate host populations. We recommend monitoring blood and organ donations for flavivirus infections. Our study sheds light on flavivirus distribution in Venezuela. However, nothing else is known about the ecology of zoonotic flaviviruses in this country. Such knowledge will be essential for designing effective surveillance and control should these viruses be shown to cause human illnesses.Locations in Venezuela sampled for West Nile virus


We present the case of a patient with a cobalamin deficiency resulting in pancytopaenia, emphasizing the importance to define, diagnose and treat cobalamin deficiency.A 52-year-old man from the Democratic Republic of Congo presented to the emergency department with shortness of breath and a sore tongue. Physical examination was unremarkable. His haemoglobin was low and the peripheral blood smear revealed pancytopaenia with a thrombotic microangiopathy. The findings were low cobalamin and folate levels, and high homocysteine and methylmalonate levels. Pernicious anaemia with chronic atrophic gastritis was confirmed by gastric biopsy and positive antiparietal cell and anti-intrinsic factor antibodies. Cobalamin with added folate was given. Six months later, the patient was asymptomatic.Cobalamin deficiency should always be ruled out in a patient with pancytopaenia. Our case report highlights a life-threatening cobalamin deficiency completely reversible after treatment. In the 1950s, Hodgkin et al. used X-ray crystallography to reveal the structure of cobalamin. Nevertheless, there is still no gold standard for diagnosis, and nor are there clear guidelines for the treatment of cobalamin deficiency [Pernicious anaemia is a common cause of cobalamin deficiency, as discussed by Biermer in 1872. Whipple, Minot and Murphy developed a treatment with a liver extract in the late 1920s, suggesting an association between an extrinsic factor found in the liver and an intrinsic factor in the gastric juice. Folkers A population survey in the United States revealed that 1.9% of people older than 60 years of age have undiagnosed pernicious anaemia. The introduction of folate fortification in the United States has left some of the population at risk for a masked cobalamin deficiency .A 52-year-old man from the Democratic Republic of Congo presented to the emergency department after 14 days of shortness of breath, general weakness, weight loss and a sore tongue. He had no cough, chest pain or fever, and there was nothing significant in his medical and family history. He did not smoke nor drink alcohol. On examination, he appeared uncomfortable, anicteric and afebrile with a respiratory rate of 28 per minute, regular pulse of 70 beats per minute and blood pressure of 111/76 mmHg. A cardiovascular examination showed normal heart sounds and his lungs were clear on auscultation. The patient's tongue was unremarkable, as were the abdominal and neurologic examinations.3. His white blood cell count was 3600/mm3 with 27% neutrophils, 63% lymphocytes and 96,000/mm3 platelets. A peripheral blood smear  was given daily for 7 days, then weekly for 4 weeks, and then prescribed at a maintenance dose of 1000 μg once a month for life [3). Six months later, the patient was asymptomatic and the peripheral blood smear was normal. His levels of cobalamin (505 pmol/l), folate , MMA , and Hcys  were also normal.Nine days after introduction of therapy, correction of his deficiencies led to marked reticulocytosis  . General% . GenerThe most common causes of cobalamin deficiency are food-cobalamin malabsorption and pernicious anaemia, mainly among older people. Other causes include dietary deficiency, malabsorption and hereditary metabolic disease . PatientDiagnosing cobalamin deficiency is straightforward when the deficiency is profound. Nevertheless, diagnosing a slight deficiency is more difficult in older patients and in patients with neurological symptoms who present a borderline cobalamin level. Classically, the diagnosis is based on a low cobalamin level, but elevated MMA and Hcys levels seem more sensitive and more specific . Renal fSolomon evaluated patients for cobalamin deficiency using the standard metabolic markers MMA and Hcys in a retrospective study of the records of patients in an ambulatory setting over a 10-year period . The levThere are also new methods available to diagnose cobalamin deficiency. In plasma, total plasma cobalamin is bound to two proteins, haptocorrin and transcobalamin. Normally, 80% of cobalamin is on plasma haptocorrin and forms a holohaptocorrin. About 20% of cobalamin is combined with transcobalamin II to form the biologically active fraction holotranscobalamin II (holoTC), which is available for cell use. HoloTC levels seem to be an early indicator of cobalamin deficiency . SeveralAnti-intrinsic factors are more specific than antiparietal cell antibodies and are found in the serum of almost 40% of patients . Both faWe present an unusual case of cobalamin deficiency resulting in severe pancytopaenia with thrombotic microangiopathy that was completely reversible after cobalamin substitution. Primary care physicians must be aware of patients at risk for cobalamin deficiencies irrespective of the ethnic origin of the patient. No single algorithm fits every patient, and there is no clear consensus on how to diagnose and treat cobalamin deficiency. Consequently, the diagnosis should rely on clinical epidemiology, clinical symptoms and biochemical markers.Hcys: homocysteine; LDH: lactate dehydrogenase; MMA: methylmalonate.Written informed consent was obtained from the patient for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests.TMC carried out the literature review and wrote the case report and is the patient's caregiver. He also obtained the written consent of the patient. BF had a significant role in data analysis and in revising the manuscript. PB had a significant role in data analysis and in the final revision of the manuscript. All authors read and approved the final manuscript.


Most infected persons shed live virus after fever abated. We estimated the proportion of persons with pandemic (H1N1) 2009 who were shedding infectious virus at diagnosis and on day 8 of illness. In households with confirmed cases, nasopharyngeal swabs were collected on all members and tested by PCR and virus culture. Of 47 cases confirmed by PCR at <7 days of illness, virus culture was positive in 92% (11/12) of febrile and 63% (22/35) of afebrile persons. Of 43 persons with PCR-confirmed pandemic (H1N1) 2009 from whom a second specimen was collected on day 8, 74% remained PCR positive and 19% were culture positive. If the 73 symptomatic household members without PCR-confirmed illness are assumed to have pandemic (H1N1) 2009, a minimum of 8% (6/73) of case-patients shed replicating virus on day 8. Self-isolation only until fever abates appears insufficient to limit transmission. Self-isolation for a week may be more effective, although some case-patients still would shed infectious virus. Since April 2009, an influenza A virus, pandemic (H1N1) 2009, has spread to most countries of the world. Widespread susceptibility of persons <60 years of age may have facilitated rapid dissemination  2009 confirmed by reverse transcription–PCR (RT-PCR). Primary case-patients were either referred by their treating physicians or identified among community contacts of persons with laboratory-confirmed cases . At the Nasopharyngeal (NP) secretions were collected from all household members, both symptomatic and asymptomatic, with NP swabs  that were inserted into 3 mL of universal transport medium (Copan Innovation). Patients with results positive by RT-PCR for pandemic (H1N1) 2009 before day 7 since disease onset had a second NP swab repeated on day 8. Persons still positive on day 8 were retested on day 11.Nucleic acids were extracted from 200 µL of specimen by using the QIAGEN Viral RNA Mini Kit . Initially, pandemic (H1N1) 2009 was detected by using a conventional RT-PCR (pH1N1 PCR) specific for the hemagglutinin gene  were referred by their treating physicians. Among the 138 other participants (household or community contacts), 73 had respiratory symptoms for <7 days at the time of enrolment and 32 (44%) of 73 were positive by pH1N1 PCR , Table 1<7 days after symptom onset and 43 had a second swab collected on day 8 (>18 years of age) , 62 were identified on day 8 . Of thes of age) . On day  of age)  Virus cu of age) . Only 5%Of the 73 symptomatic participants tested within 7 days after symptom onset, 15 of the 47 PCR–positive case-patients  2009–infected persons still shedding replicating virus on day 8 varied from 8% to 13%, with no difference between children and adults. None were still shedding infectious virus on day 11. With seasonal influenza, virus shedding may be longer in children because they have less preexisting immunity that would limit replication than in adults. However, children and adults <50 years of age appear equally susceptible to infection with pandemic (H1N1) 2009 virus, which would support our findings of comparable virus replication and shedding across age groups studied  provides the minimal positivity rate on day 8, and testing of all 73 on day 8 could only have found a proportion equal to or greater than our 8% estimate.Second, our sampling methods could have influenced positivity rates. Although collection of NP specimens with a flocked swab is one of the best methods for obtaining specimens to detect influenza, those specimens might have been improperly collected by the nurses. Suboptimal collection of swabs would have yielded false-negative PCR or cell culture results, which in turn would have underestimated the proportion of patients shedding virus on day 8.Third, PCR testing with the matrix PCR was conducted retrospectively on frozen specimens, and only 5% of those were positive by virus culture. A greater proportion of virus culture specimens might have been positive if those specimens had been processed immediately instead of going through a freeze-thaw cycle (>6 days (Our results are consistent with other reports of virus shedding in pandemic (H1N1) 2009–infected patients. In Singapore, among 70 pandemic (H1N1) 2009–infected patients treated with oseltamivir and swabbed daily until virus clearance, 37% were PCR positive on day 7 of their illness and 9% on day 10  2009 outbreak at the US Air Force Academy showed that 29% (31/106) of afebrile patients and 19% (11/58) of patients who had been symptom-free for 24 hours still shed viable pandemic (H1N1) 2009 virus. In their study, 24% of 29 swabs collected on day 7 and 13% of the 16 swabs collected on day 8 of illness were culture positive, despite the large proportion of patients prescribed antiviral drugs  and is affected by the number and proximity of contacts between a case-patient and a susceptible person. Nevertheless, our study raises concerns about current recommendations for self-isolation until only 24 hours after fever has subsided  also may be prudent.


The N-methyl-D-aspartate receptor (NMDAR), a major excitatory ligand-gated ion channel in the central nervous system (CNS), is a principal mediator of synaptic plasticity. Here we report that neuropilin tolloid-like 1 (Neto1), a complement C1r/C1s, Uegf, Bmp1 (CUB) domain-containing transmembrane protein, is a novel component of the NMDAR complex critical for maintaining the abundance of NR2A-containing NMDARs in the postsynaptic density. Neto1-null mice have depressed long-term potentiation (LTP) at Schaffer collateral-CA1 synapses, with the subunit dependency of LTP induction switching from the normal predominance of NR2A- to NR2B-NMDARs. NMDAR-dependent spatial learning and memory is depressed in Neto1-null mice, indicating that Neto1 regulates NMDA receptor-dependent synaptic plasticity and cognition. Remarkably, we also found that the deficits in LTP, learning, and memory in Neto1-null mice were rescued by the ampakine CX546 at doses without effect in wild-type. Together, our results establish the principle that auxiliary proteins are required for the normal abundance of NMDAR subunits at synapses, and demonstrate that an inherited learning defect can be rescued pharmacologically, a finding with therapeutic implications for humans. The fundamental unit for information processing in the brain is the synapse, a highly specialized site of communication between the brain's multitude of individual neurons. The strength of the communication at each synapse changes in response to neuronal activity—a process called synaptic plasticity—allowing networks of neurons to adapt and learn. How synaptic plasticity occurs is a major question in neurobiology. A central player in synaptic plasticity is an assembly of synaptic proteins called the NMDA receptor complex. Here, we discovered that the protein Neto1 is a component of the NMDA receptor complex. Neto1-deficient mice had a dramatic decrease in the number of NMDA receptors at synapses and consequently, synaptic plasticity and learning were impaired. By indirectly enhancing the function of the residual NMDA receptors in Neto1-deficient mice with a small molecule, we restored synaptic plasticity and learning to normal levels. Our findings establish the principle that inherited abnormalities of synaptic plasticity and learning due to NMDA receptor dysfunction can be pharmacologically corrected. Our discoveries also suggest that synaptic proteins that share a molecular signature, called the CUB domain, with Neto1 may be important components of synaptic receptors across species, because several CUB-domain proteins in worms have also been found to regulate synaptic receptors. Spatial learning and memory depend on the N-methyl-D-aspartic acid receptor, a synaptic ion channel regulated by Neto1. Impaired cognition due to the absence of Neto1 can be rescued pharmacologically, a finding with implications for the therapy of inherited learning defects in humans. In the mammalian central nervous system, excitatory transmission at synapses is mediated primarily by the amino acid glutamate, acting through the postsynaptic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) and N-methyl-D-aspartic acid receptors (NMDARs) . Basal sThe core NMDAR is a heterotetramer comprised of two obligate NR1 subunits and two NR2(A-D) subunits . An emerCaenorhabditis elegans has been shown to be a component of the GLR-1 glutamate receptor ). Times of exploration of the DO and NDO were recorded for 5 min and expressed as a percentage of the total time of objects investigated. In the novel object recognition session, one of the familiar NDOs was replaced with a new object (NO) at the same location and the two familiar DOs were removed. The time examining a NO or a familiar object (FO) was recorded for 5 min and was expressed as a percentage of the total time of objects investigated. Data were analyzed with ANOVA with genotype as a between-subjects factor, and object rearrangement or object replacement as a repeated measures factor. The Tukey test was used for post hoc comparisons when ANOVA yielded statistically significant main effects or interactions.To examine the effects of CX546 in spatial learning, new cohorts of Neto1-null and wild-type littermate mice were used for the water maze and displaced-object tasks. In the water maze task, a single daily intraperitoneal injection of CX546  or vehicle (25% cyclodextran) was administered 30 min prior to training. No injection was given on probe trial days. For the displaced-object task, a single intraperitoneal injection of CX546 (15 mg/kg) or vehicle was administered 30 min prior to displaced-object recognition testing.All animal procedures were conducted in accordance with the requirements of the Province of Ontario Animals for Research Act, 1971 and the Canadian Council on Animal Care .http://www.ncbi.nlm.nih.gov/Genbank) accession numbers discussed in this paper are: PSD-95 (D50621); PSD-93 (AF388675); SAP-102 (D87117); and SAP-97 (NM_007862).GenBank . Bottom: ethidium bromide staining of gel prior to blotting.Top: Adult mouse multitissue RNA blot hybridized with a (661 KB PDF)Click here for additional data file.Figure S2In the yeast two-hybrid system, the strength of the interaction between a Neto1 cytoplasmic domain (Neto1-cd) construct, or a Neto1 mutants constructs lacking the last three amino acids (Neto1-cdΔTRV), five amino acids (Neto1-cdΔ5), ten amino acids (Neto1-cdΔ10), 20 amino acids (Neto1-cdΔ20), and PSD-95 deletion constructs is shown: +++, strong interaction; ++, moderate interaction; +, weak interaction; -, no detectable interaction; nd, no data.(469 KB PDF)Click here for additional data file.Figure S3 Confocal micrographs of immunostained hippocampal slices from the CA1 region. Antibodies used are indicated in each box. Scale bar, 10 μm. Pyr, pyramidal cell layer; SR; stratum radiatum.(2.83 MB PDF)Click here for additional data file.Figure S4(A) Immunoblots of biotinylated hippocampal surface proteins. Lanes 1, 2: biotinylated proteins after initial binding to avidin beads. Lanes 3, 4: biotinylated proteins recovered from supernatant after a subsequent binding to fresh avidin beads . The lack of biotinylated protein detected in lanes 3 and 4 indicates that the binding capacity of avidin beads used in lanes 1 and 2 was not exceeded. Blots shown are representative of three separate experiments.Neto1-null (gray bar) neurons. There was no significant difference between the mean NMDA peak current density calculated in Neto1-null neurons  compared with wild-type neurons  .(B) Histogram of peak current densities evoked by NMDA 1 mM from wild-type (white bar) and Neto1-null (gray bar) neurons. There was no significant difference between mean fraction of NMDA current inhibited by ifenprodil in the wild-type  and Neto1-null neurons  . Error bars represent ± standard error of the mean (SEM).(C) Histogram depicting the mean fraction of NMDA 1 mM peak current inhibited by ifenprodil 10 μM in wild-type (white bar) and (849 KB PDF)Click here for additional data file.Figure S5+/+Neto1 (open bars) or Neto1-null (filled bars) mice. Bottom histogram shows peak amplitude of NMDAR-mediated EPSC synaptic responses recorded from CA1 pyramidal neurons from +/+ (n = 20 neurons) or tlz/tlz (n = 13 neurons) mice . Strength of Schaffer collateral stimulation is indicated on the horizontal axis.Top histogram shows peak amplitude of AMPAR EPSCs from (186 KB PDF)Click here for additional data file.Figure S6+/+Neto1 (+/+ NMDAR EPSCs) and Neto1-null (tlz/tlz NMDAR EPSCs) neurons before (INMDAR) and 40 min after (IRo-resistant) Ro25–6981 (2 μM) administration . Each EPSC is the average of six consecutive traces. IRo-resistant EPSCs are also shown scaled  to the peak of the NMDAR EPSC before Ro25–6981 administration .Example representative NMDAR EPSCs from (83 KB PDF)Click here for additional data file.Figure S7+/+Neto1 and Neto1-null mice crossed the target platform location (T) with equal frequency.(A) Number of crossings over the hidden platform location (SE) after the first acquisition phase. +/+Neto1 (+/+) and Neto1-null (tlz/tlz) mice.(B) Average swim speed was not different between F1,16 = 10.36, p < 0.01). Error bars shown are ± standard error of the mean (SEM).(C) Number of crossings over the hidden platform location (NW) after the second acquisition phase. Neto1-null mice crossed the new target platform location (NW) less frequently than wild-type mice (one-way ANOVA +/+Neto1 (n = 9) and Neto1-null mice (n = 9). (D) During the second acquisition phase in the Morris water maze, Neto1+/+ mice predominantly used spatial strategies  to navigate to the relocated hidden platform during the last 2 d of the second acquisition phase . Data in (D) and (E) represent the breakdown of each search strategy employed by each genotype during the second acquisition period  Swim search strategies used by (66 KB PDF)Click here for additional data file.Figure S8Neto1+/+ mice used nonspatial swimming strategies (chaining and scanning) in the initial trials and then switched to spatial strategies  in later trials to locate the hidden platform.(A) (B) Neto1-null mice, however, predominantly used only nonspatial strategies (chaining and scanning), throughout the task, to navigate to the hidden platform. Data represent the breakdown of each search strategy employed during days 9–12 of the delayed matching-to-place (DMP) task.(64 KB PDF)Click here for additional data file.Figure S9Neto1+/+ (n = 12) and Neto1-null (n = 12) mice both showed similar exploration of objects during the habituation session. As expected, all mice spent more time exploring the object during the first 5 min, after which their exploration of objects declined. ANOVA did not find a main effect of genotype on time spent in contact with objects during the habituation period . Analysis of repeated measures revealed the main effect of habituation of time spent in contact with objects across the testing intervals . Both Neto1+/+ and Neto1-null mice significantly decreased the time of investigation of objects .(A) Neto1+/+ mice administered vehicle (n = 7), Neto1+/+ mice administered 15 mg/kg CX546 (n = 9), Neto1-null mice administered vehicle (n = 8), and Neto1-null mice administered 15 mg/kg CX546 (n = 7). ANOVA did not find a main effect of genotype or drug treatment on time spent in contact with objects during the habituation period . Analysis of repeated measures revealed the main effect of habituation of time spent in contact with objects across the testing intervals . Vehicle- and CX546-treated Neto1+/+ and vehicle-treated Neto1-null mice significantly decreased the time of investigation of objects . Neto1-null mice administered CX546 significantly decreased their exploratory activity after 10 min of habituation (p < 0.001 in comparison with first 5 min of vehicle-treated Neto1-null mice). All groups spent more time exploring the object during the first 5 min, after which, their exploration of objects declined. Error bars represent ± standard error of the mean (SEM).(B) Habituation profile of object exploration in the DO recognition task of (46 KB PDF)Click here for additional data file.Figure S10tlz/tlz with CX546; gray circles; n = 9) and without  CX546 (25 μM). Interstimulus interval is indicated on the horizontal axis. P1, fEPSP slope first response; P2, fEPSP slope second response.(A) Paired-pulse facilitation of fEPSPs in Neto1-null slices treated with (Neto1-null (tlz/tlz) mouse. Each fEPSP is the average of six consecutive traces. Scale bars: 5 ms, 0.2 mV.(B) Representative traces show fEPSPs before (black trace) and 20–30 min after CX546  administration in a hippocampal slice from a n = 6 consecutive control fEPSPs) minus during D-APV  from a single hippocampal slice from a Neto1-null mouse. Left trace was before administering CX546 and the right trace was during bath application of CX546 (25 μM). D-APV was washed out for 40 min before administering CX546. Scale bars: 10 ms, 0.1 mV.(C) Each trace shows the average difference plots before ((150 KB PDF)Click here for additional data file.Figure S11+/+Neto1 and Neto1-null mice crossed the platform location with equal frequency in the first acquisition regardless of whether they were administered vehicle or CX546 .(A) +/+Neto1 and Neto1-null mice administered vehicle or 15 mg/kg CX546. Post hoc analysis did not indicate a difference in swim speed across groups (p > 0.6).(B) Average swim speed was not different between +/+Neto1 mice (p > 0.4). Neto1-null mice administered vehicle crossed the hidden platform location significantly fewer times compared with +/+Neto1 mice given vehicle . Error bars represent ± standard error of the mean (SEM).(C) Neto1-null mice administered CX546 crossed the platform location in the second acquisition phase with equal frequency as compared to (54 KB PDF)Click here for additional data file.Table S1(13 KB PDF)Click here for additional data file.Table S2(47 KB PDF)Click here for additional data file.


Active commuting is associated with a reduced risk of several chronic diseases, but few studies have considered institutional factors that influence it. We examined the association between cultural and physical workplace supports for active commuting and employee active commuting behavior.Data were collected from employees (N = 375) in Manhattan, Kansas, via an online survey. Physical and cultural supports for active commuting in the workplace were measured separately. Active commuting frequency was dichotomized as 0 trips versus at least 1 trip per week by foot or bicycle. Binomial logistic regression was used to predict the likelihood of commuting actively at least once per week, according to the number and type of cultural and physical workplace supports. Analyses were conducted by sex and age and for the full sample.Among the full sample, employees who reported more physical and cultural supports in the workplace for active commuting were more likely to walk or bicycle to work at least once per week. Significant, positive relationships between cultural and physical supports and active commuting were observed for women but not for men. Both younger and older adults who reported 2 or more physical supports in the workplace were more likely to actively commute, but no relationship was observed for cultural supports. The most influential types of individual supports were perceiving that other coworkers actively commute and the presence of bicycle parking and a bicycle storage policy at the workplace.The presence of workplace physical and cultural supports is related to more active commuting behavior and may especially encourage active commuting among women. Public health guidelines for physical activity recommend that adults engage in at least 150 minutes of moderate-intensity physical activity per week and that this amount can be accumulated in episodes as short as 10 minutes -5.2max, maximal treadmill time, and high-density lipoprotein cholesterol  with a 2006 population of 50,737, approximately three-quarters (76%) of which are adults aged 18 to 64 years . In 2006This study was part of a broader exploratory investigation of active commuting that was approved by the institutional review board at Kansas State University. For the larger study, an online survey was conducted from August to December 2008. Participants were recruited through links from local Web sites , e-mails to municipal and school board employees throughout the city, and fliers provided to area employers. Eligibility criteria included being aged 18 years or older, living or working full-time or part-time in Manhattan, Kansas, and being physically able to walk or bicycle. People who began the survey and reported not being employed at all (n = 20) or being physically unable to walk or bicycle (n = 12) were not included in the analyses.The online, anonymous survey  consisted of several pages of questions related primarily to active commuting behaviors. After reading the consent statement, participants were asked during the survey to indicate their sex, age, race, education level, and estimated walking time to work (dichotomized as ≤20 min or >20 min).Cultural supports for active commuting were measured with 2 Likert scale–type questions. The first asked about respondents' perceptions about the extent to which their employer encourages active commuting, and this was recoded as "absent"  versus "present" ("somewhat" to "strongly encourages"). The second question asked about the perceived number of coworkers who actively commute to work (recoded as "none" or "some"). A dichotomous variable was created that categorized participants as having neither type of cultural support or having at least 1 cultural support.Physical supports for active commuting were measured with 3 yes/no questions about the availability of bicycle parking, bicycle storage policies, and showers or lockers at the workplace. These variables were summed to designate respondents with 0, 1, or 2 or more of the 3 types of physical supports. Participants were also asked to indicate the number of times per week they walked and bicycled to or from work, and a dichotomous active commuting outcome variable was created indicating 0 trips versus at least 1 trip by foot or bicycle. All survey questions relevant to this study are provided in the Chi-square tests were used to assess differences in the prevalence of active commuting behavior and physical and cultural supports between men and women and between respondents of different ages (18-39 y vs ≥40 y). Binomial logistic regression was used to predict the likelihood of actively commuting to work at least once per week, according to the number of cultural and physical workplace supports reported, using the 0 supports category as the reference group for each model. We also examined the likelihood of active commuting on the basis of the presence or absence of the 5 types of supports. For all analyses, separate models were examined for the full sample of participants, for men and women independently, and for younger (18-39 y) and older (≥40 y) adults independently. All analyses controlled for age, race, education, perceived walking time to work, and sex (when these variables were not used to stratify the sample). Missing data were few but excluded pairwise. All analyses were conducted using SPSS version 17.0 .In total, 375 people completed the survey . The meaIn the full sample, participants who reported 2 or more physical supports for active commuting were more likely to actively commute at least once per week than those who reported none, but participants reporting only 1 physical support were not . When thIn the full sample, participants who reported 1 or more cultural supports were more than twice as likely to actively commute at least once per week as participants who reported no cultural supports . Women wWe also examined the combined and disaggregated effects of different types of supports. Among the full sample, participants who reported at least 1 cultural support and at least 1 physical support for active commuting in the workplace were more than 6 times as likely to actively commute at least once per week as participants who reported both types of supports absent . This cuFor all 5 samples examined, perceiving that the employer encourages active commuting was not related to increased odds of walking or bicycling to work at least once per week, and having showers or lockers at the workplace was related to increased odds of active commuting only for women . HoweverWe found that people with a workplace environment that had more cultural and physical supports were more likely to walk or bicycle to work. However, this relationship between workplace supports and active commuting behavior appeared to be moderated by sex, as such associations were significant only for women in our sample. For women, the presence of at least 1 cultural support, in particular their perceptions that other coworkers walk or bicycle to work, was associated with almost a threefold increase in the likelihood of active commuting at least once per week. Likewise, women who reported 1 physical support were more than 3 times as likely to actively commute, while women with 2 or 3 physical supports were 10 times as likely to do so. Furthermore, the cumulative effect of having at least 1 cultural and at least 1 physical workplace support was strongest for women in this study. To our knowledge, few, if any, studies have explored how influences on active commuting differ by sex. One study  reportedWhen the sample was stratified by age, the presence of 2 or more physical workplace supports was associated with a greater likelihood of active commuting for both younger and older adults. However, with respect to cultural supports, the significant relationship observed for the full sample failed to materialize for either the younger or older subgroups. When considered in isolation, though, greater perceptions of the number of coworkers who actively commute was related to increased active commuting behavior for both age groups. Having a larger percentage of fellow employees who walk or bicycle to work may create an environment that tolerates, fosters, or reinforces similar behavior among colleagues. Future research can shed light on the psychological  or behavioral  mechanisms behind the relevance of having coworkers who actively commute.The primary strength of our study was its novelty in considering supports in workplace settings that may influence active commuting. However, our study does have limitations. First, our data were collected from a convenience sample that was small. Because the exact number of people who were eligible and had the opportunity to complete the survey is unknown, we were unable to calculate a response rate. We also could not entirely rule out the possibility that respondents were clustered in workplaces, although some data collected in the survey (closest intersection to workplace) suggested that the participants originated from various locations. Future studies may examine our research questions with a larger, random sample of employees, potentially in a multilevel framework. Second, we used self-report measures of both the workplace environment and active commuting behavior . Third, our online survey format may have excluded potential respondents who did not have access to the Internet. Finally, our sample was well-educated and almost exclusively white, which limits the generalizability of our findings.Our results substantiate the impact of a social-ecological approach for promoting active commuting. Many studies have examined the utility of workplace interventions for promoting physical activity, with mixed results. For example, Proper et al  reviewedIn terms of health promotion, our results provide a foundation for intervention strategies, including potential physical changes to workplaces, such as the addition of showers, bicycle racks, and covered and secure bicycle parking. Likewise, a workplace culture that supports active commuting can influence mode of travel to work. The social support that results when a large number of workers actively commute, either collectively or individually, can be reinforcing . Such prOur study adds to the small body of existing literature concerning factors associated with active commuting among adults and contributes to our understanding of social-ecological influences on this behavior beyond the individual. More research is needed to evaluate interventions that aim to promote active commuting , but mak


Our analyses identified a two-locus combination - rs6939589 and rs11634386 - that proved to be significantly associated with rheumatoid arthritis under a Rec × Rec model .The goal of this paper is to search for two-locus combinations that are jointly associated with rheumatoid arthritis using the data set of Genetic Analysis Workshop 16 Problem 1. We use a two-stage strategy to reduce the computational burden associated with performing an exhaustive two-locus search across the genome. In the first stage, the full set of 531,689 single-nucleotide polymorphisms was screened using univariate testing. In the second stage, all pairs made from the 500 single-nucleotide polymorphisms with the lowest  Although the primary interest of genome-wide association studies (GWAS) is to identify single-nucleotide polymorphisms (SNPs) with detectable marginal effects, it is also of interest to identify SNPs that have an interaction effect. Testing all possible pairs of loci to identify interactions creates many practical difficulties. In this article, we use a two-stage strategy to search for two-locus joint effects, which effectively reduces the computation time. In the first stage, we only selected loci that met an initial threshold. In the second stage, any locus that met the first-stage threshold was tested under each of 17 two-locus joint effect models. By applying this two-stage analysis to Genetic Analysis Workshop 16 (GAW16) Problem 1, we successfully identified two SNPs that are jointly associated with rheumatoid arthritis (RA) and could not be identified by single-locus analysis.In this article, we propose 17 two-locus models, which include 8 epistatic models and 9 multiplicative models  = β0 + β1X, where g is the two-locus genotype; for the eight epistatic models in which the nine two-locus genotypes can be divided into high-risk group and low-risk group, X = 1 if the genotype g belongs to the high-risk group and X = 0 otherwise; for the nine multiplicative models, X = x1 + x2, where x1 is the dominant, recessive, or additive coding of the genotype at the first SNP for a dominant, recessive, or multiplicative model, respectively, and x2 is similarly defined for the second SNP. For the ith individual, let yi denote the trait value  and Xi denote the numerical code of the genotype (X in the log linear model). The score test statistic is given byThe 17 two-locus models can be described by the following log linear model log N is the sample size, X1, ..., XN, and y1, ..., yN. Under null hypothesis of no association, Tscore asymptotically follows a chi-square distribution with one degree of freedom (df). Though we use a unified test statistic Tscore for all of the models, the 17 models correspond to 17 different tests. Different models correspond to different genotype coding, and thus different values of Xi in Tscore. For example, under Rec ∩ Rec epistatic model, X = 1 if the two-locus genotype is AABB; X = 0 otherwise; under Rec× Rec multiplicative model, X = x1 + x2, where x1 = 1 if the genotype at the first SNP is AA and x1 = 0 otherwise; x2 is similarly defined for the second SNP.where P denote the smallest p-value of the three tests. We select M SNPs with the smallest P (M = 500 was used in this study). In the second step, under each of the 17 two-locus models, we apply a two-locus association test Tscore to each of the L two-locus combinations among the M selected SNPs, where L = M(M - 1)/2. In this step, we get a p-value  for each of the L two-locus combinations and each of the 17 two-locus models.The method to search for significant two-locus combinations for each of the 17 models has the following two steps. In the first step, three one-df chi-square tests  are used to test for association at each SNP. For a SNP, let pij denote the raw p-value of the lthtwo-locus combination under the jth two-locus model. A permutation procedure is then used to adjust for multiple tests and multiple models. For each permutation, we randomly shuffle the case and control status. Based on the permuted data, we redo the single-marker test for each SNP, select M SNPs with the smallest p-values, and then test each of the two-locus combinations among the M selected SNPs under each of the 17 models. For the kth permutation, let p-value of the two-locus test for the lth two-locus combinations under the jth model and let K times (we use K = 1000 in this study). Then, the overall p-value of the two-locus test for the lth two-locus combination under the jth model is given by L two-locus tests and 17 models and we also can adjust for the fact that the M SNPs to do two-locus analysis were chosen as the "top" signals from a much larger set.Let p-values of the 1000 tests is given in Figure p-values should follow a uniform distribution. Figure The two-locus analysis discussed in the last section is valid under the assumption that all sampled individuals come from a homogeneous population. Thus, we first check this assumption in the data set of GAW16 Problem 1. For this purpose, we randomly choose 1000 SNPs and perform association test at each of the 1000 SNPs using the one-df chi-square test. The histogram of the N one-df chi-square tests (single-marker test or two-locus test) with test statistics T1, T2, ..., TN. The GC method rescales Ti as i = 1, 2, ..., N), where b = median {T1, T2, ..., TN}/0.465 and considers Si following a chi-square distribution with one df.1. Genome Control (GC) method: the GC method  rescalesN one-df chi-square tests and rescale the test statistics as S1, S2, ..., SN by the GC method. Then, we carry out permutation K times. For each permutation, we perform N chi-square tests based on the permuted data and rescale the test statistics by GC method. For the kth permutation, let p-value of the ith test is given by 2. Permutation GC method: Suppose that we have p-values from the two methods are shown in Figure p-values are very similar to a uniform distribution, which indicates that both the GC and permutation GC methods can control for population stratification, at least for the data set of GAW16 Problem 1.To test whether the two methods can control for population stratification, we randomly chose 1000 SNPs from the data set of GAW16 Problem 1. The GC and permutation GC methods were used to test association for the 1000 SNPs. The histograms of the p-value of the three tests at each SNP was adjusted by Bonferroni correction for the three models and all the SNPs. From the single-marker analysis, we found one SNP (rs2476601) on chromosome 1 and 183 SNPs on chromosome 6 with adjusted p-values less than 0.05 after Bonferroni correction. The 183 SNPs that we found on chromosome 6 are in a region (29930240 to 33149012 bp) in high linkage disequilibrium with HLA-DRB1, a factor known to have a strong association with RA [PTPN22, which has been identified to be associated with RA [The GAW16 Problem 1 data set contains genotypes at 531,690 SNPs on chromosomes 1-22 (868 cases and 1194 controls). We first performed single-marker analysis. Three one-df chi-square tests  rescaled by GC method were applied to each SNP. The smallest  SNP rs246601 on cp-value of 0.045.We used the permutation GC method with 1000 permutations to carry out the two-locus analysis. In the analysis, we excluded SNPs within 10 Mbp of any of the SNPs surviving Bonferroni correction from the single-marker analysis. Then, we performed the two-stage, two-locus analysis among the remaining SNPs. For each pair of SNPs, individuals with missing genotypes at either locus were not included in that analysis. After adjusting for multiple tests and multiple SNPs, we found one two-locus combination - rs6939589 on chromosome 6 and rs11634386 on chromosome 15 - significantly associated with RA. The two-locus combination followed the Rec× Rec multiplicative model with a Although there is growing appreciation that searching for epistatic interactions in humans may be a fruitful endeavor, there are still a number of practical difficulties associated with testing all possible pair-wise comparisons in the case of GWAS, such as data storage requirements, computation time, and multiple testing. In this report we used a computationally efficient two-stage analysis to search for joint effects of genes for GAW16 Problem 1. Applying this strategy to GAW16 Problem 1, we have successfully identified two loci that are significantly associated with RA. These two loci would not have been identified as significant by single-marker analysis after correction for multiple tests.M SNPs according to the marginal effects, the loci with strong interaction effects but weak marginal effects may not be detected. Furthermore, our approach only considers 17 interaction patterns (corresponding to the 17 two-locus models). The proposed two-stage strategy may have low power to detect interactions whose patterns depart from the 17 two-locus models.Computational efficiency has imposed limits on our two-stage strategy. Because our approach only searches two-locus interaction among the top Our two-locus analysis showed that a two-locus combination, rs6939589 and rs11634386, is significantly associated with RA.GAW16: Genetic Analysis Workshop 16; GC: Genome control; GWAS: Genome-wide association study; RA: Rheumatoid arthritis; SNP: Single-nucleotide polymorphismThe authors declare that they have no competing interests.AN performed the statistical analysis and wrote the first draft of the manuscript. ZZ participated in the study design and helped to draft the manuscript. QS contributed to the design of the study and to the manuscript preparation. All authors read and approved the final manuscript.


Bemisia tabaci is known to transmit many begomoviruses, and the spread of B. tabaci, especially the B and Q ‘biotypes’, has been accompanied by the epidemics of begomoviruses. One of these identified disease-causing agents was Tomato yellow leaf curl China virus (TYLCCNV).Plant-pathogenic begomoviruses have a complex association with their insect vectors. The interactions of begomoviruses and reproduction of their vectors are poorly understood. B. tabaci, including the alien invasive B and the indigenous ZHJ1 from Zhejiang, China, feeding on either healthy or TYLCCNV-infected tobacco plants. The ovary of the whitefly was composed of 12–22 telotrophic ovarioles. According to the morphology of the oocytes and level of yolk content, oocytes in ovarioles were divided into four developmental phases (I-IV). Significantly higher proportion of immature oocytes  and mature oocytes (phase IV) was observed in ovary of females that fed on TYLCCNV-infected tobacco compared to that on healthy plants. Moreover, there was significant increase of eggs laid of B whitefly that fed on TYLCCNV-infected tobacco plants during the early developmental stages. In contrast, the proportion of oocytes of different developmental phases and eggs laid had no significant differences between ZHJ1 whiteflies feeding on TYLCCNV-infected and non-infected host plants.In this study, we compared the egg production and realized fecundity of two ‘biotypes’ or putative species of the whitefly The invasive B whitefly benefits from feeding on a begomovirus-infected plant through increased egg production and realized fecundity. Plant pathogen-vector systems are characterized by complex interactions. Some plant- pathogenic geminiviruses have caused serious problems to crops Bemisia tabaci (Hemiptera: Aleyrodidae) is genetically diverse and a vector of many begomoviruses B. tabaci indicates that the complex contains at least 24 cryptic species, some of which have been referred to as ‘biotypes’ in the last 20 years B. tabaci complex is yet to be fully resolved, the commonly used term ‘biotypes’ to refer cryptic species has been sometimes retained The whitefly B. tabaci complex, i.e., the Mediterranean and the Middle East-Minor Asia 1 Two cryptic species of the Moreover, compared to its performance on uninfected plants, B feeding on begomovirus-infected plants performed better B. tabaci species complex Tomato yellow leaf curl China virus (TYLCCNV) is a whitefly-transmitted begomovirus that has become widespread in south China for years The virus vectors of this study include the B and ZHJ1 whiteflies, which are known to belong to different putative species of the Bemisia tabaci (Gennadius) species complex were used. The B whitefly (GenBank accession no. AJ867555) population was first collected from cabbage, Brassica oleracea var. capitata L. (Cruciferae), and the indigenous ZHJ1 whitefly (GenBank accession no. AJ867556) population from cotton, Gossypium hirsutum L., in 2003 in Hangzhou, Zhejiang, China  G. hirsutum L. cv. Zhemian 1973 in separate climate chambers at 28±1 °C, 14 h light: 10 h darkness and 70±10% r. h. The purity of the cultures was monitored every 3-5 generations using the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) technique Two cryptic species of the whitefly β) constructed previously Nicotiana tabacum L. cv. NC89.Infectious clones of TYLCCNV and their satellite DNA molecules , a non-host plant of TYLCCNV, were used. Uninfected tobacco and cotton plants were grown in insect-proof cages under natural lighting and ambient temperature in screen houses as described in Jiu et al. β at the 4–5 true-leaf stage by agro-inoculated as previously described Tobacco The whitefly adults of different developmental stages reared on cotton plant were insufflated into centrifuge tube on ice. The ovaries of each female were then dissected in phosphate buffered saline (PBS) (pH 7.4) and the epidermis was removed. Different types of ovarioles were separated using a stretched microcapillary. The morphology of ovary samples and ovarioles were photographed with a phase contrast microscope  fitted with a light source. Digital images were captured using a digital camera and modified using Adobe Photoshop®.These experiments were conducted to examine the realized fecundity of adult whiteflies which were reared on cotton from egg to pupa, and then transferred upon emergence onto healthy or virus-infected tobacco plants. The experiment was carried out concurrently using the procedure as described earlier by Jiu et al. The number of eggs laid on healthy or virus-infected tobacco by the whiteflies was examined using the clip-cages described by Zang et al. © package (Version 8.01 for Windows) t-test. The effects of virus infection and age on the reproduction of B or ZHJ1 were analyzed using two-factors ANOVA and Tukey's multiple range test (MRT). The total number of oocytes and eggs laid of different cryptic species of whiteflies feeding on healthy and virus-infected plants in the first 2 d were analyzed using two-factors ANOVA and Tukey's MRT. All proportion data were transformed by arcsine root before analysis, but untransformed means are presented.All statistical analysis were conducted using the DPSThe ovaries of the whiteflies are formed by a mass of ovarioles, of which the number for a pair of ovary is ranged from 12–22 in various developmental stages. Every ovariole contains two or three oocytes in various stages of maturity . The ovaAccording to the main morphological characteristics of oocytes and the level of yolk content, the oocytes were classified into four developmental phases, separately named phase I, II, III and IV . No yolkB adults feeding on virus-infected plants had a significantly lower proportion of phase I oocytes but significant higher proportions of phase II, III, and IV oocytes, compared to the B adults feeding on healthy plants . In contDue to the short lifespan of ZHJ1 on healthy and virus-infected plants, these parameters were compared statistically at the first two days after emergence with B. The total number of oocytes in B was significantly higher than that of ZHJ1 .Compared with the ZHJ1, B whiteflies laid 5–8 times more eggs on either healthy or virus-infected plants , Table 2When the number of mature oocytes in ovaries and that of eggs laid was added, the total number in B adults was 4–6 time higher than that in ZHJ1 adults feeding on either healthy or virus-infected plants .B. tabaci belongs to the telotrophic type. This type ovariole had been reported in three species of aphid, Stomaphis quercus, Phylloxera coccinea and Phylloxera glabraB. tabaci, the ovaries were characterized by an increasing number of ovarioles and mature oocytes after eclosion and 12–22 ovarioles were observed in its adult stage. Our results are consistent with Gameel et al. B. tabaci. Previous reports had provided only a brief description of the ovarian structure and oogenesis in green house whitefly Trialeurodes vaporariorumAleurochiton aceris are composed of 5 short telotrophic ovarioles B. tabaci, each ovariole contained two or three oocytes and a germarium. Once a mature oocyte was present in ovariole, three oocytes would be observed in the same ovariole. This ovarian system, based on morphological examination of oocyte of the four developmental phases, was used to evaluate the effects of virus infection of the plants on the whitefly reproduction.The ovariole of insects is usually divided into three types: the polytrophic type, telotrophic type and the panoistic type Hamiltonella, Arsenophonus, Cardinium, Wolbachia, and Rickettsia, in a bacteriocyte of whitefly. This phenomenon may be a result of the direct enclosure of the bacteriocyte in the egg during oogenesis, providing a useful mechanism for efficient vertical transmission of endosymbiotic microorganisms. These endosymbiotic microorganisms had been reported not only in whitefly, but also in aphids and other insects S. quercus, trophocyte cytoplasm is filled with endosymbiotic microorganisms A rounded bacteriocyte sphere was observed to be transferred into the oocyte at the terminal of ovariole in oocyte III and embedded in mature oocyte. The bacteriocytes housing symbiotic bacteria had been examined in whiteflies Both B and ZHJ1 whiteflies are effective vectors of TYLCCNV B. tabaci. The yolk protein is the main composition of egg content in many insects during oogenesis. In the ovaries of the two cryptic species of the whitefly, whether the different proportions of mature oocytes were due to the synthesis and uptake of yolk protein needs to be elucidated. Further, whether feeding on the virus-infected plants could lead to structural changes of the ovaries or changes in hormone synthesis in the vectors remain to be investigated.In plant-pathogen-vector systems such as the one examined in this study, the pathogen depends on the arthropod herbivore vector for transmission and dispersal. Thus higher proportion of mature oocytes in the ovaries and consequently higher numbers of eggs will help increase vector population and this will in turn facilitate the spread of virus disease pandemic in the field. This kind of vector-virus mutualism in assisting in the spread of virus disease pandemic had been seen in other vector-virus-plant systems, such as some aphid-transmitted viruses in wheat, barley and oats


The reconstituted virus from the modified clone showed significant improvement in replication in vitro and in vivo. Excision of the BAC sequences also enhanced the pathogenicity to levels similar to that of the parental virus, as the cumulative incidence of Marek's disease in groups infected with the recombinant and the parental viruses showed no significant differences. Thus, we have been able to make significant improvements to the existing BAC clone of this highly oncogenic virus which would certainly increase its usefulness as a valuable tool for studies on identifying the oncogenic determinants of this major avian pathogen.Cloning of full length genomes of herpesviruses as bacterial artificial chromosomes (BAC) has greatly facilitated the manipulation of the genomes of several herpesviruses to identify the pathogenic determinants. We have previously reported the construction of the BAC clone (pRB-1B5) of the highly oncogenic Marek's disease virus (MDV) strain RB-1B, which has proven to be a valuable resource for elucidating several oncogenic determinants. Despite the retention of the BAC replicon within the genome, the reconstituted virus was able to induce tumours in susceptible chickens. Nevertheless, it was unclear whether the presence of the BAC influenced the full oncogenic potential of the reconstituted virus. To maximize the closeness of BAC-derived virus to the parental RB-1B strain, we modified the existing pRB-1B5 clone by restoring the Us2 and by introducing SV40- E. coli . As vRBbacteria . Once thes virus , human ces virus  and rheses virus  to generTTAATTAACGACAGACCTACTTGCTACCA) and Us3R (5'-CTCGAGGTATGGCCATGTGGTCTCTA) that contained the PacI and XhoI restriction sites respectively. The XhoI-PacI fragment containing partial mini-F sequence released from the plasmid pDS-HA1 [PacI-SalI site. The SV40-cre fragment released from pYD-C66  as EcoRI-XhoI fragment was blunt-ended and inserted into PmeI site of the above plasmid. Finally a blunt ended FRT-Kan cassette from pKD13 was further inserted into BstEII site (blunt ended) of the above plasmid. The resulting construct pPartial-F-Kan-cre-Us2-Us3 was used for making self-excisable BAC clone of RB-1B virus by lambda Red mutagenesis [Kan cassette flanked by the FRT sequences was flipped off by adding 0.2% L-arabinose for 1 hour and selecting on chloramphenicol-containing LB plates. DNA extracted from one of the chloramphenicol-resistant clones (pRB-1B*X6) was checked for infectivity by transfection into primary CEF using Lipofectamine . The virus reconstituted virus from this clone, designated vRB-1B*X6, was grown up on CEF, titrated and stored in liquid nitrogen.The modification of pRB-1B was carried out as shown in Fig  pDS-HA1  was linkagenesis  by co-trEcoRI restriction digestion (not shown) followed by Southern blot analysis using digoxigenin (DIG)-labelled DNA probes  using procedures described before [E. coli guanine phosphoribosyl transferase (gpt) sequence in the pDS-PHAI [gpt probe, specific signals were detected in pRB-1B5 (lane 1), pRB-1B*X6 (lane 2) and vRB-1B5 (lane 3), demonstrating the presence of the mini-F plasmid in these DNA samples. The absence of gpt signals in the DNA sample from vRB-1B*X6 (lane 4), similar to the wtRB-1B virus (lane 5), confirms the self-excision of the mini-F plasmid during the growth of the virus in the CEF, following the expression of the functional cre in these cells. Thus the data from the Southern blot hybridization showing the presence of MEQ and Us2 and absence of mini-F plasmid confirmed that the genome structure of the vRB-1B*X6 is similar to the wtRB-1B virus. We also examined the status of expression of Us2 by reverse transcription PCR in the cells infected with the 3 viruses. As expected, both vRB1B-X6 and wtRB-1B virus-infected cells amplified a PCR product of the expected size, cells infected with vRB1B-5 virus was negative (data not shown).The integrity of the pRB-1B*X6 clone was initially checked by d before . The mempDS-PHAI . As expepDS-PHAI . Detectiin vitro growth of the two viruses in CEF. As shown in the Fig in vitro. The wtRB-1B virus, less adapted to replicate in CEF, consistently showed lower in vitro levels than the other viruses. We then asked whether the increased in vitro replication of the vRB-1B*X6 is also reflected in the replication in vivo. For this, we monitored the MDV genome copy numbers in the peripheral blood leukocytes (PBL) of 5 line P SPF (specific-pathogen-free) chickens infected with 1000 p. f. u. of vRB-1B5 and vRB-1B*X6 viruses at different days up to 28 days post infection using real-time quantitative PCR [in vivo replication in PBL. In this respect, vRB-1B*X6 virus showed a replication kinetics almost identical to the wtRB-1B virus [Fig In order to examine whether the above modifications have given any growth advantage to the vRB-1B*X6 over vRB-1B5, we first compared its tive PCR . MDV genirus Fig . The difirus Fig .19/B19) chickens lacking maternal antibodies to MDV were infected intra-abdominally with 1000 p. f. u. of vRB-1B5, vRB-1B*X6 and wtRB-1B viruses. All procedures on experimental birds were approved by the Institute for Animal Health Ethical Committee and carried out in accordance with the Project Licence 30/2145 issued by the United Kingdom Home Office. All the groups of infected birds, together with a group of uninfected control birds, were maintained in isolation and observed for 90 days. Cumulative occurrence of MD for the different groups, based on the incidence gross or histological lesions, was used to calculate the survival rates, and the statistical differences between the different groups were calculated using Kaplin-Meier log rank test for survival [We then asked whether the increased replication ability of vRB-1B*X6 is associated with higher pathogenicity in an infection model. For this, groups (n = 10) of one-day old MD-susceptible specific-pathogen-free line P  declare that they have no competing interests.YZ contributed to design, perform the experiment and draft the manuscript. LP participated in the experiment. LPS and SB conducted animal experiments and quantitative PCR analysis. VN supervised the study and drafted the manuscript.


By analyzing vesicle fluids and crusted scabs from 136 persons with suspected monkeypox, we identified 51 cases of monkeypox by PCR, sequenced the hemagglutinin gene, and confirmed 94% of cases by virus culture. PCR demonstrated chickenpox in 61 patients. Coinfection with both viruses was found in 1 additional patient. Monkeypox (MPX) virus is an orthopoxvirus that causes human MPX, a smallpoxlike disease reported in the African rainforests. Humans acquire the virus through direct contact with infected animals or patients  were inoculated onto MA104-cells by using standard procedures in a Biosafety Level 3 laboratory. Results yielded 56 MPX virus isolates from 48 patients ; scab anDNA of all specimens was analyzed by using the RealArt Orthopox LightCycler PCR kit , which amplifies sequences of the fusion protein gene present in all orthopoxviruses, including MPX virus ; all speOf 171 specimens examined by using RealArt VZV LC PCR (QIAGEN), 78 showed specific amplification that indicated that 62 patients had VZV infection . Of 136 <0.001, t = 4.625, df = 109). That most MPX cases (94%) occurred in patients <25 years of age suggests that cross-protective immunity may still exist; a potential for continued immunity has been demonstrated  of the open reading frame of the HA gene was determined . There wFor our study, we used 2 commercially available kits to amplify orthopoxvirus and VZV sequences. The assays contain an internal control to monitor inhibition and use 4 positive controls to enable quantification of the amount of input genomic viral DNA. On the basis of our results, one can calculate that MPX lesion material usually contains several million viral genomic copies (data not shown). Whereas WHO surveillance confirmed 67% by virus culture  had laboratory-confirmed MPX infection, 61 (44.8%) had laboratory-confirmed VZV infection, and 1 (0.7%) had coinfection. MPX virus is considered to be the most important orthopoxvirus infection in humans because it causes disease clinically indistinguishable from smallpox. Recent outbreaks of MPX in the United States (


Anas crecca) from the Camargue area, in the South of France, across Europe. Based on bird-ring recoveries, local duck population sizes and prevalence of infection with these viruses, we built an individual-based spatially explicit model describing bird movements, both locally (between wintering areas) and at the flyway scale. We investigated the effects of viral excretion duration and inactivation rate in water by simulating AIV spread with varying values for these two parameters. The results indicate that an efficient AIV dispersal in space is possible only for excretion durations longer than 7 days. Virus inactivation rate in the environment appears as a key parameter in the model because it allows local persistence of AIV over several months, the interval between two migratory periods. Virus persistence in water thus represents an important component of contamination risk as ducks migrate along their flyway. Based on the present modelling exercise, we also argue that HP H5N1 AIV is unlikely to be efficiently spread by Common Teal dispersal only.Since the recent spread of highly pathogenic (HP) H5N1 subtypes, avian influenza virus (AIV) dispersal has become an increasing focus of research. As for any other bird-borne pathogen, dispersal of these viruses is related to local and migratory movements of their hosts. In this study, we investigated potential AIV spread by Common Teal ( Wild birds, especially waterbirds such as Anseriformes  and Charadriiformes , are natural hosts for influenza A viruses Wild bird migrations have been extensively studied throughout the world Anas crecca) is among the most abundant duck species in Europe Common Teal  of ducks. The aims of this study were to: (i) Build an individual-based spatially explicit model which describes duck movements in space and time. Based on the ringing dataset and duck abundance in the Camargue, we reproduced Common Teal movements in Europe, from September (beginning of the wintering period) to May (end of spring migration). (ii) Spatially represent the maximal AIV spread distance by this species . We integrated monthly AIV prevalence data recorded in the Camargue and simulated virus spread according to a large set of hypothetical viral excretion duration values . This prevalence was higher than those previously reported in the Camargue We found a clear pattern of circulation, consistent with previous studies To avoid a potential bias linked to underestimated prevalence, we also performed simulations with a prevalence of infected ducks equal, each month, to 15%. This value represents the highest infection rate recorded in the Camargue  and a hiIn this study, we assumed that infected ducks were not subjected to physiological and behavioral modifications reducing movement abilities. Under this scenario, viral excretion duration was likely to be the key parameter for long distance spread of AIV. Simulation results indeed showed clearly that long AIV excretion duration enhanced dispersal efficiency in space . This reBecause of the predominance of local movements of ducks , virus iWe simulated HP H5N1 AIV dispersal from the Camargue , accordiIn these simulations we also investigated three hypothetical contamination scenarios at stopover sites on migrations, as a function of the number of infectious virus units present in halt meshes. We assumed that when a simulated bird left the Camargue in an infected condition, it left an infectious virus unit in each halt mesh it used on its way. This infectious virus unit persisted according to the virus inactivation rate in water. For simplification we did not considered the infective dose needed to initiate an infection but considered that an infectious virus unit always led to infection in a non-infected incoming duck. This assumption overestimated the infection risk at stopover sites.When this stopover contamination risk was low , HP H5N1The results provided by simulations of duck movements through our individual-based spatially explicit model were in accordance with studies performed on the ecology of this species in Europe The first important aspect highlighted in this study is the need to consider AIV dispersion through wild bird movements according to the period of the year and their biological cycle. For Common Teal, patterns of virus spread are not the same during the wintering period and during spring migration. During winter, local ecological factors such as food availability, hunting or climatic conditions induce local duck movements Experimental studies performed on captive-reared ducks often report short viral excretion duration, sometimes with important variability between viral subtypes, bird species and individuals In the present model, we assumed that simulated ducks did not get infected during their movements (except for HP H5N1 simulations). Integration of stopover contamination risk requires a precise knowledge of local AIV circulation , but also of ecosystem functioning , virus inactivation rate in the environment This study underlines the key role of virus inactivation rate in water, not only at a local scale, but also as an important component of AIV spread over long distances and periods. For many years, water-borne transmission has been suspected to be an important component of AIV epidemiology in wild as well as in domestic ducks Furthermore, the local prevalence of AIV infection did not have an effect on the distance of virus dispersal outside our study site. In our simulations, high and constant prevalence of infection in Common Teal (15%) over time led to higher quantities of AIV at infected stopover sites and thus, are likely to increase stopover contamination risk during duck migrations. However, such high and constant levels of AIV prevalence of infection before spring migration have never been observed in the Camargue 2. Increasing contamination risk did not affect the efficiency with which HP H5N1 spread. In our simulations, high stopover contamination risk corresponded to a 100% probability of infection if one infectious virus unit was present per 2 500 km2, which is very unlikely from a biological point of view.The spread of the HP H5N1 from Asia to Europe and Africa has been the subject of an intense debate focusing of the role of wild migratory birds e.g. . BecauseDue to the rapid inactivation rate of HP H5N1 in water Cygnus columbianus bewickii) infected with LP H6 AIV. Latorre-Margalef et al. Anas platyrhynchos) than in uninfected ones, and that the amount of virus shed by infected juveniles was negatively correlated with body mass. These studies suggest that host physiology and behaviour may be affected by LP AIV viruses in subtle ways not previously envisaged, but they also raise the question of potential subtype and species related specificity linked with AIV dispersal in the wild.In this study, we assumed that infected wild ducks were not subject to behavioural modification of their movement abilities, in order to measure the maximal AIV spread. To date, physiological and behavioural effects of AIV infections in wild birds have been little studied in nature. However, van Gils et al. Long-term population studies of Common Teal have been performed in the Camargue (Southern France) between January 1952 and February 1978. In total, 59 087 ducks were ringed  among which 9 279 were recovered in Europe, mostly through hunting. In this study, we focused on migratory and wintering movements of Common Teal from this area. We considered only intra-annual ring recoveries (i.e. recovered the same season as the ringing event occurred) to avoid potential biases linked with dispersal from other wintering areas. Indeed, a bird ringed in the Camargue during a given winter may not necessarily return to the Camargue the following winter The proportion of Common Teal leaving the Camargue each month was determined based on ring recoveries. For a given month, two types of ring recoveries where distinguished: (i) recoveries performed less than 100 kilometres (km) from the ringing place, assumed to represent birds staying in the Camargue, and (ii) recoveries performed outside this area, corresponding to birds leaving the Camargue. For the later recoveries, we defined whether these flights were local (i.e. between wintering sites) or migratory. According to the biology of this species we considered that all flights more than 100 km from the ringing place, realized between September and January, corresponded to movements between wintering sites Mean abundance of Common Teal in the Camargue was computed between 1964 and 1995  were selected. We assumed that each simulated bird leaving the Camargue had a well defined destination mesh, in another wintering site. Two scenarios were thus considered: (i) the destination mesh was situated less than 300 km away from the Camargue and the simulated bird was able to reach it directly (i.e. in one single day) or (ii) the destination mesh was situated more than 300 km away from the Camargue and at least one intermediate mesh should be defined (corresponding to a stopover for the bird). In the latter case, the intermediate mesh was determined as for migratory movements. We randomly chose a flight distance in a uniform distribution between 100 and 300 km with an interval of ±50 km. An interval of ±20° around the flight direction (flight direction being the azimuth from the present bird location to the destination mesh) was also determined. In the same manner as for migratory movement, an accessible (A) area was thus defined in which a preferred (P) mesh was determined. The simulated bird movement stopped when the individual reached its destination mesh.AIV dispersal by Common Teal was included in the modeling exercise according to three parameters: (i) monthly prevalence of AIV infection in this duck species in the Camargue, (ii) viral excretion duration and (iii) virus inactivation rate in water.We sampled 799 Common Teal from September 2007 to February 2008. Freshly killed birds were sampled in seven private hunting marshes (85% of our sampling). Live birds were also caught daily with funnel traps placed at the periphery of wintering marshes in the private natural reserve of “La Tour du Valat”. Cloacal swabs were performed to collect fecal samples and birds were marked with a steel ring before being released.5′-CTT CTA ACC GAG GTC GAA ACG-3′ and M/253/− 5′-AGG GCA TTT TGG ACA AAK CGT CTA-3′5′-[FAM]-CCT CAA AGC CGA GAT CGC GCA-[BHQ1]-3′, using the following cycling conditions: 15 min at 45°C, 3 min at 95°C, then 10 s at 95°C, 10 s at 55°c and 20 s at 72°C repeated fifty times and finally 30 s at 40°C. Positive samples were tested for highly pathogenic HP H5N1 AIV using a q-RT-PCR technique and molecular sequencing of the hemagglutinin cleavage site.Cloacal swabs were collected using the Viral Pack kit  and kept at −80°C until RNA extraction was performed. Automatic RNA extraction was performed using the BioRobot Mdx workstation and QIAamp Virus BioRobot MDX kit (QIAGEN GmbH) according to the manufacturer's instructions. The presence of influenza viruses was detected by quantitative real-time polymerase chain reaction (q-RT-PCR) targeting the Matrix gene segment, using a LightCycler 480 (Roche). Amplification was performed on 2.5 µL RNA with SuperScript III Platinum One Step Quantitative RT-PCR System (Invitrogen) in the presence of oligonucleotides (0.5 µM) M/52/+ Logistic regressions by General Linear Models  were fitted to the data to investigate the effect of the sampling period in infection status.When a simulated bird left the Camargue, its probability of being infected was assigned as the prevalence of AIV infection recorded during the month of its movement. To avoid potential bias linked with underestimated prevalences, we also performed simulations with a prevalence of infected birds equal each month to 15%. This value represents the highest infection rate recorded at our study site since we initiated AIV survey in wild birds in 2005 see .For viral excretion duration and virus inactivation rate in water, we investigated a large range of hypothetical values corresponding to possible biologically extreme cases. The following values were tested: viral excretion duration: 2, 7, 10, 15, 20, 25 and 30 days In our model, we assumed that simulated birds did not get infected during their movements. When a simulated bird left the Camargue being in a healthy state, it moved to its destination mesh (breeding area or other wintering site) without contamination at stopovers. This assumption is the main limitation in our model and led to the further assumptions that AIV circulation did not previously occur in halt meshes, or that infected birds became immunized and cannot be re-infected during their movements. Although the stopover contamination risk was not possible to predict in our general AIV model , we investigated the case of HP H5N1 dispersal, using fixed values of viral excretion duration and virus inactivation rate in water, corresponding to those provided in the literature.We investigated dispersal of HP H5N1 AIV by Common Teal by considering a hypothetical circulation in the Camargue and integrating recent data concerning viral excretion duration and virus inactivation rate in water. For viral excretion duration we used the maximal excretion length (6 days) recently provided by Keawcharoen et al., on European Common Teal Because excretion duration and inactivation rates in water were fixed for HP H5N1, we were able to investigate a hypothetical contamination risk at stopovers. We assumed that when a simulated bird left the Camargue in an infected condition, it left an infectious virus unit in each halt mesh it used on its way. This infectious virus unit persisted according to the virus inactivation rate in water. We thus considered for each mesh a stopover contamination risk depending on the number of infectious virus units in each mesh (i.e. the number of infected birds which had stopped in a given mesh before and the virus inactivation rate). We considered that an infectious virus unit always led to infection in a non-infected bird.2 (i.e. 25 000 infectious virus units per mesh), (ii) medium contamination risk  and (iii) a high, or extreme contamination risk .We defined the stopover contamination probability as a linear function of the number of infectious virus units present in each mesh at the arrival date of a simulated bird. The more infected birds use a given mesh, the higher the density of infectious virus units and thus stopover contamination probability is higher. We tested 3 hypothetical scenarios, corresponding to 3 densities of infectious virus unit thresholds in halt meshes: (i) low contamination risk: a simulated healthy bird gets infected if the density of infectious virus units in the halt mesh is equal to 1 per 100 000 mFigure S1Avian influenza virus prevalence in Common Teal in the Camargue, during winter 2007–2008 .(0.02 MB PDF)Click here for additional data file.Figure S2Effect of local prevalence of infection on AIV dispersal during the wintering period  and spring migration . Maps represent AIV circulation with prevalence measured during the 2007–2008 season  and with a hypothetical constant monthly prevalence of 15% . Colours represent the maximum number of infectious virus units per mesh: intermediate blue: 1, 2 to 10, 11 to 49, 50 to 99, 100 to 200; black: more than 200 infectious virus units.(7.57 MB EPS)Click here for additional data file.Figure S3Common Teal ring recoveries (September to May) recorded in Europe between January 1952 and February 1978. Green scale color represents the number of ring recoveries recorded on each mesh: light green: 1 to 4; intermediate green: 5 to 29; dark green: more than 30 ring recoveries. The white star represents the geographic location of the Camargue.(2.17 MB EPS)Click here for additional data file.Figure S4Mean abundance (and standard deviation) of Common Teal in the Camargue, computed between 1964 and 1995.(0.02 MB PDF)Click here for additional data file.


Serum samples from 22 hantavirus pulmonary syndrome (HPS) patients were tested for Sin Nombre virus (SNV)-reactive antibodies. In the acute phase of HPS, 100% and 67% of the samples tested positive for SNV-specific immunoglobulin (Ig) M and IgA, respectively. Among the virus-specific IgG antibodies, the most prevalent were IgG3 (in 97% of samples), followed by IgG1 (70%), IgG2 (30%), and IgG4 (3%).


Adequate self-care in diabetes improves quality of life and decreases the number of inpatient cases. The health locus of control theory is used to assess adherence to diabetes regimen in some studies in developed countries. The primary purpose of this cross-sectional study is to determine the status of diabetes locus of control in a sample of diabetic patients in Iran as a developing country. We investigated selected factors contributing to locus of control and adherence to diabetes regimen.This cross-sectional study was carried out on 120 patients referred to Yazd Diabetes Research Center. The Iranian versions of Diabetes Locus of Control scale and Diabetes Self-care Activities scale were used for data collection.Men revealed more internal locus of control and women revealed more chance locus of control. The attributions of external locus of control increased by age, while the internal locus of control increased by education level and chance locus of control decreased by education level. A positive association between internal locus of control and adherence to diabetes regimen was found and there was a negative association between chance locus of control and adherence to diabetes regimen.Findings suggest that interventions aimed at improving internal locus of control may improve adherence to diabetes regimen but different diabetic patients have different attribution styles and interventional programs to enhance diabetes self-care will be more successful if patient's locus of control is addressed. The diabetes regimen is extremely complex. It is geAdherence has been defined as the degree to which a patient's voluntary behavior corresponds with the clinical recommendations of health care providers. AdherencVarious psychosocial variables have been previously examined to determine their influence on adherence to the diabetes regimen. Predictors such as personality, family behaviors, health beliefs, demographic characteristics and beliThe locus of control theory was developed by Rotter. The concRotter's Social L14Health locus of control is defined as a generalized expectation about whether one's health is controlled by one's own behavior or forces external to oneself. Health lThe health locus of control theory is used to assess adherence to diabetes regimen in some studies. According to Rodin, an indivet al.[et al.[In a study, Macrodimitris et al. examinedl.[et al. investigAlthough the above studies have shown an association between LOC and adherence to diabetes regimen, some others have found no link between them.23–25 MorThis was a cross-sectional study. The non-probability sample consisted of 120 diabetic patients at Yazd Diabetes Research Center (DRC) in central Iran.A recognized diabetic patient for at least three months,Patient having a medical file at Yazd DRC,Able to speak Farsi andNo severe mental disorders and Alzheimer's disease.t-test, ANOVA with Tukey's HSD post hoc and linear regression with Enter method.A power analysis showed that 100 was the sufficient sample size to achieve a power of 0.80. To ensurThe survey instruments consisted of a demographic data form, Diabetes Locus of Control scale and diabThe Diabetes Locus of Control scale was deveThe instrument was then pilot-tested with a group of diabetic patients (n = 30) to collect data to examine the internal consistency of the scales. Specifically, Cronbach's Coefficient Alpha was computed for each of the scales. The reliability coefficients were 0.76, 0.67 and 0.79 for the internal locus of control, powerful others locus of control and chance locus of control scales, respectively. For the actual study, these indices were 0.80, 0.65 and 0.82, respectively.Adherence to regimen was measured, using the Diabetes Self-care Activities scale. This meaThe participants circle how many of the past seven days they have adhered to their prescribed regimen on each of the above behaviors. Mean scores are collected for each self-care behavior and a total adherence score can be obtained by summing the mean subscale scores. In this The 120 study participants ranged in age from 17 to 73 . Majority (60.8%) were female, all were married. Their education levels were 33.3% illiterate, 17.5% had reading and writing ability, 27.5% had primary school education and 21.7% had higher than primary school education. Most of the participants were housewives (60.8%). The overwhelming majority of the participants (82.5%) had type 2 diabetic and the rest had type 1 diabetic. The duration of diabetes ranged from three months to 30 years, with 9.8 as the average age and 6.8 as the standard deviation.The mean score for Adherence to regimen was 48.4 (SD = 10.0). With regard to the LOC dimensions, Internal LOC solicited the highest score with an average of 26.6 (SD = 3.2), followed by Powerful others LOC and chance LOC with the means of 23.2 (SD = 2.6) and 9.4 (SD = 6.6) respectively.t-test for independent samples showed that 1) men significantly outscored women on the basis of internal locus of control, 2) women significantly outscored men on the basis of chance locus of control and 3) gender differences on the basis of powerful other locus of control were not statistically significant. None of the type of diabetes differences on the basis of locus of control scales scores was statistically significant. A series of one-way analysis of variance showed that type of job differences on the basis of internal and chance locus of control scales scores were statistically significant, and in both cases, Tukey's HSD post hoc procedure showed that the differences between housewives and self-employed were statistically significant. Results are summarized in A series of Pearson Product Moment Correlation Coefficient (Pearson r) was used to describe the magnitude and direction of the bivariate associations between Adherence to regimen scores and locus of control subscales scores. Results are summarized in P < .05) and a statistically significant negative correlation between chance locus of control and level of education Spearman rank order correlation coefficient showed a statistically significant positive correlation between internal locus of control and level of education . Additionally, when type 1 diabetic patients were excluded, statistically significant correlations between adherence to regimen and internal and chance locus of control were observed .Regression analysis was performed to explain variation on adherence to diabetes regimen on the basis of internal and chance locus of control. As powerful others locus of control was not significantly associated with adherence to diabetes regimen, it was not included in regression analysis. The two variables together accounted for 9.8% of the variation. However, internal locus of control was the only statistically significant predictor of adherence to diabetes regimen. Results are summarized in This study evaluated the status of diabetes locus of control among diabetic patients and its related factors and also quality of its association with adherence to diabetes regimen in a cross-sectional design in Iran as a developing country.Subjects displayed internal locus of control, followed by powerful others and chance locus of control. These finding indicate that the participants considered themselves to be the greatest influence on their adherence to diabetes regimen. The findings, are similar to some studies in developed countries.32–33et al.[22The study's male participants demonstrated more internal locus of control, whereas the females displayed evidence of chance locus of control. Aalto and Uutela did not et al. the younet al.–2235 in Despite the cultural and economic differences between developed and developing countries, it seems that there is not major difference regarding the status of LOC as well as its role in determining adherence to diabetes regimen. Low level of prediction of variances in adherence to diabetes regimen by LOC attribution style indicated that the phenomenon of adherence is extremely complex and other psychosocial variables should be examined together with LOC to compare their influence on adherence to the diabetes regimen.Due to correlational nature of the study, the reader is cautioned that no causal inferences are drawn. Moreover, the study population had low education level and most of them were housewives. Therefore, the results may not necessarily apply to all populations in developing countries. We recommend further studies in developing countriesThese findings suggest that interventions aimed at improving internal locus of control may improve adherence to diabetes regimen but different diabetic patients have different attribution style. Counselors and educators should attend to the locus of control in their interventional courses and programs. The following activities will enhance internal locus of control attribution and could be used in interventional programs.Providing situations which may encourage diabetic patients for adherence to regimen.Providing positive feedback to patients for their small successes, as any feeling of success may make them feel that they are in control of their illness.