Diarrhea prevalence increases from around the time that complementary foods are introduced. Improving caregiver’s hand hygiene during food preparation could reduce complementary food contamination and enteric pathogen transmission. Washing hands with soap is more common when water and soap are together at a convenient location. We conducted a three-month pilot intervention to evaluate two options for setting up handwashing stations: i) provide a handwashing station, or ii) help the family to make their own from available materials. Additionally, we assessed the feasibility of this intervention to be integrated with a child feeding program.We conducted the intervention among two groups; 40 households received a free of cost handwashing station and another 40 households were motivated to place their own soap/soapy-water and water vessel near the food preparation and child feeding area. Community health workers encouraged caregivers to wash hands with soap/soapy-water before food preparation and feeding a child. They either assisted study participants to install the study-provided handwashing station at the recommended place or encouraged caregivers to develop their own. Field researchers assessed placement and composition of handwashing stations and the feasibility of integrating handwashing and nutrition messages.By end of the trial, 39/40 households developed their own handwashing station, comprising a bucket, mug and bar soap/soapy-water of which 60% (6/10) households were observed with a functional and complete handwashing station set. Observed handwashing with soap was detected among 8/10 households from the study-provided handwashing station group and 5/10 among households who had made their own handwashing station. Sixty-seven of the 76 caregivers recalled integrated intervention messages on social and health benefits of infant and young child feeding correctly; and all recalled key handwashing with soap times, before food preparation and feeding a child.Encouraging households to develop their own handwashing station with soap and water to place at a food preparation/child feeding location is feasible over the short term. In the absence of large-scale provision of handwashing stations, caregivers can be encouraged to create and use their own. Integrating handwashing with soap into a nutrition intervention was feasible and acceptable and should be considered by policy makers. Diarrhea and pneumonia continue to be leading causes of death in low income countries , 2 and bChildren in rural Bangladesh whose mothers were observed to wash hands before preparing food, had less diarrhea over the next 2 years compared to children of mother who did not wash hands during food preparation . HoweverWashing hands with soap is more common when water and soap are together at a convenient location . In BangWe conducted a pilot study to compare the effectiveness of two different strategies that integrated handwashing with soap into a nutrition intervention for the improvement of complementary feeding practices: one group received a study-provided model handwashing station positioned near the area for feeding children and the food preparation area; the other group was encouraged to develop their own handwashing facilities. We also evaluated the feasibility of integrating handwashing with soap and the nutrition intervention.upazilas (sub-districts) from the two districts from the Bangladesh Population Census 2001 [Pouroshova) from the upazila list and, in order to select our sample proportional to the population, we randomly selected one upazila from Manikgonj district and two upazilas from Dinajpur. From the three upazilas we randomly selected four unions . From each of the four unions we randomly selected one village; thus 4 villages in total were selected from the 3 upazilas based on the population size. Within each selected village the field workers identified eligible households based on the criterion of having a child aged between 6 and 23 months who was being fed complementary food. Twenty households from each of the 4 villages were selected through a computer generated random number; 80 households in total.This behavior change intervention trial was conducted in two rural districts; Manikgonj  and Dinajpur (northern Bangladesh) Fig. . Two sitsus 2001 . First wAlive & Thrive worked to develop a scaled-up intervention for preventing child undernutrition by improving infant & young child feeding (IYCF) practices . Alive &Alive & Thrive recruited CHWs to implement the program activities. CHWs belonged to the same community as the primary audience groups and were accepted by the community. They were at least 25 years of age, with those with some experience in delivering health messages were prioritized. Selected CHWs participated in a 3 day training program on IYCF, handwashing and handwashing stations, home visits, and community meetings and forums. A one day practical session involved demonstrating how to construct a handwashing station (with both water and handwashing agent placed together). All eligible households received behavior change promotion about complementary feeding practices and handwashing with soap before food preparation and before feeding the child. The intervention behavior recommendations were: (i) wash hands with soap/soapy-water after defecation, cleaning a child anus, before food preparation and before feeding a child, (ii) maintain and use handwashing station near the food preparation and child feeding area, and (iii) follow the recommendations of age specific quantity, frequency and varieties of food in addition to continued breast feeding.For 40 households field staff provided a model handwashing station that comprised a 40 l plastic bucket with tap and lid, a bowl to collect residual rinse water and a stool to use as a stand for the bucket. We also provided a 1.5 l plastic bottle and detergent powder free of charge to make and store soapy-water , 24  and encouraged use. They also demonstrated soapy-water making in these households. CHWs encouraged mothers in the self-made handwashing station group (self-made HWS group) to develop their own handwashing station from available household resources and demonstrated soapy-water making. CHWs asked caregivers to place some type of water vessel but didn’t suggest the need to install a tap as part of their own handwashing station. They encouraged households to place either bar soap or soapy-water near the water vessel Fig. . CHWs woAs part of the integrated IYCF intervention, CHWs distributed handwashing stickers and placed them near the handwashing stations Fig.  as a remTV advertisements that encouraged mothers to wash hands with soap and water before food preparation and child feeding, asked to put water and soap near the place, and messages on complementary feeding were broadcast throughout the intervention period between late September and mid-December, 2011.A team of data collectors (field researchers) from icddr,b, separate from the CHWs, conducted qualitative assessments including interviews, observations and group discussions over three rounds at days 20, 58 and 83 after the intervention commenced.The field research team conducted informal discussions with caregivers at each visit and from each study group. They took detailed notes for each question asked to document their responses. When appropriate, the field researchers included verbatim quotations in the notes that illustrated a particular perspective. The interviewer asked open-ended questions to all eligible caregivers about their perceptions and practices regarding complementary feeding of the child aged 6–24 months, handwashing with soap at the recommended key times, regular use and maintenance of the handwashing station near the food preparation and child feeding area(s), and motivations and barriers to using the handwashing station they received or made themselves.Field researchers conducted unstructured observations in 20 randomly selected households (10 households from each HWS group) from within the study population during each of the three follow-up visits. Randomization was done by the lead investigator at icddr,b through computer generated random numbers. Field researchers observed caregiver’s handwashing practices before food preparation and before feeding a child and the use of handwashing stations and soap/soapy-water. Each observation took place from early morning (6 AM) and each session lasted for 2–4 h to cover both recommended key times; e.g. food preparation and child feeding by the family. Field researchers took detailed notes to record actual behaviors.Field researchers conducted 12 group discussions, four at each of the assessment times with equal numbers of family members from each handwashing station group and included one to two elders from the selected households. During the group discussions field researchers discussed family member’s perceptions about handwashing with soap, their experience with handwashing stations and their intention to practice the recommended behaviors.Field researchers used an open-ended questionnaire for data collection at each assessment. The data from group discussions were audio recorded and transcribed. Field researchers conducted interviews and discussions in Bengali. On the day of data collection, field researchers translated data to English and entered them into a table with pre-designated headings using Excel, based on the questionnaire. For each question the field researchers included as much information as possible in the table, and we (led by DB and FN) determined the number of persons providing these responses. This was done by including a column for counts . On subsequent data collection days, if a response category did not exist for a question, the field researchers added this to the listed responses. We analyzed the interview and group discussion data according to the different behavioral determinants based on the study objectives, at the community level, for handwashing to identify the number of caregivers who were practicing the recommended behavior.We considered regular handwashing practice (observed) as instances when the household caregiver washed their hands at each time point of the total handwashing opportunities that occurred during the observation period. Similarly, if a complete handwashing station (both water and soap/soapy-water) was found near the recommended places during the observation period and the caregiver used the handwashing station to wash her hand at least once, then we considered this household as a regular user of handwashing station.We also summarized respondent recall of the IYCF messages for children aged 6–24 months that they heard during the intervention period. Findings from informal interviews, group discussions and observations were compared for consistency and were triangulated. We summarized and compared the responses from the study HWS group and the self-made HWS group. Through comparison and summary, the analysis identified key areas of behavior change and intervention uptake.p = 0.16).Informal discussions were commonly conducted with mothers, the primary caregivers; 93% (74/80) during the first assessment, 98% (78/80) during the second assessment and 95% (76/80) for the final assessment. The average age of respondents was 24 years in the study HWS group and 23 years in the self-made HWS group. The reported average household income was lower in the study HWS group  than for the self-made HWS group   visits our para (neighborhood) very frequently and observes whether I placed water or not, even if somehow I met her outside my home she has just one question to me that whether I am using that bucket and washing my hands or not. So it helps me a lot to become habituated day by day". One barrier for developing a self-made handwashing station was lack of available resources in the households. Some caregivers reported having old vessels that they thought were not suitable for storing water and they used these functional buckets for other purposes. Due to financial difficulties they were unable to buy new vessels to use for handwashing.By end of the trial, 39/40 households developed their own handwashing station (both water vessel and soap placed together). The self-made handwashing station developed by the community members often comprised an existing household bucket for water storage, a jug or mug to pour water over hands, and a plastic bottle for making and storing soapy-water  of caregivers in the study-HWS group and 50% (5/10) from the self-made HWS group at the final assessment. Although there is increasing trends of handwashing with soap in the study-HWS group, greater than for the self-made HWS group, the small sample size did not permit a rigorous statistical assessment. During group discussions and interviews, for both HWS group categories, study participants explained that the main motivation to wash hands with soap was child’s health and having water and soap in one place near the food preparation and child feeding area facilitated the practice. Mothers explained that nutritious food that they were providing to their children will not provide nourishment if they do not wash their hands before feeding their child. For example one mother from the self-made HWS group said- "At the final assessment 90% (9/10) of households in the study-HWS group and 60% (6/10) households in the self-made HWS group were observed to have a complete handwashing station (both water and soap/soapy-water) that was functional and caregivers used at least once during the observation period. However, soapy-water was detected among 100% of the observed households in both HWS groups.handwashing station makes my handwashing easier because we get water and soap near my cooking place and no need to go to the tube well frequently. I can wash my hands by sitting at my cooking place.” Another caregiver said “We have no tubewell at our home. Before getting this handwashing station we had to go outside  for washing our hands and utensils. Moreover many households use one tube well and no one keeps soap beside tube well. Now we get water and soapy-water together and we don’t need to go outside and we can wash our hands whenever we need".Respondents from both study groups reported that a handwashing station was easy to use, and that the presence and placement of the handwashing station motivated people to regularly use and maintain it, encouraging handwashing at promoted times. Study participants explained that having water and soap in one place near food preparation and child feeding areas reduced the need to go to a distant, inconvenient location to wash hands with soap. For example one caregiver explained that “Similarly, caregivers in the self-made HWS group described the advantages and benefits of keeping water and soap together and close at hand as they didn’t need to go far if they maintain a stocked handwashing station near the food preparation and child feeding area(s). Community members of both intervention groups reported a preference for soapy-water over bar soap as it is easier to use than bar soap.During the interviews caregivers in the study HWS group also reported that they liked the appearance and free provision of the handwashing station and that it acted as a reminder to wash hands.we could not set up a tap with our water vessel by ourselves because we need to spend money to buy a tap to hire a mechanic to set it on the bucket. Also the tap can become cracked after a certain moment".Study participants reported some strengths of the handwashing stations that influenced regular use. Study participants in the study HWS group explained that the tap on the bucket increased convenience in providing running water while washing their hands. They reported that the station facilitated handwashing among children providing readily available water by turning the tap, compared to the alternative where they need to pump water from tube wells, which can be strenuous and thus act as a barrier to handwashing. Although, in the self-made HWS group nobody had a tap on the water vessel they arranged, they used a mug to pour water over hands. Study participants reported that this arrangement had an advantage of the tap. They explained that, "The bowl with the study-provided bucket was considered a strength as it held residual rinse water and reduced the risk of water remaining on the floor. Many participants in the self-made HWS group placed a bowl at their own handwashing station for this purpose. In addition, the lid on the study provided bucket kept water clean and the study participants reported that children can’t waste water when the lid is kept locked. They also reported that the lid enhanced safety by preventing children from falling into the water. On the other hand, lack of a lid for the self-made handwashing stations acted was considered a weakness. Study participants from the self-made HWS group mentioned that their handwashing water vessel that was uncovered encouraged children to play with the water. None from the self-made HWS group used an existing bucket that had a lid. Caregivers feared that playing with water may cause a child to develop a cough or a cold. They also described a risk for young child to fall into the uncovered water vessel. Although, the main reported barrier for both intervention group categories was wastage as children frequently play with the water and soap/soapy-water which was more frequently reported by respondents from the self-made HWS group.My son and his father can also wash their hands from this handwashing station. Now we don’t need to go outside at night for washing hands and utensils.” However, in the self-made HWS group field researchers didn’t observe any water vessel of a similar size to the bucket provided with model handwashing station. Study participants from both groups explained that the handwashing station was convenient for other purposes related to cooking and feeding/eating, not just for handwashing.Participants from the study HWS group liked the large size of the bucket they received, so that they could store water for all household members for a full day. They reported that they kept the handwashing station inside their living room at night so that they can wash their hands whenever necessary. For example one mother said, "“Our tube well is located so far and in an open place. If I keep soap (bar soap) in that place, sometimes a crow will take away the soap, and sometimes it becomes spoiled. But the soapy-water is better than soap. It’s easy to use, and is less likely to be damaged.” Caregivers in the self-made HWS group reported that soapy-water was inexpensive to make; most of the observed households from this group made their own soapy-water, even though they didn’t receive a bottle or detergent. For example, one caregiver from Dinajpur said, "Soap (bar soap) is expensive. Detergent powder remains available in my home all the time for washing my clothes; and I can use a small amount for making soapy-water. So I don't need to expend extra money for making soapy-water. Even some of my neighbors prepared and are using soapy-water by their own initiative even though they don't have children under 2 years of age (were not part of the intervention)." This was detected during the final assessment, when field researchers observed neighbors that were not included in promotion activities prepared their own soapy-water.Another strength of handwashing intervention was provision of soapy-water. Dispensing soapy-water from the bottle with a hole in the cap onto the palm of their hands reduced the duration of handwashing and this product was less likely to be wasted by children. For example one mother from Manikgonj said, "we are giving good food to my child for his good health, but if we do not wash our hands before feeding him then the food will not work at all".The majority of participants (caregivers) recalled intervention messages on social and health benefits of infant and young child feeding correctly. They described appropriate frequency, amount and variety of complementary food and how this helps a child’s physical and mental growth. They also reported the health risk if they did not follow recommended practices. In addition to the IYCF messages, all of the caregivers recalled the importance of handwashing with soap before food preparation and feeding a child. One caregiver explained that In Bangladesh, preparing foods and feeding a child with bare hands is common. Food preparation and feeding a child with unwashed hands can contaminate food and can be a source of diarrheal pathogens , 26. A sAmong households from the self-made HWS group, more than half of them were observed to have a handwashing station at a place convenient to cooking and child feeding even when no products were provided. However, those who received study provided handwashing stations were more likely to have them placed convenient to cooking and child feeding. We observed that many mothers who were not supplied with hardware were sufficiently motivated to develop and place their own handwashing station and prepare soapy water. It is likely that having a water source close to the house, near the cooking/feeding area(s) increased convenience, not only for handwashing but for other domestic tasks. Soapy-water was a popular handwashing agent as it was viewed as being less expense than bar soap, easy to maintain and less likely to be wasted, as described among hardware recipients previously .Among the self-made HWS group, many of the caregivers used hardware that was not dedicated for handwashing and could be removed for other purposes thereby leaving the cooking area without a water source. Engaging male members of the family while explaining the importance of maintaining the handwashing stations in the specified location is an important step. Future interventions that promote development of handwashing stations from households’ resources could address this by providing guidance on hardware composition and stressing the convenience of dedicating the hardware for handwashing and food preparation. The handwashing stations developed by household members did not have a lid and participants described their concerns of having open buckets of water in the household area and the danger that presented. A lid can address concerns about the potential for water wastage, concerns about children catching a cold and prevent children falling into the water. Future program should include not only dedicated devices for self-made handwashing station, but promoting the advantages of a lid and encourage purchase and use of a cover to reduce caregiver concerns. The tap, while useful, was not seen as essential, therefore a bucket with lid and jug would be a low cost, readily available model. Also, lack of facilities to capture waste water made the courtyard wet and muddy, a potential reason for lower observed handwashing with soap in the self-made HWS group. Hence, recommendation for a container to collect waste water should be included as a component of self-made handwashing station. However, an encouragingly large number of households were observed to place self-made handwashing station and soapy water bottles through behavior change promotion. Preparing and placing a soapy-water bottle by those outside the promotion audience indicates that this is likely a feasible solution for keeping soap near water (for other key handwashing times such as post defecation), and near cooking and feeding areas.Stunting is a marker of chronic malnutrition and is associated with an increased risk of diarrhea. Integration of handwashing into a nutrition program has the potential to prevent diarrheal diseases. Our study suggests that it is possible to integrate handwashing messages into an IYCF intervention. The majority of the study participants understood the importance of complementary food and could connect the link between unwashed hands and food contamination which ultimately increased their handwashing behavior. We suggest further research to investigate the potential impact of an integrated handwashing intervention and child feeding intervention on the occurrence of diarrhea.Our study also suggests that CHWs have a positive impact on improving handwashing behavior. This finding is consistent with other studies that concluded that CHWs can improve health-service use and child health outcomes . HoweverA limitation of the study is that we conducted the study in only two rural sites, however, these were typical of rural Bangladeshi communities. As it was a pilot study it was conducted on a small scale for a short period, yet it provided useful insights for future interventions. Sustained practice can be assessed during longer duration trials.Encouraging caregivers to create and use their own handwashing station was feasible and effective in increasing handwashing behavior before food preparation and before feeding a child. In the absence of large-scale provision of handwashing stations, caregivers can be encouraged to create and use their own. Soapy water was a popular alternative to bar soap and should be considered for promotion at scale. Strategies to promote own handwashing station placement should includes motivation through behavior change communication, creating a designated place for handwashing at the recommended key times, through regular home visits by community health workers that provide supports in designing own handwashing stations by using available household resources. Integrating handwashing with soap into a nutrition intervention was feasible and acceptable and should be considered by policy makers.


After taking detailed medical history and performing physical examination including the PT, TT and CCT, all the data were gathered in a checklist for each patient. EDX studies were conducted and definitive diagnosis of CTS was based on the results (by EDX criteria). Patients were divided into two groups of CTS and non-CTS. Statistical analysis was performed using independent-sample t and chi-square tests via SPSS software . P-value of less than 0.05 was accepted as statistically significant. The sensitivities and specificities of the PT, TT and CCT were gained via comparison with the electrodiagnostic tests, as gold standard. Of 89 patients, CTS was diagnosed in 80.9%. In terms of gender, age and employment, there was no statistical significant difference between the CTS and non-CTS groups . The frequency of CCT-positive patients was statistically greater in CTS group (80.6%) than non-CTS one (47.1%) (P = 0.008). The frequency of the PT-positive patients was greater in non-CTS group (59.7 vs. 64.7%) with no statistically significant difference (P = 0.464). The calculated sensitivity and specificity were 80.6 and 52.9% for CCT, 59.7 and 35.3% for PT, and 65.3 and 47.1% for TT, respectively. In receiver operating characteristics (ROC) curve analysis, the area under the curve (AUC) was 0.67  for CCT, 0.56 for TT and 0.48 for PT (P > 0.050 for both) . In non-CTS group, there was a moderate negative agreement between the TT and CCT . There was a moderate positive agreement between the PT and TT in CTS group .4,5 Our results confirm the importance of CCT in diagnosis of CTS.In our study, TT and CCT were more positive in CTS group and PT was more positive in non-CTS group but just positive CCT was statistically different between CTS and non-CTS groups. ROC cure analysis showed the accuracy of CCT for diagnosis of CTS is higher than TT and PT. AUC for TT and PT were around 0.5 which means that these tests cannot help us in depicting patients with CTS. Previous studies reported contradictory results about the sensitivity and specificity of CCT, but in most of them, they were greater than the sensitivities and specificities of the TT and PT.One another point is that the moderate negative agreement between the TT and CCT in non-CTS group means that both the tests would rarely be positive among them. In other words, if both CCT and TT were positive, the diagnosis is more likely to be CTS. In addition, the positive agreement between the PT and TT in CTS group means that performing one of these tests is sufficient for patients suspected to suffer from CTS; because if one of them is positive, it is very probable that the other one is also positive and vice versa.In conclusion, as the sensitivity and specificity of the CCT are greater than those of the TT and PT, we recommend routine use of CCT for screening the patients suspected to CTS.


The lack of sauropod body fossils from the 20 My-long mid-Cenomanian to the late Campanian interval of the Late Cretaceous in Europe is referred to as the ‘sauropod hiatus’, with only a few footprints reported from the Apulian microplate (i.e. the southern part of the European archipelago). Here we describe a single tooth from the Santonian continental beds of Iharkút, Hungary, that represents the first European body fossil evidence of a sauropod from this critical time interval. The mosaic of derived and plesiomorphic features documented by the tooth crown morphology points to a basal titanosauriform affinity suggesting the occurrence of a clade of sauropods in the Upper Cretaceous of Europe that is quite different from the previously known Campano-Maastrichtian titanosaurs. Along with the footprints coming from shallow marine sediments, this tooth further strengthens the view that the extreme rarity of sauropod remains from this period of Europe is the result of sampling bias related to the dominance of coastal over inland sediments, in the latter of which sauropod fossils usually occur. This is also in line with the hypothesis that sauropods preferred inland habitats to swampy environments. Almost all of these sporadic remains, both skeletal elements and footprints, have been discovered in Cenomanian localities13 with some of these even possibly reworked from older, Albian sediments. Accordingly, the late Cenomanian to late Campanian time period, an approximately 20 My long interval14, was long thought to represent a hiatus in the European sauropod record15. The discovery of some Turonian-Coniacian sauropod footprints in Croatia16 and a trackway of a probable small sauropod from the Santonian of Italy17, however, seem to challenge this view, and suggest a sampling bias instead18, mainly due to the “rarity of inland sediments and dominance of coastal deposits” (Mannion and Upchurch1 2011:529) in the European Upper Cretaceous.Sauropod dinosaurs were important elements of different Late Cretaceous continental vertebrate communities in Europe. Their record comes, however, mainly from upper Campanian to upper Maastrichtian sediments, and only a very few isolated and fragmentary remains are known from older Upper Cretaceous depositsHere we report a sauropod dinosaur tooth from the Santonian of Iharkút, Hungary, an unexpected discovery that represents the first body fossil of the clade known from this poorly sampled period of the sauropod fossil record in the European Cretaceous.The isolated tooth (MTM PAL 2017.1.1.) described here was collected in the Iharkút vertebrate locality (western Hungary) and is housed in the Vertebrate Paleontological Collection of the Hungarian Natural History Museum, Budapest. The specimen was prepared mechanically in the lab of the Hungarian Natural History Museum and the fragmentary margins of the tooth were fixed by cyanoacrylic glue.19. Quantitative shape descriptors such as Slenderness Index 20 and Compression Index 2 were also calculated.The description of the tooth follows the dental terminology proposed by Smith and Dodson21. Bones at the site are accumulated in bonebeds, among which the most productive one  the tooth has exactly the same dark brownish colour  and general textural pattern as that of the other vertebrate remains from the site; 2) the pulp cavity is lined with a crust of early diagenetic pyrite, in a pattern that is characteristic for many teeth and bones from Iharkút, especially those that have extensive internal cavities, such as theropod and pterosaur bones27;3) the tooth is completely void of any signs of abrasion that would have eventually resulted from the interaction between sediment particles and tooth during reworking, 4) the tooth surface is pristine, well-preserved and shows ornamentation as well as features generated only by tooth-food contact. Taken together, these taphonomic features indicate that, similarly to the other teeth and bones preserved in site Sz-6 from Iharkút, the primary depositional setting of MTM PAL 2017.1.1. is represented by the bone-yielding beds of this site.Since this tooth represents the only indication of sauropod dinosaurs in Iharkút up to now, it raises the question whether this specimen might have been reworked from older deposits, as teeth are known to survive relatively long-distance transport and reworking without significant damageTeeth of almost all dentulous vertebrate taxa discovered at Iharkút (from fish to enantiornithine birds) are known from the locality, and MTM PAL 2017.1.1. differs markedly from all of these 30 are not present on the preserved part of the crown, making the assessment of tooth-tooth occlusion details impossible. The specimen displays a mosaic of basal and advanced dental features within Eusauropoda. It retains the lingual concavity and a D-shaped cross section, but the tooth crown is narrow and not markedly expanded relative to the root, the labial grooves are absent, and no denticulate mesial and distal margins are present.Among sauropods, the tooth MTM PAL 2017.1.1. can be referred to eusauropods based on the possession of a concave lingual surface and a D-shaped crown cross-sectionDiplodocus32, and the spatulated, mesiodistally wide teeth of non-titanosauriform eusauropods 33, although the crown curvature in mesial/distal view and the lingual concavity are similar to those seen in Mamenchisaurus34. MTM PAL 2017.1.1. is similar to a brachiosaurine tooth from the Lower Cretaceous of Galve, Spain35 in having a D-shaped cross-section, concave lingual surface, and parallel, non-carinated mesial and distal margins, although the details of the crown curvature differ slightly. The general form and cross-section of the crown is reminiscent of the premaxillary teeth of the Early Cretaceous North American brachiosaurid Abydosaurus23 as well. Some similarities can also be pointed out with the teeth of somphospondylan Euhelopus38, and those of some indeterminate basal titanosauriforms from the Lower Cretaceous of Japan39 that also have parallel-sided crowns with concave lingual surface and relatively low SI values. Nevertheless, they differ from MTM PAL 2017.1.1. in their simple lingual apical curvature, as well as in the presence of a midline ridge within the lingual concavity and of basal lingual buttresses. On the other hand, the tooth markedly differs from the subcylindrical or cylindrical teeth of derived lithostrotian titanosaurs such as Rapetosaurus40 or Nemegtosaurus42 in having a much lower SI value and a morphologically more complex crown. Indeed, according to the character list of Mannion et al.43, the Hungarian tooth does not represent a lithostrotian, since it lacks synapomorphies of this clade such as the high-angled planar wear facets (C105) and the cylindrical tooth crown (C109) with a convex lingual surface (C110). The only lithostrotian character present in MTM PAL 2017.1.1. is the absence of an apicobasally orientated lingual ridge (C111).The tooth differs from the peg-like teeth of diplodocoids, such as Ampelosaurus, Lirainosaurus, Atsinganosaurus, Lohuecotitan, Magyarosaurus, and Paludititan), among which the first three genera preserve teeth as well44, and further isolated, indeterminate titanosaur tooth morphotypes are also reported from different localities from Spain24, southern France45 and western Romania (pers. observ.). Isolated titanosaur teeth from the Haţeg Basin, Romania, possibly referable to either Magyarosaurus or Paludititan, are very simple, cylindrical and peg-like, with a mildly convex lingual surface and a high SI value (~5) making these markedly different from the Iharkút tooth. The single known tooth referred to Ampelosaurus, and found in a bonebed from southern France47, is labiolingually flattened, mesiodistally expanded with mesially and distally positioned longitudinal grooves, again, being clearly distinct from MTM PAL 2017.1.1. Whereas the French taxon Atsinganosaurus has gracile, spatulate teeth with a cylindrical crown and mesial and distal ridges extending from the apex to the middle of the crown, the teeth of Lirainosaurus from northern Spain are simple cylindrical with a circular cross section48 - both of these morphologies are also very different from that of the Iharkút specimen. Besides these three Iberoarmorican taxa, Díez Díaz and colleagues24 described two additional morphotypes from the Spanish locality of Lo Hueco. Among them, ‘morphotype B’ is more similar to the Iharkút tooth in having mesiodistally parallel sided crown and shallow ridge-like margins mesially and distally; however, crown curvature and cross section are different. Finally, the ‘Massecaps’ titanosaur tooth morphotype reported by Díez Díaz et al.2 from southern France and described as ‘robust spatulate’ has a flat lingual surface, without the complex morphology shown by the Iharkút specimen, and lacks the labial bend of the crown in mesial/distal view.New discoveries of European latest Cretaceous titanosaurs document an increasing diversity with at least six different taxa  basal somphospondylan Sarmientosaurus from South America50. Although details of the morphology are different, the teeth of Sarmientosaurus also show moderate SI values , a D-shaped cross-section of the crown, and more particularly the labially leaning crown at mid-height, below a lingually recurved apical part.Interestingly, MTM PAL 2017.1.1. bears some resemblance to the isolated and indeterminate sauropod teeth reported from the mid-Lower Cretaceous of western FranceEuhelopus38 and Sarmientosaurus50. Thus, we suggest a non-titanosaur titanosauriform affinity for this specimen, pending discovery of further material that might reveal its more precise taxonomic status.To sum up, specimen MTM PAL 2017.1.1. is certainly a tooth composed of an extensive pulp cavity and dentine covered by heavily worn enamel that shows a number of parallel, elongate scratches along the entire crown. Its morphology, being an elongate non-carinated, spatula-like and pointed tooth, is most closely reminiscent of those of certain sauropods. The mosaic of derived and plesiomorphic characters displayed by the Iharkút tooth clearly suggests a neosauropod affinity. It markedly differs from the peg-like diplodocoid and chisel-like derived titanosaurian teeth (including most titanosaur morphotypes reported previously from the uppermost Cretaceous of Europe), instead being more similar to some brachiosaurid teeth or to those of the basal somphospondylan titanosauriform Despite being a single piece of evidence, the sauropod tooth from the Santonian of Hungary is of great importance for at least two reasons. First, this specimen is the first sauropod body fossil from a 20 My long hiatus in the fossil record of this clade in Europe, extending from the mid-Cenomanian to the late Campanian interval. Second, the mosaic of derived and plesiomorphic features documented by the crown morphology points to a basal titanosauriform affinity and suggests the occurrence of a clade of sauropods in the Upper Cretaceous of Europe that is markedly different from that encompassing the previously known Campano-Maastrichtian titanosaurs.51 to account for the absence of sauropod fossils for the largest part of the mid to Late Cretaceous interval in North America, Le Loeuff 8 and Le Loeuff and Buffetaut15 suggested that the fossil record supports the absence of sauropods from the Cenomanian to late Campanian continental vertebrate record of Europe. This assertion was based on the fact that until the end of the 1990’s not even a single bone or footprint, certainly referable to this group, was known from the, admittedly few, European vertebrate localities representing this time period. The discovery of tracks identified as belonging to small sauropods from the Santonian of southern Italy52 and trackways of larger sauropods16 (probably titanosaurs)1 from the upper Turonian–lower Coniacian of Dalmatia, Croatia, however, indicates that sauropods were present in the Cenomanian to Coniacian continental ecosystems of Europe as well3. The sauropod tooth from Iharkút further strengthens this view, filling in the previously hypothesized Late Cretaceous gap in the sauropod fossil record, and shows that instead of their disappearance, the absence of sauropod fossils in European Late Cretaceous assemblages is probably in part the by-product of sampling bias.Similarly to the ‘sauropod hiatus’ hypothesis proposed by Lucas and Hunt53 (2011:534) convincingly demonstrated “the abundance of titanosaurs during the Early and latest Cretaceous and their apparent absence during the mid-Cretaceous” in Europe, and pointed out a positive correlation between the abundance (or lack) of sauropod remains and the amount of terrestrial sediment deposition during the Cretaceous. The Iharkút sauropod tooth came from the deposits of a flash flood event that was formed on a low-lying alluvial floodplain developed not far from swampy/deltaic environments that existed under humid conditions21. Accordingly, this landscape was probably more similar to a ‘coastal’ environment than to the much drier and open inland habitats likely preferred by the titanosaur sauropods53. The fact that this tooth represents the only fossil of a sauropod discovered so far among more than 50.000 bones and teeth of the Iharkút assemblage fits well into this environmental scenario, but also confirms that sauropods existed in pre-Campanian times within the European archipelago. In addition, the Santonian sauropod fossil evidence from southern Italy and from Iharkút reveals their presence in both the southern17 and northern21 parts of the Apulian microplate, and suggests their more widespread existence in this region.Mannion and Upchurch55. If this suggested affinity is upheld by future discoveries, the presence of the Iharkút titanosauriform expands the apparently cryptic sauropod diversity in Europe during the Late Cretaceous, from where only lithostrotian titanosaurs58 have been reported before. It further supports the endemic and relictual nature of these latest Cretaceous European assemblages, highlighted by the presence of a basal titanosauriform sauropod clade that most probably went extinct by Santonian times in most other landmasses59.The basal titanosauriform affinity of the Iharkút tooth, as assessed based on its mosaic features, might further suggest that the Santonian-aged Iharkút sauropod apparently represented a lineage different from, and more basal than, that of the known European Campano-Maastrichtian sauropods15, or it is a relict form that survived in a geographically limited refugium within the European Cretaceous archipelago, a biogeographical phenomenon already pointed out in the case of many other latest Cretaceous continental vertebrates61. Certain morphological similarities with the Hauterivian-Barremian aged sauropod teeth from Charentes, western France might support the second scenario, while possible affinities with the ‘mid’-Cretaceous Argentinian Sarmientosaurus would rather argue for a southern immigrant. Hopefully further material of the enigmatic Iharkút sauropod will be discovered and will help clarifying this problematic aspect of the Late Cretaceous European biogeography as well.However, the uncertain taxonomic status of the specimen does not allow a more precise clarification of its affinities and relationships. As such, it also remains unknown whether this form represents an immigrant from Gondwana or Asia, as suggested for some Late Cretaceous European titanosaursSupplementary_information 1


There is limited knowledge about the perspective of patients undergoing allogeneic haematopoietic stem cell transplantation (HSCT) about having a sibling as donor. It is essential to understand the main concerns of stem cell recipients in order to enable nurses to provide person-centred care.The study aim was to explore patients’ main concerns about having a sibling stem cell donor and how the patients handle them, from immediately before until one year after transplantation.Twenty-eight interviews were performed prospectively during one year with ten adult sibling stem cell recipients with a mean age of 52 years (range 19-68 years). The interviews were analyzed by the Grounded Theory method.Invest, Compensate and Celebrate. Recompensation is defined as a lasting compensation given by the recipient to the sibling donor for the loss or harm suffered or effort made. The sense of having to reward, protect, appreciate, maintain peace and work on the relationship with the sibling donor at the same time as having to accept a serious illness, cope with their situation and promote their own recovery is strenuous for the recipients.The core category Recompensation summarises the process in the generated grounded theory including the three main categories; The main concern for stem cell recipients during their first post-transplant year is to recompensate the sibling donor by investing, compensating and celebrating her/him. Although there is a positive aspect of recompensation, it can also imply pressure and guilt. It is essential to understand the main concerns of stem cell recipients in order to enable health professionals to provide person-centred care.-established treatment that offers a potential cure for patients with haematological malignancies. One essential prerequisite for HSCT is finding a donor with a reasonably close Human leukocyte antigen (HLA) match to increase the patient’s chances of survival [Allogeneic HSCT is a wellsurvival . The donsurvival  and worlsurvival . The onesurvival .Although HSCT is established as a successful treatment for haematological malignancies, there is a significant risk of acute complications, Late side effects and mortality , 6. The Only one study has truly explored patients’ experiences of having a sibling donor in detail . That stThe sibling donors’ situation has also received little attention. These donors are in a vulnerable situation and both negative and positive experiences are reported, such as anxiety, pain, guilt, happiness, an increased sense of self-worth and pride , 14. A r2The starting point for this study was patients’ main concerns about having a sibling stem cell donor from immediately before HSCT until one year afterward. Grounded Theory (GT) according to Charmaz  was the 2.1The approval of the study was made by the Regional Ethical Review Board of Southern Sweden (Dnr 541/2007) and performed in accordance with the Declaration of Helsinki . The pot2.2Patients planned for HSCT with a sibling donor at a University Hospital in Sweden between March 2011 and December 2012 were consecutively asked to participate in the study. All recipients and their sibling donors were ≥ 18 years and had good ability in understanding and speaking Swedish. Ten patients who met the inclusion criteria were asked to participate in the study and all ten agreed to do so. At the recipients’ appointment for medical investigation and information pre-transplantation, they were informed and asked about participation in the study by the first author (AK). We decided from the start to follow the included ten participants during one year. Thus, the methodological step involving true theoretical selection was abandoned. Instead, we ensured that the sample would reflect clinical reality. After agreement to participate, none of the recipients were excluded or decided to withdraw from the study. All donors of the recipients participated in a corresponding interview study .2.3The method chosen for data collection was face-to-face interviews performed by the first author (AK) at a time and location decided by the informants. The first author (AK) is a nurse specialist and has extensive experience from nursing experience from recipients of stem cells and from stem cell donor care, but did not take part in the care of the informants in this study. Altogether, 28 interviews were carried out: immediately before transplantation , and three months and one year after HSCT. One recipient only participated in the first interview as she, unfortunately, died due to complications from the transplantation less than three months afterward. All interviews began with the open question: “Can you tell me about your thoughts and feelings when you became aware that you needed a stem cell donor for transplantation?” followed by: “What did you feel and think when you were told that a sibling could become your donor?” and “Can you please tell me now, three months/one year after transplantation, what being transplanted with stem cells from your sister/brother was like?” Additional questions were asked to make the recipients elaborate their answers about their experiences and thoughts about having a sibling donor, and also to extend the answers given in prior interviews. The interviews were digitally recorded and lasted between 14 and 121 minutes (median 53 min). After finalizing each interview they were transcribed verbatim.2.4A search of the literature revealed that no study had been performed within this particular context, which is in line with the recommendations of Hallberg  and Glas33.11. The mean age of the sibling donors was 49 years (26-66 years). The gender constellation was: male donor to male patient (3), male donor to female patient (2), female donor to male patient (1), female donor to female patient (4).A total of ten adult patients undergoing HSCT, four men, and six women, participated in the study. Their mean age at the time of the first interview was 52 years (19-68 years). Demographics and characteristics of the recipients are presented in Table 3.2Invest, Compensate and Celebrate. Recompensation is defined as a lasting compensation given by the recipient to the sibling donor for the loss or harm suffered or effort made, i.e., the stem cell donation.The essence of the results derived from the grounded theory analysis of the interviews shows the core category Recompensation and the three main categories; i.e., knowing the origin of the stem cells, the donor’s previous lifestyle and what kind of person the donor is. The alternative, i.e., receiving stem cells from a stranger, a registry donor, meant a great sense of uncertainty about the origin of the stem cells and the unfamiliar donor’s life and personality. The consequence of donation from a stranger was perceived as difficult to grasp. In one way receiving stem cells from an unrelated donor would be easier because the sense of guilt towards the donor would be less. However, the recipients preferred a sibling donor to an unrelated one, Even though, it meant a sense of guilt about the sibling. This sense of guilt entailed a need to compensate the donor for her/his effort, which became evident by the recipients’ recompensation. The recompensation was, manifested in taking great responsibility for her/his well-being and possible pain, harm and fear, which started even before the donation and transplantation. The process of recompensation from before until one year after transplantation clearly demonstrates the recipients’ efforts to protect the donor and show their gratitude, even though they are unable to alleviate the donor’s suffering. Furthermore, the three main categories comprise nine sub-categories illustrating the strategies used by the recipients to recompensate, . The core of the process is recompensation towards the sibling donor, while the driving force is the balance between guilt and compensation.For the recipients, the most important aspect was to recover from the serious illness. In order for this to happen, they were painfully aware of the need for a donor, while at the same time they did not wish to harm anyone. Receiving stem cells from a sibling implied a sense of security for the recipient, Working on the relationship with the sibling donor was an important strategy for all of the recipients throughout the first year, from immediately before until one year after HSCT, irrespective of the closeness in relation with the sibling donor. All recipients made efforts, regardless of their capacity and situation, to keep in contact with the sibling donor in the year following transplantation and wished to preserve the relationship. When the relationship was good their efforts concerned maintaining the proximity and companionship. When the quality of the relationship was poor or even non-existent the informants tried to increase proximity in various ways or at least make the donor aware of their gratitude by enlisting the help of other family members to convey their message. Thus, the recipients hoped for survival and possibly better health. Regardless of the level of proximity, geographical distance or quality of the relationship they tried to recompensate their sibling for her/his loss, harm or effort due to the donation. The main categories and sub-categories will be presented in bold italics illustrating the strategies used by the recipients during the first year.3.3i.e., the project that would ensure the recipient’s survival. Thus, if the recipient prepared her/himself, coped and worked on various relationships, in particular, the sibling relationship, she/he thereby optimized the circumstances so that there would be less harm to compensate afterward.Investment is the starting point for the recipient’s recompensation towards the sibling donor. The efforts made in this phase were intended as investments in the transplantation project, collaborate was a way of investing in themselves by self-optimization. This involved having a positive attitude, preparing themselves practically, socially and professionally and taking care of themselves, e.g., eating well, exercising and not smoking. They also chose to trust and rely on health care and healthcare professionals. Some donors expressed this as: helping the healthcare professionals to help them by making themselves deserving of the donation.The interviews revealed that for the recipients, the pre-transplantation phase lasted from the time the donor was identified until the planned donation. To “Being away from home for such a long time and not knowing if I would ever come home again, or what shape I would be in…it demands a lot of long-term planning.” “try to be strong, try to eat even when I have no appetite and try to drink even if I can’t stand it. Everything to help you cure me, so to speak.” cope in various ways to invest in the situation. They adopted different coping strategies and played either an active role by establishing goals for the future and confronting the situation, e.g., by reading, planning, thinking, discussing, informing relatives and friends, or a passive role by avoiding and distancing themselves from the situation.Before transplantation, the recipients made different attempts to “Now I have these goals… things that will happen during this year, that I know I want to be a part of. First, there is the sailing and then I will become a grandfather.” “My family knows. But I haven’t told my friends or colleagues at work… There are too many questions, they speculate and I’m not interested. They can’t help me.” work on the relationship with the sibling donor to ensure the donation. This included taking responsibility for the relationship by informing themselves about the donation procedure, increasing the amount of contact with the sibling donor and paying extra attention to her/him. The recipients trusted the donor to complete the donation and some even offered the sibling donor an opportunity to withdraw.A common approach among all informants, regardless of the closeness of the relationship, was to “They said that she would experience the bone marrow harvest like being kicked in the back or on the behind by a horse. But we haven’t told her that. So she’s probably not aware of what is coming. I am worried because she is a bit worried about it and when she is extra worried I chat with her every day.” work on family relationships by taking responsibility for the relationship between various members of the family. This was done by, for example, involving the family in the transplantation procedure, listening to and talking more with different family members than before and playing down the risks of the transplantation to reduce their worries. The aim of this strategy was to cause no conflicts, thereby preventing the risk of doubt or regrets on the part of the donor, her/his spouse/partner or significant others. The investment was all about not jeopardizing the “transplantation project” or what was at stake.Another way to ensure the donation was to ”My focus is on his wife and children. I feel I need to talk more with my sister in law… I want to acknowledge the fact that she is making an effort too.” accept the serious illness and that the transplantation would be hard and strenuous. All recipients managed to practice the art of mindfulness and took one day at a time, reasoning that whatever happens will happen. However, at the same time as they expressed that they mainly relied on faith in a fatalistic manner, They directed all their efforts to ensure the donation from their sibling in a very concrete way.A part of the investment was also to “I have to be very enthusiastic and very grateful… I simply have to put her first…. I need to keep her in a very good mood now.” 3.4cope three months after transplantation meant managing the situation of being recently transplanted with stem cells from a sibling donor and having to recover. Although the sequence in which the coping strategies were employed differed between individuals, they all made use of the same strategies at some stage of the period from before transplantation until three months afterward. Either the recipients played an active role by confronting the situation, processing what they had gone through, prioritizing themselves and establishing goals for the future, or a passive role by avoiding and distancing themselves by not talking or thinking about the situation very much.Compensation, defined as repaying the sibling donor, started three months after transplantation by means of a two-fold approach: firstly, coping with the situation and recovering and secondly, thanking the donor and protecting the relationship with her/him. To “I try to avoid thinking about it as much as possible and do as many things as possible to be active and on the go.” “I make no plans but simply try to take each day as it comes.” promote recovery in various ways, depending on the individual and on the outcome of the transplantation. They accepted the situation and their level of well-being in order not to disturb the recovery process. Many of the recipients focused on the new stem cells and some even talked to the cells and ate sweets that the donor liked to feed the stem cells in a way they were used to.During this phase, they “This might sound weird, but I talk to my new stem cells every day. I have tried, this is a part of my pep talk. Therefore we, my stem cells and I, will make this work. I tell them that they are welcome to my body to remove my sick cells.” The recipients who were feeling well enjoyed being out of the hospital, tried to create new routines in their everyday life and started to plan their future. Those who were not feeling so well tried to appreciate the positive aspects of their situation and did their best not to show their malaise to family and friends.” I try to eat fatty food, using butter and cream, eating carbohydrates and sugar. I try to keep to that diet and eat as much as I can. I simply try to do my best. This is my only option. Simply bite the bullet and look happy.” appreciate the sibling donor. They thanked their sibling donor by verbally expressing their gratefulness, by not showing any impatience in their contact with the donor and telling her/him what a great contribution she/he has made. Most of the recipients were planning some kind of reward for the donor, e.g., offer a holiday or a visit to the theatre.An essential strategy three months post-donation was to “I will try to compensate if he suggests something. He would love to travel with me sometime. So, I feel I will do it for him, if that is his wish.” maintain peace with the donor. They thanked the sibling donor for the gift by ensuring that no conflict arose with the donor or her/his family. Peace was also maintained by increased contact with the sibling donor and her/his family, in the course of which the recipient treated them with greater understanding and patience.It was very obvious that the recipients worked to “Maybe you’re… more careful to never show impatience when he phones me. Sometimes I have been very tired and he wants to talk forever…” 3.5enrich everyday life and to do the things they enjoyed, but it was dependent on their illness burden, which was rather extensive for some of the informants. One recipient who was doing very well decided to retire from work to have more time for joy. She allowed herself various new things and made plans to travel. Those who experienced health lived a good life and made plans for the future.Celebration meant paying tribute to the donor and to oneself, as well as keeping the stem cells working. The celebration involved enriching everyday life, rewarding and protecting the donor. The ways of celebrating depended to a large extent on the recipient’s well-being. One year after transplantation all recipients had changed their perspective on life and the majority took advantage of the time they had been given. They tried to “I am content that I have retired. To do things I want to do, living a good life. As I say, it is better to live in the present, spend your money. I travel wherever I want and do all the shopping I want now.” e.g., painting pictures, sewing or simply sitting on the balcony. This served as a way of paying tribute to oneself and even suppressed the fact that they were doing badly. However, one recipient had no strategies whatsoever one year after transplantation and experienced only suffering and disappointment. For this particular recipient, there was no celebration at all.Those who suffered from severe illness due to complications and set-backs tried to make the best of everyday life by undertaking light activities at home, “I can say one thing. If I had known that I would feel this poorly I would never have accepted the treatment. It is not worth it.” reward the sibling donor. They made plans to reward the donor and her/his partner by, for example, celebrating the first anniversary of the donation and transplantation.The recipients who experienced health one year after the transplantation used different strategies to “I had this huge celebration on my birthday and E (the donor) was there. He was sort of a prince and that was simply right. The donor should be in the spotlight and one should not forget that, it is extremely important.” Appreciation was verbally expressed to the sibling donor by a simple ‘Thank you’, but also by describing how the good quality of the stem cells had contributed to excellent well-being. One recipient expressed that he will never ever be able to become angry with the sibling donor again. As the donor had saved his life he will be forever indebted to him.“I will probably not be pissed off with him, at least not for a while. Because it feels like no matter what he does it is a mere trifle in the context.” protect the sibling donor from knowing. They avoided talking to the donor about their malaise and attempted to conceal how they were doing.The recipients who suffered from severe illness lacked conditions or had few opportunities to reward the donor. These recipients suffered a great deal one year after transplantation and realised that they would probably not survive. Thus, they prepared themselves in different ways for their death, while at the same time trying to “Unfortunately, I haven’t had the strength to keep in contact lately. I mostly use text messages or the computer, or suchlike.” The wish to protect the donor was handled in different ways by the recipients. One example is the recipient who eventually needed a second donation from her sister. However, the sister was pregnant and therefore could not donate again. The recipient did not want to burden the donor and also felt guilty due to being unable to rejoice about her sister’s pregnancy. Another example was the recipient who was suffering from severe chronic GvHD and did not want to burden the donor with the knowledge of her condition. At the same time, she experienced a sense of guilt because she was aware that the sibling donor had suffered during the donation procedure.“ I felt very selfish…. no, you can’t be pregnant. I felt like I ruined her happiness about the pregnancy. I needed her help and she wanted to start a family.” In conclusion, throughout the first year after stem cell transplantation, in sickness and in health, the stem cell recipients strived to recompensate their sibling donor for the loss, harm or effort caused by the donation, while at the same time coping with their own suffering and losses.44.1No prospective grounded theory studies of recipients of stem cells from sibling donors have been previously performed, as far as we know. We chose to use the criteria for rigour in Grounded Theory studies by Charmaz . Credibii.e. with similar health care as in Western countries. This generated Grounded Theory has to be further tested to secure the applicability to recipients of living organ donors, e.g., kidney donors.To confirm and optimise the result, the codes, sub and the main categories were all along the analysis checked against the transcripts of the interviews. We believe that the result from this study is relevant to the recipients of stem cells from sibling donors in similar contexts as in this study; One limitation of this study is the possibility of transferability since this study was conducted in only one country, Sweden, with only Swedish speaking recipients. The study sample is small; however, it is representative of adult recipients of stem cells from adult sibling donors; the median age of 61 years and about the same number of women and men. By choosing in this context a unique prospective design we did not fulfil the true methodological step where sampling is performed until theoretical saturation. However, the 28 interviews enabled an in-depth understanding of the transplantation process.4.2i.e., that of the recipient and the donor.To the best of our knowledge, this is the first study of the main concerns of recipients transplanted with stem cells from a sibling donor, making it a unique contribution to the field of stem cell transplantation. This study and the recent grounded theory study on adult sibling donors’ main concerns  show thei.e., the stem cell donation, can partly explain the recipients’ decreased social well-being over time. For many recipients, the sense that they have to reward, protect, appreciate, maintain peace with and work on the relationship with the sibling donor, while at the same time having to accept a serious illness, cope with their situation, promote their own recovery enrich their everyday life is most certainly strenuous. In addition, the result shows that recipients of stem cells from a sibling donor experience that they have to work on family relationships, especially the relationship with the sibling donor. Therefore, one assumption is that in some ways it might be easier for the recipient to receive stem cells from an anonymous registry donor than from a sibling donor. The theoretical framework of recompensation among patients transplanted with stem cells from a sibling donor may be of value in supporting patients undergoing allogeneic haematopoietic stem cell transplantation (HSCT) with a sibling donor and to enable stem cell transplant nurses to facilitate person centred caring interventions.The quality of life (QoL) of patients treated with HSCT has been explored in several previous studies , 20-22 mThe main concern for stem cell recipients during their first post-transplant year is to recompensate the sibling donor by investing, compensating and celebrating her/him.Although there is a positive aspect of recompensation, it can also imply pressure and guilt.This knowledge of the perspectives of both the recipient and the donor could allow transplant professionals to consider trying to influence the situation. One way of doing this might be to organize a formal occasion for celebration and closure. This could prevent the long-term pressure of recompensation among recipients suffering from various side-effects and complications, as they already have enough problems to handle. Patient information to recipients could include a paragraph stating that the donor usually does not wish to be celebrated, which would help to ease the recipients’ burden.


Since the concentration of p-TA in dog cerebrospinal fluid (CSF) has been reported to be greater than that in plasma, it is proposed that clearance of cerebral p-TA is important for normal function. The purpose of this study was to examine the role of the blood–brain barrier and blood-cerebrospinal fluid barrier (BCSFB) in p-TA clearance from the brain.3H]p-TA elimination from rat cerebral cortex and from CSF was examined after intracerebral and intracerebroventricular administration, respectively. To evaluate BCSFB-mediated p-TA transport, [3H]p-TA uptake by isolated rat choroid plexus and conditionally immortalized rat choroid plexus epithelial cells, TR-CSFB3 cells, was performed.In vivo [3H]p-TA elimination from rat CSF was found to be 2.9 min, which is 62-fold faster than that from rat cerebral cortex. In addition, this [3H]p-TA elimination from the CSF was significantly inhibited by co-injection of excess unlabeled p-TA. Thus, carrier-mediated p-TA transport process(es) are assumed to take part in p-TA elimination from the CSF. Since it is known that transporters at the BCSFB participate in compound elimination from the CSF, [3H]p-TA transport in ex vivo and in vitro models of rat BCSFB was examined. The [3H]p-TA uptake by isolated rat choroid plexus and TR-CSFB3 cells was time-dependent and was inhibited by unlabeled p-TA, indicating carrier-mediated p-TA transport at the BCSFB. The p-TA uptake by isolated choroid plexus and TR-CSFB3 cells was not reduced in the absence of extracellular Na+ and Cl−, and in the presence of substrates of typical organic cation transporters. However, this p-TA uptake was significantly inhibited by cationic drugs such as propranolol, imipramine, amantadine, verapamil, and pyrilamine. Moreover, p-TA uptake by TR-CSFB3 cells took place in an oppositely-directed H+ gradient manner. Therefore, this suggested that p-TA transport at the BCSFB involves cationic drug-sensitive transport systems which are distinct from typical plasma membrane organic cation transporters.The half-life of [p-TA elimination from the CSF is greater than that from the cerebral cortex. Moreover, it is suggested that cationic drug-sensitive transport systems in the BCSFB participate in this p-TA elimination from the CSF.Our study indicates that  Amo in part . Regardi in part . In addi in part , 20. Tak/Slc6a3) –24 and, p-TA concentration, the purpose of this study was to examine the role of the brain barriers in p-TA clearance from the brain. To examine p-TA elimination across the BBB, we used an intracerebral microinjection technique. To evaluate BCSFB-mediated p-TA efflux transport, in vivo intracerebroventricular administration and ex vivo transport studies using isolated choroid plexus were performed. In addition, the properties of p-TA uptake were examined using an in vitro model of rat BCSFB, conditionally immortalized rat choroid plexus epithelial cells (TR-CSFB3 cells).To increase our knowledge of the homeostatic mechanism(s) governing the cerebral n [1-14C] , mannitol, d-[1-14C] , and tyramine hydrochloride, - were obtained from American Radiolabeled Chemicals . All other reagents were commercially available.Butanol, Wistar rats  were purchased from Japan SLC  and maintained in a controlled environment.3, 10 mM d-glucose, and 10 mM 2-[4-(2-hydroxyethyl)-1-piperazinyl]ethansulfonic acid (HEPES)-NaOH, 3 mM KCl, 1.4 mM CaCl2, 1.2 mM MgSO4, 0.4 mM K2HPO4, pH 7.4) containing [3H]p-TA  and [14C]d-mannitol  was injected into the parietal cortex area two region of the brain over a period of 1 min. [14C]d-Mannitol, a BBB-impermeable marker, was used to normalize the actual injection volume. At a designated time, each rat was decapitated, and the ipsilateral cerebrum was removed. During the procedure, the behavior of rats and abnormal morphology of the brain was monitored, and there was no apparent change in the anesthetized conditions and brain structure. The tissue was dissolved in 2 N NaOH (2 mL) at 55 °C for 3 h and then mixed with 10 mL Hionic-Fluor . The radioactivity was measured in a liquid scintillation counter equipped with an appropriate crossover correction for 3H and 14C . The percentage of [3H]p-TA remaining in the ipsilateral cerebrum (%) was obtained from Eq. Anesthetized rats, obtained by intraperitoneally injecting pentobarbital sodium solution (50 mg/kg), were fixed in a stereotaxic apparatus . Physiological buffer A  and [14C]d-mannitol (5 nCi), with or without unlabeled 75 mM p-TA, was injected into the left lateral ventricle over a period of 20 s. At designated times, CSF (50 μL) was withdrawn by cisternal puncture and the levels of 3H and 14C in the CSF and injectate were measured in a liquid scintillation counter . The rats exhibited no abnormal behavior during this technique.Rats were anesthetized by intraperitoneally injecting pentobarbital solution (50 mg/kg), and the head was fixed in the SR-5R (Narishige). A hole was drilled in the skull, 1.5 mm to the left and 0.5 mm posterior to the bregma, into which a needle was fixed as a cannula for injection. Then, 10 μL buffer A containing p-TA or [14C]d-mannitol.The kinetic parameters of this study were obtained using a one-compartmental model Eq. , where CCCSF(t)/dose × 100” is the percentage of the residual concentration of the compound in the CSF normalized by the amount of injectate (% of dose/mL CSF). By multiplying ke by Vd,CSF, the apparent elimination clearance from the CSF (CLCSF) was obtained.3H]p-TA and 0.05 μCi [14C]n-butanol, a marker of water space in the choroid plexus l-proline in TR-CSFB3 cells seeded onto a Transwell® was similar to the uptake value of [3H]l-proline in TR-CSFB3 cells seeded onto a culture plate p-TA-containing buffer A (0.75 μCi/mL) was applied. Na+- or Cl−-free buffer A was prepared as described in the previous section. To stop the uptake reaction, cells were rinsed three times with 4 °C buffer A. The cells were subsequently solubilized with 1 N NaOH solution and incubated at room temperature for 12 h. After neutralization with 1 N HCl, the cellular protein and [3H]p-TA-derived radioactivity were determined using a detergent-compatible protein assay kit  and liquid scintillation counting, respectively.TR-CSFB3 cells were cultured at 33 °C as described previously. Kitazawa et al. have determined the apical localization of Nass slide . It has re plate . The cel3H]p-TA in isolated rat choroid plexus and TR-CSFB3 cells, namely the tissue/medium ratio and cell/medium ratio, respectively, was obtained from the following equations  of [3H]p-TA uptake by isolated rat choroid plexus from 0.25 min to 1.0 min was found to be 0.981, which was greater than that from 0.25 min to 2.0 min (r2 = 0.822). This suggested that the initial [3H]p-TA uptake by isolated rat choroid plexus is reflected in the data up to 1 min. Regarding the uptake of [3H]p-TA by TR-CSFB3 cells, the r2 of the [3H]p-TA uptake from 0.25 min to 2.0 min was calculated as 0.989, which is similar to that of [3H]p-TA uptake by isolated rat choroid plexus from 0.25 min to 1.0 min and is close to 1. This result suggests that the cell/medium ratio at all time points reflects the initial [3H]p-TA uptake process.The equation was fitted using the nonlinear least-squares regression analysis program, MULTI . To examp-TA uptake by TR-CSFB3 cells were obtained from Eq. V is the uptake rate of p-TA, [S] is the p-TA concentration in buffer, Km is the Michaelis–Menten constant, and Kd is the non-saturable transport clearance. The equation was fitted using the MULTI program [t test for two groups and one-way analysis of variance followed by Dunnett’s test for more than three groups.The kinetic parameters for  program . The kin3H]p-TA remaining in rat ipsilateral cerebrum at 5, 10, 20, and 40 min after micro-administration into the parietal cortex area 2 is shown in Fig. p = 0.143), although a tendency for a time-dependent decrease was observed. As a reference value, the half-life (t1/2) of the percentage of cerebral [3H]p-TA remaining after microinjection into the cerebral cortex was calculated to be 178 ± 68 min.The time-course of the percentage [CCSF(t)/dose × 100” values of [3H]p-TA and [14C]d-mannitol after intracerebroventricular injection. The residual concentration in CSF of [3H]p-TA at 1.0, 1.5, and 2.0 min was significantly lower than that of [14C]d-mannitol. [3H]p-TA was eliminated from the CSF with a t1/2 of 2.85 ± 0.83 min, which is 3.0-fold faster than the t1/2 of [14C]d-mannitol. From the Vd,CSF and ke,CSF values of [3H]p-TA , the CLCSF of [3H]p-TA was found to be 38.6 ± 12.0 μL/rat. The kinetic parameters for [14C]d-mannitol, Vd,CSF, ke,CSF, and CLCSF, were calculated to be 135 ± 11 μL/rat, 8.07 × 10−2 ± 6.09 × 10−2 min−1, and 10.9 ± 8.3 μL/rat, respectively. To examine the possible contribution of carrier-mediated p-TA elimination system(s) from the CSF, the effect of unlabeled excess p-TA on the concentration ratio of [3H]p-TA and [14C]d-mannitol was examined. The [3H]p-TA/[14C]d-mannitol residual concentration ratio at 2 min was significantly (1.1-fold) increased by co-administration of 75 mM unlabeled p-TA into the rat cerebroventricle compared with that in the control  and 0.895 ± 0.223 μL/μL ChP, respectively  on the choroid plexus, the compound/drug-sensitivities to [3H]p-TA uptake were examined , MPP+ , tetraethylammonium , and p-aminohippurate .In order to examine whether p-TA transport at the BCSFB, a [3H]p-TA transport study using an in vitro model of rat BCSFB, TR-CSFB3 cells, was performed. TR-CSFB3 cells exhibited time-dependent [3H]p-TA uptake with an initial uptake rate and an initial distribution volume of 2.93 ± 0.32 μL/(min mg protein) and 2.25 ± 0.24 μL/mg protein , and 9.78 × 10−1 ± 0.66 × 10−1 μL/(min mg protein), respectively , pyrimethamine (a substrate of MATE), norepinephrine (a substrate of norepinephrine transporter), PAH, serotonin, and cimetidine (a substrate of OCT and MATE). In addition, the [3H]p-TA uptake by TR-CSFB3 cells was significantly increased, but not decreased, by the co-presence of tyrosine (a precursor of p-TA), choline (a substrate of OCT), MPP+, and TEA , was 62-fold longer than that of the residual concentration of [3H]p-TA in the CSF  since the total volume of rat choroid plexus has been reported to be 6 μL (7.14 μL/(min rat) = 1.19 μL/μL ChP × 6 μL/rat) p-TA uptake clearance values obtained from the uptake studies of isolated rat choroid plexus (7.14 μL/(min rat) and TR-CSFB3 cells (11.0 μL/(min rat)) were 18.5 and 28.5% of the in vivo total [3H]p-TA elimination clearance from rat CSF (38.6 μL/(min rat), Fig. 14C]d-mannitol from rat CSF was found to be 10.9 μL , the incorporation of p-TA into the neural cells facing the cerebroventricles, including the ependymal cells, is a possibility. Further analysis to check the radioactivities in choroid plexus and neural cells surrounding the cerebroventricle could help these cells contribute to p-TA elimination from the CSF. Nevertheless, it is considered that the BCSFB is involved in the carrier-mediated p-TA elimination from the CSF, at least in part, since an inhibitory effect of unlabeled excess p-TA on [3H]p-TA uptake by isolated rat choroid plexus  was found to be 2.09 μL/(min mg protein), which is 2.1-fold higher than that of the non-saturable process . This result indicates that the transporter-mediated process makes a major contribution to apical p-TA transport at the BCSFB. It has been reported that human OCT3 and PMAT accept p-TA as a substrate with a Km of 0.281 and 283 μM, respectively p-TA transport into TR-CSFB3 cells and/or isolated rat choroid plexus was observed in the presence of these transporter inhibitors, such as MPP+, TEA, l-carnitine, pyrimethamine, and cimetidine  involve blood-to-brain transport across the BBB of cationic compounds and drugs, such as pyrilamine, oxycodone, and nicotine, although the molecular identification of the system(s) has not been reported [p-TA uptake by isolated rat choroid plexus  which also recognize cationic drugs are needed, since this could help improve the pharmacotherapy of these patients in addition to increasing our understanding of the role of the BCSFB in homeostasis of p-TA levels in the brain. Moreover, it is considered that our findings about the in vitro characteristics of tyramine transport at the BCSFB , are helpful for identifying the responsible molecule(s) for tyramine transport at the BCSFB.In patients with Parkinson’s disease and depression, monoamine oxidase inhibitors, such as selegiline, are regularly prescribed. It is widely known that patients who take hibitors , 45. As s Tables , 2. ThusB Tables , 2, suchp-TA elimination from the CSF to the clearance of p-TA from the brain parenchyma in vivo. From the transport studies using isolated choroid plexus and TR-CSFB3 cells, p-TA efflux transport at the BCSFB was shown to involve cationic drug-sensitive transport systems. These are distinct from typical plasma membrane organic cation transporters in the BCSFB. This study provides a new view of the role of the BCSFB in homeostasis of trace amines in the brain, including p-TA.Our study demonstrates the greater contribution of


Chlorocebus aethiops sabaeus) is a potentially valuable animal model of neurodegenerative disease. However, the trajectory of aging in vervets and its relationship to human disease is incompletely understood.The Caribbean vervet monkey (CSF) concentrations of Aβ1–40, Aβ1–42, total tau, and p‐tau181 in 329 members of a multigenerational pedigree. Linkage and genome‐wide association were used to elucidate a genetic contribution to these traits.To characterize biomarkers associated with neurodegeneration, we measured cerebrospinal fluid  was found, though suggestive linkage was highlighted on chromosomes 4 and 12. Genome‐wide association identified a suggestive locus near the chromosome 4 linkage peak.Aβ1–40 and p‐tau181 as potentially valuable biomarkers of these processes.Overall, these results support the vervet as a non‐human primate model of amyloid‐related neurodegeneration, such as Alzheimer's disease and cerebral amyloid angiopathy, and highlight Aβ Vervets display AD‐like pathology, including accumulation of Aβ‐containing cerebral plaques associated with reactive astrocytosis, dystrophic neurites, and vascular Aβ immunoreactivity resembling cerebral amyloid angiopathy (CAA), but unlike humans have relatively preserved cognition that does not appear correlated with plaque burden and demonstrate scant phosphorylated tau aggregates  and residues 1–42 (Aβ1–42). Aβ1–40 is the most abundant isoform, but CSF concentration differences of Aβ1–40 are generally subtle in AD  harbors useful biomarkers of neurodegenerative disease. CSF levels of Aβ and tau change with AD. Aβ exists in several isoforms, predominantly composed of residues 1–40 (AβThe Vervet Research Colony (VRC) was established from vervets captured in St. Kitts and Nevis, and maintained as a single extended pedigree , with several exceptions. During 2004 to 2008, animals were fed LabDiet 5052  as previously described  solid‐phase enzyme immunoassay . Briefly, the amyloid peptides are first captured by the 2G3 monoclonal antibody, and quantified with a biotinylated monoclonal antibody, 3D6. The protein is then detected using a peroxidase‐labeled streptavidin readout. Concentrations of CSF Aβ1–42, tau, and phosphorylated tau (p‐tau181) were measured simultaneously with the INNO‐BIA AlzBio3 fluorimetric immunoassay . Briefly, the proteins are first captured on beads by specific monoclonal antibodies – 4D7A3 for Aβ1–42, AT120 for tau, and AT270 for p‐tau181. Additional biotinylated antibodies are then used to quantify the proteins – 3D6 for Aβ1–42, and HT7 for tau and p‐tau181. The proteins are then detected using a phycoerythrin‐labeled streptavidin readout. Calibration of the standard curve was performed as described in the manufacturer's directions using six accompanying standards provided in the kit. All assays were run in duplicate. Measurements with coefficient of variation (CV) >25% or bead count <30 were excluded in quality control.CSF was collected from the cisterna magna of each animal under a protocol approved by the Animal Research Committee at the University of California, Los Angeles, as previously described  program ‘mri_ca_train’ to generate a probabilistic anatomical map (https://surfer.nmr.mgh.harvard.edu/fswiki/mri_ca_train) that was then used to segment the entire dataset. The Freesurfer segmented images were then inspected and manually adjusted to correct gross segmentation errors.Details of the six brain phenotypes investigated in the initial association analysis have been described previously . SOLAR uses the pedigree structure to adjust for the dependency structure among the subjects that would otherwise violate assumptions for standard linear regression methods. For the neuroimaging measures, age and sex were controlled as covariates in a linear regression model; the residuals were then associated with CSF traits. The procedure of Benjamini and Hochberg was used to correct for multiple testing to identify significant associations in the initial set of six neuroimaging traits with the four CSF traits (24 tests), using a false discovery rate (FDR) threshold of 0.05. In the secondary analysis of regional cortical measures of thickness and surface area with Aβ1–40 and p‐tau181 traits, the procedure of Benjamini and Hochberg was similarly used, applied to each of the two sets of association tests independently (29 tests per set).Brain‐CSF associations were tested using linear regression as implemented with the Sequential Oligogenic Linkage Analysis Routines (SOLAR) software package ; a p‐value for this estimate; and the proportion of variance attributable to covariates. Genetic mapping was performed using multipoint linkage analysis implemented in SOLAR. First, the multipoint identity‐by‐descent (IBD) relationships were computed for the vervet pedigree as previously described . EMMAX uses a variance component model to account for high relatedness among the vervet subjects . A linear mixed model was then fit to the data and used to calculate the association. We used standard genome‐wide significant thresholds from humans  were obtained in a total of 329 animals , while levels of Aβ1–42, tau, and p‐tau181 were not .Animals were selected from a complex, nine‐generation‐deep vervet pedigree with a high degree of consanguineous mating. Following quality control procedures see , CSF coned Table . Levels 3.2APP (encoding the amyloid precursor protein) and MAPT (encoding tau) could affect the performance of the antibodies.The antibody‐based CSF assays were designed and validated for humans; therefore, we evaluated whether species‐specific differences or polymorphisms in the vervet orthologs of Species‐specific differences between vervet and human in the antibody epitope regions were first identified. Across the Aβ peptide the human and vervet proteins were identical, including the 2G3, 4D7A3, and 3D6 epitope regions Figure . DiffereCoding polymorphisms across vervet subjects were then identified to determine whether inter‐individual differences in the polypeptide sequence of Aβ or tau could confound their quantification. In our cohort, polymorphisms were identified in APP (Ala15Ser) and MAPT (Gly148Arg and Leu213Pro). None overlapped with the antibody epitope regions; furthermore, no associations between these polymorphisms and quantified CSF concentrations were observed Figure .3.31–40, Aβ1–42, tau, and p‐tau181 with neuroimaging phenotypes, we initially calculated the correlations of CSF biomarker concentrations with six global measures , Aβ1–40 was correlated with corpus callosum cross‐sectional area (p = .024), hippocampal volume (p = .013), total gray matter thickness (p = .007), and total surface area (p = .006). The correlation with total gray matter thickness and total surface area was significant (controlling FDR at 0.05) after applying the Benjamini‐Hochberg correction for multiple comparisons. The direction of each regression coefficient was positive; higher concentrations of Aβ1–40 were associated with larger brain size. Likewise, p‐tau181 concentrations were correlated with cerebral volume (p = .001), cerebellar volume (p = .039), corpus callosum cross‐sectional area (p = .018), hippocampal volume (p = .039), and total surface area (p = .008). The correlation with cerebral volume and total surface area was significant after the Benjamini‐Hochberg correction. In contrast with Aβ1–40, the sign of each regression coefficient was negative; that is, higher concentrations of p‐tau181 tended to be associated with smaller brain sizes.Brain imaging phenotypes and genetic data had been collected as part of a previous study  that control for the false discovery rate. Concentrations of Aβ1–40 were correlated with increased gray matter thickness across many brain regions, including the orbital frontal gyri , lingual gyrus , inferior frontal gyrus , precuneus , angular gyrus , middle frontal gyrus , inferior precentral gyrus , superior precentral gyrus , superior frontal gyrus , postcentral gyrus , and lateral occipital gyri . Concentrations of p‐tau181 were correlated with decreased gray matter thickness in the fusiform gyrus , and surface area of the corpus callosum  and inferior occipital gyri . The direction of relationship was largely consistent with those in the global brain measures; Aβ1–40 was generally correlated with increased gray matter thickness and surface area, while p‐tau181 was generally correlated with decreased gray matter thickness and surface area of brain structures. Of these brain regions, only the gray matter thickness in the fusiform gyrus was significantly correlated with age, after correction for multiple comparisons (uncorrected p = 3.68 × 10−14). Fitting a linear regression model for fusiform gyrus gray matter thickness using both CSF p‐tau181 and age at MRI scan as covariates, we found that both age  and p‐tau181  were strong predictors. This pattern remained even after including age at CSF draw (p = .00057 for p‐tau181), suggesting that the contributions of CSF Aβ1–40 and p‐tau181 on regional brain volume measurements that we have identified are independent of age.Because we observed suggestive association between Aβex Table . MRI mea3.41–40, Aβ1–42, tau, and p‐tau181 concentrations. Narrow‐sense polygenic heritability (h2) was estimated using SOLAR, taking into account age and sex as possible covariates. After correcting for relatedness, CSF Aβ1–40 (p = .001) and p‐tau181 (p = .01) were significantly correlated with age, but not Aβ1–42 (p = .46) or tau (p = .31). CSF p‐tau181 (p = .007) but not Aβ1–40 (p = .32), Aβ1–42 (p = .25), or tau (p = .14) was significantly correlated with sex. The CSF concentration of Aβ1–40 was found to be significantly heritable . CSF concentrations of Aβ1–42, tau, and p‐tau181 were not heritable .We first sought to quantify the heritability of CSF Aβ1–40 concentration, we then attempted to map genetic loci that could play a role in the phenotypic variation using genome‐wide multipoint linkage analysis. At a LOD score threshold of 3.3, no markers reached genome‐wide significance  corresponding to cerebral atrophy in the vervet. Though previous studies have found limited phospho‐tau immunoreactivity and few neurofibrillary tangles in the vervet brain  mirror the distribution of CAA and subsequent neurodegeneration in humans . The lack of association with CSF Aβ1–42 with age and with brain regions may reflect inconsistent amyloid plaque burden in the aged vervet, compared to CAA pathology.These findings are reminiscent of the observed relationship between CSF biomarkers and neuroimaging measurements in aging and neurodegenerative disease in humans. Previous work has demonstrated associations between longitudinal changes in Aβ1–40 concentrations in the vervet. Linkage analysis identified two suggestive linkage peaks: a wide 9 cM peak on chromosome 4, and a narrow 6 cM peak on chromosome 12. The lack of statistical significance in linkage analysis despite the moderate heritability of CSF Aβ1–40 concentration may suggest a highly polygenic contribution to the phenotype. While the signal did not reach genome‐wide significance and spanned a wide genomic range, precluding identification of a single causal gene or variant, our work provides a basis for further fine mapping of these regions. Potential candidate genes in the region include PTCD2 and MAP1B, at the proximal end of the linkage peak and approximately 330 and 470 kilobases from the top associated marker. The pentatricopeptide repeat domain 2 gene, encoded by PTCD2, has been observed to be accumulated in AD brain in citrullinated form, and autoantibodies against PTCD2 have been observed in AD patient serum . Interestingly, neither cerebral volume or p‐tau181 were associated with age across the wide range of ages in the vervet cohort; however, the strongest association was detected between p‐tau181 and cerebral volume, suggesting that at least some of the associations were not driven by dependency on age. Furthermore, frank cognitive deficits were not observed or ascertained in the vervet subjects, and the study design was cross‐sectional; correlation with measures of brain structure and function in future longitudinal studies may provide additional insight into the extent of neurodegeneration.Another limitation of the study is the fact that neuroimages and CSF samples were not acquired simultaneously; the interval between imaging and CSF collection was variable, but imaging was performed approximately 5 years before CSF collection for each sample. Somewhat mitigating the possibility for extensive confounding, we had previously demonstrated high heritability of neuroimaging phenotypes, in some instances approaching 100%; therefore, environmental influences are likely to be low (Fears et al., 1–40 decreases with advancing age, correlates with global and regional measures of brain size, and has a large heritable component, suggesting its utility as a biomarker of AD‐ or CAA‐like changes in this potentially valuable animal model. Aβ1–40 was most closely associated with decreases in gray matter thickness in the frontal lobe and hippocampus, mirroring the regional vulnerability patterns of AD. However, structural associations were also observed in the occipital lobe and precuneus, regions that are generally spared early in Alzheimer's disease and more reminiscent of the distribution of pathology in CAA. Age‐related cerebral atrophy in the vervet likely involves β‐amyloid pathological processes, and Aβ1–40 and p‐tau181 appear to be robust biomarkers of these processes, though their sensitivities and correlations to disease pathology remain important open questions. We have also mapped suggestive linkage peaks on vervet chromosomes 4 and 12 that correspond to CSF Aβ1–40 concentration and may represent a modifier of neurodegeneration. The aged vervet therefore has the potential to advance our understanding of the genetics, pathogenesis, and treatment of neurodegenerative disease.In vervets, CSF AβNone declared. Click here for additional data file.