Kidney transplantation in low- and middle-income countries: the Transplant Links experience.

Paediatric kidney failure is a global problem responsible for significant childhood morbidity and mortality. The gold-standard treatment is kidney transplantation. However, the availability of kidney transplantation remains limited in some low- and middle-income countries (LMICs). Transplant Links Community (TLC) is a UK-based charity that mentors units in LMICs wishing to start kidney transplantation; the ultimate goal is for these units to become self-sufficient. TLC provides this support through in-person training visits and skill transfer, plus direct mentorship from the UK that is maintained over many years. From such mentoring programmes, it is evident that there are numerous challenges in the initial establishment and long-term maintenance of kidney transplant services, with specific and unique barriers applying to setting up paediatric transplant programmes compared to their adult counterparts. This review summarises TLC's first-hand experience of developing paediatric kidney transplantation services in LMICs over the past 15 years, the challenges encountered, and the major ongoing barriers that must be addressed to facilitate further progress in delivering transplantation services to children globally.

Pediatric peritoneal dialysis training program and its relationship to peritonitis: a study of the International Pediatric Peritoneal Dialysis Network.

BACKGROUND: The guidelines for training of patients and caregivers to perform home peritoneal dialysis (PD) uniformly include recommendations pertaining to the prevention of peritonitis. The objective of this study conducted by the International Pediatric Peritoneal Dialysis Network (IPPN) was to investigate the training practices for pediatric PD and to evaluate the impact of these practices on the peritonitis and exit-site infection (ESI) rate. METHODS: A questionnaire regarding details of the PD program and training practices was distributed to IPPN member centers, while peritonitis and ESI rates were either derived from the IPPN registry or obtained directly from the centers. Poisson univariate and multivariate regression was used to determine the training-related peritonitis and ESI risk factors. RESULTS: Sixty-two of 137 centers responded. Information on peritonitis and ESI rates were available from fifty centers. Training was conducted by a PD nurse in 93.5% of centers, most commonly (50%) as an in-hospital program. The median total training time was 24 hours, with a formal assessment conducted in 88.7% and skills demonstration in 71% of centers. Home visits were performed by 58% of centers. Shorter (< 20 hours) training duration and lower number of training tools (both p < 0.02) were associated with higher peritonitis rate, after adjustment for proportion of treated infants and income of country of residence. CONCLUSIONS: An association between training duration and the number of training tools represent potentially modifiable risk factors to reduce peritonitis rates within the pediatric PD population. A higher resolution version of the Graphical abstract is available as Supplementary information.

Reference Range of Plasma Fatty Acids in North Indian Pregnant Population.

Reference values for Fatty Acids (FAs) are not well defined in the Indian population. Therefore, it is critical to establish FAs reference range for the healthy non-pregnant and pregnant Indian population. The present multi-centric, and cross-sectional study determines the 95% reference interval for FAs in an apparently pregnant Indian population and compare it to the healthy non-pregnant women. Physicians identified 164 reference individuals as healthy (56 non-pregnant and 108 pregnant) at various government and private hospitals of northern India. The 95th and 97.5th percentile reference limits were used to estimate the 95 percentile of the reference distribution. The reference ranges observed for Alpha-linolenic acid (0.29-0.42%; 0.36-0.58%), Docosahexaenoic-acid (3.38-4.23%; 3.8-4.55%), Eicosapentaenoic-acid (1.24-1.76%; 1.09-1.62%), Docosapentaenoic-acid-3 (0.61-0.69%; 0.65-0.76%), Linoleic-acid (18.44-20.75%; 19.51-21.88%), gamma-linolenic-acid (0.24-0.35%; 0.32-0.42%), Eicosatrienoic-acid (0.26-0.32%; 0.34-0.39%), Arachidonic-acid (9.29-11.02%; 10.02-11.56%), Docosatetraenoic-acid (0.62-0.89%; 0.79-1.09%), Docosapentaenoic-acid-6 (0.23-0.31%; 0.33-0.41%), Eicosatrienoic-acid (1.17-1.41%; 1.43-1.74%), Eicosenoic-acid (0.28-0.38%; 0.37-0.49%), Nervonic-acid (1.39-1.69%; 1.41-1.74%), Palmitoleic-acid (1.17-1.58%; 2-2.66%), Oleic-acid (19.8-22.26%; 19.68-22.94%), Myristic-acid (1.16-1.68%; 0.82-1.3%), Palmitic-acid (20.05-21.8%; 20.7-22.43%), Stearic-acid (11.34-12.56%; 10.29-11.02%), Arachidic-acid (0.17-0.2%; 0.18-0.23%), Lignoceric-acid (0.81-1.08%; 0.77-1.08%), trans-palmitoleic-acid (0.22-0.29%; 0.26-0.37%), trans-oleic-acid (0.55-0.72%; 0.68-0.84%), trans-linoleic-acid (0.38-0.54%; 0.42-0.59%) respectively for non-pregnant and pregnant women. Furthermore, total FAs were significantly (p ≤ 0:05) higher in women aged 31-45 years than in women aged 16-30 years. whereas, there was no significant change in total FAs profile based on omega-supplementation, diet category, preterm-birth history, and gestation period. Thus, the current study provides information about an individual who is deficient in FAs and the dose required to increase FA concentrations in the body.

PPP2R5D-Related Neurodevelopmental Disorder or Developmental and Epileptic Encephalopathy?: A Novel Phenotypic Description and Review of Published Cases.

BACKGROUND: Protein phosphatase 2 regulatory subunit B' delta (PPP2R5D)-related neurodevelopmental disorder is caused by pathogenic variations in the PPP2R5D gene, product of which is involved in dephosphorylation. This is a rare disorder with description limited to case reports. Its phenotypic spectrum has expanded over the last decade. METHODS: We report a child with a developmental and epileptic encephalopathy phenotype with a pathogenic PPP2R5D variant. This phenotype has not been previously reported. We also reviewed the previously published reports of patients with this disorder. RESULTS: Including the index child, 28 cases (15 girls) were identified from nine relevant research items for analysis. All patients had developmental delay. History of seizures was observed in seven patients while macrocephaly was seen in nearly 80% of patients. Nonneurological manifestations were observed in 13 patients with the most common one being ophthalmological manifestations. The most common genetic variation was c.G592A (p.E198K). The common phenotypic associations of this variation were developmental delay, macrocephaly (11/15), and epilepsy (6/15). CONCLUSION: PPP2R5D gene variations should be suspected in children with developmental delay, autistic features, macrocephaly with or without epilepsy in the absence of any clear etiology. Dysmorphic features might provide a diagnostic clue. DEE phenotype may also be the presenting feature and might be an underreported entity.

Routine serum biomarkers, but not dual-energy X-ray absorptiometry, correlate with cortical bone mineral density in children and young adults with chronic kidney disease.

BACKGROUND: Biomarkers and dual-energy X-ray absorptiometry (DXA) are thought to be poor predictors of bone mineral density (BMD). The Kidney Disease: Improving Global Outcomes guidelines suggest using DXA if the results will affect patient management, but this has not been studied in children or young adults in whom bone mineral accretion continues to 30 years of age. We studied the clinical utility of DXA and serum biomarkers against tibial cortical BMD (CortBMD) measured by peripheral quantitative computed tomography, expressed as Z-score CortBMD, which predicts fracture risk. METHODS: This was a cross-sectional multicentre study in 26 patients with CKD4 and 5 and 77 on dialysis. RESULTS: Significant bone pain that hindered activities of daily living was present in 58%, and 10% had at least one low-trauma fracture. CortBMD and cortical mineral content Z-scores were lower in dialysis compared with CKD patients (P = 0.004 and P = 0.02). DXA BMD hip and lumbar spine Z-scores did not correlate with CortBMD or biomarkers. CortBMD was negatively associated with parathyroid hormone (PTH; r = -0.44, P < 0.0001) and alkaline phosphatase (ALP; r = -0.22, P = 0.03) and positively with calcium (Ca; r = 0.33, P = 0.001). At PTH <3 times upper limit of normal, none of the patients had a CortBMD below -2 SD (odds ratio 95% confidence interval 7.331 to infinity). On multivariable linear regression PTH (ß = -0.43 , P < 0.0001), ALP (ß = -0.36, P < 0.0001) and Ca (ß = 0.21, P = 0.005) together predicted 57% of variability in CortBMD. DXA measures did not improve this model. CONCLUSIONS: Taken together, routinely used biomarkers, PTH, ALP and Ca, but not DXA, are moderate predictors of cortical BMD. DXA is not clinically useful and should not be routinely performed in children and young adults with CKD 4-5D.

Neuroimaging Spectrum of Severe Hypernatremia in Infants with Neurological Manifestations.

BACKGROUND: Variable neurological manifestations and imaging findings have been described in children with severe hypernatremia. We aimed to describe the spectrum of neuroimaging changes in infants with severe hypernatremia. METHODS: This retrospective study included infants with severe hypernatremia (serum sodium >160 mEq/L), abnormal neurological examination, and an abnormal magnetic resonance imaging (MRI) of the brain over a period of 2 years in a tertiary care hospital. Relevant clinical data, including the feeding practices, clinical features, complications, and biochemical and radiological parameters, were entered in a structured pro forma. MRI findings were classified as vascular (hemorrhages and cerebral sinus venous thrombosis), osmotic demyelination syndrome (pontine and extrapontine myelinolyses), and white matter changes. RESULTS: The common clinical features in the neonates were poor feeding (n = 4) and decreased urine output (n = 4); the older infants presented with gastrointestinal losses (n = 5). All cases had dehydration with encephalopathy. The patterns of radiological injury were vascular (hemorrhages, n = 5 and venous thrombosis, n = 3), osmotic demyelination (n = 8), and white matter changes (n = 7). Coagulopathy was correlated with the vascular complications (r = 0.8, p < 0.0001); the degree of dehydration was correlated with the venous thrombosis (r = 0.7, p < 0.04) and acute kidney injury (r = 0.8, p < 0.001). Neurological sequelae were seen in four cases and correlated with hypernatremia (r = 0.6, p = 0.03) and hyperosmolarity (r = 0.6, p = 0.03). CONCLUSION: Characteristic neuroimaging findings are vascular changes in the form of venous thrombosis and hemorrhages, osmotic demyelination and white matter tract injury, and/or mostly combinations of these findings. Severe hypernatremia and resulting hyperosmolarity frequently cause neurological sequelae in neonates and infants.

May Measurement Month 2019: an analysis of blood pressure screening results from India.

AIMS: May Measurement Month is a global screening campaign to raise awareness regarding elevated blood pressure (BP). With the growing burden of hypertension, it is imperative to regularly assess the disease's prevalence, risk factors, and awareness levels in a country. The current prevalence of hypertension in India as per the National Family Health Survey Data stands at 25.3%. May Measurement Month mobilizes healthcare professionals and sensitizes them to regularly measure BP, and impart lifestyle modification advice to the community. It also complements the deficiency in screening programmes at a national and international level. METHODS AND RESULTS: May Measurement Month was carried out in May 2019 as an opportunistic screening campaign for adults (≥18 years). It was carried out by over 5000 trained volunteers across approximately 1000 screening sites (hospitals, public places, pharmacies, villages, and malls) in India. A total of 362 708 (57% males and 42.7% females) people were screened, among whom 68.1% had never measured their BP, and 29.4% (n = 106 522) were found to have hypertension. Of these, only 42.0% were on antihypertensive medication and 23.3% had controlled hypertension. CONCLUSION: Almost a third of the screened population had hypertension, and less than half of those with hypertension were aware of it or on treatment for it. Among those on antihypertensive drugs, BP was controlled in only half of them. These results support the need for greater impetus on BP screening initiatives to detect hypertension early in the community and prevent complications due to uncontrolled BP.

Corrigendum to: May Measurement Month 2019: an analysis of blood pressure screening results from India.

[This corrects the article DOI: 10.1093/eurheartjsupp/suab047.].

Cost analysis of pediatric intensive care: a low-middle income country perspective.

BACKGROUND: Globally, Pediatric Intensive Care Unit (PICU) admissions are amongst the most expensive. In low middle-income countries, out of pocket expenditure (OOP) constitutes a major portion of the total expenditure. This makes it important to gain insights into the cost of pediatric intensive care. We undertook this study to calculate the health system cost and out of pocket expenditure incurred per patient during PICU stay. METHODS: Prospective study conducted in a state of the art tertiary level PICU of a teaching and referral hospital. Bottom-up micro costing methods were used to assess the health system cost. Annual data regarding hospital resources used for PICU care was collected from January to December 2018. Data regarding OOP was collected from 299 patients admitted from July 2017 to December 2018. The latter period was divided into four intervals, each of four and a half months duration and data was collected for 1 month in each interval. Per patient and per bed day costs for treatment were estimated both from health system and patient's perspective. RESULTS: The median (inter-quartile range, IQR) length of PICU stay was 5(3-8) days. Mean ± SD Pediatric Risk of Mortality Score (PRISM III) score of the study cohort was 22.23 ± 7.3. Of the total patients, 55.9% (167) were ventilated. Mean cost per patient treated was US$ 2078(₹ 144,566). Of this, health system cost and OOP expenditure per patient were US$ 1731 (₹ 120,425) and 352 (₹ 24,535) respectively. OOP expenditure of a ventilated child was twice that of a non- ventilated child. CONCLUSIONS: The fixed cost of PICU care was 3.8 times more than variable costs. Major portion of cost was borne by the hospital. Severe illness, longer ICU stay and ventilation were associated with increased costs. This study can be used to set the reimbursement package rates under Ayushman Bharat - Pradhan Mantri Jan Arogya Yojana (AB-PMJAY). Tertiary level intensive care in a public sector teaching hospital in India is far less expensive than developed countries.

Pre-emptive rituximab and plasma exchange does not prevent disease recurrence following living donor renal transplantation in high-risk idiopathic SRNS.

BACKGROUND: Children with non-genetic steroid-resistant nephrotic syndrome (SRNS) are at high risk of disease recurrence (DR) and graft loss following renal transplant (RT). Although pre-emptive plasma exchange (PE) and rituximab have been suggested to prevent DR, there is insufficient published data to support this practice. The aim is to study the role of pre-emptive PE and rituximab in the prevention of DR in children with non-genetic SRNS undergoing living donor (LD) RT. METHODS: Prospective single-centre study of four consecutive children (age 6-17 years) with non-genetic SRNS (including two with previous graft loss due to DR) who underwent LD RT between July 2014 and September 2016. All patients received a single dose of rituximab 375 mg/m2 2-4 weeks prior to the RT and four sessions of PE in the week prior to RT. All patients had previously undergone bilateral native nephrectomies. RESULTS: All children had early DR (2-26 days) following LD RT. Following early initiation of PE, three children achieved partial remission (PR) or complete remission (CR) 5-22 days after commencing treatment. One child continued to have heavy proteinuria along with graft dysfunction despite 52 sessions of PE and lost the graft 5 months after RT. At the latest follow-up of 36-60 months following RT, one child remains in CR and two are in PR. The latest eGFR was 45-104 ml/min/1.73m2. CONCLUSIONS: Pre-emptive rituximab and PE does not prevent DR in high-risk non-genetic SRNS. Prompt initiation of PE following DR appears to achieve PR or CR in the majority of patients.

Bilateral native nephrectomies for severe hypertension in children with stage 5 chronic kidney disease leads to improved BP control following transplantation.

BACKGROUND: Hypertension is a common problem in stage 5 chronic kidney disease (CKD 5) and following kidney transplantation (KT). There is limited data on the outcome of children with CKD 5 who undergo bilateral native nephrectomies (BNN) for the management of hypertension. METHOD: Retrospective review of 134 children who underwent KT at a single centre over a 10-year period and had a minimum follow up period of 1 year. Children who had undergone BNN for hypertension prior to, and after, KT were identified and their outcome with regard to blood pressure (BP), anti-hypertensive medications and graft function was compared with that of the rest of the cohort. RESULTS: Eleven children (8.2%) underwent BNN, including 2 performed after KT, due to poorly controlled BP despite a median of 3 anti-hypertensive medications. The median age at BNN was 7 years (range 0.5-17 years). All 9 children who underwent BNN prior to KT discontinued anti-hypertensive medication after a median of 6 months and remained normotensive post KT. After a median follow up of 5 years following KT, there was a trend towards lower prevalence of hypertension in children who underwent BNN compared with that of the rest of the cohort (9.1% vs 25%, p 0.23). None of the children who underwent BNN had any evidence of proteinuria, and the median eGFR was 74 ml/min/1.73 m 2 after KT. CONCLUSION: BNN for severe hypertension in CKD 5 is associated with resolution of hypertension prior to KT. It is also associated with a trend towards lower prevalence of hypertension and good graft function following KT.

Clinical Efficacy of Topical Tranexamic Acid With Microneedling in Melasma.

BACKGROUND: Melasma is a common, recurrent, and refractory cause of facial pigmentation resulting in cosmetic disfigurement. Tranexamic acid (TA) has been used systemically and locally for clearance of pigmentation. OBJECTIVE: To assess the clinical efficacy of topical TA (10%) with microneedling in melasma. METHODS: A split face, prospective, randomized, open-label study with a sample size of 40. Left or right side of the face was chosen randomly and microneedling was done on both the sides, followed by 10% TA solution application on one side of the face (test side) and distilled water on the other side of face (control). The procedure was done at 2 weekly intervals (0, 2, 4, and 6 weeks). Clinical images were taken at each visit including modified Melasma Area and Severity Index (mMASI) scoring of each half of the face to assess the clinical response along with patient satisfaction scores and side effects. RESULTS: On the test side, there was 65.92% improvement in the mean mMASI score compared with 20.75% on the control side of the face at the end of 8 weeks. CONCLUSION: Tranexamic acid may be a promising therapeutic agent in melasma and the topical solution along with microneedling seems to be efficacious.

Quantification of human C1 esterase inhibitor protein using an automated turbidimetric immunoassay.

BACKGROUND: Impaired levels or function of C1 inhibitor (C1-INH) results in angioedema due to increased bradykinin. It is important to distinguish between angioedema related to C1-INH deficiency and that caused by other mechanisms, as treatment options are different. In hereditary (HAE) and acquired (AAE) angioedema, C1-INH concentration is measured to aid patient diagnosis. Here, we describe an automated turbidimetric assay to measure C1-INH concentration on the Optilite® analyzer. METHODS: Linearity, precision, and interference were established over a range of C1-INH concentrations. The 95th percentile reference interval was generated from 120 healthy adult donors. To compare the Optilite C1-INH assay with a predicate assay used in a clinical laboratory, samples sent for C1-INH investigation were used. The predicate results were provided to allow comparison. RESULTS: The Optilite C1-INH assay was linear across the measuring range at the standard sample dilution. Intra and interassay variability was <6%. The 95th percentile adult reference interval for the assay was 0.21-0.38 g/L. There was a strong correlation between the Optilite concentrations and those generated with the predicate assay (R2  = 0.94, P < 0.0001, slope y = 0.83x). All patients with Type I HAE (n = 24) and AAE (n = 3) tested had concentrations below the measuring range in both assays, while all patients with unspecified angioedema (UAE), not diagnosed with HAE or AAE had values within the reference range. CONCLUSION: The Optilite assay allows the automated and precise quantification of C1-INH concentrations in patient samples. It could therefore be used as a tool to aid the investigation of patients with angioedema.

Active modified atmosphere packaging of yellow bell pepper for retention of physico-chemical quality attributes.

This investigation was carried out to evaluate the effect of active and passive modified atmosphere packaging on quality and shelf life of yellow bell pepper fruits. Yellow bell pepper fruits were packaged in 150 gauge LDPE packages with oxygen absorbers for active modification and without oxygen absorber for passive modification of headspace and were stored at different temperatures i.e. 5, 10 and 15 °C and RH of 85 ± 5%. Headspace gas concentration within the packages was monitored regularly. The quality of packaged fruits was studied in terms of physiological loss in weight, firmness, total colour difference antioxidant capacity and total phenolic content. The actively modified packages attained steady state levels of 4.8% O2 and 7.1% CO2 on 4th day of storage as compared to passively modified packages in which steady state was not attained even at end of storage period of 12 days. The retention of quality attributes was observed to be higher in active packages than in passive packages. Moreover, the shelf life of actively packaged fruits was enhanced to 28 days as compared to 12 days for passively packaged fruits. The in-pack atmosphere attained in active packages hence proved beneficial in retarding the senescence thereby extending the shelf life.

Topical delivery of tetrahydrocurcumin lipid nanoparticles effectively inhibits skin inflammation: in vitro and in vivo study.

Tetrahydrocurcumin (THC) also referred to as 'white curcumin', is a stable colorless hydrogenated product of curcumin with superior antioxidant and anti-inflammatory properties. The present study is an attempt to elevate the topical bioavailability of THC, post-incorporation into a nano-carrier system with its final dosage as a hydrogel. Lipid nanoparticles of THC (THC-SLNs) prepared by microemulsification technique were ellipsoidal in shape (revealed in transmission electron microscopy) with a mean particle size of 96.6 nm and zeta potential of -22 mV. Total drug content and entrapment efficiency of THC-SLNs was 94.51% ± 2.15% and 69.56% ± 1.35%, respectively. Differential scanning calorimetry and X-ray diffraction studies confirmed the formation of THC-SLNs. In vitro drug release studies showed the drug release from THC-SLNs gel to follow Higuchi's equation revealing a Fickian diffusion. Ex vivo permeation studies indicated a 17 times (approximately) higher skin permeation of THC-SLNs gel as compared with the free THC gel. Skin irritation, occlusion, and stability studies indicated the formulation to be nonirritating, and stable with a desired occlusivity. Pharmacodynamic evaluation in an excision wound mice model clearly revealed the enhanced anti-inflammatory activity of THC-SLNs gel and was further confirmed using biochemical and histopathological studies. It is noteworthy to report here that THC-SLNs gel showed significantly better (p ≤ 0.001) activity than free THC in gel. As inflammation is innate to all the skin disorders, the developed product opens up new therapeutic avenues for several skin diseases. To the best of our knowledge, this is the first paper elaborating the therapeutic usefulness of white curcumin-loaded lipidic nanoparticles for skin inflammation.

Physico-chemical properties, bioactive compounds and color parameters of coriander puree: effect of pretreatments and freezing.

Coriander, one of the most widely utilized herb, is a short duration herb having very short shelf life. The present investigation involved development of an innovative process for enhancing the utilization and the availability of this herb. This process involves pretreatment, blanching and freezing to form ready to use frozen coriander puree tablets. Coriander was subjected to different pretreatments viz. T1: 0.25% potassium metabisulphite, T2: 0.5% citric acid and T3: 0.2% potassium metabisulphite + 0.1% magnesium chloride + 0.1% sodium bicarbonate. The pretreated coriander was blanched in hot water for 2 min and processed into puree having total soluble solids (TSS) as 2.5, 3.5 and 4.5°Brix. Pretreatments significantly affected the all quality parameters except moisture content. TSS had non significant effect on a* value, chroma, pH ascorbic acid and total flavonoid content of processed coriander puree. Coriander puree obtained with T1 pretreatment exhibited better quality than with other pretreatments. T1 pretreated and blanched puree was then frozen in silicon moulds using air blast freezing and deep freezing. Frozen puree tablets were evaluated for quality. The best quality coriander puree obtained from T1 pretreated, blanched puree having TSS 4.5°Brix frozen by continuous air-blast freezing (at a conveyor speed of 4.40 × 10-3m/s) comparable results to deep freezing having. All quality parameters, except pH, were significantly affected by TSS level of puree as well as different freezing treatments.

Factor H autoantibody is associated with atypical hemolytic uremic syndrome in children in the United Kingdom and Ireland.

Factor H autoantibodies can impair complement regulation, resulting in atypical hemolytic uremic syndrome, predominantly in childhood. There are no trials investigating treatment, and clinical practice is only informed by retrospective cohort analysis. Here we examined 175 children presenting with atypical hemolytic uremic syndrome in the United Kingdom and Ireland for factor H autoantibodies that included 17 children with titers above the international standard. Of the 17, seven had a concomitant rare genetic variant in a gene encoding a complement pathway component or regulator. Two children received supportive treatment; both developed established renal failure. Plasma exchange was associated with a poor rate of renal recovery in seven of 11 treated. Six patients treated with eculizumab recovered renal function. Contrary to global practice, immunosuppressive therapy to prevent relapse in plasma exchange-treated patients was not adopted due to concerns over treatment-associated complications. Without immunosuppression, the relapse rate was high (five of seven). However, reintroduction of treatment resulted in recovery of renal function. All patients treated with eculizumab achieved sustained remission. Five patients received renal transplants without specific factor H autoantibody-targeted treatment with recurrence in one who also had a functionally significant CFI mutation. Thus, our current practice is to initiate eculizumab therapy for treatment of factor H autoantibody-mediated atypical hemolytic uremic syndrome rather than plasma exchange with or without immunosuppression. Based on this retrospective analysis we see no suggestion of inferior treatment, albeit the strength of our conclusions is limited by the small sample size.