Serum Trypsinogen Research Articles

Applicability of the Pancreatic Exocrine Insufficiency Test (PEI-TEST) in Pediatric Patients.

In mild cases, it is difficult to diagnose pancreatic exocrine insufficiency (PEI). There is no gold standard method for the diagnosis of PEI. A reliable method is needed for preliminary diagnosis of PEI. The PEI-TEST was applied to the patients with nonspecific gastrointestinal complaints. Serum amylase, lipase, serum trypsinogen, and fecal elastase 1 (FE-1) were analyzed from each patient. According to the PEI-TEST, PEI was present in 42 (47.7%) and PEI was not present in 46 (52.3%) patients. No significant difference was observed between the 2 groups with regard to age, gender and amylase, lipase, serum trypsinogen, and FE-1. When an FE-1 value of <200 µg/dL was considered as indicating PEI, the sensitivity and specificity of the test were found to be 47.4% and 52.2%, respectively. Although it is promising that PEI-TEST is a validated test in our country and suitable for our society, it is not suitable for pediatric patients.

RF16 | PSUN203 Deep Characterization of Pancreas Volume of New-Onset Type 1 Diabetes Patients Reveals Puberty-Specific Patterns and New Topographic Correlations with Pancreatic Functions.

Abstract There is a current understanding of type 1 diabetes (T1D) as being the result of a heterogeneous disease of whole pancreas. So far, descriptions of pancreas alterations suggested a global involvement of inflammation throughout the organ, characterized by abnormal age-specific patterns (i.e. &amp;gt; or &amp;lt;7 yo). In our DIATAG study, we thoroughly characterized pancreatic structure modifications in new-onset T1D patients and investigated whether these correlated with pancreatic functions during the first postdiagnosis year. Pediatric patients with new-onset T1D (n=31; 11.2±2.9 years) underwent abdominal MRI scan at diagnosis (i.e. 40±37 days postdiagnosis). Exocrine function (i.e. serum trypsinogen) and residual endocrine secretion estimates (i.e. fasten = CPEPBASAL, stimulated model = CPEPEST (1)) were evaluated on fasten blood tests at diagnosis (Δ) and Δ+3, +12 months, respectively. Clinical parameters of glucose homeostasis (i.e. A1C, insulin daily dose, insulin-dose adjusted A1c [IDAA1C]) were collected at Δ+3, +6, +9 and +12 months. We compared pancreas volumes (i.e. PV, PVHEAD, PVBODY, PVTAIL) and ratios (pancreas volume ratio [PVR] = PVHEAD/PVBODY+TAIL; pancreas index [PI] = PV/body weight) to retrospectively identify age-, BMI- and sex-matched controls (n=29). Compared to control group, T1D patients demonstrated a homogeneous (PVR, 1.39±0.7 vs 1.46±0.4, p&amp;gt;0.05) decrease of PI (0.6±0.2 vs 1.1±0.3 mL/kg, p&amp;lt;0.0001) independently of pancreatic subregion. Interestingly, prepubertal children with T1D (n=15) exhibited greater pancreas atrophy of both the body-tail (10±5.6%, p&amp;lt;0.05) and head (7.2±3%, p&amp;lt;0.05) subregions compared to pubertal group (n=16; PI difference of mean, 0.64 vs 0.48 mL/kg). Next, we evaluated correlations between pancreatic volumes and functions during the natural evolution of T1D using multivariate and univariate linear regressions. PV and PI correlated moderately with exocrine function (trypsinogen; R=0.46, R=0.52, p&amp;lt;0.001) but not with residual endocrine function evaluated at Δ+3 months. Interestingly, we observed using topographical analysis that PVHEAD correlated with trypsinogen (R=0.5, p&amp;lt;0.001) but not with any endocrine estimates (p&amp;gt;0.05). PVTAIL positively correlated with CPEPEST and CPEPBASAL at Δ+3 months (R=0.55, p=0.002; R=0.47, p&amp;lt;0.01) and not with clinical parameters or trypsinogen (p&amp;gt;0.05). PVBODY did not correlate with any pancreatic function. Finally, we identified multivariate models that included PV and predicted pancreatic endocrine function at Δ+6 and +12 months (i.e. IDAA1C, HbA1C). MRI performed early after T1D onset helped us define specific pancreas alterations and demonstrate for the first time: (1) the increased pancreas atrophy in prepubertal children; (2) correlations between pancreatic subregion volumes and their respective residual functions early after diagnosis and (3) the improvement of endocrine function prediction models by adding PV estimates as predictors. (1) Wentworth et al. 2019;62(1): 33–40 Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m., Sunday, June 12, 2022 1:00 p.m. - 1:05 p.m.

Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells (huMSCs) Carrying MicroRNA-342 Relieve Protect Severe Acute Pancreatitis Injury (SAP)

To investigate whether exosomes (exo) derived from human umbilical cord mesenchymal stem cells (huMSCs) and microRNA (miRNA)-342 have a protective effect on severe acute pancreatitis (SAP). Human umbilical cord blood was collected to extract huMSC-exo. With sham-operated mice as control group (n = 10), the other mice were induced to SAP model (n = 20), while 10 of the SAP mice received treatment with huMSC-exo. ELISA was performed to determine amylase and TAP level as well as inflammatory factors and HE staining to evaluate pathological changes of pancreatic tissue. The expression of miR-342 and Shh, Ptchl, and Smo in the Hh signal pathway was detected using RT-qPCR. The expression of miR-342 and the mRNA expression of Shh, Ptchl, and Smo was higher than that in model group (p &lt; 0.05). The level of serum amylase, trypsinogen, and IFN-γ,Fasl, and IL-6 was upregulated in pancreas tissues of SAP mice relative to healthy mice, but their levels were decreased upon treatment with huMSC-exo and slightly higher than those of the control group, just not significantly. Collectively, the huMSC-exo may activate the Hh signaling pathway by regulating the expression of miR-342 increasing the expression of Shh, Ptchl, and Smo, and thereby healing of damaged pancreatic tissues in SAP.

Biochemical markers in differentiating gallstones pancreatitis from non-gallstone pancreatitis

Acute pancreatitis has many causes; however, the most common cause of pancreatitis is gallstones globally. We have searched for relevant studies in this field aiming at establishing a short review about the usefulness of biomarkers in the diagnosis of acute pancreatitis and the relevant biomarkers and tests that can differentiate acute gallstone pancreatitis from other etiologies. It is worth mentioning that for the diagnosis and prognosis of biliary pancreatitis, radiological and laboratory diagnosis plays a major role in this field. Three biomarkers including serum amylase, lipase, and trypsinogen are the most important and most frequently noticed biomarkers in association with acute pancreatitis. Serum amylase has a specificity rate of 95% and a sensitivity of 61% when it measures more than 1000 IU/l which is three times as high as the normal level. However, it can be also found in other intrabdominal inflammatory conditions, and therefore, it cannot be used alone in the diagnosis. It has been reported that the specificity and sensitivity were 95% and 94%, respectively, for detecting high trypsinogen-2 levels in the patients’ urinary samples, which were indicative of acute pancreatitis. For the diagnosis of biliary pancreatitis, liver function tests should be assessed. Although they might be specific, they are not always diagnostic in some cases, and therefore, other approaches for detecting gallstones as ultrasonography and MRCP should be considered together with the liver enzymes for an appropriate diagnosis.

Exocrine Pancreatic Enzymes Are a Serological Biomarker for Type 1 Diabetes Staging and Pancreas Size.

Exocrine pancreas abnormalities are increasingly recognized as features of type 1 diabetes. We previously reported reduced serum trypsinogen levels and in a separate study, smaller pancreata at and before disease onset. We hypothesized that three pancreas enzymes (amylase, lipase, and trypsinogen) might serve as serological biomarkers of pancreas volume and risk for type 1 diabetes. Amylase, lipase, and trypsinogen were measured from two independent cohorts, together comprising 800 serum samples from single-autoantibody-positive (1AAb+) and multiple-AAb+ (≥2AAb+) subjects, individuals with recent-onset or established type 1 diabetes, their AAb-negative (AAb-) first-degree relatives, and AAb- control subjects. Lipase and trypsinogen were significantly reduced in ≥2AAb+, recent-onset, and established type 1 diabetes subjects versus control subjects and 1AAb+, while amylase was reduced only in established type 1 diabetes. Logistic regression models demonstrated trypsinogen plus lipase (area under the receiver operating characteristic curve [AUROC] = 81.4%) performed equivalently to all three enzymes (AUROC = 81.4%) in categorizing ≥2AAb+ versus 1AAb+ subjects. For cohort 2 (n = 246), linear regression demonstrated lipase and trypsinogen levels could individually and collectively serve as indicators of BMI-normalized relative pancreas volume (RPVBMI, P < 0.001), previously measured by MRI. Serum lipase and trypsinogen levels together provide the most sensitive serological biomarker of RPVBMI and may improve disease staging in pretype 1 diabetes.

Low serum trypsinogen levels in chronic pancreatitis: Correlation with parenchymal loss, exocrine pancreatic insufficiency, and diabetes but not CT-based cambridge severity scores for fibrosis

BackgroundChronic pancreatitis (CP) is a complex inflammatory disorder of the pancreas affecting acinar cells, duct cells, islet cells and inflammatory cells including fibrosis-producing stellate cells. Serum trypsinogen is a biomarkers of acinar cell function. AimTo define the degree of correlation between low trypsinogen levels as a marker of acinar cell function and variable features of CP. MethodsSerum samples from previously ascertained and well phenotyped case and control subjects from the North American Pancreatitis Study II (NAPS2) were used to measure serum trypsinogen levels in a commercial laboratory. Control samples were used to define normal ranges and compared with levels in CP patients with defined features. ResultsA final cohort of 279 CP patients and 262 controls from the NAPS2 studies were evaluated. In controls trypsinogen had a mean of 34.96 ng/ml and SD = 11.99. Cut-off values for low trypsinogen ranged from <20 to 10 ng/ml and very low trypsinogen at <10 ng/ml. Compared to controls, CP was associated with very low trypsinogen levels (p < 0.0001). Within CP, very low trypsinogen levels correlated with parenchymal loss (pancreatic surgery [p < 0.05]; atrophy with calcifications, [p < 0.001]), EPI (p < 0.01, trend p < 0.001) and diabetes (trend p < 0.01) but not CT-based criteria for fibrosis (pancreatic duct dilation, irregularity, strictures). ConclusionsVery low serum trypsinogen levels correlate with measures of acinar cell loss including surgical resection, atrophic-calcific CP, diabetes and functional symptoms EPI but not duct morphology criteria. Serum trypsinogen levels correlate with decreased acinar cell function and therefore have biomarker utility clinical management.

Serum Trypsinogen and Lipase as Biomarkers of Exocrine Pancreatic Function in Newly Diagnosed Children and Adolescents with Type 1 Diabetes Mellitus

Type 1 diabetes mellitus (T1DM) in literature has usually been associated with laboratory decrease in pancreatic exocrine enzymes levels. Nevertheless, the exact onset of decreased pancreatic enzymes levels in sera of patients is still unknown. We measured pancreatic serum levels of trypsinogen and lipase in Egyptian children and adolescents with newly diagnosed T1DM to reveal their role in diagnosis and their relation to HbA1c as a marker of glycemic control, lipid profile and fasting c-peptide as a marker of endocrine pancreatic insufficiency.

Hydrostatin-SN10 Ameliorates Pancreatitis-Induced Lung Injury by Affecting IL-6-Induced JAK2/STAT3-Associated Inflammation and Oxidative Stress

Hydrostatin-SN1 (peptide sequence, DEQHLETELHTLTSVLTANGFQ), a kind of peptides extracted from snake venom, has been reported to have anti-inflammatory effect, but its truncated mutant hydrostatin-SN10 (peptide sequence, DEQHLETELH) on pancreatitis-induced acute lung injury has not been well documented. Interleukin- (IL-) 6-induced Janus Kinase 2/Signal Transducer and Activator of Transcription 3 (JAK2/STAT3) pathway is involved with inflammatory and oxidative stress activities and may be associated with the pathogenesis of lung injury, and related molecules were measured. Taurocholate-induced pancreatitis associated with acute lung injury was established and treated with hydrostatin-SN10. Pancreatitis was confirmed by measuring the serum levels of amylase, lipase, and trypsinogen and urinary amylase. Lung injury was determined by histologically assessing acinar cell changes. The related molecules of IL-6-induced JAK2/STAT3-associated inflammation and oxidative stress were quantitated by real time-PCR, Western blot, and/or immunochemical assay. Hydrostatin-SN10 reduced the levels of serum amylase, lipase, and trypsinogen and urinary amylase when compared with the model group (p < 0.05). Hydrostatin-SN10 significantly inhibited the IL-6-stimulated JAK2/STAT3 pathway and reduced the number of apoptotic cells via the downregulation of caspase 3 and BAX (proapoptotic) and upregulation of Bcl2 (antiapoptotic) (p < 0.05). IL-6 induced the increase in the levels of JAK2 and STAT3, which was reversed by hydrostatin-SN10 treatment (p < 0.05). In addition, hydrostatin-SN10 reduced the expression of IL-6 and TNF- (tumor necrosis factor-) α and increased the level of IL-10 (p < 0.05). On the other hand, hydrostatin-SN10 treatment increased the levels of superoxide dismutase (SOD) and reduced glutathione (GSH) and the levels of malondialdehyde (MDA) and alanine aminotransferase (ALT) (p < 0.05). These results suggest that hydrostatin-SN10 may inhibit pancreatitis-induced acute lung injury by affecting IL-6-mediated JAK2/STAT3 pathway-associated inflammation and oxidative stress.

Secondhand Smoke Exposure and Serum Trypsinogen in Cystic Fibrosis Carriers.

The objective of this study was to determine if infants carrying 1 cystic fibrosis transmembrane receptor (CFTR) mutation demonstrate pancreatic inflammation in response to tobacco exposure. Cystic fibrosis carrier infants aged 4 to 16 weeks were prospectively enrolled. Tobacco exposure was assessed by survey and maternal hair nicotine analysis. Serum immunoreactive trypsinogen (IRT) levels at birth and at the time of recruitment were analyzed relative to the presence or absence of tobacco exposure. The effect of the severity of the CFTR mutation carried by the infant on the tobacco-IRT relationship was also analyzed. Forty-eight infants completed the study. Newborn screen and follow-up IRT levels were not different between exposed infants (19 by hair analysis) and nonexposed infants (29 by hair analysis). Follow-up IRT levels were lower in infants with more severe CFTR mutations (P = 0.005). There was no difference in follow-up IRT based on CFTR mutation severity in exposed infants. Nonexposed infants with milder CFTR mutations had higher median IRT values on follow-up testing than those with more severe CFTR mutations (P < 0.05). The pancreas of cystic fibrosis carrier infants is affected by tobacco exposure, and those carrying less severe CFTR mutations may be more susceptible to tobacco effects.

Method Performance of Bicarbonate and Electrolytes by Use of Duodenal Fluid for Assessment of Exocrine Pancreatic Function.

Evaluation of exocrine pancreatic insufficiency is challenging for both clinicians and laboratories. Indirect pancreatic function tests such as serum trypsinogen, fecal elastase, and fecal fat measurements are moderately sensitive for diagnosis of advanced chronic pancreatitis but show reduced sensitivity and specificity for diagnosis of early disease. An alternative is the endoscopic pancreatic function test, which uses duodenal secretions after administration of IV secretin. Samples are collected at various times via the endoscopic tube and then analyzed for bicarbonate, sodium, potassium, and chloride. Precision, linearity, method comparison, and stability studies were performed on the Beckman Coulter AU5822 chemistry analyzer with duodenal fluid. Comparison with the Vitros 4600 dry slide chemistry instrument was used to interrogate differences between methods. All assays produced a CV <2% without any measurable effects from the endoscopy fluid matrix and showed acceptable imprecision near the limit of detection (CV < 5%). All analytes showed linear dilution across the analytical measuring range. All the calculated error biases from dilutions were within 50% of the CLIA-allowable error for serum for each of the respective analytes. The calculated slopes ranged from 0.841 to 1.274 when compared to the Vitros 4600. Stability studies demonstrated that sodium, potassium, chloride, and bicarbonate remained stable after storage at -20 °C and after multiple freeze-thaw cycles. The percent change for all analytes was <5% mmol/L. The AU5800 series demonstrated adequate performance for the analysis of bicarbonate in duodenal fluid and therefore can be used for assessment of exocrine pancreatic function. However, notable discrepancies were observed for sodium, potassium, and chloride between the AU5800 series and the Vitros 4600.

Exocrine Pancreatic Function as a Novel Biomarker in Pre-T1D

Recent studies demonstrated reduced serum trypsinogen levels as well as pancreatic weight/volume in patients with type 1 diabetes (T1D) and nondiabetic/autoantibody (AAb)-positive subjects. Histological defects including increased CD8+ T cell infiltration and elevated C4d deposition have been observed in the exocrine T1D pancreas. Like trypsinogen, amylase and lipase are produced by the exocrine pancreas and serve as more common markers of exocrine function. We hypothesized that amylase and lipase levels would be lower in patients with T1D as well as in subjects with multiple (i.e., ≥2) T1D-related AAb compared with controls. To test this, serum amylase and lipase levels were determined in 70 patients with recent-onset T1D (duration &amp;lt;3 mo), 57 patients with established T1D (duration &amp;gt;3 mo), 56 single AAb positive (1AAb+) subjects, 42 multiple AAb positive (≥2AAb+) subjects, 96 AAb negative (AAb-) relatives of T1D patients, and 110 AAb- control subjects. Amylase levels, which are independent of age (r = -0.02, P = 0.7), were significantly lower in patients with T1D (mean ± SD 39.98 ± 17.8 ng/mL; P &amp;lt; 0.005) vs. AAb- subjects (47.40 ± 20.17 ng/mL). Lipase levels, which are positively correlated with age (r = 0.30, P &amp;lt; 0.0001), were significantly lower in T1D patients (25.95 ± 21.ng/mL; P &amp;lt; 0.001) as well as ≥2AAb+ subjects (22.59 ± 11.12 ng/mL; P &amp;lt; 0.001) compared with 1AAb+ individuals (37.60 ± 16.79 ng/mL). These results provide further evidence that exocrine pancreas abnormalities exist in patients with T1D and ≥2AAb+ subjects considered to have early-stage T1D with high risk of progressing to clinical diagnosis. These data also support the possibility that an inherent or acquired defect in exocrine pancreas development and/or function may contribute to disease pathogenesis, but cause and effect cannot yet be ascertained. Importantly, these exocrine pancreas enzymes may serve as biomarkers of T1D to better predict disease progression and/or stratify patients according to etiology. Disclosure J.J. Ross: None. C. Wasserfall: None. D.J. Perry: None. K.M. McGrail: None. A.L. Posgai: None. T.M. Brusko: Stock/Shareholder; Self; OneVax, LLC. Advisory Panel; Self; Caladrius Biosciences, Inc.. Consultant; Self; Merck &amp; Co., Inc., Sanofi-Aventis. D. Schatz: None. M.J. Haller: None. M.A. Atkinson: Other Relationship; Self; Patent Issued.

Serum Trypsinogen Levels in Type 1 Diabetes.

OBJECTIVEThe pancreas in type 1 diabetes exhibits decreased size (weight/volume) and abnormal exocrine morphology. Serum trypsinogen levels are an established marker of pancreatic exocrine function. As such, we hypothesized that trypsinogen levels may be reduced in patients with pre–type 1 diabetes and type 1 diabetes compared with healthy control subjects.RESEARCH DESIGN AND METHODSSerum trypsinogen levels were determined in 100 persons with type 1 diabetes (72 new-onset, 28 established), 99 autoantibody-positive (AAb+) subjects at varying levels of risk for developing this disease, 87 AAb-negative (AAb−) control subjects, 91 AAb− relatives with type 1 diabetes, and 18 patients with type 2 diabetes.RESULTSTrypsinogen levels increased significantly with age in control subjects (r = 0.71; P < 0.0001) and were significantly lower in patients with new-onset (mean ± SD 14.5 ± 6.1 ng/mL; P < 0.0001) and established type 1 diabetes (16.7 ± 6.9 ng/mL; P < 0.05) versus AAb− control subjects (25.3 ± 11.2 ng/mL), AAb− relatives (29.3 ± 15.0 ng/mL), AAb+ subjects (26.5 ± 12.1 ng/mL), and patients with type 2 diabetes (31.5 ± 17.3 ng/mL). Multivariate analysis revealed reduced trypsinogen in multiple-AAb+ subjects (P < 0.05) and patients with type 1 diabetes (P < 0.0001) compared with AAb− subjects (control subjects and relatives combined) and single-AAb+ (P < 0.01) subjects when considering age and BMI.CONCLUSIONSThese findings further support the interplay between pancreatic endocrine and exocrine dysfunction. Longitudinal studies are warranted to validate trypsinogen as a predictive biomarker of type 1 diabetes progression.

Both Exocrine Pancreatic Insufficiency and Signs of Pancreatic Inflammation Are Prevalent in Children with Complicated Severe Acute Malnutrition: An Observational Study

To assess whether pancreatic function is impaired in children with severe acute malnutrition, is different between edematous vs nonedematous malnutrition, and improves by nutritional rehabilitation. We followed 89 children with severe acute malnutrition admitted to Queen Elizabeth Central Hospital in Blantyre, Malawi. Stool and blood samples were taken on admission and 3days after initial stabilization to determine exocrine pancreatic function via fecal elastase-1 (FE-1) and serum trypsinogen and amylase levels. A total of 33 children (37.1%) had nonedematous severe acute malnutrition, whereas 56 (62.9%) had edematous severe acute malnutrition. On admission, 92% of patients showed evidence of pancreatic insufficiency as measured by FE-1 <200μg/g of stool. Patients with edematous severe acute malnutrition were more likely to have low FE-1 (98% vs 82.8%, P=.026). FE-1 levels remained low in these individuals throughout the assessment period. Serum trypsinogen was elevated (>57ng/mL) in 28% and amylase in 21% (>110 U/L) of children, suggesting pancreatic inflammation. Exocrine pancreatic insufficiency is prevalent in children with severe acute malnutrition and especially in children with edematous severe acute malnutrition. In addition, biochemical signs suggestive of pancreatitis are common in children with severe acute malnutrition. These results have implications for standard rehabilitation treatment of children with severe acute malnutrition who may benefit from pancreatic enzyme replacement therapy. ISRCTN.com:13916953.

The North American Shwachman-Diamond Syndrome Registry: Genetically Undefined Shwachman-Diamond Syndrome

Shwachman-Diamond syndrome (SDS) is an inherited marrow failure syndrome associated with exocrine pancreatic dysfunction and an increased risk of myelodysplasia and leukemia. The majority of individuals with SDS carry biallelic SBDS gene mutations, however a subset of patients remain genetically undefined. The objective of this study was to compare the clinical characteristics of patients with and without SBDS mutations. To address these questions, we conducted a retrospective study of patients enrolled on the North American Shwachman-Diamond Syndrome Registry (SDSR). Clinical data from the SDSR were available for 55 individuals with biallelic SBDS mutations and 16 individuals who fulfilled clinical diagnostic criteria for SDS but lacked biallelic mutations in the SBDS gene. Study subject ages for SBDS mutation positive and negative cohorts span 2-52.4 and 2.8-21.4 years with median ages of 12.4 and 10.9 years respectively. Cytopenias were present for both SBDS mutation positive and negative cohorts, with neutropenia the most common event in 94% and 81% respectively. Bone marrow hypocellularity was reported in 91% of those with SBDS mutations and 69% of those without. Marrow dysplasia was reported in 65% of those with SBDS mutations and none of those without. Clonal abnormalities were present in 44% and 25% of those with and without SBDS mutations with median age of initial appearance at 9 years (0.8-45.1) and 7 years (1.2-14) respectively. Abnormalities included del7q and del20q in both groups as well as iso7q, trisomy 8 and others in the SBDS mutation positive group. Clonal abnormalities were all transient in the SBDS mutation negative cohort. One SBDS mutation positive individual developed AML. None of the SBDS mutation negative individuals developed malignancy or progressed to require HSCT thus far. Pancreatic dysfunction determined by low serum trypsinogen or pancreatic isoamylase was similar in both cohorts 79% vs 80%. However, only 27% (15/55) of SBDS mutation positive individuals reported requiring enzyme therapy with 33% (18/55) documenting failure to thrive, in contrast to 75% (12/16) of SBDS mutation negative individuals with 73% (11/15) having failure to thrive. A broad spectrum of congenital anomalies were reported in 55% and 56% of SBDS mutation positive and negative individuals respectively, with skeletal anomalies being the most common in both groups. Medical comorbidities commonly reported in both groups included eczema and endocrinopathies. Elevated liver transaminases were seen in 27% of SBDS mutation positive individuals but this was not seen in the SBDS mutation negative cohort.Conclusion: Patients with genetically undefined (SBDS mutation negative) SDS share clinical characteristics with SBDS mutation positive patients; however, the risk of leukemia in the genetically undefined patients remains unclear due to low patient numbers with short follow-up. Further studies of this young cohort are required to inform medical management and to advance our understanding of genetic etiology, mechanism, disease pathophysiology and treatment of these marrow failure disorders. DisclosuresDale:Amgen: Consultancy, Honoraria, Research Funding, Speakers Bureau.

Inconclusive diagnosis of cystic fibrosis after newborn screening.

To prospectively study infants with an inconclusive diagnosis of cystic fibrosis (CF) identified by newborn screening (NBS; "CF screen positive, inconclusive diagnosis" [CFSPID]) for disease manifestations. Infants with CFSPID and CF based on NBS from 8 CF centers were prospectively evaluated and monitored. Genotype, phenotype, repeat sweat test, serum trypsinogen, and microbiology data were compared between subjects with CF and CFSPID and between subjects with CFSPID who did (CFSPID→CF) and did not (CFSPID→CFSPID) fulfill the criteria for CF during the first 3 years of life. Eighty-two subjects with CFSPID and 80 subjects with CF were enrolled. The ratio of CFSPID to CF ranged from 1:1.4 to 1:2.9 in different centers. CFTR mutation rates did not differ between groups; 96% of subjects with CFSPID and 93% of subjects with CF had 2 mutations. Subjects with CFSPID had significantly lower NBS immunoreactive trypsinogen (median [interquartile range]:77 [61-106] vs 144 [105-199] μg/L; P < .0001) than did subjects with CF. Pseudomonas aeruginosa and Stenotrophomonas maltophilia were isolated in 12% and 5%, respectively, of subjects with CFSPID. CF was diagnosed in 9 of 82 (11%) subjects with CFSPID (genotype and abnormal sweat chloride = 3; genotype alone = 4; abnormal sweat chloride only = 2). Sweat chloride was abnormal in CFSPID→CF patients at a mean (SD) age of 21.3 (13.8) months. CFSPID→CF patients had significantly higher serial sweat chloride (P < .0001) and serum trypsinogen (P = .009) levels than did CFSPID→CFSPID patients. A proportion of infants with CFSPID will be diagnosed with CF within the first 3 years. These findings underscore the need for clinical monitoring, repeat sweat testing at age 2 to 3 years, and extensive genotyping.

COST-EFFECTIVE AND EFFICIENT APPARATUS FOR ELECTROELUTION OF MICRO- AND MACROGRAM QUANTITIES OF PROTEINS FROM POLYACRYLAMIDE GELS

Protein recovery from gel electrophoresis plays a significant role in functional genomics and proteomics. To assist in this, a simple, cost-effective, and efficient apparatus for electroelution of proteins has been designed. The performance of the apparatus was demonstrated using the proteins bovine serum albumin (BSA), phosphorylase, ovalbumin, pepsin, and trypsinogen. In all the cases the yield of elution was found to be consistently greater than 85% and the proteins could be eluted without degradation in less than 15 min. The utility of this method can be extended to protein elution from denatured and native polyacrylamide gels, DNA purification from agarose gels, and oligomeric primers purification from polyacrylamide gels. In addition to this, the method offers an effortless purification and characterization of microbial extracellular proteins. The eluted proteins can be directly used in N-terminal amino acid sequencing, and in amino acid and proteomics analyses.

Risk factors for acute pancreatitis in patients with severe motor and intellectual disabilities

Acute pancreatitis in patients with severe motor and intellectual disability (SMID) is a rare but life-threatening condition. Possible causes of acute pancreatitis in these patients including valproic acid therapy, hypothermia and nasoduodenal tube feeding, have not yet been investigated in detail. The aim of this study was therefore to investigate the risk factors for acute pancreatitis in patients with SMID. Five SMID patients with acute pancreatitis and 15 SMID patients without acute pancreatitis were reviewed. Age; serum total cholesterol, triglyceride, total protein, and albumin; height; bodyweight; body surface area; body mass index; daily calorie intake; daily calorie intake per unit of body mass surface area; daily calorie intake per kilogram bodyweight; and valproic acid usage were examined. A statistically significant difference was observed in serum albumin level between the two groups (P = 0.026). The mechanism of acute pancreatitis in these patients was considered as pancreatic morphological change, acinar damage, and elevated serum trypsinogen level caused by malnutrition. It is likely that acute pancreatitis in patients with SMID occurs due to the same mechanism as in anorexia nervosa and malnourished patients. To prevent acute pancreatitis in these patients, it is important to maintain adequate nutritional status.

Variable Clinical Presentation of Shwachman-Diamond Syndrome: Update From the North-American Shwachman-Diamond Syndrome Registry.

AbstractAbstract 2367 Background:Shwachman-Diamond syndrome (SDS) is an autosomal recessively inherited marrow failure syndrome associated with exocrine pancreatic dysfunction and an increased leukemia risk. SDS is caused by biallelic mutations in the SBDS gene. Timely diagnosis prior to the development of life-threatening complications is essential for optimal medical management and outcomes. SDS hematologic complications such as severe marrow failure or leukemia are treated with a hematopoietic stem cell transplant. SDS patients require reduced intensity conditioning regimens to avoid undue regimen-related toxicities, thus the recognition of the underlying diagnosis of SDS is critical. The common constellation of clinical characteristics reported for SDS includes neutropenia, steatorrhea, and failure to thrive. Hypothesis:With the advent of genetic testing, diagnosis is now possible for patients with non-classical presentations of the inherited marrow failure syndromes. The aim of this study was to investigate the range of clinical presentations for SDS with the long-term goal of improving diagnosis. Methods:This study was a retrospective review of medical records obtained by the North American Shwachman-Diamond Syndrome Registry (SDSR) which was founded in 2008. The SDS Registry works in partnership with the Severe Chronic Neutropenia International Registry. Results:Genetic reports of biallelic SBDS mutations confirming the diagnosis of SDS were available for 31 patients. This SDS study cohort included 19 male and 12 female patients. Median patient age was 10 years with a range of 2 – 49 years. Radiologic reports were available for 23 patients. Pancreatic lipomatosis was noted in 22/23 patients. One patient had an atrophic pancreas without lipomatosis on CT scan at age 6.9 years. Two patients initially had normal early pancreatic studies by CT or ultrasound at ages 1.3 years and 2.6 years, but subsequently were found to have pancreatic lipomatosis by the same imaging modalities later in life. Fecal elastase has not been previously evaluated as a screen for SDS. 14/17 patients (82%) had low fecal elastase levels while 3/17 patients had normal fecal elastase levels. Serum trypsinogen or pancreatic isoamylase were low in all 18 patients tested. 26/31 patients (84%) presented with failure to thrive. 17/31 (54%) patients presented with diarrhea or steatorrhea. 17/31 patients (55%) had congenital anomalies. Neutropenia was the most common hematologic abnormality at presentation (77%). Strikingly, 2 patients presented with isolated thrombocytopenia, 3 patients presented with severe anemia requiring transfusion support, and 5 patients presented without any cytopenias. One patient presented with the initial diagnosis of MDS with a del 20q clone. One patient presented with short stature and a sibling who died from AML. Interestingly, one patient had very short telomeres (<1st percentile for age) across 3 lymphocyte subsets as well as in total lymphocytes, a pattern typically associated with dyskeratosis congenita. Conclusion:Data from the North-American Shwachman Diamond Syndrome Registry reveals an unexpected range of clinical presentation for this rare disorder that expands beyond that which is currently classically recognized. The diagnosis of SDS should be considered even in the absence of neutropenia or diarrhea. In our cohort, clues to the underlying diagnosis of SDS included other cytopenias, congenital anomalies, family history, and the del20q clonal marrow abnormality. Reliance on classical clinical descriptions of SDS would miss or delay diagnosis of a significant subset of SDS patients. Disclosures:Dale:AMGEN: Consultancy.

Reference Intervals for and Validation of Recalibrated Immunoassays for Trypsinogen-1 and Trypsinogen-2

To the Editor: Serum trypsinogen assays are used as diagnostic and prognostic tools for cystic fibrosis and acute pancreatitis (AP) (1, 2). Calibrator stability is a challenge in these assays, because trypsinogen readily autoactivates and subsequently autodegrades. Furthermore, the reference intervals described thus far have been based on samples from a limited number of individuals (1, 3). We recalibrated a new immunoassay for trypsinogen-2 and a previously described assay for trypsinogen-1 (1) with stable calibrators (2, 4) and report reference intervals for these 2 analytes in serum. We have produced new monoclonal antibodies and developed a time-resolved immunofluorometric assay for trypsinogen-2, as previously described (1). Monoclonal antibodies F87–9E6 and F88–8F7 were used as capture antibody and tracer antibody, respectively. To prevent activation, we produced stable mutated (Lys23Gln) trypsinogen-1 and trypsinogen-2 by recombinant technology (2, 4) and used them to recalibrate both the previously described trypsinogen-1 assay (1) and the new trypsinogen-2 assay. In this assay, the calibrators covered the concentration interval of 1.0 to 1000 μg/L, the limit of detection was 0.24 μg/L, and …

703 Case Report - A Patient with Bone Formation Defects During Prenatal Life Diagnosed as ‘Shwachman-Diamond Syndrome’

CaseReportShwachman-Diamond syndrome(SDS) is an autosomal recessive disorder with clinical features that include pancreatic exocrine insufficiency, hematological dysfunction and skeletal abnormalities during development after birth and varies with age. The patient...