Histochemical Diagnosis of Hirschsprung Disease

PEDIATRICS ◽  
1982 ◽  
Vol 69 (6) ◽  
pp. 755-761
Author(s):  
Carolyn Coker Huntley ◽  
Louis deS. Shaffner ◽  
Venkata R. Challa ◽  
Anne D. Lyerly
Keyword(s):  
Lamina Propria ◽  
False Positive ◽  
Ganglion Cells ◽  
False Negative ◽  
Hirschsprung Disease ◽  
Nerve Fibers ◽  
Infants And Children ◽  
Muscularis Mucosa ◽  
False Positive Reactions ◽  
In Infants And Children

A histochemical staining technique for detection of acetylcholinesterase (AChE) in rectal suction biopsies was compared with the presence or absence of ganglion cells in full-thickness or suction biopsies for the diagnosis of Hirschsprung disease (HD) in infants and children. Biopsies from 55 of 58 children were adequate for both the AChE assay and routine pathologic examination for ganglion cells. Two patterns of AChE staining were noted. With pattern A, prominent nerve fibers staining for AChE were seen throughout the muscularis mucosa and the lamina propria. With pattern B, similar fibers were seen only in the muscularis mucosa and the areas of lamina propria that were immediately adjacent. No "false-negative" AChE staining reactions were found in patients with HD. No "false-positive" reactions showing pattern A were found. This pattern was diagnostic for HD. Three false-positive reactions were found showing pattern B in patients with conditions other than HD. Among 22 patients with HD, 19 were males and three were females. Pattern A occurred in all age groups and in both sexes. Pattern B in patients with HD was seen exclusively in male infants 1 month of age or less. Experience suggests that the AChE staining of rectal suction biopsies is an excellent screening test for HD in infants and children. If pattern B is encountered, however, the specimen should be examined by routine pathologic techniques for the presence of submucosal ganglion cells.

Total Colonic Aganglionosis: Case Report, Practical Diagnostic Approach and Pitfalls

2010 ◽  
Vol 134 (10) ◽  
pp. 1467-1473 ◽  
Author(s):  
Raja Rabah
Keyword(s):  
Transition Zone ◽  
Ganglion Cells ◽  
Hirschsprung Disease ◽  
Rectal Biopsy ◽  
Nerve Fibers ◽  
Submucosal Plexus ◽  
Total Colonic Aganglionosis ◽  
Biopsy Specimens ◽  
Suction Rectal Biopsy ◽  
Acetylcholinesterase Staining

Abstract Hirschsprung disease remains a challenging diagnosis for many pathologists. The disease is characterized by a lack of ganglion cells in the myenteric and submucosal plexus, associated with increased numbers of acetylcholinesterase-positive nerve fibers. Hypertrophic nerve fibers are present in most but not all patients. Total colonic aganglionosis (TCA) is an uncommon form of Hirschsprung disease with clinical, histologic, and genetic differences and is even more difficult to diagnose and manage. This case illustrates some of the difficulties frequently faced by the pathologists dealing with total colonic aganglionosis. Suction rectal biopsy specimens often lack significant nerve hypertrophy and positive acetylcholinesterase staining, which aid in the diagnosis. Pathologists have to depend mainly on the lack of ganglion cells in adequate submucosa to establish the diagnosis. Transition zone is often long in total colonic aganglionosis and interpretation of frozen sections can be difficult. The presence of several uniformly distributed clusters of mature ganglion cells and lack of nerve hypertrophy are required to avoid connections at the transition zone.

Comparative Efficiency of Brilliant Green, Bismuth Sulfite, Salmonella-Shigella, Hektoen Enteric, and Xylose Lysine Desoxycholate Agars for the Recovery of Salmonella from Foods: Collaborative Study

1981 ◽  
Vol 64 (4) ◽  
pp. 899-928
Author(s):  
Wallace H Andrews ◽  
Paul L Poelma ◽  
Clyde R Wilson ◽  
◽  
C Abeyta ◽  
...  
Keyword(s):  
Statistical Analysis ◽  
False Positive ◽  
Relative Efficiency ◽  
Brilliant Green ◽  
Collaborative Study ◽  
False Negative ◽  
Reaction Rates ◽  
Negative Reaction ◽  
Comparative Efficiency ◽  
False Positive Reactions

Abstract The relative efficiency of brilliant green (BG), bismuth sulfite (BS), Salmonella-Shigella (SS), xylose lysine desoxycholate (XLD), and Hektoen enteric (HE) agars for the recovery of Salmonella from 5 foods was collaboratively studied in 11 laboratories. The analytical efficiency of various paired combinations of the 5 agars was statistically compared according to 3 parameters: (1) productivity or recovery of Salmonella, (2) rate of enumeration of cultures that were false positive for Salmonella, and (3) rate of enumeration of false-negative reactions. In descending order of productivity, the sequential rankingwasBS, XLD, HE, BG, and SS agars. In ascending order, the rates of false-positive reactions based on a statistical analysis of paired agar combinations was HE, BS, BG and XLD (tie), and SS agars. Analogously, in ascending order, the sequence of false-negative reaction rates was BS, XLD, HE, BG, and SS agars. The combination of BS, XLD, and HE agars is more efficient for recovery of Salmonella from foods than is the present official combination of BG, BS, and SS agars. The revision of official final action method 46.054 to replace the combination of BG, BS, and SS agars with a combination of BS, XLD, and HE agars has been adopted official first action.

Ulnar nerve conduction velocity and H-reflex in infants and children

1960 ◽  
Vol 15 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Juergen E. Thomas ◽  
Edward H. Lambert
Keyword(s):  
Ulnar Nerve ◽  
Conduction Velocity ◽  
Premature Infants ◽  
Newborn Infants ◽  
Nerve Fibers ◽  
H Reflex ◽  
Full Term ◽  
Infants And Children ◽  
Term Newborn ◽  
In Infants And Children

Measurements of conduction velocity of the motor fibers of the ulnar nerve in the segment between the upper part of the arm and the wrist were made in 6 premature infants, 42 full-term newborn infants and 98 children up through the age of 14 years. In premature infants, 21 – 40 days before full term, values ranged from 18 to 22 meters per second with a mean of 21. Full-term newborn infants had velocities that ranged from 21 to 33 m.p.s., with a mean of 28. The values in the newborn were about one-half those of normal young adults, which range from 47 to 73 m.p.s. (mean, 60). By the age of 3 years almost all values were in the lower part of the adult range and, at 5 years, the velocities were not significantly different from those of the adult. These observations are in harmony with information about the diameter of nerve fibers during growth in man. The H-reflex could be elicited by stimulation of the ulnar nerve in almost every newborn infant. By the age of 1 year and thereafter it could rarely be elicited. Between the elbow and wrist, afferent fibers subserving this reflex conducted with a mean velocity of 30 m.p.s. Data on the action potential of the hypothenar muscles, residual latency of conduction in the ulnar nerve, latency of the H-reflex and conduction in the peroneal nerve in infants and children are compared with those on adults. Submitted on July 15, 1959

Laparotomic Versus Laparoscopic Duhamel Pull-Through for Hirschsprung Disease in Infants And Children

2007 ◽  
Vol 17 (1) ◽  
pp. 119-123 ◽  
Author(s):  
Vittorina Ghirardo ◽  
Pietro Betalli ◽  
Guendalina Mognato ◽  
Piergiorgio Gamba
Keyword(s):  
Hirschsprung Disease ◽  
Infants And Children ◽  
Pull Through ◽  
In Infants And Children

Hirschsprung Disease and Use of Calretinin in Inadequate Rectal Suction Biopsies

2013 ◽  
Vol 137 (8) ◽  
pp. 1099-1102 ◽  
Author(s):  
David Hernandez Gonzalo ◽  
Thomas Plesec
Keyword(s):  
Gold Standard ◽  
Clinical Course ◽  
Ganglion Cells ◽  
Hirschsprung Disease ◽  
Disease Status ◽  
Nerve Fibers ◽  
Clinical Suspicion ◽  
Muscularis Mucosae ◽  
Hematoxylin Eosin

Context.—Up to 17% of all rectal suction biopsies performed in the evaluation of Hirschsprung disease are considered inadequate. In most instances, inadequate biopsies contain too little submucosa or are taken within the anal transition zone. Objectives.—To examine the utility of calretinin stain in the workup of inadequate biopsies for patients with clinical suspicion of Hirschsprung disease. Design.—A retrospective analysis was conducted of all rectal suction biopsies performed in the evaluation of Hirschsprung disease during the previous 12 years that were considered “inadequate.” Seventeen cases were identified, and Hirschsprung disease status was determined by clinical or surgical follow-up. Immunohistochemistry for calretinin was performed for all cases containing columnar mucosa, which were evaluated without knowledge of clinical course. Results.—All 12 patients without Hirschsprung disease had calretinin-positive nerve fibers in the lamina propria or muscularis mucosae, and all 5 patients with Hirschsprung disease had no calretinin staining of nerves. Conclusions.—In this retrospective series, calretinin immunohistochemistry correctly predicted outcome in all instances. Although the gold standard for the diagnosis of Hirschsprung disease in rectal suction biopsies remains the evaluation of ganglion cells in a hematoxylin-eosin staining with sufficient submucosa, calretinin immunohistochemistry is quite helpful in triaging further workup based on clinical suspicion.

Characterization of the Transition Zone in Short Segment Hirschsprung Disease Using Calretinin Immunostaining

2021 ◽  
pp. 109352662110539
Author(s):  
Franziska Righini-Grunder ◽  
Dorothée Bouron-Dal Soglio ◽  
Lara Hart ◽  
Ann Aspirot ◽  
Christophe Faure ◽  
...  
Keyword(s):  
Transition Zone ◽  
Ganglion Cells ◽  
Short Form ◽  
Hirschsprung Disease ◽  
Nerve Fibers ◽  
Normal Density ◽  
Short Segment ◽  
Pull Through ◽  
Distal Border ◽  
Proximal Border

Introduction: The detailed expression pattern of calretinin immunohistochemistry in the transition zone (TZ) of Hirschsprung disease (HSCR) has not yet been reported. This study aims to examine the value of calretinin immunohistochemistry for more accurately determining the distal and proximal border of the TZ in short segment HSCR. Methods: Specimens of pull-through surgery from 51 patients with short form of HSCR were analyzed on two longitudinal strips using hematoxylin and eosin (H&E) staining and calretinin immunohistochemistry. Results: In all but two patients, the first appearance of calretinin expression was seen on mucosal nerve fibers before the appearance of any ganglion cells, indicating the distal border of the TZ. The maximum distance between the distal border of the TZ and the proximal border of the TZ, defined by ganglion cells in a normal density on H&E stained sections, a strong calretinin expression on mucosal nerve fibers and in >80% of submucosal and myenteric ganglion cells, with no nerve hypertrophy and absence of ganglionitis was 60 mm. Conclusion: The distal border of the TZ is characterized by calretinin positive intramucosal neurites in nearly all of short form of HSCR and not by calretinin expression on ganglion cells.

Ganglion Cells Are Frequently Present in Pediatric Mucosal Colorectal Biopsies: H&E Sections and Calretinin Immunohistochemistry in the Evaluation of Children With Chronic Constipation

2017 ◽  
Vol 21 (1) ◽  
pp. 48-53
Author(s):  
Alexandra E Kovach ◽  
M Cristina Pacheco
Keyword(s):  
Chronic Constipation ◽  
Ganglion Cells ◽  
Single Case ◽  
Hirschsprung Disease ◽  
Clinical Indication ◽  
Muscularis Mucosa ◽  
Older Children ◽  
Anatomic Site ◽  
Colorectal Biopsies ◽  
Work Up

Hirschsprung disease (HD) rarely presents as chronic constipation after the newborn period. At our institution, calretinin immunohistochemistry (CAL) is frequently requested by clinicians on rectal mucosal biopsies (RMBs) taken during colonoscopy in older children in whom suspicion for HD is low. We hypothesized that review of these biopsies would frequently reveal ganglion cells (GCs). We reviewed features of mucosal biopsies (November 2013 to September 2015) from children ≥1 year of age on which clinicians had requested CAL on at least one specimen. A total of 93 biopsies with paired CAL from 83 patients were suitable for study (ages 1–18 years, M:F 1.2). Submitted clinical indication was constipation in 62 patients (75%). GCs were found within or subjacent to muscularis mucosa in 63 biopsies (68%), 12 (19%) of which were designated from a specific anatomic site, eg, 2 or 3 cm. In 25 of 63 (40%) cases, GCs were identified on one of the first 3 sections (median 5th, range 1st–54th). Forty-six cases (73%) contained no or <0.5 mm of submucosa (SM, range 0–2 mm). All but one case (62/63, 98%) with identified GCs showed positive CAL staining; a single case showed equivocal staining. Among the 30 biopsies with no observed GCs, none (0%) had >1 mm of SM, and 21 (70%) had no SM. CAL was positive in 28 (93%) and equivocal/weak in 2 (7%); no additional work-up for HD was pursued. The data suggest that H&E sections of RMBs can exclude HD at a specified site in many cases and provide the basis for a future study examining the utility of CAL in RMBs without SM as a means for excluding HD.

Expression of bcl-2 in Enteric Neurons in Normal Human Bowel and Hirschsprung Disease

1999 ◽  
Vol 123 (12) ◽  
pp. 1264-1268 ◽  
Author(s):  
Tomas Wester ◽  
Yngve Olsson ◽  
Leif Olsen
Keyword(s):  
Ganglion Cells ◽  
Neuron Specific Enolase ◽  
Hirschsprung Disease ◽  
Immunohistochemical Analysis ◽  
Nerve Fibers ◽  
Enteric Neurons ◽  
Normal Colon ◽  
Normal Human ◽  
Immunohistochemical Methods

Abstract Objective.—The bcl-2 protein has the functional role of blocking apoptosis, ie, programmed cell death. This protein is widely expressed in the developing central and peripheral nervous systems. The purpose of this study was to map bcl-2 expression in the human enteric nervous system, as this has not previously been done. Methods.—Rectal specimens were obtained at autopsy of 13 fetuses at 13 to 31 weeks of gestation. Normal colon was also obtained from 5 children and 2 adults, and, in addition, ganglionic and aganglionic bowel resected in 11 patients with Hirschsprung disease was examined. Specimens were fixed in formalin, embedded in paraffin, and analyzed with immunohistochemical methods, using antibodies raised against bcl-2 and neuron-specific enolase (NSE). Results.—The bcl-2 protein was expressed in myenteric and submucous ganglion cells in fetuses, children, and adults. Nerve fibers of the enteric plexuses that were bcl-2 immunoreactive were few compared with the number of NSE-immunoreactive nerve fibers. In aganglionic bowel no bcl-2–or NSE-immunoreactive ganglion cells were revealed. Results of NSE immunohistochemistry showed clearly stained hypertrophic nerve bundles, known to be of extrinsic origin, which were only weakly bcl-2 immunoreactive. Conclusion.—Expression of bcl-2 in enteric ganglion cells of the myenteric and submucous plexuses is displayed in the fetus and during childhood and is also retained in adult bowel. Immunohistochemical analysis of bcl-2 provides a good marker for identification of ganglion cells in Hirschsprung disease and may also be valuable for the diagnosis of disorders characterized by hypoganglionosis or hyperganglionosis.

scholarly journals Intercurrence of Paratuberculosis in Intradermal Tuberculin Test Reactive Cattle

2020 ◽  
Vol 48 ◽  
Author(s):  
Mariana Assunção De Souza ◽  
Nicolle Pereira Soares ◽  
Alessandra Aparecida Medeiros-Ronchi ◽  
Brendhal Almeida Silva ◽  
Pedro Paulo Feitosa De Albuquerque ◽  
...  
Keyword(s):  
Lymph Nodes ◽  
False Positive ◽  
False Negative ◽  
Complex Mixture ◽  
Pathological Finding ◽  
Intradermal Test ◽  
Mesenteric Lymph Nodes ◽  
Serological Tests ◽  
Mesenteric Lymph ◽  
False Positive Reactions

Background: Bovine tuberculosis control programs are based on a standard diagnostic method, the intradermal test with purified protein derivatives, which is used to identify and eliminate diseased animals. Currently none of the tests available allow complete differentiation between infected and uninfected animals. The main limitations of the tests available are related to diagnostic sensitivity and specificity, which results in false positive reactions due to the existence of cross infections, and also false negative, inherent to the state of energy of some animals. The aim of this work was to study the intercurrence of paratuberculosis in tuberculosis reactive cattle by the comparative cervical test.Materials, Methods & Results: Three hundred and thirty four cattle were evaluated using the comparative cervical test (CCT) and serology for tuberculosis (TB) and paratuberculosis (PTB) ELISA IDEXX®. All of the animals testing positive by CCT were euthanized and necropsied. Fragments of lymph node, lung and intestine were collected and analyzed using histopathological techniques, with staining by Hematoxylin-Eosin (HE). Samples of lung and lymph nodes (retropharyngeal, submandibular, cervical and mediastinal) of the animals testing positive by CCT were evaluated using qPRC for M. bovis, and intestinal and mesenteric lymph nodes using PCR for PTB. Of the 334 cattle evaluated using the comparative cervical test, 16 were considered positive. No lesions suggestive of tuberculosis were found in the macroscopic inspection of the carcasses. The most evident anatomical and pathological finding was a thickening of intestinal mucosa, found in 12 of the 16 cattle submitted to necropsy. No microscopic lesions suggestive of TB were identified nor was the presence of M. bovis detected by qPCR. The main histopathological findings were observed in the small intestine and mesenteric lymph nodes and identified as enteritis, lymphangitis, lymphangiectasia and granulomatous lymphadenitis. In the intestine the changes are characterized by dilated and inflamed lymphatic vessels and intense inflammatory infiltrate on the mucosa and submucosa. Of the 334 serum samples evaluated, the M. bovis ELISA Antibody Test (IDEXX®) identified 17 positive animals. All the cattle considered positive by M. bovis ELISA were considered negative by CCT. In the samples from nine animals (9/16), DNA from M. avium subsp. paratuberculosis (MAP) was identified and in twelve carcasses (12/16) lesions characteristic of PTB were found, which were subsequently confirmed by histopathological techniques. In another nine animals of the herd anti-MAP antibodies were detected. None of those that tested positive by PTB ELISA were reactive by CCT.Discussion: Animals considered positive by TB ELISA that were not positive in the intradermal test does not mischaracterize the clinical picture of the disease. Considering the inverse relationship between cell-mediated and humoral responses to M. bovis, the intradermal test and the serological tests are designed to measure different immunological responses, which develop during different stages of infection. The progress of the cellular immunological response to humoral immunity occurs in the most advanced stages of tuberculosis. Of the 16 cattle considered positive by CCT, 12 animals presented macroscopic and histological lesions suggestive of PTB and DNA from MAP was detected in nine. Although it is the official test for the control of TB in different countries, the intradermal test with PPD has presented limitations, primarily related to specificity. M. avium subsp. Paratuberculosis is considered the main cause of false positive reactions in the intradermal test. The PPD bacterial extract is a complex mixture of proteins, lipids, sugars and nucleic acids, and many of these components are also shared by numerous species of mycobacteria (tuberculous or not). 

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