Ultrastructural injury to interstitial cells of Cajal and communication with mast cells in Crohn's disease.

Crohn's disease associated dysmotility has been attributed to fibrosis and damage to enteric nerves but injury to interstitial cells of Cajal (ICC) could also be involved. We assessed ICC in specimens obtained from patients with Crohn's disease and determined the relation between ICC and the inflammatory infiltrate, particularly mast cells (MC) using quantitative immunohistochemistry and electron microscopy. Ultrastructural injury to ICC was patchy in all ICC subtypes but ICC-Auerbach's plexus (AP) showed damage more frequently, i.e. swelling of mitochondria, decreased electron density, autophagosomes and partial depletion of the cytoplasm. Light microscopy confirmed a significant decrease in c-kit immunoreactivity for ICC-AP and an increased number of MC in the muscularis externa. Electron microscopy showed MC exhibiting piecemeal degranulation and making frequent and selective membrane-to-membrane contact with all types of injured ICC which suggests chronic release of granule content to affect ICC. Extent of ICC injury was not associated with duration of the disease. In conclusion, ultrastructural injury and loss of ICC-AP is evident in Crohn's disease. Epidemiological and morphological data suggest that ICC have the capacity to regenerate in spite of the chronic insult. The muscularis hosts a marked number of MC that exhibit piecemeal degranulation associated with ICC and may facilitate ICC maintenance.

Quantitative assessment of human muscle glycogen granules size and number in subcellular locations during recovery from prolonged exercise.

Although data relating to muscle glycogen are interpreted as showing it is homogenous when quantified biochemically, it is actually in granules in specific subcellular locations. We hypothesized that postexercise restoration of muscle glycogen would occur initially by an increase in granule number followed by an increase in size, and also that restoration would differ in various subcellular locations. Five men performed prolonged exercise and had muscle biopsies taken at 0, 4, 24 and 48 h of recovery. We quantified granule number and size as well as the total volume of glycogen in the subsarcolemmal and the intra- and intermyofibrillar regions, using transmission electron microscopy. Muscle glycogen was reduced to 36 +/- 8.3 mmol glucosyl units (kg dry weight)(-1) at exhaustion, and was preferentially depleted and subsequently repleted in the intramyofibrillar space. The repletion rate was greatest in the first 4 h; this was associated with a 186% increase in number (P < or = 0.05) and no change in particle size (P > or = 0.05). From 4 h to 48 h, there was an increase in particle size (P < or = 0.05) but not number (P > or = 0.05). Net rate of G volume synthesis per unit area was 50% greater (P < or = 0.05) in the subsarcolemmal than the myofibrillar compartment. Conversely, the net rate of single-particle volume synthesis was greater (P < or = 0.05) in the myofibrillar than the subsarcolemmal compartment. Glycogen granules varied in size and number depending on location, and in all compartments resynthesis of glycogen was characterized initially by an increase in granule number and later by an increase in size.

Novel SCO2 mutation (G1521A) presenting as a spinal muscular atrophy type I phenotype.

Rare cases of suspected spinal muscular atrophy (SMA) have been found to have cytochrome c oxidase (COX) deficiency. To date, four cases with SMA features have been reported in children with mutations in the synthesis of cytochrome oxidase 2 (SCO2) gene. We report a male neonate who was born hypotonic, with persistent lactic acidosis, spontaneous activity with EMG testing, development of respiratory distress in the first few hours of life, and died at 30 days of age with progressive cardiomyopathy. Testing for survival motor neurone (smn) and NAIP deletions were negative and a skeletal muscle biopsy showed neurogenic features with severe reductions of COX enzymatic and histochemical staining intensity. Post-mortem muscle, heart, and liver biopsies showed severe, moderate, and mild reductions in COX activity, respectively, with parallel findings in the protein content for the mitochondrial DNA (COII) and nuclear DNA (COIV) encoded subunits. DNA sequencing of exon 2 of the SCO2 gene revealed compound heterozygosity with mutations at G1541A (common mutation, E140K) and also at a novel site in the copper binding region (G1521A in the current case (converting a highly conserved cysteine to tyrosine [corrected] (C133Y) [corrected]); mother heterozygous for G1521A; and father heterozygous for G1541A). This case provides strong support that SCO2 mutations can result in neonatal hypotonia with an SMA 1 phenotype. SCO2 mutations should be screened in suspected SMA cases with normal smn mutation analysis and any one of; cardiomyopathy, lactic acidosis, or COX deficiency in muscle.

Pathology of skeletal muscle in mitochondrial disorders.

Muscle histology is an essential component of the diagnostic work-up for mitochondrial cytopathies and is very important in both ruling in disease as well as ruling out the disease (i.e., alternate diagnoses). A muscle biopsy method must provide tissue for histology, electron microscopy, enzymes and DNA and this can be obtained with a suction-modified 5 mm needle. Proper embedding and processing is important for optimal diagnostic yield. The essential stains for mitochondrial histology remain the modified Gomori trichrome, cytochrome oxidase, succinate dehydrogenase, and NADH. Electron microscopy can be helpful in selected cases, however, the decision to perform this on all samples remains a contentious issue. Some cases of mitochondrial cytopathy may show no abnormalities on histology or electron microscopy (i.e., LHON), whereas, other conditions can mimic mitochondrial disease through secondary mitochondrial changes (i.e., inclusion body myositis). Athletes show evidence of increased mitochondrial numbers but do not normally develop ragged red fibers or paracrystalline inclusions. Aging is associated with an accumulation of mitochondrial abnormalities and is an important factor to consider in the interpretation of the sample. For example, biopsies in young children with mitochondrial disease may be normal at the histological level and otherwise healthy older adults can show mitochondrial changes such as ragged red and COX-negative fibers.

Histological assessment of intermediate- and long-term creatine monohydrate supplementation in mice and rats.

Creatine monohydrate (CrM) supplementation appears to be relatively safe based on data from short-term and intermediate-term human studies and results from several therapeutic trials. The purpose of the current study was to characterize pathological changes after intermediate-term and long-term CrM supplementation in mice [healthy control and SOD1 (G93A) transgenic] and rats (prednisolone and nonprednisolone treated). Histological assessment (18-20 organs/tissues) was performed on G93A mice after 159 days, and in Sprague-Dawley rats after 365 days, of CrM supplementation (2% wt/wt) compared with control feed. Liver histology was also evaluated in CD-1 mice after 300 days of low-dose CrM supplementation (0.025 and 0.05 g x kg-1x day-1) and in Sprague-Dawley rats after 52 days of CrM supplementation (2% wt/wt) with and without prednisolone. Areas of hepatitis were observed in the livers of the CrM-supplemented G93A mice (P < 0.05), with no significant inflammatory lesions in any of the other 18-20 tissues/organs that were evaluated. The CD-1 mice also showed significant hepatic inflammatory lesions (P < 0.05), yet there was no negative effect of CrM on liver histology in the Sprague-Dawley rats after intermediate-term or long-term supplementation nor was inflammation seen in any other tissues/organs (P = not significant). Dietary CrM supplementation can induce inflammatory changes in the liver of mice, but not rats. The observed inflammatory changes in the murine liver must be considered in the evaluation of hepatic metabolism in CrM-supplemented mice. Species differences must be considered in the evaluation of toxicological and physiological studies.

Diaphragm kinetics during pneumatic belt respiratory assistance: a sonographic study in Duchenne muscular dystrophy.

The principal aim of this study was to demonstrate the usefulness of M-mode sonography as a noninvasive technique to evaluate diaphragm excursion. The secondary aim was to assess the efficacy of pneumatic abdomino-diaphragmatic belt ventilation in patients with Duchenne muscular dystrophy. Using M-mode sonography, we measured the amplitude of diaphragm excursion in seven patients with Duchenne muscular dystrophy in various positions (0 degrees, 45 degrees, 75 degrees ) with and without pneumatic abdomino-diaphragmatic belt respiratory assistance. The belt significantly increased mean amplitude of diaphragm excursion by 62% at 45 degrees and by 55% at 75 degrees, and increased mean tidal volume by 43.5% at 45 degrees and by 49% at 75 degrees. Two patients were unable to tolerate the horizontal position (0 degrees ) During quiet breathing without the belt, amplitude of diaphragm excursion and tidal volume were positively correlated at 45 degrees (r=0.81; P=0.027) and 75 degrees (r=0.75; P=0.05). There was a significant intra-individual correlation between these two parameters during belt use but no inter-individual correlation. Without the belt, thoracic posture had no significant effect on amplitude of diaphragm excursion, either in quiet or deep breathing. After overnight respiratory assistance, arterial oxygen pressure and arterial oxygen saturation increased significantly, and arterial carbon dioxide pressure decreased from 52+/-6.4 to 46.4+/-4 mmHg. The pneumatic abdomino-diaphragmatic belt significantly improved gas exchanges and ventilation by increasing diaphragm excursion, as was clearly shown by noninvasive M-mode sonography. Indeed, M-mode sonography may be helpful in pneumatic abdomino-diaphragmatic belt pressure adjustment.

Diaphragm movement before and after cholecystectomy: a sonographic study.

UNLABELLED: Respiratory disorders after abdominal surgery are commonly explained by changes in diaphragmatic movement that are difficult to demonstrate and quantify. Our aim was thus to quantify these changes using a noninvasive method. We used M-mode sonography for the prospective study to measure diaphragmatic amplitude in 14 patients before and after cholecystectomy. During quiet breathing, the diaphragm inspiratory amplitude (DIA) was significantly decreased after surgery from 1.4 +/- 0.2 cm to 1 +/- 0.1 cm and from 1.6 +/- 0.3 cm to 1.2 +/- 0.3 cm in the Laparoscopic and Open Cholecystectomy groups, respectively. The total time cycle of diaphragmatic motion decreased significantly in the two groups. The DIA also decreased significantly during deep breathing after cholecystectomy from 6.0 +/- 0.8 cm to 3.0 +/- 1.8 cm and from 6.1 +/- 1.3 cm to 3.1 +/- 1.6 cm in the Laparoscopic and Open Cholecystectomy groups, respectively. The six patients who underwent spirometric examination showed, during quiet breathing, a significant decrease in DIA without change in tidal volume, i.e., 0.51 +/- 0.08 L to 0.45 +/- 0.08 L. We found a significant decrease in DIA after cholecystectomy and a significant interindividual correlation between DIA during deep inspiration and inspiratory capacity. Using M-mode sonography techniques, we were able to demonstrate changes in diaphragmatic mobility after laparoscopic or open cholecystectomy. IMPLICATIONS: Cholecystectomy at times results in impaired respiratory and diaphragmatic functions. The techniques currently used to study these repercussions are both laborious and invasive. Our sonographic technique is completely noninvasive and can be used to study diaphragm morphology and movement in real time.

Plexiform neurofibroma of the submandibular salivary gland in a child.

Plexiform neurofibromas in major salivary glands are rarely described. In the literature, most reported tumours have been present in the parotid gland region. A three-year-old boy with a family history of neurofibromatosis presented with a rapidly growing left submandibular mass. The clinical diagnosis was that of a neurofibroma rather than a primary salivary gland tumour. Resection of the lesion revealed a plexiform neurofibroma involving the submandibular gland. Although these tumours have a neurogenic rather than a salivary gland origin, they must be considered in the differential diagnosis of a salivary gland lesion in a patient with a history of neurofibromatosis.

Gender differences in muscle inflammation after eccentric exercise.

Unaccustomed exercise is followed by delayed-onset muscle soreness and morphological changes in skeletal muscle. Animal studies have demonstrated that women have an attenuated response to muscle damage. We studied the effect of eccentric exercise in untrained male (n = 8) and female (n = 8) subjects using a unilateral exercise design [exercise (Ex) and control (Con) legs]. Plasma granulocyte counts [before (Pre) and 48 h after exercise (+48h)] and creatine kinase activity [Pre, 24 h after exercise (+24h), +48h, and 6 days after exercise (+6d)] were determined before (Pre) and after (+24h, +48h, +6d) exercise, with biopsies taken from the vastus lateralis of each leg at +48h for determination of muscle damage and/or inflammation. Plasma granulocyte counts increased for men and decreased for women at +48h (P < 0.05), and creatine kinase activity increased for both genders at +48h and +6d (P < 0.01). There were significantly greater areas of both focal (P < 0.001) and extensive (P < 0.01) damage in the Ex vs. Con leg for both genders, which was assessed by using toluidine blue staining. The number of leukocyte common antigen-positive cells/mm(2) tissue increased with exercise (P < 0.05), and men tended to show more in their Ex vs. Con leg compared with women (P = 0.052). Men had a greater total (Ex and Con legs) number of bcl-2-positive cells/mm(2) tissue vs. women (P < 0.05). Atrophic fibers with homogeneous bcl-2-positive staining were seen only in men (n = 3). We conclude that muscle damage is similar between genders, yet the inflammatory response is attenuated in women vs. men. Finally, exercise may stimulate the expression of proteins involved in apoptosis in skeletal muscle.

Molecular detection of the ETV6-NTRK3 gene fusion differentiates congenital fibrosarcoma from other childhood spindle cell tumors.

Congenital fibrosarcoma (CFS) is a pediatric spindle cell tumor of the soft tissues that usually presents before the age of 2 years. Although these tumors display histologic features of malignancy and frequently recur, they have a relatively good prognosis and only rarely metastasize. CFS must therefore be differentiated from more aggressive spindle cell sarcomas that occur during childhood, particularly adult-type fibrosarcoma (ATFS), which can have an identical morphology. CFS must also be distinguished from benign but cellular fibroblastic lesions of the same age group, including infantile fibromatosis (IFB) and myofibromatosis (MFB). Unfortunately, standard pathologic examination often does not differentiate CFS from these other conditions. The authors recently identified a novel chromosomal translocation in CFS, t(12;15)(p13;q25), which gives rise to an ETV6-NTRK3 gene fusion. They subsequently developed reverse transcription-polymerase chain reaction (RT-PCR) assays that can detect ETV6-NTRK3 fusion transcripts in CFS frozen or paraffin-embedded tumor specimens. To confirm the use of this assay in the differential diagnosis of CFS, they have screened a larger series of childhood pediatric spindle cell lesions for ETV6-NTRK3 gene fusions, including 11 cases of CFS, 13 malignant spindle cell tumors (including ATFS), and 38 benign spindle cell tumors (including IFB and MFB). Of the 11 cases diagnosed as CFS, 10 showed the ETV6-NTRK3 gene fusion, whereas none of the 51 other malignant or benign spindle cell tumors demonstrated this fusion gene. They also compared their RT-PCR findings with those of conventional cytogenetics and with immunohistochemical detection of the ETV6-NTRK3 protein using antisera to NTRK3. They conclude that RT-PCR analysis is superior to these techniques for the detection of the ETV6-NTRK3 gene fusion in pediatric spindle cell tumors, and it is a reliable and specific modality for the diagnosis of CFS.

Antenatal and postnatal findings in encephalocraniocutaneous lipomatosis.

Encephalocraniocutaneous lipomatosis (ECCL) is a congenital neurocutaneous syndrome that comprises unilateral porencephalic cysts with cortical atrophy; ipsilateral hamartomas of scalp, eyelid, and outer part of the eye; lipomas of the central nervous system (CNS); cranial asymmetry; and characteristic cutaneous lesions. We report on the antenatal and postnatal sonographic and radiologic, and on the postnatal pathologic findings of an infant diagnosed with ECCL at birth. The antenatal sonogram at 28 weeks of gestation showed normal intracranial structures; the sonogram at 38 weeks of gestation showed asymmetry of the cerebral hemispheres and ventriculomegaly. Magnetic resonance imaging at age 6 weeks demonstrated a porencephalic cyst on the left, hemiatrophy of the left cerebrum with cortical dystrophy, and a lipoma in the middle cranial fossa. Histologic findings of the resected cutaneous lesion confirmed the presence of fibrolipoma hamartoma. This case offers a unique insight into the antenatal and postnatal development of the hamartoneoplastic lesions of ECCL, and it highlights the difficulties inherent in the antenatal sonographic diagnosis of hamartoneoplastic syndromes.

Pallister-Killian syndrome [i(12p)]: first pre-natal diagnosis using cordocentesis in the second trimester confirmed by in situ hybridization.

Pallister Killian syndrome (PKS) is the most frequent form of partial autosomal tetrasomy 12p in humans. Sufferers have a mosaic of isochromosome 12p [i(12p)]. We report the first pre-natal diagnosis on fetal blood cells after cordocentesis during the second trimester. The extra chromosome was first diagnosed by in situ hybridization. Fluorescence in situ hybridization (FISH) was used to count the interphase and/or metaphase cells containing the isochromosome. A review of the literature identified 27 other reports of PKS diagnosed pre-natally. We showed that the most consistent pre-natal ultrasound findings include hypertelorism, broad neck, shorts limbs, abnormal hands or feet, diaphragmatic hernia and hydramnios. Recognition of this congenital malformation pattern pre-natally may allow utilization of FISH.

Non-invasive quantification of diaphragm kinetics using m-mode sonography.

PURPOSE: The standard conditions of spirometry (i.e., wearing a noseclip and breathing through a mouthpiece and a pneumotachograph) are likely to alter the ventilatory pattern. We used "time motion" mode (M-mode) sonography to assess the changes in diaphragm kinetics induced by spirometry during quiet breathing. METHODS: An M-mode sonographic study of the right diaphragm was performed before and during standard spirometry in eight patients without respiratory disease (age 34 to 68 yr). RESULTS: During spirometry, the diaphragm inspiratory amplitude (DIA) increased from 1.34 +/- 0.18 cm to 1.80 +/- 0.18 cm (P = 0.007), whereas the diaphragmatic inspiratory (T1 diaph) increased from 1.27 +/- 0.15 to 1.53 +/- 0.23 sec, (P = 0.015, without change in diaphragmatic total time interval (Ttot diaph). Therefore, the diaphragm duty cycle (T1 diaph/Ttot diaph) increased from 38% +/- 1% to 44% +/- 4% (P = 0.023). The diaphragm inspiratory (DIV) and expiratory (DEV) motion velocity (P = 0.007). CONCLUSION: M-mode sonography enabled us to demonstrate that the wearing of a nose clip and breathing through a mouthpiece and a pneumotachograph induce measurable changes in diaphragm kinetics.

[Right diaphragmatic kinetics measured by TM-mode ultrasonography with concomitant spirometry in normal subjects and asthmatic patients. Preliminary results]. / Cinétique diaphragmatique droite mesurée par ultrasonographie en mode TM avec spirométrie concomitante chez le sujet normal et asthmatique. Résultats préliminaires.

The purpose of this study was to assess the diaphragmatic excursion using the TM-mode ultrasonography with concomitant pneumotachography in eight normal and five asthmatic subjects before and after salbutamol. We report the results in normal and asthmatic patients of this procedure. Particular findings were demonstrated in asthma. Different significant correlations were found between sonographic and respiratory measurements. In view of the safety of this direct and real-time diaphragmatic investigation, we conclude that this new method could offer an effective and reliable procedure in the evaluation of patients with diaphragmatic dysfunction. However, our study was limited by the small number of patients included and definite conclusions should await further investigations.

[Comparative evaluation of clinical and ultrasonographic screening of hip dislocation in Breton and Languedoc populations]. / Evaluation comparée du dépistage clinique et échographique de la luxation de la hanche dans une population bretonne et languedocinne.

BACKGROUND: Congenital dislocation of the hip varies greatly in incidence among different regions. This work is aimed at comparing results of clinical investigation and ultrasonography of the hip in Languedoc-Roussillon and Brittany. POPULATION AND METHODS: Two thousand eight hundred and twelve and 2,809 neonates admitted to Nîmes and Vannes hospitals respectively, were enrolled in a prospective study. Clinical examinations were made according to the protocol established by the "Groupe d' études en orthopédie pédiatrique". Ultrasound investigations were performed in every risk case. RESULTS: No dislocation occurred in the 4946 neonates without risk factors. In the 675 neonates with risk factors, 213 ultrasonographic examinations were abnormal, more frequently in the Brittany group (P < 0.001); nine dislocations were observed. A familial history of hip dysplasia (P < 0.001) and the addition of two risk factors (P < 0.001) were more frequent in Brittany. One hundred and six cases required treatment, more frequently in Brittany (P < 0.001). CONCLUSIONS: Some clinical risk factors and delayed ossification or hip dysplasia at ultrasound examination are significantly more frequent in Brittany. Clinical examination with selective ultrasonography is a reliable method, allowing early diagnosis and treatment of delayed dislocations.

Sonographic lung surface evaluation in pulmonary sarcoidosis: preliminary results.

The value of sonography in lung diseases such as sarcoidosis is not established. Twelve patients with pulmonary sarcoidosis and eight control subjects underwent a sonographic examination using a high frequency probe (7.5 MHz). The normal lung surface showed smooth and regular hyperechoic patterns, respiratory motions, and occasionally minute interruptions with comet tails. All patients with sarcoidosis showed various surface abnormalities, which appeared irregular and rough with coarse interruptions, producing an increase of artifacts. Nodular patterns were found in nine of 12 patients (75%). Sonography also revealed abnormalities not diagnosed by chest radiographs (n = 2). These findings could offer an complementary approach for evaluating sarcoidosis.

[Preliminary study of the feasibility of intraoperative ultrasonography of the biliary tract during cholecystectomy under celioscopy]. / Etude préliminaire de la faisabilité de l'échographie peropératoire des voies biliaires au cours des cholécystectomies par coelioscopie.

Peroperative echography and cholangiography provide similar information during the exploration of the biliary tree for conventional surgery for gall stones. Peroperative cholangiography is more difficult to perform via the coeolioscope with a failure rate of 10% in the most skillful teams. We investigated the technical feasibility of peroperative echography during coeliosurgery for gall stones. The technique was performed in 13 patients using a pulsed Doppler 7.5 MHz scan laparoscope. The entire biliary tree was successfully imaged in 12 patients and the three structures of the hepatic pedicle were identified in all cases. The exploration took less than 10 minutes and there was no morbidity. One gall stone was discovered and endoscopic sphincterotomy was performed immediately. None of the patients presented signs suggesting residual lithiasis with a mean follow-up of 2 months. This preliminary study suggests that the technique is very favourably feasible and may play an important role in the exploration of the bile tract during coelioscopic cholecystectomy.

Peripheral pulmonary lesions: ultrasonic features and ultrasonically guided fine needle aspiration biopsy.

When transthoracic biopsy is required for diagnosing lung diseases, radiographic procedures are the methods of choice, sonographic application being still limited. Sixty-four consecutive patients with pulmonary lesions adjacent to the chest wall underwent sonography. Findings showed hypoechoic homogeneous lesions (56 cases) with posterior regular margin (57 cases). Increasing echoes deep to the mass and sonographic interruption of hyperechoic surface were seen in all patients. In two cases, chest wall invasion had been diagnosed. Histologic diagnosis was made in 55 of the 64 patients (85.9%). Two pneumothoraces occurred. Sonography is a useful, accurate, and safe technique for diagnosing selected pulmonary lesions.

Ultrasonic signs of pneumothorax: preliminary work.

Ultrasonography is considered to have limited application in respiratory diseases because air reflects sound waves. Twenty-four patients with radiologically confirmed pneumothorax and 100 healthy subjects underwent sonography. In all normal subjects, the hyperechoic pulmonary interface showed respiratory motions termed the "gliding sign" with some comet-tail artifacts. Sonographic signs were shown in all pneumothoraces: disappearance of the gliding sign and no comet tails. The extent of collapse cannot be evaluated, but it is possible to determine its area in partial pneumothorax (N = 5). The follow-up (N = 8) showed the reappearance of the gliding sign. Ultrasonography may be helpful in diagnosing pneumothorax in certain cases.

Ultrasonically guided aspiration biopsy in osteolytic bone lesions of the chest wall.

Sixteen consecutive patients with one or more osteolytic bone lesions of the chest wall radiologically confirmed underwent ultrasonically guided aspiration biopsy. Nine patients (56.2 percent) had bronchogenic carcinoma with a direct extension. Other diagnostic techniques had failed to diagnose disease in these patients. The lesion showed heterogeneous echogenicity (n = 16) caused by the hyperechoic signals of bone fragments. The interruption of the cortex bone was detected in all cases and extraosseous tumor portion in 14 of 16 patients (87.5 percent). No respiratory motions of the lesion could be demonstrated (n = 16). Definitive histologic diagnosis was made in 14 of the 16 patients (87.5 percent). In malignancy, diagnosis was established in 13 of 14 patients (92.8 percent). Of two confirmed benign lesions, one diagnosis of tuberculosis was obtained. No complication occurred. Sonography and consequently ultrasonically guided aspiration biopsy are a useful, accurate, safe, and low-cost technique for osteolytic lesions in thoracic diseases.