Another observation with VATER association and a complex IV respiratory chain deficiency

The VATER association of vertebral anomalies (V), anal atresia (A), esophageal atresia and/or tracheo-esophageal fistula (TE), radial and renal anomalies (R) is a common congenital association of unknown origin with probably heterogeneous causes. Here, we report on a girl presenting with pre- and postnatal growth retardation, esophageal atresia, vertebral and costal anomalies and a unilateral radial defect, consistent with the diagnosis of VATER association. In the first month of life, she presented with failure to thrive, severe episodes of hypoglycemia, liver dysfunction and high levels of lactate, which prompted us to perform metabolic screening. A complex IV respiratory chain deficiency (RCD) was diagnosed on a liver biopsy. The respiratory chain defect was not observed in skin fibroblasts. No mtDNA point mutation or deletion was identified. The girl is now 9 years old and has a normal mental development but persistent feeding difficulties and moderate hyperlactatemia. To our knowledge, this is the second report of VATER association with mitochondrial disorder. In a previous report, a VACTERL association was observed in a girl with the mitochondrial A3243G point mutation. The association of VATER phenotype with a mitochondrial disorder may be coincidental but could also suggest that the presence of multiple malformations is the result of the antenatal expression of RCD.     

Internal quality control of histological diagnosis in pathology. A nine-year experience

Internal quality control (IQC) is a necessary component of total quality management. We report our experience with an internal audit scheme focusing on the histological diagnosis. We outline other strategies of IQC and analyze the causes of errors and ways to prevent them. Some practical guidelines to initiate this type of procedure are presented. Our audit was designed to check the accuracy of diagnosis, the clarity and completeness of the report, the quality of the documents leading to the diagnosis, and the turn-around time. It consisted of a retrospective analysis of 4185 randomly selected cases (representing 2% of all cases), over nine years. The control took place once a week and was done by two pathologists working as a team. The mean time spent by each pathologist was 45 minutes per week. Errors were scored using a 3-level grading scheme depending on their potential harm or impact on patient care. The overall rate of errors was 1.1%, and 0.1% of errors were potentially harmful to the patients. A single case (0.02%), in which a cancer was missed, had a real impact on patient care. Retrospective analysis of randomly selected cases mirrors the overall activity of a surgical pathology department. Nevertheless, each lab has to develop its own strategy of IQC, based on its size, its functioning, and its objectives. Although it may be difficult to initiate quality assurance when medical time is already limited, it is a helpful procedure in a more and more demanding medical and societal context and a pragmatic step towards "culture of quality".     

Defibrotide for Sinusoidal Obstruction Syndrome/Veno-Occlusive Disease Prophylaxis in High-Risk Adult Patients: A Single-Center Experience Study

Sinusoidal obstruction syndrome (SOS), also known as hepatic veno-occlusive disease (VOD), is a serious complication after hematopoietic stem cell transplantation (HSCT). SOS/VOD usually occurs within 3 weeks of HSCT, but the 2016 European Society for Blood and Marrow Transplantation diagnosis criteria have been revised to include late forms. Prophylactic use of defibrotide is recommended in the pediatric setting, but its value remains uncertain in the adult population. We report here a single-center series of 63 adult patients considered at high risk for SOS/VOD who received defibrotide prophylaxis in combination with ursodeoxycholic acid between May 2012 and August 2016. The median duration of defibrotide therapy was 23 days. Bleeding occurred in 14 patients (21.5%). Defibrotide prophylaxis was discontinued in 7 patients (10.8%): 4 cases (6.3%) due to bleeding and 3 cases (4.6%) because of the need for antithrombotic therapy. Overall, SOS/VOD occurred in 4 cases (6.3%) within 21 days after HSCT (days 13 and 14) in 2 cases and late-onset SOS/VOD (days 57 and 58) in the other 2 cases. SOS/VOD was moderate in 1 case, very severe in 3 cases, with 2 deaths related to SOS/VOD. Cumulative incidence of grades II to IV acute graft-versus-host disease and transplant-associated thrombotic microangiopathy were 22.2% and 3.2%, respectively. With a median follow-up of 31 months (range, 10.7 to 60.3), the rates of 2-year overall survival, progression-free survival, incidence of relapse, and nonrelapse mortality were 56.5%, 49%, 28.7%, and 22.3%, respectively. In our experience defibrotide prophylaxis is associated with a low incidence of SOS/VOD after allogeneic HSCT in a high-risk adult population with an acceptable safety profile.     

Quantitation of HHV-7 genome by real-time polymerase chain reaction assay using MGB probe technology

A real-time PCR assay was developed to quantify human herpesvirus-7 (HHV-7) genome based on TaqMan technology using the new MGB probe. Primers and probe were chosen in the conserved U100 gene. Plasmid containing the sequence of interest was constructed for the standardisation of the method and to assess its sensitivity. This HHV-7 genomic quantitation assay has a threshold sensitivity of fourteen equivalent genome copy number (EqCop) per reaction. This method was applied to the quantitation of HHV-7 in the peripheral blood mononuclear cells (PBMCs) obtained from 31 healthy subjects. Eighty seven per cent had HHV-7 positive detection in the PBMCs with a viral load ranging from 275 to 14545 EqCop per million of cells. This method presents interesting characteristics with a wide range of quantitation, a good sensitivity, and constitutes a new tool for the study of HHV-7 infection in vivo and in vitro.     

Adaptation of a CXCR4-using human immunodeficiency type 1 NDK virus in intestinal cells is associated with CD4-independent replication

Infection of epithelial colon carcinoma cell line HT29 with human immunodeficiency virus type 1 (HIV-1) NDK, a subtype D virus highly cytopathic for CD4-positive lymphocytes, results in the selection of HIV-1 variants, 1000 times more infectious for CD4(-) intestinal cells than the parental virus. Here, we demonstrate that the envelope gene of intestinal cell-adapted virus conferred to recombinant clone HIV-1 iNDK the ability to utilize CXCR4 without CD4 while retaining its tropism for CD4 lymphocytes. Among the major genetic changes required for infection of intestinal cells and CD4 independence, two potential N-glycosylation sites appeared as a result of the extension of five amino acids in the V1/V2 region and three amino acid changes ((296)KYT --> (296)NNI) were identified in the V3 loop of HIV-1 iNDK gp120. Our studies suggest that CD4-independent use of CXCR4 can be mediated by different adaptive changes related to the microenvironment of CD4(-) cell.     

Combination of WAGR and Potocki-Shaffer contiguous deletion syndromes in a patient with an 11p11.2-p14 deletion

Aniridia, Wilms tumor, genitourinary abnormalities, growth and mental retardation are the cardinal features of the WAGR 11p13 deletion syndrome. The Potocki-Schaffer syndrome or proximal 11p deletion syndrome (previously DEFECT11 syndrome) is a contiguous gene syndrome associated with deletions in 11p11.2, principal features of which are multiple exostoses and enlarged parietal foramina. Mental handicap, facial dysmorphism and craniosynostosis may also be associated. We report a patient with combined WAGR and Potocki-Shaffer syndromes, and obesity. She presented with aniridia, cataract, nystagmus, corneal ulcers and bilateral congenital ptosis. A left nephroblastoma was detected at 15 months. Other features included moderate developmental delay, growth deficiency, facial dysmorphism, multiple exostoses and cranial lacunae. High-resolution and molecular cytogenetics confirmed a del(11)(p11.2p14.1) deletion with a proximal breakpoint between the cosmid DO8153 and the BAC RP11-104M24 to a distal breakpoint between cosmids CO8160 (D11S151) and F1238 (D11S1446). The deletion therefore includes EXT2, ALX4, WT1 and PAX6. This case appears to be the second patient reported with this combined deletion syndrome and confirms the association of obesity in the WAGR spectrum, a feature previously reported in four cases, and for which the acronym WAGRO has been suggested. Molecular and follow-up data on the original WAGRO case are briefly presented.     

Keratinocyte growth factor expression by fibroblasts in pulmonary fibrosis: poor response to interleukin-1beta

Keratinocyte growth factor (KGF) is secreted by fibroblasts and protects from pulmonary fibrosis in animal models. Interleukin (IL)-1beta is the most potent inducer of KGF in fibroblasts, acting through the c-Jun pathway. We evaluated in vitro KGF production by human lung fibroblasts from patients with idiopathic pulmonary fibrosis (IPF, n = 10) and from control subjects (n = 7) at baseline and after IL-1beta stimulation. Basal KGF secretion by IPF fibroblasts was similar to controls. In fibroblasts from control subjects, IL-1beta increased c-Jun expression, c-Jun activation, and KGF secretion. SP600125, a specific c-Jun N-terminal kinase (JNK) inhibitor, inhibited the effect of IL-1beta. By contrast, in IPF fibroblasts, IL-1beta did not increase c-Jun expression and c-Jun activation, and weakly increased KGF secretion, whereas SP600125 had no effect. IL-1beta similarly increased JunB expression in fibroblasts from patients with IPF and control subjects. Total JNK content was not different in either unstimulated or IL-1beta-stimulated IPF and control fibroblasts. IL-1beta increased phosphorylated JNK in control and IPF fibroblasts, but this increase was weaker and heterogeneous in IPF. Altogether, our results demonstrate a dysregulation of KGF secretion by IPF fibroblasts. The weak response to IL-1beta is associated with a defect of c-Jun expression and activation and a defect of JNK activation.     

Compensation for distal impairments of grasping in adults with hemiparesis

Previous studies have shown that patients with arm and hand paresis following stroke recruit an additional degree of freedom (the trunk) to transport the hand during reaching and use alternative strategies for grasping. The few studies of grasping parameters of the impaired hand have been case studies mainly focusing on describing grasping in the presence of particular impairments such as hemi-neglect or optic ataxia and have not focussed on the role of the trunk in prehension. We hypothesized that the trunk movement not only ensures the transport of the hand to the object, but it also assists in orienting the hand for grasping when distal deficits are present. Nineteen patients with chronic hemiparesis and seven healthy subjects participated in the study. Patients had sustained a stroke of non-traumatic origin 6–82 months previously (31±22 months) and had mild or moderate to severe arm paresis. Using a whole hand grasp, subjects reached and grasped a cylinder (35 mm) that was placed sagittally (T1) or at a 45° angle to the sagittal midline in the ipsilateral workspace (T2), both at about 90% arms length (10 trials per target). Eight infrared emitting diodes were placed on bony landmarks of the hand, arm and trunk and kinematic data were recorded by an optical motion analysis system (Optotrak) for 2–5 s at 120 Hz. Hand position and orientation were recorded by a Fastrack Polhemus system. Our results show that during goal-directed prehension tasks, individuals with hemiparesis oriented the hand more frontally for grasping and used more trunk anterior displacement or rotation to transport the hand to the target compared to healthy subjects. Despite these changes, the major characteristics of reaching and grasping such as grip aperture size, temporal coordination between hand transport and aperture formation and the relative timing of grip aperture were largely preserved. For patients with more severe distal impairments, the amount of trunk displacement was also correlated with a more frontal hand orientation for grasping. Furthermore, in healthy subjects and patients without distal impairments, the trunk movement was mostly related to proximal arm movements while in those with distal impairments, trunk movement was related to both proximal and distal arm movements. Data support the hypothesis that the trunk movement is used to assist both arm transport and hand orientation for grasping when distal deficits are present.