Real-world EGFR testing practices for non-small-cell lung cancer by thoracic pathology laboratories across Europe.

BACKGROUND: Testing for epidermal growth factor receptor (EGFR) mutations is an essential recommendation in guidelines for metastatic non-squamous non-small-cell lung cancer, and is considered mandatory in European countries. However, in practice, challenges are often faced when carrying out routine biomarker testing, including access to testing, inadequate tissue samples and long turnaround times (TATs). MATERIALS AND METHODS: To evaluate the real-world EGFR testing practices of European pathology laboratories, an online survey was set up and validated by the Pulmonary Pathology Working Group of the European Society of Pathology and distributed to 64 expert testing laboratories. The retrospective survey focussed on laboratory organisation and daily EGFR testing practice of pathologists and molecular biologists between 2018 and 2021. RESULTS: TATs varied greatly both between and within countries. These discrepancies may be partly due to reflex testing practices, as 20.8% of laboratories carried out EGFR testing only at the request of the clinician. Many laboratories across Europe still favour single-test sequencing as a primary method of EGFR mutation identification; 32.7% indicated that they only used targeted techniques and 45.1% used single-gene testing followed by next-generation sequencing (NGS), depending on the case. Reported testing rates were consistent over time with no significant decrease in the number of EGFR tests carried out in 2020, despite the increased pressure faced by testing facilities during the COVID-19 pandemic. ISO 15189 accreditation was reported by 42.0% of molecular biology laboratories for single-test sequencing, and by 42.3% for NGS. 92.5% of laboratories indicated they regularly participate in an external quality assessment scheme. CONCLUSIONS: These results highlight the strong heterogeneity of EGFR testing that still occurs within thoracic pathology and molecular biology laboratories across Europe. Even among expert testing facilities there is variability in testing capabilities, TAT, reflex testing practice and laboratory accreditation, stressing the need to harmonise reimbursement technologies and decision-making algorithms in Europe.

Clinical and molecular practice of European thoracic pathology laboratories during the COVID-19 pandemic. The past and the near future.

BACKGROUND: This study evaluated the consequences in Europe of the COVID-19 outbreak on pathology laboratories orientated toward the diagnosis of thoracic diseases. MATERIALS AND METHODS: A survey was sent to 71 pathology laboratories from 21 European countries. The questionnaire requested information concerning the organization of biosafety, the clinical and molecular pathology, the biobanking, the workload, the associated research into COVID-19, and the organization of education and training during the COVID-19 crisis, from 15 March to 31 May 2020, compared with the same period in 2019. RESULTS: Questionnaires were returned from 53/71 (75%) laboratories from 18 European countries. The biosafety procedures were heterogeneous. The workload in clinical and molecular pathology decreased dramatically by 31% (range, 3%-55%) and 26% (range, 7%-62%), respectively. According to the professional category, between 28% and 41% of the staff members were not present in the laboratories but did teleworking. A total of 70% of the laboratories developed virtual meetings for the training of residents and junior pathologists. During the period of study, none of the staff members with confirmed COVID-19 became infected as a result of handling samples. CONCLUSIONS: The COVID-19 pandemic has had a strong impact on most of the European pathology laboratories included in this study. Urgent implementation of several changes to the organization of most of these laboratories, notably to better harmonize biosafety procedures, was noted at the onset of the pandemic and maintained in the event of a new wave of infection occurring in Europe.

Protein tyrosine phosphatase-PEST regulates focal adhesion disassembly, migration, and cytokinesis in fibroblasts.

In this article, we show that, in transfected COS-1 cells, protein tyrosine phosphatase (PTP)-PEST translocates to the membrane periphery following stimulation by the extracellular matrix protein fibronectin. When plated on fibronectin, PTP-PEST (-/-) fibroblasts display a strong defect in motility. 3 h after plating on fibronectin, the number and size of vinculin containing focal adhesions were greatly increased in the homozygous PTP-PEST mutant cells as compared with heterozygous cells. This phenomenon appears to be due in part to a constitutive increase in tyrosine phosphorylation of p130(CAS), a known PTP-PEST substrate, paxillin, which associates with PTP-PEST in vitro, and focal adhesion kinase (FAK). Another effect of this constitutive hyperphosphorylation, consistent with the focal adhesion regulation defect, is that (-/-) cells spread faster than the control cell line when plated on fibronectin. In the PTP-PEST (-/-) cells, an increase in affinity for the SH2 domains of Src and Crk towards p130(CAS) was also observed. In (-/-) cells, we found a significant increase in the level of tyrosine phosphorylation of PSTPIP, a cleavage furrow-associated protein that interacts physically with all PEST family members. An effect of PSTPIP hyperphosphorylation appears to be that some cells remain attached at the site of the cleavage furrow for an extended period of time. In conclusion, our data suggest PTP-PEST plays a dual role in cell cytoskeleton organization, by promoting the turnover of focal adhesions required for cell migration, and by directly or indirectly regulating the proline, serine, threonine phosphatase interacting protein (PSTPIP) tyrosine phosphorylation level which may be involved in regulating cleavage furrow formation or disassembly during normal cell division.

Radiofrequency ablation of retained placenta accreta after conservative management: preliminary evaluation in the pregnant ewe and in normal human placenta in vitro.

OBJECTIVE: To evaluate radiofrequency (RF) efficiency and safety for the ablation of retained placenta in humans, using a pregnant sheep model. DESIGN: Experimental study. SETTING: Laboratory of Surgery School, Nancy, France. POPULATION/SAMPLE: Three pregnant ewes/ten human placentas. METHODS: Various RF procedures were tested in pregnant ewes on 50 placentomes (individual placental units). Reproducibility of the best procedure was then evaluated in a further 20 placentomes and on ten human term placentas in vitro after delivery. MAIN OUTCOME MEASURES: Placental tissues destruction, lesions' size, myometrial lesions. RESULTS: Low power (100 W) and low target temperatures (60 degrees C) lead to homogenous tissue destruction, without myometrial lesion. No significant difference was observed in terms of lesion size and procedure duration for in the placentomes of pregnant ewe in vivo and in human placentas in vitro. The diameter of the ablation could be correlated with the tines deployment. CONCLUSION: The placental tissue structure is very permissive to RF energy, which suggests that RF could be used for the ablation of retained placenta, providing optimal control of tissue destruction. These results call for further experimental evaluations.

Adipose-tissue engineering: taking advantage of the properties of human adipose-derived stem/stromal cells.

Adipose tissue is now recognized as an important source of postnatal mesenchymal stem cells for regenerative medicine applications. For example, adipose-tissue engineering is an emerging approach that enables the development of autologous substitutes that could be used as an alternative to fat transplantation methods currently yielding variable outcomes for the long-term repair of soft-tissue defects. Here, we describe the production of unique tissue-engineered adipose tissues devoid of exogenous biomaterials produced from human adipose-derived stem/stromal cells. Our strategy is based on the dual self-assembly of extracellular components secreted and organized by the adipose-derived stromal cells after ascorbic acid stimulation, as well as their concomitant differentiation into adipocytes after adipogenic induction. When compared to stromal cells isolated from resected fat, lipoaspirated fat-derived cells featured an increased adipogenic potential and the enhanced ability to recreate three-dimensional adipose substitutes in vitro. These substitutes were histologically similar to native adipose tissue. They featured lipid-filled adipocytes embedded into an extracellular matrix rich in fibronectin as well as collagens I and V. On a functional level, the reconstructed adipose tissues expressed adipocyte-related transcripts and secreted adipokines typical of adipose tissue, such as leptin. Finally, the successful in vitro production of human adipose substitutes featuring an increased surface area (>30cm2) is described, reinforcing the notion that customized autologous reconstructed adipose tissues could be produced in the future to repair a wide range of soft-tissue defects.

[Malignant mixed müllerian tumor of the uterus following tamoxifen for breast cancer: case report]. / Tumeur müllérienne mixte de l'utérus après traitement par tamoxifène pour cancer du sein: à propos d'un cas.

Tamoxifen is an antioestrogen widely used in the breast cancer treatment. Its paradoxical antioestrogenic action on breast and its oestrogenic action on the endometer is well-known. Since 1988, few cases of malignant mixed müllerian tumors following tamoxifen were described. We report a new case of uterine malignant mixed müllerian tumor four years after the end of tamoxifen for breast cancer.

The Cdc42/Rac1 regulator CdGAP is a novel E-cadherin transcriptional co-repressor with Zeb2 in breast cancer.

The loss of E-cadherin causes dysfunction of the cell-cell junction machinery, which is an initial step in epithelial-to-mesenchymal transition (EMT), facilitating cancer cell invasion and the formation of metastases. A set of transcriptional repressors of E-cadherin (CDH1) gene expression, including Snail1, Snail2 and Zeb2 mediate E-cadherin downregulation in breast cancer. However, the molecular mechanisms underlying the control of E-cadherin expression in breast cancer progression remain largely unknown. Here, by using global gene expression approaches, we uncover a novel function for Cdc42 GTPase-activating protein (CdGAP) in the regulation of expression of genes involved in EMT. We found that CdGAP used its proline-rich domain to form a functional complex with Zeb2 to mediate the repression of E-cadherin expression in ErbB2-transformed breast cancer cells. Conversely, knockdown of CdGAP expression led to a decrease of the transcriptional repressors Snail1 and Zeb2, and this correlated with an increase in E-cadherin levels, restoration of cell-cell junctions, and epithelial-like morphological changes. In vivo, loss of CdGAP in ErbB2-transformed breast cancer cells impaired tumor growth and suppressed metastasis to lungs. Finally, CdGAP was highly expressed in basal-type breast cancer cells, and its strong expression correlated with poor prognosis in breast cancer patients. Together, these data support a previously unknown nuclear function for CdGAP where it cooperates in a GAP-independent manner with transcriptional repressors to function as a critical modulator of breast cancer through repression of E-cadherin transcription. Targeting Zeb2-CdGAP interactions may represent novel therapeutic opportunities for breast cancer treatment.

Comparison of semi-automated and manual methods to measure the volume of prostate cancer on magnetic resonance imaging.

PURPOSE: The purpose of this study was to determine the accuracy of manual semi-automated and volumetric measurements to assess prostate cancer volume on multiparametric magnetic resonance imaging (MP-MRI) using whole-mount histopathology for validation. MATERIALS AND METHODS: We evaluated 30 consecutive men (median age, 65.7 years; interquartile range [IQR], 61.5-70.9 years) with a median prostatic specific antigen of 8.5ng/dL (IQR, 5.5-10.5ng/dL), who underwent MP-MRI before radical prostatectomy. Index tumor volume was determined prospectively and independently on the basis of MRI and whole-mount section volumetric assessment using the maximum histologic diameter (MHD) and the histologic volume (HV). The MRI index tumor volume was determined by two independent radiologists using a single measurement of the maximum tumor dimension (MTD), a simplified MR ellipsoid volume (MREV) calculation and a MR region of interest volume (MROV) segmentation displayed by a commercially available OsiriX®. MTD was compared to MHD, whereas MREV and MROV were compared to HV. RESULTS: Thirty index lesions (median HV, 1.514 cm3; IQR, 0.05-3.780 cm3) were analyzed. The MREV, MROV and HD were significantly correlated with each other (r>0.5). Inter-observer agreement for measurements was good for each method (r>0.780). The MTD was the best predictor of maximum histologic diameter (r=0.980 and 0.791) and had an excellent inter-variability correlation (P<0.0001). CONCLUSION: Prostate cancer histologic volume can be assessed using MREV or MROV with a good accuracy and low inter-observer variability. MTD has the lowest inter-observer variability and provides best degrees of correlation with MHD. MTD should be used on MRI for selecting and following patients for active surveillance and staging before focal treatment of prostate cancer.

Nitric oxide synthase in cat brain: cofactors--enzyme-substrate interaction.

Nitric oxide, derived from L-arginine by the enzyme nitric oxide synthase, is an activator of the soluble guanylate cyclase and a cellular messenger. This work demonstrates that, in cat brain, the neuronal constitutive nitric oxide synthase activity is a) NADPH/calcium dependent, b) independent upon exogenous calmodulin in crude brain supernatant, c) significantly enhanced by exogenous FAD and tetrahydrobiopterin (Vmax: 118 instead of 59.4 pmol of citrulline formed .mg of prot.-1 min-1, d) inhibited by calcium chelators and calmodulin antagonist, and e) present in several neuroanatomical structures. Moreover, the Km value for L-arginine was of 11 microM instead of 41 microM in the presence of FAD and tetrahydrobiopterin in the incubation mixture, thus demonstrating that these cofactors are able to stabilize the enzyme-substrate interactions.

Collagenous gastritis revealed by severe anemia in a child.

Collagenous gastritis is a rare histopathological disorder of unknown origin, characterized by a subepithelial collagen deposit greater than 10 microm thick, associated with an inflammatory infiltrate of the gastric mucosa. This report describes a second pediatric case of collagenous gastritis, revealed by severe anemia caused by gastric bleeding, as was the first case. Unlike the adult cases of collagenous gastritis, lesions were limited to the stomach, and remained unchanged on six series of biopsies taken during a 30 month follow-up, despite treatment with omeprazole, sucralfate and corticosteroids. An immunohistochemical study showed signs of local immune activation on all biopsy specimens, including overexpression of HLA-DR by epithelial cells, increased numbers of CD3+ intraepithelial lymphocytes, and CD25+ cells in the lamina propria. Although the cause of the disease remains unclear, our findings suggest that the histopathological lesions of collagenous gastritis may result from a local immune process.

Twenty years of experience with dairy herd health in Ontario.

This article expresses observations on planned herd health for dairy cattle, based on experience gained in the Ambulatory Clinic practice of the Ontario Veterinary College. The author and his colleagues, especially Dr. R.A. Curtis, have initiated and delivered a preventive medicine approach to veterinary practice in the teaching program and teaching practice for the past 20 years. In addition, herd health presentations have been made to veterinary associations in every province in Canada and to many breed associations and producer organizations. The Canadian food animal veterinarian and his clients have been informed at meetings and by the media of the need, objectives, methods and benefits of dairy herd health and many veterinary practices now offer programs to their clients. Herd health has become a household word in Canada's dairy practices and dairy farms.A formal herd health program is an important step to achieving total health management; but maximum returns on investment can only be realized after three or four generations of cattle have been reared on the program. In conclusion, herd health practice has been a very satisfying aspect of veterinary medicine and a profitable and valued service for our clients. Maintenance of health involves the application of all knowledge and procedures which veterinarians have to offer.

Effectiveness of an ivermectin liquid formulation given by nasogastric tube against strongyles in horses.

Twenty horses were treated with ivermectin either by nasogastric tube with a liquid formulation for sheep or per os with a paste formulation for horses at a dosage of 200 mug/kg of body weight. Fecal samples were collected from these horses and from ten untreated horses at the time of treatment and every 2 wk thereafter for up to 10 wk. The samples were examined for nematode eggs using the Cornell-McMaster dilution and the Cornell-Wisconsin Double Centrifugation procedures.There were no signs of toxicosis in horses treated with ivermectin. Strongyle eggs were found in the feces of all horses before treatment. Subsequently, they were found in untreated horses, but not in treated horses at 2 wk nor in most of them for up to 8 wk after treatment. At 10 wk most of these horses had strongyle eggs in their feces, but in general fewer than at pretreatment.

Effectiveness of ivermectin paste for removal of nematodes in the horse.

Thirteen Standardbred horses, two to five years of age, were treated with ivermectin paste per os at 200 mug/kg of body weight and 13 were untreated. Two weeks after treatment, previously untreated horses were given the paste. Fecal samples were collected from all horses at the time of treatment and periodically thereafter up to 14 weeks and were examined for nematode eggs using the Cornell-McMaster dilution and the Cornell-Wisconsin double centrifugation procedures.All horses consumed the paste readily and had no signs of toxicosis. Strongyle eggs were found in the feces of all horses before treatment but not at two to three weeks after treatment. At five to six weeks after treatment only two horses had eggs in the feces. At eight, ten, 12 and 14 weeks after treatment 27, 69, 88 and 100% of the horses examined, respectively, had a few strongyle eggs but these were no greater than 18% of that of the pretreatment samples. Ivermectin oral paste, therefore, appeared to be highly effective against both adult and immature strongyles.

Effectiveness of oxibendazole against benzimidazole-resistant strongyles in horses.

Twenty-eight horses with a residual burden of strongyle eggs in the faces after treatment with mebendazole (MBZ) paste were treated with a suspension of either MBZ or oxibendazole (OBZ). Fecal samples were collected before and 14 days after these treatments. The number of strongyle eggs/g (epg) of feces for each horse was estimated using the Cornell-McMaster dilution and the Cornell-Wisconsin double centrifugation procedures. The epg for each horse was transformed using log (x + 1) and in an analysis of variance of the reduction in egg count for each horse on the logarithmic scale, there was a highly significant difference between the treatments. The mean epg was increased in the MBZ-treated horses and reduced in the OBZ-treated horses, but the reduction was only by 82% with an upper confidence limit of 89%. Subsequently, the horses were retreated with MBZ and OBZ suspensions without significant reduction in the mean epg for OBZ-treated horses.

The effect of retained placenta and metritis complex on reproductive performance in dairy cattle -- a case control study.

A study of the computer stored records of 293 dairy cows and 652 calvings reveals the effects of retained placenta and metritis complex on reproductive performance. The overall incidence rate of retained placenta was 11.2%. Retained placenta was 4.6 times more likely to occur following twin births than following single births. Most cases of retained placenta occurred during the fall. Forty-five percent of that seasonal increase was explained by an increased number of calvings. Metritis complex was diagnosed following 54.8% of retained placenta cases. Retained placenta alone did not significantly impair reproductive performance. Metritis complex, in the presence or absence of retained placenta, caused a significant (P

[Is sentinel lymph node mapping relevant for colon cancer?: a feasibility study]. / Intérêt de l'analyse du ganglion sentinelle pour les cancers coliques : étude de faisabilité.

BACKGROUND: Sentinel lymph node (sln) technique using blue injection is controversial for colon cancer. The aim of this study was to evaluate the feasibility and interest of sentinel node detection to identify the ultrastaging rate detecting occult nodal micrometastases missed on routine H&E examination. METHODS: During surgery blue dye was injected subserously around the tumor in 30 patients operated for a colon cancer. The first lymph nodes to turn blue were noted as sln. For each sln three examination levels were performed; if no tumor was detected by H&E examination, a cytokeratine immunohistochemistry study was performed. RESULTS: For each case, one or more sln were found (100%). The median number of lymph nodes examined and of sln found was, respectively, 23 (range 10-55) and 2 (1-4). There were 21 pN0 tumors, among which we found two cases (9%) with a micrometastasis and one case of isolated tumor cells detected, resulting in a 14% (3/21) ultrastaging for pTxN0. The sln was positive in five patients out of nine with a N+ disease. CONCLUSIONS: Sln detection was a successful technique when there was no evident lymph node involvement, no primary large lesion or no associated liver metastasis. Focused examination of the sln identified 10-20% of additional ultrastaging disease for staged pT1, 2, 3N0M0 tumor. This may have an important implication for follow-up and adjuvant treatment in future protocols.

[Adenomatoid hamartoma of the ethmoid sinus: one case report]. / L'hamartome adénomatoide naso-sinusien: à propos d'un cas.

INTRODUCTION: Hamartomas are non-malignant malformations or inborn errors of tissue development. In the head and neck region, especially in the nasal cavity and the ethmoid sinuses, they are relatively rare. PATIENTS AND METHODS: A case of respiratory epithelial adenomatoid hamartoma of the ethmoid sinus and a review of literature are reported in order to describe the diagnostic and therapeutic management of this tumour. RESULTS: A 74-year-old man complaining for unilateral nasal obstruction for years was referred to our institution. Clinical and radiological studies revealed a large intra-nasal tumour, osteolytic in nature, arising from the right ethmoidal sinus. Fifteen months after a complete excision of the tumour using an endoscopic procedure, the nasal cavity was free of tumour. DISCUSSION AND CONCLUSION: Very rare and classified as non-malignant tumours, hamartomas are composed of excessive proliferation of one or more cellular components specific to a given tissue. They can grow out of any part of the body for example the surface epithelium, seromucous glands and vessels. Hamartomas commonly originate from the lung, kidney and intestine. Their localisation in the nasal cavity, especially in the ethmoid sinus, is unusual, but it is really important to be known to distinguish hamartomas from papillomas and adenocarcinomas not to perform useless and destructive surgery.

Impaired cell death and mammary gland involution in the absence of Dock1 and Rac1 signaling.

Throughout life, the tight equilibrium between cell death and the prompt clearance of dead corpses is required to maintain a proper tissue homeostasis and prevent inflammation. Following lactation, mammary gland involution is triggered and results in the death of excessive epithelial cells that are rapidly cleared by phagocytes to ensure that the gland returns to its prepregnant state. Orthologs of Dock1 (dedicator of cytokinesis 1), Elmo and Rac1 (ras-related C3 botulinum toxin substrate 1) in Caenorhabditis elegans are part of a signaling module in phagocytes that is linking apoptotic cell recognition to cytoskeletal reorganization required for engulfment. In mammals, Elmo1 was shown to interact with the phosphatidylserine receptor Bai1 and relay signals to promote phagocytosis of apoptotic cells. Still, the role of the RacGEF Dock1 in the clearance of dying cells in mammals was never directly addressed. We generated two mouse models with conditional inactivation of Dock1 and Rac1 and revealed that the expression of these genes is not essential in the mammary gland during puberty, pregnancy and lactation. We induced mammary gland involution in these mice to investigate the role of Dock1/Rac1 signaling in the engulfment of cell corpses. Unpredictably, activation of Stat3 (signal transducer and activator of transcription 3), a key regulator of mammary gland involution, was impaired in the absence of Rac1 and Dock1 expression. Likewise, failure to activate properly Stat3 was coinciding with a significant delay in the initiation and progression of mammary gland involution in mutant animals. By using an in vitro phagocytosis assay, we observed that Dock1 and Rac1 are essential to mediate engulfment in epithelial phagocytes. In vivo, cell corpses accumulated at late time points of involution in Dock1 and Rac1 mutant mammary glands. Overall, our study demonstrated an unsuspected role for Dock1/Rac1 signaling in the initiation of mammary gland involution, and also suggested a role for this pathway in the clearance of dead cells by epithelial phagocytes.