The Advantage of Targeted Next-Generation Sequencing over qPCR in Testing for Druggable EGFR Variants in Non-Small-Cell Lung Cancer.

The emergence of targeted therapies in non-small-cell lung cancer (NSCLC), including inhibitors of epidermal growth factor receptor (EGFR) tyrosine kinase, has increased the need for robust companion diagnostic tests. Nowadays, detection of actionable variants in exons 18-21 of the EGFR gene by qPCR and direct DNA sequencing is often replaced by next-generation sequencing (NGS). In this study, we evaluated the diagnostic usefulness of targeted NGS for druggable EGFR variants testing in clinical NSCLC material previously analyzed by the IVD-certified qPCR test with respect to DNA reference material. We tested 59 NSCLC tissue and cytology specimens for EGFR variants using the NGS 'TruSight Tumor 15' assay (Illumina) and the qPCR 'cobas EGFR mutation test v2' (Roche Diagnostics). The sensitivity and specificity of targeted NGS assay were evaluated using the biosynthetic and biological DNA reference material with known allelic frequencies (VAF) of EGFR variants. NGS demonstrated a sufficient lower detection limit for diagnostic applications (VAF < 5%) in DNA reference material; all EGFR variants were correctly identified. NGS showed high repeatability of VAF assessment between runs (CV% from 0.02 to 3.98). In clinical material, the overall concordance between NGS and qPCR was 76.14% (Cohen's Kappa = 0.5933). The majority of discordant results concerned false-positive detection of EGFR exon 20 insertions by qPCR. A total of 9 out of 59 (15%) clinical samples showed discordant results for one or more EGFR variants in both assays. Additionally, we observed TP53 to be a frequently co-mutated gene in EGFR-positive NSCLC patients. In conclusion, targeted NGS showed a number of superior features over qPCR in EGFR variant detection (exact identification of variants, calculation of allelic frequency, high analytical sensitivity), which might enhance the basic diagnostic report.

Outcomes and experience of anatomical partial lobectomy.

OBJECTIVE: The use of limited anatomic resection for early-stage lung cancer is widely practiced worldwide. However, most studies have focused on standard segmentectomy or subsegmentectomy, and data on the short-term outcomes after anatomic sublobar resection in China are lacking. METHODS: In 2014, the use of anatomic partial lobectomy (APL), which is defined as lesion-centered resection of anatomical sublobular parts, was proposed by the National Cancer Center in China. We retrospectively evaluated all consecutive patients who underwent APL between November 2013 and October 2019 from our database, and the operative techniques and short-term outcomes were analyzed. RESULTS: A total of 3336 patients with a median age of 56 years underwent APL during the study period. Benign lesions were present in 8.5% of all patients and decreased across time. The mean total operation time was 127.3 minutes, the mean overall number of nodal sampling/dissections was 13, and the mean number of stations sampled was 4. Postoperative complications (grade ≥2) developed in 359 patients (10.8%), and no mortality occurred in the 30 days after surgery. Multivariate analysis showed that smoking, surgeon's early experience, thoracotomy or unplanned conversion to thoracotomy, and complex cases were risk factors for the occurrence of postoperative complications. CONCLUSIONS: Despite the increasing proportion of complex cases treated with APL, the incidence of postoperative complications decreased as our center accumulated surgical experience. APL procedures are safe and feasible when conducted in a specialized center.

Ligaments of the os trigonum: an anatomical study.

PURPOSE: The aim of the study was to examine the ligaments of the os trigonum. METHODS: The ankle joint magnetic resonance imaging (MRI) of 104 patients with the os trigonum (experimental group) and 104 patients without the os trigonum (control group) were re-reviewed. The connections of the os trigonum and posterior talofibular ligament (PTFL), the fibulotalocalcaneal ligament (FTCL), the paratenon of the Achilles tendon, the posterior talocalcaneal ligament (PTCL), the osteofibrous tunnel of the flexor hallucis longus (OF-FHL) and the flexor retinaculum (FR) were studied. RESULTS: The os trigonum is connected to structures. The posterior part of the PTFL inserted on the os trigonum in 85.6% of patients, whereas in all patients in the control group, the posterior part of the PTFL inserted on the posterior talar process (p < 0.05). The connection of the PTCL was seen in 94.2% of patients in the experimental group, while it was seen in 90.4% of patients in the control group (p > 0.05). The connection to the FTCL in the experimental group was 89.4%, while in the control group, it was 91.3% (p > 0.05). The communication with the paratenon was seen more often in the control group compared to that in the experimental group (31.7% vs. 63.8%, p < 0.001). The FTCL was prolonged medially into the FR in 85.6% of patients in the experimental group and in 87.5% of patients in the control group (p > 0.05). The flexor hallucis longus (FHL) run at the level of articulation between the os trigonum 63.5% and the posterior process of the talus 25% and less often on the os trigonum 11.5%. CONCLUSION: The os trigonum is connected with all posterior ankle structures and more connections than previously reported.

The Kager's fat pad radiological anatomy revised.

PURPOSE: The aim of the study was to map connections within the Kager's fat pad between the structures which limit it. METHODS: A retrospective re-review of 200 ankle magnetic resonance imaging (MRI) examination was conducted. Connections within the Kager's fat pad between the superior peroneal retinaculum, the fibulotalocalcaneal ligament, the posterior talocalcaneal ligament, the flexor hallucis longus, the paratenon of the Achilles tendon, the flexor retinaculum and bones were studied and a model of the connections was constructed. RESULTS: The superior peroneal retinaculum was directly connected with the fibulotalocalcaneal ligament in 85.5% of cases, the lateral part of the paratenon in 82.5%, the processus posterior tali in 78.5%, the posterior talofibular ligament in 32%, the flexor retinaculum in 29.5% and the anterior talofibular ligament in 9%. The fibulotalocalcaneal ligament was connected with the paratenon (on the medial side 88.5%, on the lateral side 68.5%), the flexor retinaculum in 70%, the posterior process of the talus in 79%, the osteofibrosus tunnel for the flexor hallucis longus in 53%, the posterior talofibular ligament in 43.5% and the calcaneofibular ligament in 10.5%. The posterior talocalcaneal ligament was connected with the fibulotalocalcaneal ligament in 71%, with the osteofibrosus tunnel for the flexor hallucis longus in 76.5%, with the flexor retinaculum in 70%. The plantaris tendon showed projection to the crural fascia in 34 of % cases. CONCLUSION: In the Kager's fat pad there are present more connections than previously reported. All the connections unit at the level of the posterior process of the talus.

The double fascicular variations of the anterior talofibular ligament and the calcaneofibular ligament correlate with interconnections between lateral ankle structures revealed on magnetic resonance imaging.

The anterior talofibular ligament and the calcaneofibular ligament are the most commonly injured ankle ligaments. This study aimed to investigate if the double fascicular anterior talofibular ligament and the calcaneofibular ligament are associated with the presence of interconnections between those two ligaments and connections with non-ligamentous structures. A retrospective re-evaluation of 198 magnetic resonance imaging examinations of the ankle joint was conducted. The correlation between the double fascicular anterior talofibular ligament and calcaneofibular ligament and connections with the superior peroneal retinaculum, the peroneal tendon sheath, the tibiofibular ligaments, and the inferior extensor retinaculum was studied. The relationships between the anterior talofibular ligament's and the calcaneofibular ligament's diameters with the presence of connections were investigated. Most of the connections were visible in a group of double fascicular ligaments. Most often, one was between the anterior talofibular ligament and calcaneofibular ligament (74.7%). Statistically significant differences between groups of single and double fascicular ligaments were visible in groups of connections between the anterior talofibular ligament and the peroneal tendon sheath (p < 0.001) as well as the calcaneofibular ligament and the posterior tibiofibular ligament (p < 0.05), superior peroneal retinaculum (p < 0.001), and peroneal tendon sheath (p < 0.001). Differences between the thickness of the anterior talofibular ligament and the calcaneofibular ligament (p < 0.001), the diameter of the fibular insertion of the anterior talofibular ligament (p < 0.001), the diameter of calcaneal attachment of the calcaneofibular ligament (p < 0.05), and tibiocalcaneal angle (p < 0.01) were statistically significant. The presence of the double fascicular anterior talofibular ligament and the calcaneofibular ligament fascicles correlate with connections to adjacent structures.

Distribution of the subtendons in the midportion of the Achilles tendon revealed in vivo on MRI.

The aim of the study was to check if the subtendons of the Achilles tendon can be identified in vivo on MRI in the midportion of the tendon. The relation of the plantaris tendon to the Achilles tendon was also examined. A retrospective study of 200 MRI of ankle joints including the Achilles tendon was conducted. Statistical analysis of the correlation between the possibility of identifying the subtendons and the side, gender, presence of the central soleus tendon and plantaris tendon variation was performed. The inter-observer agreement between two reviewers in their evaluation of the subtendons was assessed using kappa statistics. The subtendon from the lateral head of the gastrocnemius muscle was identified in 65% (k = 0.63) and was located in the anterior part of the Achilles tendon. The subtendon from the soleus muscle was recognized in 12% (k = 0.75) comprising anterior part of the tendon. In 6% the subtendon from the medial head of the gastrocnemius muscle was identified (k = 0.58). The central soleus tendon was identified in 85% of cases. Statistical analysis shows the weak correlation of the presence of the central soleus tendon and the possibility of identifying the subtendon from the soleus muscle. The plantaris tendon was directly related to the insertion of the Achilles tendon in 42.5%. Identification of the subtendons of the Achilles tendon on MRI is challenging, and most often it is only possible to find the subtendon of the lateral head of the gastrocnemius muscle.

How to optimize the treatment strategy for patients of pulmonary sequestration with an elevated risk of fatal hemorrhage during operation: case discussion.

Video-assisted thoracoscopic surgery (VATS) lobectomy, especially uniportal VATS, is increasingly used for pulmonary sequestration (PS). However, there are few descriptions of safe handling of the aberrant artery with atherosclerosis, especially the diameter of arteries exceeds than 2.0 cm, under uniportal VATS approach. Here we report a 56-year-old man who was diagnosed with pulmonary sequestration following trauma. The patient had a long history of cough with purulent sputum. One month before the trauma, he had copious expectoration with foul smell again. A contrast CT scan revealed a 7.5 cm mass in his right lower lobe. The mass was supplied by a thick aberrant atherosclerotic artery (over than 2 cm in diameter), which stemmed from the thoracic aorta with multiple calcifications on both. After adequate preoperative evaluation, we performed a right lower lobectomy under uniportal VATS approach. No surgical-associated complications occurred, and the patient was discharged on the 5th days after the operation. We organized an iMDT (international multidisciplinary team) to discuss the reasonability and optimal treatment pattern for this patient. We found that fully assess the quality of the aberrant arteries of PS following blocking and cutting off in an appropriate way are crucial to avoid the happening of fatal bleeding during the operation.

Pulmonary veins variations with potential impact in thoracic surgery: a computed-tomography-based atlas.

BACKGROUND: Pulmonary veins (PVs) are important during segmentectomy. Many case reports prove that they may be the source of bleeding during surgery, especially when anatomical variants are present. We decided to describe venous variations and prepare a computed tomography based atlas of our observations. METHODS: The study was conducted using 135 chest computed tomography studies with intra venous iodine contrast injection. The study population contained 86 females and 49 males, mean age was 60. Thirteen people had atrial fibrillation. Images were analysed using radiological workstation. RESULTS: The variations were divided into three categories: atypical topography of the PV, atypical venous outflow to the left atrium (LA), atypical venous vascularization of the lung bronchopulmonary segment. Retrobronchial course of the vein of the posterior segment of the right upper lobe was observed in 8.15%. The most common variant of atrial venous outflow was the direct outflow of the middle lobe vein, observed in 25.19% of cases and the long common trunk of left PVs in 11.11%. The split drainage from the middle lobe into the right superior pulmonary vein (RSPV) and the right inferior pulmonary vein (RIPV) was observed in 9.63% as the full drainage into the RIPV in 2.96%. CONCLUSIONS: Long common trunk of left PVs and numerous variants of venous vascularisation of the middle lobe are the variations that may pose potential problems during thoracic surgeries. The frequency is high enough to justify the routine assessment of pulmonary vessels with computed tomography before surgery.

Polymetastatic testicular cancer turns out to be secondary syphilis: a case report.

Syphilis is a sexually transmissible infection, with increasing rates of infection worldwide. The differential diagnosis of syphilis should include various diseases, not excluding cancer. Making the right diagnosis can protect the patient against life-threatening complications and the repercussions of a misdiagnosis, as in the present case (orchidectomy).

How to increase the accuracy of the diagnosis of the accessory bone of the foot?

PURPOSE: The study was conducted to search for confident radiological signs in symptomatic cases of accessory bones. A normal accessory bone appearance on X-ray does not exclude that the accessory bone is the source of the discomfort; because of this, MRI examination can later be applied as part of the diagnosis. METHODS: We retrospectively analysed cases of 64 patients with recognized 70 symptomatic accessory bones of the foot. The average age was 29.2 (range 8-42) years. We included only patients with X-ray and MRI examinations. We investigated the following radiological features of the bone (structural and signal) in relation to soft tissue. RESULTS: The most constant symptoms identified in our study were bone marrow oedema (93%) and soft tissue oedema (77%). Changes in structures in which accessory bones were located or in adjacent structures to accessory bone were identified: tendon changes 51%, fluid adjacent to bone 51% and tenosynovitis 46%. MRI revealed changes in bone structure that are not seen on X-ray, including changes in contour (28%), sclerosis (3%) or osteonecrosis (3%). CONCLUSIONS: MRI plays an important role in determining whether accessory bones cause symptoms because it shows specific and accurate changes in accessory bone and/or in adjacent soft tissue.

Morphology and morphometry of pulmonary veins and the left atrium in multi-slice computed tomography.

PURPOSE: Pulmonary veins are involved in pathogenesis and treatment of atrial fibrillation and structures at risk during thoracic surgeries. There is lack of data regarding pulmonary vein morphology and morphometry in normal population. METHODS: The study was conducted using 135 chest computed tomography studies with intra-venous iodine contrast injection. The study population contained 86 females and 49 males, mean age was 60. 13 had atrial fibrillation.The studies were analyzed using radiological workstation. RESULTS: Mean dimensions of the left atrium: transverse 52 mm, coronal 49 mm, and sagittal 35 mm. The mean volume of the left atrium was 93 cm3. The mean volume of the left atrium in patients with atrial fibrillation was 176 cm3. The sagittal dimension and the volume of the left atrium were correlated with age, r = 0.43 and r = 0.42, respectively. Surface area of the left inferior pulmonary vein ostium was 136 mm2, significantly less than the surface area of other ostia of pulmonary veins. The mean distance between two pulmonary veins was 5.42 mm on the right and 4.02 mm on the left side. 13 types of pulmonary veins outflow patterns were described on the right side and 5 types on the left side. 66.7% of right pulmonary veins and 82% of the left pulmonary veins emptied into the left atrium with two venous trunks on each side (the typical pattern). CONCLUSIONS: Morphological features of pulmonary veins and morphometry of the left atrium and pulmonary veins are important for clinical purposes and are in accordance with previous papers.

Clinical presentation and characteristics of 25 adult cases of pulmonary sequestration.

BACKGROUND: Pulmonary sequestration (PS) is a rare congenital abnormality of lung tissue. Only few series of adult cases are reported. The aim was to describe clinical characteristics in adult cases of PS and to compare outcomes in different clinical situations. METHODS: Using MSD engine we searched for cases of PS that have been diagnosed between Jan 1st, 2005 and Dec 31st, 2015. Clinical data was retrospectively gathered. Statistica v.12 (StatSoft, Inc.) was used for statistical analyses. RESULTS: We found 25 cases (18 females, 7 males), which underwent surgery and were histologically proven. There were 22 cases of intralobar PS. 7 cases were asymptomatic, 12 had infectious history (including 3 cases of lung abscess and pleural empyema), 4 presented with hemoptysis, 2 with chest pain. The average age to undergo surgery was 38.24, in the asymptomatic group 34, in symptomatic 39.89. In the latter the symptoms preceded the surgery for 2.45-year. Great majority of sequestrations was located in lower lobes (96%), 52% on the left. Symptomatic cases were at higher than expected risk of surgical complications, comparing to asymptomatic (chi2, P=0.04). In most cases there were surgical and histological signs of infection, only in 9 cases etiological factor was determined: in 5 cases it was A. fumigatus. A 0.53-day longer post-surgical hospital stay was observed in the symptomatic group, no statistical significance was found (U-test, P=0.45). CONCLUSIONS: Surgical treatment of symptomatic cases of PS is characterized by slightly longer post-surgical hospital stay and higher risk of surgical complications. Fungal infections are the most likely to occur in PS.

Fire-eater's lung.

Fire eater's lung (FEL) is an acute hydrocarbon pneumonitis caused by aspiration or inhalation into airways of liquid hydrocarbons. This disorder is classified into distinct form of chemical toxic pneumonitis. An amateur fire-eater is presented in this work. He accidentally aspirated into airways about 1/3 of glass of grill lighter fluid composed of mixture of liquid hydrocarbons. A few hours after this incident he had severe symptoms like weakness, high temperature, midsternal pleuritic chest pain, myalgia of the back, shortness of breath, and dry cough. Radiologic examination revealed consolidations with well-defined cavitary lesions (pneumatoceles) in lower lobes mainly in the left lower lobe. After one week of this event clinical improvement was observed. The lesions resolved nearly completly during three months. The review of the literature connected with fire-eater's lung is also presented.

Organizing pneumonia appearing in B-cell chronic leukemia malignancy progression - a case report.

Patients with chronic lymphocytic leukemia or non-Hodgkin's lymphoma are at risk of infectious diseases of respiratory system because of immunodeficiency. Occurrence of organizing pneumonia in leukemic patients is most commonly correlated with bone marrow transplant or treatment with antimitotic agents. There have been only four reported cases of organizing pneumonia related solitarily to leukemia or lymphoma. We present a case of 65-year old gentlemen, diagnosed 8 months earlier with B-cell chronic lymphocytic leukemia with no previous hematologic treatment, who presented symptoms of persistent pneumonia with no significant reaction to antibiotics. Chest computed tomography scans showed well-localized consolidation with ground glass opacities and some air bronchogram, suggesting infectious disease. All results of microbiological examinations were negative. Due to radiological progression of parenchymal consolidation despite two intravenous courses of antibiotics open lung biopsy was performed. The histologic examination of lung specimen revealed structures typical for organizing pneumonia pattern. There was no evidence for leukemic involvement in lung tissue, as no sign for infectious factors from histological staining was observed. In the inferior mediastinal lymph node sample progression of chronic lymphatic leukemia to mixed cell lymphoma was diagnosed. Patient was commenced on prednisone 60 mg/daily with fast improvement. We believe that this is the first case of organizing pneumonia as a reaction to the conversion of B-cell chronic lymphocytic leukemia progression to more malignant stage.

[Pneumocystis pneumonia in HIV-infected patients with cytomegalovirus co-infection. Two case reports and a literature review]. / Pneumocystozowe zapalenie pluc u zakazonych HIV przy wspólistniejacym zakazeniu cytomegalowirusem. Opis dwóch przypadków i przeglad literatury.

Pneumocystis pneumonia (PCP) is one of the infectious lung diseases diagnosed in HIV-infected patients. The pathogen responsible for the development of this opportunistic infection is an atypical fungus called Pneumocystis jiroveci. PCP remains the most common disorder diagnosed at the onset of acquired immunodeficiency syndrome (AIDS), especially in individuals not aware of their HIV infection. The most important risk factor of PCP development in HIV-infected person is the decrease of T CD4+ cell number below 200/mcL. Clinical symptoms consist of: chronic cough, dyspnoea and weakness. Arterial blood gas analysis often reveals the presence of hypoxaemia, and high-resolution computed tomography imaging shows diffuse ground glass opacities. Treatment is based on intravenous administration of trimetoprim-sulfamethoxasole. In patients with moderate and severe symptoms of PCP it is recommended that corticosteroids are used. Co-infection with cytomegalovirus (CMV) is a poor prognostic sign in PCP. CMV-related organ disease should be suspected in patients with T CD4+ cell number lower than 50/mcL. It is arguable whether CMV infection in symptomatic PCP patients should be treated. There are suggestions that corticosteroids used in PCP patients with CMV co-infection could promote the development of CMV pneumonia. In the present paper we present two patients with PCP, unaware of their HIV infection. In both cases a CMV co-infection was found.

How does Adamkiewicz artery influence blood supply to the fetal spinal cord?

Adamkiewicz artery became important in clinical practice since it was noticed that its damage during aorta aneurysm repair surgery can sometimes lead to distal spinal cord ischemia. The complexity of anatomical variations can be related to the development of spinal cord arteries. The aim was to describe topography of Adamkiewicz artery and its relations to the anterior spinal artery in fetuses. The study was carried on 4 Batson's resin corrosion casts and 24 formalin-fixed fetuses injected with dyed gelatin or latex aged 15-24 weeks gestational age. In fixed specimens vertebral canals were dissected, the anterior spinal artery was traced and Adamkiewicz artery localized. Arteries were photographed and digitally measured. Data were afterwards statistically analyzed. Anterior spinal artery was duplicated in 3/28 cases. There were from 1 to 3 Adamkiewicz arteries per specimen, mean 1.71. No relation was found between the number of Adamkiewicz artery and age. In 37/48 cases Adamkiewicz artery emptied into the anterior spinal artery on the left side. Mean degree of narrowing in anterior spinal artery (diameter of the anterior spinal artery above junction with Adamkiewicz artery divided by its diameter under that junction) was 76.74%. The diameter of Adamkiewicz artery was also correlated linearly with the degree of narrowing of anterior spinal artery (r=0.68; p<0.05). The arteries of the anterior aspect of thoracolumbar spinal cord in the 2nd trimester of pregnancy represent the adult pattern. A potentially great impact of Adamkiewicz artery on the distal spinal cord circulation may be postulated on the basis of these morphological data.