Detection of ubiquitous and heterogeneous mutations in cell-free DNA from patients with early-stage non-small-cell lung cancer.

BACKGROUND: The aim of this pilot study was to assess whether both ubiquitous and heterogeneous somatic mutations could be detected in cell-free DNA (cfDNA) from patients with early-stage non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Three stage I and one stage II primary NSCLC tumors were subjected to multiregion whole-exome sequencing (WES) and validated with AmpliSeq. A subset of ubiquitous and heterogeneous single-nucleotide variants (SNVs) were chosen. Multiplexed PCR using custom-designed primers, coupled with next-generation sequencing (mPCR-NGS), was used to detect these SNVs in both tumor DNA and cfDNA isolated from plasma obtained before surgical resection of the tumors. The limit of detection for each assay was determined using cfDNA from 48 presumed-normal healthy volunteers. RESULTS: Tumor DNA and plasma-derived cfDNA was successfully amplified and sequenced for 37/50 (74%) SNVs using the mPCR-NGS method. Twenty-five (68%) were ubiquitous and 12 (32%) were heterogeneous SNVs. Variant detection by mPCR-NGS and WES-AmpliSeq in tumor tissue was well correlated (R(2) = 0.8722, P < 0.0001). Sixteen (43%) out of 37 SNVs were detected in cfDNA. Twelve of these were ubiquitous SNVs with a variant allele frequency (VAF) range of 0.15-23.25%, and four of these were heterogeneous SNVs with a VAF range of 0.28-1.71%. There was a statistically significant linear relationship between the VAFs for tumor and cfDNA (R(2) = 0.5144; P = 0.0018). For all four patients, at least two variants were detected in plasma. The estimated number of copies of variant DNA present in each sample ranged from 5 to 524. The average number of variant copies required for detection (VCRD) was 3.16 (range: 0.2-7.6 copies). CONCLUSIONS: The mPCR-NGS method revealed intratumor heterogeneity in early-stage NSCLC tumors, and was able to detect both ubiquitous and heterogeneous SNVs in cfDNA. Further validation of mPCR-NGS in cfDNA is required to define its potential use in clinical practice.

Ultrasonography in the diagnosis of gallbladder carcinoma.

Sonographic examination of the gallbladder (GB) was performed in 21.646 patients. A histopathologically documented gallbladder carcinoma (GBC) was present in 59 patients (0.27%). Five different sonographic patterns were revealed with differences in number of associated findings, echo pattern and correct preoperative diagnosis. The sonographic diagnosis was correct in 59% and retrospective diagnosis in 65%. Most of the sonographically detectable GBC were inoperable (90%). 15 fine-needle biopsies were positive in 100% and reduced the number of explorative laparotomies. The survival rate was 3 days to 4 years after the first sonographic examination.

Movement of the epiglottis during deglutition. A cineradiographic study.

The movements of epiglottis during swallowing of barium were studied by high-speed cineradiography in 150 volunteers who had no dysphagia. In 137 individuals the epiglottis tilted down in a two-step fashion during deglutition. The first movement from an upright to transverse position was accomplished by elevation of the larynx and approximation of the hyoid bone and thyroid cartilage. This first movement is evidently a passive one and induced by the muscles that lift the hyoid bone. The second movement of the epiglottis, from transverse to an inverted position, occurs later in swallowing and seems related to contraction of the thyroepiglottic muscle. The second epiglottic movement was absent in 7 individuals, and 6 others demonstrated obliquity of their epiglottis (30-90 degrees) when studied in AP projection.

Ultrasonographic examinations of muscle hematomas in hemophiliacs.

Twenty-one ultrasonographic examinations were performed in 14 hemophiliacs with suspected intramuscular bleeding. In all patients who had an ultrasonographic examination within the first days after the onset of the symptoms, a hematoma was identified, and its resolution during transfusion therapy was possible to follow. In patients who had their first examination 3 to 5 weeks after the onset of symptoms and who had received transfusion therapy during this period, no hematoma was detectable. It is concluded that ultrasonography provides all necessary information needed for diagnosis and follow-up of treatment. Being a safe method, and giving no radiation, it should be the method of choice in the evaluation and follow-up of muscle hematomas in this patient group.

Recognition of gas in gallstones in routine computed tomograms of the abdomen.

In a consecutive group of 1,067 noncholecystectomized patients referred for computed tomography including the upper abdomen, gallstones were seen in 13%. In 4% of the patients with gallstones, gas-containing calculi were found. On reviewing the conventional plain films such stones were revealed in less than one-half of the patients.

[Arteriovenous fistula after partial gastrectomy. A case report.].

Portal arteriovenous fistulas are uncommon and are most often seen following trauma to the abdomen. In a few cases they have occurred as complications of abdominal surgery. In this report we present a 74 year old man with a fistula between the left gastric artery and vein, secondary to a Billroth I partial gastrectomy that was performed 32 years earlier. The patient complained of a diffuse abdominal pain for two weeks and mild cardiac failure symptoms. An epigastric bruit was heard and the abdomen was distended with transudatic ascitic fluid. The fistula was diagnosed by Doppler-ultrasound and the diagnosis confirmed by angiography. At laparotomy the fistula was excised and the patient's symptoms subsequently subsided.

Reliability of ultrasound-guided fine-needle biopsy of pancreatic masses.

Fine-needle aspiration biopsy (FNB) was performed with ultrasound guidance in 79 patients in whom sonography had revealed a mass suggesting pancreatic malignancy. The final diagnosis (surgery, autopsy and clinical course) in 69 of these 79 patients was a malignancy closely related to the pancreas while in the remaining 10 patients benign disease was confirmed. A correct diagnosis of malignancy was attained by FNB in 59 of the 69 patients with a malignant tumour while in 10 it failed to confirm the diagnosis. FNB yielded a true negative result in 10 patients with benign disease. The accuracy of sonographically guided FNB in the present investigation was 87 per cent. Ultrasound-guided fine-needle biopsy is considered the method of choice for further evaluation of pancreatic masses.

Acute pancreatitis: a prospective study of its incidence, aetiology, severity, and mortality in Iceland.

OBJECTIVE: To evaluate the incidence, aetiology, severity and mortality of patients with acute pancreatitis. DESIGN: Prospective study. SETTING: University hospital, Iceland. PATIENTS AND METHODS: All 50 patients diagnosed with acute pancreatitis during the one-year period October 1998-September 1999 inclusive. MAIN OUTCOME MEASURES: APACHE II, and Ranson and Imrie scores, and C-reactive protein (CRP) concentrations. The Balthazar-Ranson criteria were used for scoring of computed tomograms (CT). RESULTS: 27 of the 50 patients were male. The median age of the whole series was 60 years (range 19-85). The estimated incidence was 32/100000 for the first attack of acute pancreatitis. The causes were; gallstones 21 (42%), alcohol 16 (32%), miscellaneous 12 (24%), and idiopathic 1 (2%). 15 (33%) of the patients had APACHE II scores > or = 9, 17 (38%) had Ranson scores of > or = 3, 23 (50%) had Imrie scores of > or = 3, and 16 (34%) had CRP concentrations over 210 mg/L during the first 4 days or > 120 mg/L during the first week. Seven patients had severe pancreatitis. 2 patients in the whole group died, and both had clinically severe pancreatitis. CONCLUSIONS: This study indicates that the incidence of less severe acute pancreatitis is rising. Prospective assessment makes it possible to evaluate the aetiological factors more accurately. Measurement of the CRP concentration is an attractive and simple alternative to the severity scoring systems currently in use.

Aneurysms of the popliteal artery.

A retrospective study of 13 patients with 18 popliteal artery aneurysms has been undertaken. The patients were seen under a period of 12 years. Six of the patients had extrapopliteal aneurysms as well. Seven of the popliteal aneurysms presented as vascular emergencies, due to occlusion of the popliteal artery. In this group the amputation rate was 57%. In the elective group no amputation was necessary nor did any patient suffer from claudication during follow-up. Ultrasonography is a good tool for demonstrating the nature of the disease. Usually a short exclusion bypass is enough to reconstruct the area. A popliteal aneurysm, if not occluded and asymptomatic, is an indication to operate because of the threat to the limb in case of embolism or thrombosis.

Intravenous computed tomographic cholangiography in acute cholecystitis. A comparison with conventional cholangiography.

Forty-nine patients with clinical signs of acute cholecystitis underwent conventional and computed tomographic cholangiography. Among 39 patients with signs of contrast medium in the biliary system at both examinations there was a diagnostic discrepancy in only one patient. Conventional radiography demonstrated cholecystopathy in this patient while contrast medium in the gallbladder and an acute pancreatitis were found at computed tomography. Ten patients with an indeterminate conventional cholangiography had a conclusive computed tomographic examination. Twenty of 30 patients with an abnormal computed tomographic cholangiography underwent cholecystectomy and all had diseased gallbladders. All 17 patients with histopathologically confirmed acute cholecystitis had signs of subserosal edema and/or changes in the omental fatty tissue adjacent to the gallbladder at computed tomography. A layer of tissue of water-density adjacent to the gallbladder and/or changes in omental fatty tissue were also seen in one patient with congestive heart failure and in one with a penetrating duodenal ulcer. None of the 19 patients with a normal computed tomographic cholangiography had a proven acute cholecystitis.