Search

New therapies are being developed for breast cancer and in this process some “old” biomarkers are re-utilized and given a new purpose. It is not always recognized that, by changing a biomarker’s intended use, a new biomarker assay is created. The Ki-67 biomarker is typically assessed by immunohistochemistry (IHC) to provide a proliferative index in breast cancer. Canadian laboratories assessed the analytical performance and diagnostic accuracy of their Ki-67 IHC laboratory developed tests (LDTs), of relevance for the LDTs’ clinical utility.

We consider a mathematical model for the group size determination by the intra-reactions, self-growth, ostracism and fission within a group, and by the inter-reactions, immigration and fusion between two groups. In some group reactions, a conflict between two groups occurs about the reaction to change the group size. We construct a mathematical model to consider such conflict, taking into account the inclusive fitnessof members in each group. In the conflict about the fusion between two groups, our analysis shows that the smaller group wants to fuse, while the larger does not. Also the criterion to resolve the conflict is discussed, and some numerical examples are given, too. It is concluded that, depending on the deviation in the total cost paid for the conflict by counterparts, the group reactions could result in a terminal group size different from that reached only by a sequence of outsider's immigrations into a group.We consider a mathematical model for the group size determination by the intra-reactions, self-growth, ostracism and fission within a group, and by the inter-reactions, immigration and fusion between two groups. In some group reactions, a conflict between two groups occurs about the reaction to change the group size. We construct a mathematical model to consider such conflict, taking into account the inclusive fitnessof members in each group. In the conflict about the fusion between two groups, our analysis shows that the smaller group wants to fuse, while the larger does not. Also the criterion to resolve the conflict is discussed, and some numerical examples are given, too. It is concluded that, depending on the deviation in the total cost paid for the conflict by counterparts, the group reactions could result in a terminal group size different from that reached only by a sequence of outsider's immigrations into a group.

Background & Aims: Although a significant amount of regenerating gene (REG) I@a protein is present not only in normal gastric mucosa but also in gastric cancer tissues, its pathophysiologic role in gastric cancer development remains unclear. We investigated REG I@a protein expression in early gastric cancers, and examined whether cytokines are responsible for REG I@a gene expression and whether REG I@a protein has a trophic and/or an antiapoptotic effect on gastric cancer cells. Methods: Early gastric cancer specimens were analyzed histologically using immunohistochemistry for REG I@a protein and proliferating cell nuclear antigen (PCNA). The effects of cytokines on REG I@a promoter activity and its messenger RNA (mRNA) expression in AGS (a kind of gastric cancer cell line) cells were examined by luciferase reporter assay and Northern blot analysis, respectively. Effects of REG I@a protein on cell growth and H"2O"2-induced apoptosis in AGS cells were examined by 3,-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphatase nick-end labeling (TUNEL) assays, respectively. Results: REG I@a-positive early gastric cancers showed a significantly higher PCNA labeling index and more severe inflammatory cell infiltration in adjacent gastric mucosa than the negative cancers. REG I@a gene expression and its promoter activity were enhanced by interferon (IFN)-@c and interleukin (IL)-6. REG I@a protein promoted cell growth and cell resistance to H"2O"2-induced apoptosis in AGS cells. These effects were abolished by concomitant treatment with anti-REG I@a antibody. REG I@a protein enhanced Akt phosphorylation and Bcl-xL expression in AGS cells. Conclusions: REG I@a gene is inducible by cytokine stimulation and its gene product may function as a mitogenic and/or an antiapoptotic factor in the development of early gastric cancer.

Background & Aims: Endoscopic variceal ligation (EVL) therapy has been performed widely to treat or prevent variceal bleeding. We sought to examine the influence of EVL for esophageal varices on collateral vessels in the vicinity of gastric cardia. Methods: In 42 patients with esophagogastric varices, conventional endoscopy and endoscopic ultrasonography with a 20-MHz probe (CUP-EUS) were performed before and at every 3 months after EVL for esophageal varices. By using conventional endoscopy, cardial variceal sizes were divided into 3 grades: F0, F1, and F2. The sizes of submucosal, perforating, and paracardial vessels at the cardia also were classified into 3 grades according to CUP-EUS findings. Results: Conventional endoscopy showed cardial varices in 33 (79%) patients before and 23 (55%) patients at 3 months after the treatment (P < 0.05). CUP EUS showed that 29 (69%) patients had severe grade cardial submucosal vessels before EVL, but only 13 (31%) patients did after the treatment (P < 0.01). Nineteen (45%) patients had severe grade cardial perforating vessels before EVL, but only 4 (10%) patients did after the treatment (P < 0.001). Furthermore, patients with severe grade residual submucosal or perforating vessels at the cardia had shorter recurrence-free times of esophageal varices (P < 0.01, 0.05, respectively). Conclusions: Collateral vessels in the vicinity of gastric cardia were improved significantly after EVL, indicating that esophageal varices can be treated by EVL even though they connect with cardial varices. Furthermore, eradication of such collateral vessels by EVL may lead to longer recurrence-free status of esophageal varices.

AbstractBackground: In the COOPERATE trial, the combination treatment of the angiotensin-II receptor blocker losartan and the angiotensin-converting-enzyme inhibitor trandolapril significantly retarded progression of non-diabetic kidney disease compared with each monotherapy. The benefit could be greatly attributable to the potent reduction of proteinuria, because the three treatment groups showed the same reductions of office blood pressure (OBP). Ambulatory blood pressure (ABP) is reported to be better than OBP in predicting progression of kidney disease. Methods: Ninety-two patients enrolled in the COOPERATE trial underwent 24-hour ABP monitoring at randomization and at month 6, year 1, year 2 and year 3 on randomized treatment. Results: Both OBP and ABP were similarly reduced among the three groups at all measurement points (p = NS) and throughout the whole study period (p = NS). No significant correlation between the change in 24-hour ABP and the change in proteinuria was seen (p = NS). A Cox-multivariable analysis showed that covariates affecting the renal outcomes (a doubling serum-Cr level and/or end-stage renal failure) were the change in proteinuria (hazard ratio 0.49, 95% CI 0.34–0.78, p = 0.01) and treatments (0.58, 0.45–0.99, 0.03), but not 24-hour ABP (0.98, 0.89–2.01, 0.17). Conclusion: The better renoprotective effect of the combination treatment is attributed to BP-independent mechanisms by more complete renin-angiotensin system blockade.Copyright © 2004 S. Karger AG, Basel

A curious autopsy case following the car crash of a 20-year-old male, in which self-strangulation and lung collapse were observed, is presented. His motor vehicle had crashed into a restaurant as a result of self-strangulation using an electrical cord wound four times around his neck. At autopsy, we found small rupture holes of spontaneous bullae in both lung apices, which had probably taken place upon collision during driving, petechial hemorrhages in the face skin and the absence of severe injuries. Since it seemed unlikely that the small holes in both lung apices caused fatal pneumothorax instantly, the cause of his death was judged to be asphyxia due to self-strangulation. It is not clear whether the self-strangulation was suicidal or autoerotic, because neither traces of suicidal intent nor circumstances suggesting either of them were disclosed.

Background: This study assessed the risk of recurrence of esophageal varices by evaluating the severity of cardia vascular structures in patients with portal hypertension by EUS with a catheter US probe before endoscopic variceal ligation. Methods: Thirty consecutive patients with esophageal varices at high risk for bleeding were studied. Simultaneous conventional endoscopy and EUS with a 20 MHz catheter US probe were performed before endoscopic variceal ligation. By catheter US probe EUS findings, vascular structures in the gastric cardia were classified into 2 grades, mild and severe, and the relationship between the catheter US probe EUS findings and the recurrence rate of esophageal varices was analyzed. Results: Catheter US probe EUS before endoscopic variceal ligation demonstrated cardial submucosal varices in all patients, whereas conventional endoscopy revealed cardial varices in only 21 patients (70.0%, NS). Patients with recurrent esophageal varices after endoscopic variceal ligation were more likely to have severe-grade perforating veins before treatment than those without recurrence (71.4% vs. 12.5%, p < 0.01). Patients with severe as opposed to mild-grade perforating veins before treatment had a significantly higher recurrence rate (90.9% vs. 21.0%, p < 0.01%). Conclusions: Catheter US probe EUS findings for cardial vascular structures before treatment are useful for predicting the likelihood of recurrence of esophageal varices. (Gastrointest Endosc 2002;55:197-203.)

Chloroform and methylene chloride were extracted from human whole blood and urine by headspace solid phase microextraction (SPME) using a Carboxen/polydimethylsiloxane (Carboxen/PDMS) fibre before capillary gas chromatography (GC)/flame ionization detection (FID). Whole blood or urine was placed in a vial and mixed with distilled water. The vial was heated at 30°C, and the solvents were extracted from the headspace by SPME. The extraction efficiencies of chloroform and methylene chloride from whole blood were 40.3% and 35.8%, respectively; those for urine were 57.0% and 43.6%, respectively. The calibration curves for chloroform were linear in the range of 1–8 μg/ml for blood and urine samples, and those for methylene chloride in the range of 0.5–8 μg/ml. The detection limits for both compounds were 0.3 μg/ml for whole blood, and 0.2 μg/ml for urine. The headspace SPME with Carboxen/PDMS fibre coupled with GC/FID seems useful for analyses of chloroform and methylene chloride in forensic toxicology and environmental chemistry.

A simple and rapid method for isolation of nine barbiturates with Sep-Pak C18cartridges from human urine, plasma and whole blood, is presented; the detection of drugs was made by wide-bore capillary gas chromatography (GC) with flame ionization. The drug-containing samples, after mixing with dilute acid solution, were directly applied to the cartridges and eluted with either chloroform/methanol (9:1) or acetonitrile. Separation of the nine drugs was satisfactory with use of an intermediately polar HP-17 capillary column. Recoveries of most compounds were excellent for both chloroform/methanol and acetonitrile as elution solvents. However, backgrounds were cleaner and evaporation time was much shorter for the chloroform/methanol system. The present isolation method with use of Sep-Pak C18cartridges and wide-bore capillary GC seem very useful in the fields of forensic chemistry, clinical toxicology and clinical pharmacology.

Summary A simple and rapid method for isolation of seven antiepileptics (2 hydantoin, 2 oxazolidin, and 3 suximide derivatives) from urine and plasma is presented. Urine and plasma (1ml) samples containing seven antiepileptics were mixed with distilled water (4ml), and the sample solution was poured into a pretreated Sep-Pak C18 cartridge; this was washed with water and chloroform/methanol was passed through it to elute the antiepileptics. The eluate was mixed with isoamyl acetate and evaporated under a stream of N2. The drugs were detected by gas chromatography with fused silica capillary columns, splitless injection and flame ionization detection. Separation of the seven antiepileptics from each other and from impurities was satisfactory with the use of an SPB-1 capillary column. The detection limit for the seven antiepileptics with the present method was 0.1–l.0μg/ml urine or plasma. The recovery of the drugs from urine and plasma was more than 70% and 50%, respectively.

A three-square-flap procedure is described for surgical treatment of minor syndactyly. The average follow-up period in 58 cases (48 patients) was 2 years 4 months. In the three-square-flap method, the interdigital space is regarded as a cube and flaps are designed on each surface of the cube (dorsal side, interdigital surface, and volar side); the flap on the dorsal side forms the basal surface of the new interdigital portion. This technique is most suitable for reconstruction of minor three-dimensional contracture syndactyly. This method does not require skin grafting; the design is uncomplicated and easy to understand, and the operative procedure is simple.

We have cloned a eDNA which encoded for rat complement regulatory protein, 5I2 antigen. The deduced amino acid sequence of 497 residues predicted essentially the same domain structure as the mouse complement regulatory protein, Crry/p65, composed of five short consensus repeat (SCR) domains, a transmembrane domain and a cytoplasmic domain. 5I2 antigen, however, had an additional SCR which showed a close similarity to the sixth SCR of mouse complement receptor 1 (CR1) or to the sixth, thirteenth, twentieth and twenty seventh SCR of human CR1. Corresponding domains of 5I2 antigen and mouse Crry/p65 showed 70%-86% amino acid identity, indicating that 5I2 antigen is the rat counterpart of mouse Crry/p65.Copyright 1994, 1999 Academic Press

Local anaesthetics have been shown to be extractable from human whole blood samples by direct immersion (DI)-solid phase micro extraction (SPME). After deproteinization with perchloric acid, the pH of the clear supernatants of human whole blood samples containing the drugs were adjusted to about 7 with 10 M NaOH in the presence of NaCl; a polydimethylsiloxanecoated SPME fiber was then immersed directly into the sample solution to allow adsorption of the drugs before capillary gas chromatography (GC) with flame ionization detection. The DI-SPME for 1-mL whole blood gave peaks for all the drugs; only a small amount of background noise appeared and this gave no problems in the detection of the drugs. Recoveries of the ten drugs from human whole blood was 0.74–19.7 %. The calibration curves for seven drugs showed linearity in the range of 0.25–12 μg mL−1 whole blood, with detection limits of 54–158 ng mL−1.

Ethanol has been found extractable from human whole blood and urine samples by headspace solid-phase micro extraction (SPME) with a Carbowax/divinylbenzene-coated fiber. After heating a vial containing the body fluid sample with ethanol, and isobutanol as internal standard (IS) at 70°C in the presence of (NH4)2SO4, a Carbowax/divinylbenzene-coated SPME fiber was exposed in the headspace of the vial to allow adsorption of the compounds. The fiber needle was then injected into a middle-bore capillary gas chromatography (GC) port. The headspace SPME-GC gave intense peaks for both compounds; a small amount of background noises appeared, but did not interfere with the detection of the compounds. Recoveries of ethanol and IS were 0.049 and 0.026% for whole blood, respectively, and 0.054 and 0.085% for urine, respectively. The calibration curves for ethanol showed excellent linearity in the range of 80–5000 mg L−1 for whole blood and 40–5000 mg L−1 for urine; the detection limits for both samples were 20 and 10 mg L−1, respectively. The data on actual determination of ethanol after the drinking of beer are also presented for two subjects.

A rapid method for isolation of various benzodiazepines from human samples with Sep-Pak® C18cartridges before wide-bore capillary gas chromatography is described. The drugs dissolved in alkaline samples were directly applied to the cartridges and eluted with hexane/isopropanol (9:1). The recoveries were excellent for all drugs in urine samples, but were somewhat lower for some drugs in plasma samples. The latter problem could be easily circumvented by using a deproteinization process before their application to the cartridge. We can recommend the Sep-Pak C18cartridges for isolation of benzodiazepines because of their simplicity and rapidity.

A simple and rapid method for isolation of five butyrophenones with Sep-Pak C18cartridges from human samples, and their wide-bore capillary gas chromatography (GC), are presented. The GC was made by both flame ionization and electron capture detections. The drugs contained in alkaline samples were directly applied to the cartridges and eluted with chloroform/isopropanol (9:1). The recoveries with use of the cartridges were excellent for most drugs in both urine and plasma samples. We can recommend the Sep-Pak C18cartridges for isolation of butyrophenones because of simplicity and rapidity, and also wide-bore capillary GC because of high sensitivity and low decomposition of drugs during passage through the column. Es wird eine einfache und rasche Methode zur Isolierung von fünf Butyrophenonderivaten mittels Sep-Pak C18Säulen aus Humanproben sowie deren gaschromatographische Bestimmung mit wide-bore Kapillaren beschrieben. Bei der Gaschromatographie kamen die FID-und ECD-Detektion zum Einsatz. Die im alkalisierten Untersuchungsmaterial enthaltenen Substanzen wurden direkt auf die Säulen gegeben und mit Chloroform/iso-Propanol (9:1) eluiert. Die mit den Säulen erzielten Wiederfindungsraten waren für die meisten Arzneistoffe, sowohl beim Einsatz von Urin als auch bei Plasmaproben, hervorragend. Die Sep-Pak C18Säulen können wegen ihrer einfachen und schnellen Anwendung für die Isolierung von Butyrophenonderivaten empfohlen werden, ebenso wie die wide-bore Kapillar-Gaschromatographie mit ihrer hohen Empfindlichkeit und geringen Gefahr der Zersetzung der Proben während des Trennvorganges.

The levels of pyrroloquinoline quinone (PQQ) in eggs, skim milk and milk was re-examined by the use of gas chromatography/mass spectrometry. PQQ was extracted from the samples, after addition of [U-13C]PQQ as internal standard, with n-butanol and Sep-Pak C₁₈ cartridges. After derivatization of PQQ with phenyltrimethylanunonium hydroxide, molecular peaks at m/z 448 and 462 were used for detection of PQQ and [U-13C]PQQ, respectively, by selected ion monitoring. PQQ could be detected in eggs, skim milk and milk, but their levels were very low (e.g., egg yolk: 7.0 ng/ml; skim milk: 2.5 ng/g dry weight). Our data for eggs and skim milk are f at below those measured by the redox cycling method of Gallop's group.Copyright 1993, 1999 Academic Press

We report the case of a 62-year-old man who was administered sodium valproate (VPA) and who subsequently developed the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). He had been taking VPA for treatment of idiopathic generalized tonic-clonic convulsions since he was 56 years old. After substituting VPA with zonisamide, the serum sodium level returned to normal. We consider this episode of SIADH to be the result of a combination of factors including a weakness of the central nervous system and the long-term administration of VPA.

Phencyclidine (PCP) was found to be extractable by headspace solid-phase microextraction (SPME) from human whole blood and urine. Sample solutions were heated at 90°C in the presence of NaOH and K2CO3, and an SPME fiber was exposed in the headspace of a vial for 30 min. Immediately after withdrawal of the fiber, it was analyzed by gas chromatography with surface ionization detection (GC-SID). Recoveries of PCP were approximately 9.3–10.8% and 39.8–47.8% for whole blood and urine samples, respectively. The calibration curve for PCP showed good linearity in the range 2.5–100 ng mL−1 whole blood and 0.5–100 ng mL−1 urine. The detection limits were approximately 1.0 ng mL−1 for whole blood and 0.25 ng mL−1 for urine.

Strychnine was found measurable with high sensitivity by gas chromatography (GC) with surface ionization detection (SID) and using brucine as the internal standard. The calibration curve was linear in the range of 62.5 to 1000 pg, on column. The detection limit of strychnine was about 50 pg on column (2.5 ng/mL sample); the sensitivity was 70 times higher than that reported by high-performance liquid chromatography. A detailed procedure for extraction of strychnine from human whole blood and urine was established using Sep-Pak C18cartridges; the recovery of strychnine and brucine, which had been spiked in both body fluids, was about 90%.

The levels of free pyrroloquinoline quinone (PQQ) in various foods were examined by the use of gas chromatography-mass spectrometry. PQQ was extracted from the samples, after addition of [U-13C]PQQ as internal standard, with n-butanol and Sep-Pak C18 cartridges. After derivatization of PQQ with phenyltrimethylammonium hydroxide, molecular peaks at m/z 448 and 462 were used for detection of PQQ and [U-13C]PQQ respectively, by selected ion monitoring. Free PQQ could be detected in every sample in the range 3.7-61 ng/g or ng/ml. Since its levels in human tissues and body fluids are 5-10 times lower than those found in foods, it is probable that PQQ existing in human tissues is derived, at least partly, from the diet.