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The consumption of a pufferfish, Takifugu flavipterus or komonfugu in Japanese, formerly known as Takifugu poecilonotus, is popular in Japan. However, T. flavipterus is frequently involved in cases of tetrodotoxin (TTX) poisoning in Japan. Although victims have usually consumed inedible parts, some cases are related to consumption of flesh. To improve the risk management of pufferfish poisoning, we studied TTX level in the flesh and skin of T. flavipterus. Ninety-seven specimens obtained from the Seto Inland Sea and landed in Fukuoka Prefecture were analyzed by liquid chromatography-tandem mass spectrometry. The flesh from six specimens was toxic (>10 MU/g = 2.2 mg/kg): one was in poor condition (not freeze–thawed); three were freeze–thawed before sample preparation; and two freshly prepared and in good condition (not freeze–thawed). The fillets were divided into outer and inner portions; the TTX levels in the outer portions were notably higher. The skin of the six specimens was moderately to extremely toxic: 165 MU/g (36.3 mg/kg) in the fresh specimen not in good condition, 600–950 MU/g (132–200 mg/kg) in freeze–thawed specimens, and 4500 and 6000 MU/g (990 and 1320 mg/kg) in the two fresh specimens. We concluded that TTX in the flesh migrated from the highly toxic skin. In addition, TTX levels in the skin appeared to be regionally specific. We recommend that toxic portions of T. flavipterus are removed as soon as possible after individuals are caught, and that fish from known highly toxic areas are not consumed.

Specimens of Ligidium sp. (Crustacea, Isopoda, Oniscidea) collected from Kanagawa Prefecture in Japan were found to be infected with parasitic nematodes. We have obtained two Mermithidae (Nematoda) by dissecting the fresh specimen and from the dead specimen of Ligidium, which marks the second known discovery of a mermithid infection in the order Isopoda. Herein, a report on the nuclear 28S rDNA and 18S rDNA sequence of the isolated Mermithidae nematodes and on the morphology of the juveniles is provided.

The rare squaretail Tetragonurus pacificus Abe 1953 was described on the basis of a single poor-condition specimen obtained from the stomach of a yellowfin tuna collected from west of the Solomon Islands. A fresh specimen of T. pacificus was recently collected from southwestern Taiwan and its morphology and the fresh coloration are herein described in detail. In addition, a juvenile specimen collected from the southern Pacific Ocean was found in the museum collection. The species is characterized by dorsal-fin spines 10–11; longitudinal scale rows along body axis 72–80; and predorsal scales 17–20. The present specimen collected from off southwestern Taiwan represents the northernmost record of the species from the Pacific Ocean.

Amphicnemis rigiketit sp. nov. is described from southwestern Sarawak (holotype ♂ in forest around stream, near road from Kota Samarahan to Siburan, Samarahan Division, Sarawak, Malaysian Borneo, 21 ii 2018, deposited in the Natural History Museum, London). The new species is allied to A. remiger Laidlaw, 1912. The female of A. remiger is described for the first time and a description of the male from a fresh specimen is provided along with notes on variation.

Purpose A critical indicator of the overall survival of patients with high-grade glioma is the successful isolation of tumor mesenchymal stem-like cells (tMSLCs), which play important roles in glioma progression. However, attempts to isolate tMSLCs from surgical specimens have not always been successful, and the reasons for this remain unclear. Considering that the amount of surgical high-grade glioma specimens varies, we hypothesized that larger surgical specimens would be better for tMSLC isolation. Materials and methods We assessed 51 fresh, high-grade glioma specimens and divided them into two groups according to the success or failure of tMSLC isolation. The success of tMSLC isolation was confirmed by plastic adherence, presenting antigens, tri-lineage differentiation, and non-tumorigenicity. Differences in characteristics between the two groups were tested using independent two sample t-tests, chi-square tests, or Kaplan-Meier survival analysis. Results The mean specimen weights of the groups differed from each other (tMSLC-negative group: 469.9±341.9 mg, tMSLC positive group: 546.7±618.9 mg), but the difference was not statistically significant. The optimal cut-off value of specimen weight was 180 mg, and the area under the curve value was 0.599. Conclusion Our results suggested a minimum criterion for specimen collection, and found that the specimen amount was not deeply related to tMSLC detection. Collectively, our findings imply that the ability to isolate tMSLCs is determined by factors other than the specimen amount.

Objective To perform an MR imaging-anatomical correlation to elucidate the anatomy of muscles, tendons, and ligaments about the hallux as seen on MR imaging. Materials and methods Four fresh and 4 embalmed cadaver specimens were used for this study. The embalmed specimens and one fresh specimen were dissected by three investigators. The fresh specimens were preserved deep frozen. They were thawed and imaged with a 3 T MR system. Proton density weighted images were obtained. The specimens were refrozen and sectioned with a band saw into 3 mm thick slices. Results Musculotendinous structures were equally well seen in the fresh and embalmed specimens. The capsular ligaments could best be studied in the fresh specimens. Proximal to the sesamoids the following muscle and tendon anatomy was delineated: the abductor hallucis tendon inserted on the medial sesamoid bone together with the medial belly of the flexor digitorum brevis; the adductor hallucis (transverse and oblique heads) inserted on the lateral sesamoid together with the lateral head of the flexor hallucis brevis. At the level of the sesamoid bones and distally, the ligaments making up the plantar plate were delineated, including the presesamoid band, the flexor hallucis longus pulley, the sesamometatarsal ligaments, the sesamophalangeal ligaments, and the capsular ligaments. Conclusion MRI allows for accurate assessment of the complex anatomy of the hallux. At the level proximal to the sesamoids the muscle and tendon insertions are well demonstrated. At the level of the sesamoids and distally, the ligaments making up ‘the plantar plate’ are well demonstrated.

A rare sandperch, Parapercis fuscolineata Fourmanoir, 1985 , is reported for the first time from Japan, based on a single specimen (36.8 mm SL) collected from Tosa Bay, Kochi Prefecture, Shikoku Island, at a depth of 200–203 m. This species was previously known only from the Philippines (type locality) and the tropical southwest Pacific region (Solomon Islands, Vanuatu, and New Caledonia); thus the Japanese specimen represents the northernmost record of the species. The unique coloration of P. fuscolineata, including a dark continuous stripe extending from the hind rim of the orbit to the caudal peduncle, a large black ocellus on the upper caudal-fin base, and about 12 diagonal yellowish bars on the lower half of the body, readily distinguishes this species from its congeners. The Japanese specimen is fully described, with color photographs of the fresh specimen. A new Japanese name, “Kuroobi-toragisu”, is proposed for P. fuscolineata.

The purpose is to demonstrate the optical charactering concerning nasopharyngeal tissue of pig by fresh sections and frozen correlating sections with optical coherence tomography (OCT). After being imaged on a fresh specimen, samples are then stored in low temperature refrigerators (−80°C) for one year for the second OCT measurement. The OCT structure of the epithelium, lamina propria, and the basement membrane are still resolvable; the median scattering coefficients and anisotropy factors fitting from OCT images based on the multiple scattering effects for epithelium are 27.6 mm−1 [interquartile range (IQR) 23.6 to 29.3 mm−1] versus 22.5 mm−1 (IQR 20.5 to 24.4 mm−1), 0.86 (IQR 0.81 to 0.9) versus 0.88 (IQR 0.87 to 0.9) for fresh and frozen tissue, respectively; and 10.2 mm−1 (IQR 8.1 to 13.6 mm−1) versus 9.6 mm−1 (IQR 8.1 to 13.8 mm−1), 0.96 (IQR 0.93 to 0.98) versus 0.92 (IQR 0.9 to 0.98) for lamina propria, respectively. The results show that the frozen storage method can be used for OCT research.

Background: Urine excreted by the body has a variable composition in different physiological and pathological conditions. The cells that come from the renal pelvis, ureters, bladder, and urethra are carried by the urine, and therefore, they can be observed in fresh samples and in smears with Giemsa and Papanicolaou stain. Aim: The aim of this study was to show that high correlation that exists between the cytological examination of fresh urine samples and smears stained with Papanicolaou and Giemsa methods. Materials and Methods: A total of 45 cases with no tumor of the urinary tract and 36 patients with lower urinary tract neoplasms were included in the study (20: Low-grade urothelial tumors; 16: High-grade urothelial tumors, squamous carcinomas, and adenocarcinomas). The sediments in the urine samples were observed in fresh specimen and in smears stained with Papanicolaou method. Results: The meticulous observation of fresh urinary sediments allowed identification of diverse cellular types associated with varied pathologies. Conclusions: The cytological examination of urinary samples in fresh smears, and its later diagnostic confirmation with the Papanicolaou stain is important not only as a diagnostic procedure of tumoral or non-tumoral pathologies, but also as a method for the 'screening' of pre-cancerous lesions or carcinoma in situ, especially in high-risk populations.

Owing to the peculiar morphology of the goose barnacle Pollicipes pollicipes (Gmelin, 1789), many different biometric variables have been used in its study. The lack of a standard morphological variable makes it difficult to compare results from different authors. This aspect takes on even greater importance, since the goose barnacle is a commercially exploited species and must be regulated with clear-cut technical measures related to its biology. The need to measure a large quantity of specimens in the management and control of the fishery calls for a biometric variable that can be processed easily and requires little training. The base of the rostrum and the apex of the subcarina form two conspicuous clefts in the capitulum of the goose barnacle, allowing a calliper to be fitted quickly and easily. The results of this study showed that the diameter of the base of the capitulum (DBC), defined as the distance between these two clefts, may be considered a consistent variable both when the same specimen is measured by two different technicians and when the measurements taken from a fresh specimen are compared with the measurements taken after the specimen has been frozen and then thawed. In order to make comparisons easier, two linear equations have been found to relate DBC and two of the more usual biometric variables: rostral-carinal distance (RC) and capitulum height (CH). The mean length at sexual maturity was also calculated in terms of DBC (13.35 mm) for specimens collected on the Galician coast (NW Spain).

The most important margin in determining the prognosis of rectal cancer is circumferential resection margin (CRM). But, the type of surgery is determined by distal rectal margin (DRM), whether sphincter saving procedure is possible or patient needs an abdominoperineal resection. There are no standardized uniform guidelines for measurement of DRM. The purpose of this study is to assess the distal microscopic spread beyond gross margin after neoadjuvant concurrent chemoradiation (CCRT) in rectal cancers, the factors influencing the distal microscopic spread, the shrinkage of the distal margin in pinned and unpinned fresh and fixed specimen, and to find out the best method of measurement of distal rectal margin. A prospective analytical study was conducted from May 2013 through February 2015 in 47 cases of carcinoma rectum (both AR and APR) who had received neoadjuvant CCRT. Fresh specimen was collected within 30 min of specimen retrieval and a longitudinal cut was made in the distal margin of all specimens. One side of the specimen was pinned onto a cork board and the other side was left unpinned. Measurements were made from the distal end of clinical gross tumor. DRM was determined in both pinned and unpinned sides in fresh and fixed specimen. Of the 47 patients, 2 patients (4.2%) had small focus of tumor beyond gross margins, 1 at 6 mm and another at 3.5 mm on the unpinned side. The average margin for fresh and fixed pinned specimens was 3.67 and 3.47 cm, respectively, with percentage shrinkage of 5.4% for the pinned specimens. The average margin for fresh and fixed unpinned specimens was 3.32 and 2.84 cm, respectively, with percentage shrinkage of 14.4% for the unpinned specimens. Six patients (12.7%) had complete pathological response. Correlation of distal margin was better in pinned specimen. A correction factor of 15% for shrinkage needs to be taken into account while assessing unpinned specimen. Only in 4.2% of patients, there was distal submucosal spread beyond gross margin. Long-term follow up is required for assessing adequacy of DRM post neoadjuvant CCRT.

The compaction characteristics of a lateritic soil modified with a low-calcium fly ash and lime was investigated in the laboratory. The maximum amount of soil replaced with fly ash was limited to 50%. Lime was added in quantities varying from 2% to 4% of the dry weight of soil-fly ash mix. Moisture-dry unit weight relationships were determined for the mixes by using a fresh specimen for each point even after a delay period extending up to 3 days. Fresh specimens were also subjected further to three re-compaction cycles without any delay. The test results indicate that the addition of both fly ash and lime cause immediate reduction in the maximum dry unit weight. Compaction delay leads to a further decrease only for mixes containing lime. However, there is an increase in maximum dry unit weight after re-compaction of the soil-fly ash mixes, and the effect is more pronounced for mixes with lime added.

Pancreatic cancer primary tumor size measurements are often discordant between computed tomography (CT) and pathologic specimen after resection. Dimensions of the primary tumor are increasingly relevant in an era of highly conformal radiotherapy.We retrospectively evaluated 97 consecutive patients with resected pancreatic cancer at two Boston hospitals. All patients had CT scans before surgical resection. Primary endpoints were maximum dimension (in millimeters) of the primary tumor in any direction as reported by the radiologist on CT and by the pathologist for the resected gross fresh specimen. Endoscopic ultrasound (EUS) findings were analyzed if available.Of the patients, 87 (90%) had preoperative CT scans available for review and 46 (47%) had EUS. Among proximal tumors (n = 69), 40 (58%) had pathologic duodenal invasion, which was seen on CT in only 3 cases. The pathologic tumor size was a median of 7 mm larger compared with CT size for the same patient (range, -15 to 43 mm; p0.0001), with 73 patients (84%) having a primary tumor larger on pathology than CT. Endoscopic ultrasound was somewhat more accurate, with pathologic tumor size being a median of only 5 mm larger compared with EUS size (range, -15 to 35 mm; p = 0.0003).Computed tomography scans significantly under-represent pancreatic cancer tumor size compared with pathologic specimens in resectable cases. We propose a clinical target volume expansion formula for the primary tumor based on our data. The high rate of pathologic duodenal invasion suggests a risk of duodenal under-coverage with highly conformal radiotherapy.

Aims: To develop a method for preparing diagnostic-quality, whole-mount serial sections of breast specimens while preserving 3-D conformation. This required supporting the fresh specimen prior to breadloafing and refining the conventional tissue processing method. The overall goal is to use digital images of whole-specimen histopathology to improve the estimation of extent of disease. Methods and results: To maintain a 3-D conformation, the specimen is suspended in 3.5% agar at 55°C. The block is sliced at 5-mm intervals. Sectioning is performed after extended fixation in 4% formaldehyde from paraformaldehyde in 0.1 m Millonig's buffer, followed by paraffin processing using a non-routine schedule and extended paraffin infiltration. Whole-mount serial breast sections are produced with features of equal or superior quality to that which can be achieved using conventional methods. The method is compatible with some immunohistochemical stains but requires further optimization for others. Conclusions: The technique is currently suitable for research applications. With the reduction in processing time achievable with microwave-assisted processing, there is the potential for its use as a routine clinical method. This tool may improve the accuracy of margin estimates and identification of multifocality in breast cancer; further evaluation is necessary.

There is no reliable method to assess tumour thickness preoperatively or intraoperatively in cases of oral tongue squamous cell carcinoma (SCC). The purpose of this study was to evaluate the efficacy of a macroscopic technique to measure tumour thickness. This was a prospective study of 51 consecutive patients with T1/T2 primary SCC of the oral tongue. All patients underwent primary resection with ipsilateral neck dissection. Thickness measurements were obtained using Vernier calipers on the fresh specimen. The technique was correlated with the microscopic evaluation statistically using (1) Pearson's correlation coefficient, (2) intra-class correlation, and (3) Bland-Altman plot with 95% confidence intervals. On comparing the macroscopic technique to the microscopic evaluation, Pearson's correlation (r) was 0.915 (P

Background Most studies have directly established the optimal perioperative in situ clearance margin in surgery for rectal cancer from the histologically observed extent of distal spread, neglecting the tissue variability that occurs after resection and fixation of the rectal specimen. Purpose To measure the length of the distal resection margin in the fresh and fixed specimen following partial mesorectal excision for rectal cancer using magnetic resonance imaging (MRI) to document tissue shrinkage after surgical removal and fixation. Material and Methods The length of the distal resection margin was measured by MRI of the fresh and fixed specimen and at histopathological examination of the fixed specimen in 10 patients who underwent surgery for upper rectal cancer. In addition, tissue shrinkage was estimated by measuring the total length of the fresh and fixed specimen and distance from the peritoneal reflection anteriorly to the distal cut edge of the specimen. Results Measured by MRI, the distal resection margin was in the range of 0.6–10.2 cm (mean, 4.6 cm) in the fresh specimen, and 0.5–6.2 cm (mean, 3.2 cm) in the fixed specimen. The tissue shrinkage ratio was a mean of 69% (interquartile range, 61–77%). Taking all ratios from MRI and histopathological examination of tissue shrinkage into account, the collective tissue shrinkage ratio was 70% (95% confidence interval, 67–73%) Conclusion The length of the distal resection margin was reduced by 30% after surgical removal and fixation of the specimen.

A patient with nonspecific complaints had four previous venous blood samples showing elevated methemoglobin fractions of 15.6-20.1%. Cooximetry on a fresh specimen revealed a methemoglobin fraction of 0.8%, while that reported by the original laboratory on the simultaneously collected specimen was 14.9%. The laboratory assayed the specimen after holding it in frozen storage. Venous blood from a healthy volunteer was assayed by cooximetry after storage under conditions of room temperature (22-24 degrees C), refrigeration (1-4 degrees C), and freezing (-14 to -12 degrees C). Methemoglobin level in frozen-thawed specimens rose over time from 1.8% (0.29 g/dL) at 6 hour to 10.9% (1.71 g/dL) after 6 days. With the exception of a single specimen stored in an EDTA-containing tube at room temperature for 6 days, methemoglobin in nonfrozen specimens never exceeded 0.8% (0.12 g/dL).

Many taxonomic distinctions are made or refined on the basis of herbarium material that is either dried or preserved in spirit medium. Hitherto, examination of internal structure has only been possible by the destructive sectioning of the preserved material. In this paper, the use of nuclear magnetic resonance (NMR) imaging for the non-destructive, non-invasive, complete three-dimensional structural examination of herbarium material is demonstrated for the first time. The experimental materials were the fruiting structures of two species of Southern Hemisphere Podocarpaceae: Acmopyle pancheri and Podocarpus nivalis. Material dried in accordance with standard herbarium techniques was used, as well as material preserved in spirit and freshly gathered fruits. The dried material was subsequently rehydrated using standard techniques, and protocols established for the specimens. Appropriate selection of NMR imaging parameters allowed a variety of anatomical features to be highlighted on a single specimen. Fresh specimens from living material gave the best NMR signals. Dry specimens gave no signal except from the lipid in the seed, but when rehydrated the images yielded almost as much information about internal structure as did a fresh specimen of the same taxon. Thus, NMR imaging has great potential value as a non-invasive method for obtaining details of the internal structure of fruits and seeds and is particularly useful when, as in the case of Acmopyle, the sclerotesta of the seed is too lignified for sectioning by conventional methods.

Besides their main function of assisting in breathing, the intercostal muscles also play an important role in maintaining the balance of forces acting on the thoracic cage including the thoracic spine. Since it is virtually impossible to conduct a morphometric evaluation of these muscles, a study was undertaken to standardize an ultrasound method for accurate determination of the area of the intercostal muscles and hence, indirectly, their function. In a pilot study, the area of the intercostal muscles was determined on the torso of a fresh specimen of a grown-up lamb, using high-resolution ultrasound and CT, and by direct measurements of the intercostal space at two points equidistant from the midline on the left and the right sides of the back of the specimen. The size of the intercostal muscles was determined either by tracing or from the perpendiculars of the area of the muscles both on sonographs and on CT scans. The results showed that measurements derived from the perpendiculars of the muscle area on the sonographs give better estimates than those derived from CT scans, and were in good accordance with the direct measurements of the corresponding intercostal space on the specimen. To evaluate the applicability of the method in vivo the area of the intercostal muscles at maximal inhalation and exhalation was determined in one adult person. It was found that measurements at maximal inhalation were more accurate than those taken at maximal exhalation. It is concluded that ultrasonography is a reliable, safe, easy to apply and high-resolution method for measurements of the area and, indirectly, of the activity of the intercostal muscles in humans, and that the measurements are more accurate at maximal inhalation.

Anatomy of the thyroid has been extensively studied but the presence of pyramidal lobe varies in percentages from 15% to 75% of cases according to different authors. We therefore investigated systematically this peculiar anatomical aspect. From January 2001 to December 2011, 1002 patients underwent total thyroidectomy in our Division of General Surgery. We analyzed the data collected on the latest 200 thyroidectomies: for all patients pyramidal lobe was measured at removal of the specimen then dimension of the pyramidal lobe after fixation was checked. We found the pyramidal lobe in all cases. In most cases, it was approximately 2 cm (range 1 - 8 cm on fresh specimen). At histology it was described in 72% of cases, after fixation size decreased by a third approximatively. This is the first systematic intraoperative study to verify the prevalence of the thyroid pyramidal lobe. In our experience, the pyramidal lobe is always present and the thyroglossal duct is identifiable.

Polymerase chain reaction methods are becoming the standard for the diagnosis of herpes simplex virus (HSV) encephalitis. Little is known, however, about the stability of HSV DNA in cerebrospinal fluid (CSF) specimens. Our results demonstrate that HSV DNA is extremely stable in CSF specimens containing lymphocytes and monocytes. We observed little DNA degradation in specimens stored for 30 days at room temperature (20-23 degrees C), refrigerator temperature (2-8 degrees C), or freezer temperatures (-18 to -22 degrees C and -69 to -72 degrees C). Specimen storage conditions are therefore not so critical for HSV encephalitis detection as for other viral illnesses. It is therefore not necessary to perform a second lumbar puncture to obtain a fresh specimen for the detection of HSV DNA.

The aim of this paper is to identify advantages that could be obtained by orientation of the biopsy specimen using the marking technique. We reviewed our experience (4,500 cases) and the published literature. The peripheral (proximal) end of the fresh specimen is marked with ink soon after needle delivering in a few minutes. It is performed easily in association with pre-embedding method. Five potential clinical advantages were identified: (1) tumor localization, (2) atypical lesions localization and planning rebiopsy strategy, (3) planning surgical strategy, (4) selection criteria for focal therapy and active surveillance, and (5) cost reduction. Peripheral end marking is low cost, easy and reproducible. It drives several potential advantages in cancer diagnosis or isolated atypical lesions, in particular, spatial localization within the biopsy (transition versus peripheral zone, anterior versus posterior, subcapsular versus intraparenchima, and extraprostatic extension) should be easy and reliable. We can add a new pathological parameter: pathological orientation or biopsy polarity.

Background: Tumor size in breast cancer is a key factor for staging, prognosticating, and deciding the choice of treatment. Currently, there are no standard rules for measuring the T status in breast cancer. The purpose of this study was to determine an accurate method to evaluate the T status by various parameters compared with the actual size in fresh specimens. Materials and Methods: This prospective study was conducted on 134 breast cancer patients scheduled to undergo a modified radical mastectomy. The paired t-test was used for analyses. Results: Using a paired t-test, the differences in tumor size as measured by physical examination (P ≤ 0.001) and in the formalin-fixed specimen (P ≤ 0.001) when compared with the postoperative fresh specimen were highly significant. These differences indicated that the physical examination and formalin-fixed specimen measurements were inaccurate in estimating tumor size. Tumor size, as measured by a mammogram and ultrasonogram when compared with the referenced P= 0.077 and 0.149, respectively, showed that the ultrasonogram is the most accurate method of determining tumor size in vivo. The mean percentage decrease in size of the formalin-fixed specimen was 7.8, which was significant enough to downstage two patients from T2 to T1 and seven patients from T3 to T2. Conclusion: An ultrasonogram is the most accurate way of defining tumor size in vivo as measured in postoperative fresh specimens. Tumor shrinkage with formalin fixation may give a false T status.

Objectives: The existence of tumour cells on the surgical margin can be shown in frozen sections. However, there has been no method to obtain information about surgical clearance intraoperatively. We propose a simple method to confirm the surgical clearance of tongue carcinomas during surgery. Methods: Just after the resection of a tumour in a conventional manner, the resected fresh specimen was immersed in a gelatine solution, maintaining its original shape and orientation, and refrigerated for solidification of the gelatine. Then ultrasound observation of the gelatine-embedded specimen was performed from the surface of the gelatine. Results: Very fine images could be obtained easily and without any specialist skills. Conclusions: With this method, the operator can easily obtain information about surgical clearance during the operation.

The assessment and management of congestive heart failure relies increasingly on the measurement of B‐type natriuretic peptide (BNP). However, the effective contribution of this biochemical test in the clinical decision making is influenced by reliability of the measure, which also depends on several preanalytical issues. Since there is controversy on the influence of the matrix and the storage conditions on BNP measurement, we compared results of BNP in serum, K2 ethylene diamine tetra‐acetic acid (EDTA) plasma and lithium heparin plasma fresh samples and in matching samples stored at −20 and −80°C for 1 week. BNP measured on the Bayer Advia Centaur was systematically underestimated in heparin plasma (−47%) and serum (−62%) when compared to K2 EDTA plasma. According to the established 100 ng/L cutoff value, 25% and 37% of the fresh samples collected in heparin plasma or serum were misclassified from the reference K2 EDTA fresh specimen, respectively. When compared to the fresh specimens, the mean and interindividual bias observed for samples stored at either −20°C or −80°C was, overall, modest for K2 EDTA plasma (−2%) and heparin plasma (+6% and −4%, respectively), though it appeared clinically meaningful in serum (+47% and +28%, respectively). Although we can not rule out that other BNP assays using different antibodies may be not affected from degradation during storage to the same extent, results of our investigation demonstrate that K2 EDTA plasma is the most suitable specimens for BNP testing on fresh and frozen samples stored at either −20°C or −80°C for up to 1 week. J. Clin. Lab. Anal. 21:293–297, 2007. © 2007 Wiley‐Liss, Inc.

Background: In the kidney, the trachea, and the colon, nitric oxide (NO) can modulate transepithelial fluid transport. This study investigates whether isolated human and porcine ciliary processes produce NO.Methods: Porcine ciliary processes and iris were used either fresh or thawed after storage at –70°C. Post-mortem (8–12 h) human ciliary processes were used thawed after storage at –70°C. NO was measured by placing a nafion-coated polymeric porphyrinic microsensor (differential pulse voltammetry) on the surface of the tissue. Measurements were conducted in the absence or in the presence of the NO formation inhibitor N G-nitro-l-arginine methyl ester (L-NAME; 0.2 mM, 1 mM) or its biologically inactive d-enantiomer N G-nitro-d-arginine methyl ester (D-NAME; 1 mM). Results: NO concentrations in porcine ciliary processes (1.27±0.25 µM) were higher (P=0.001) than those in the iris (0.00±0.02 µM) and were significantly (P

Background and Study Aims: Histopathological examination for superficial gastrointestinal lesions has been mainly based upon the light microscopic examination of thin-slice specimens with hematoxylin and eosin (H&E) staining. However, it takes at least a couple of days to create a slide-glass for microscopic study. In order to obtain immediate microscopic images for untreated specimens, the authors used laser-scanning confocal microscopy (LCM) to study fresh samples of gastrointestinal mucosa. Materials and Methods: Fresh untreated mucosal specimens from the esophagus, stomach, and colon, obtained by endoscopic pinch biopsy, polypectomy, or endoscopic mucosal resection (EMR), were fixed in normal saline and examined by LCM collecting the reflective light of a 488-nm wavelength argon laser beam. Findings from the LCM image were compared with those of conventional H&E staining in all specimens. For objective evaluation of the similarity of both pictures, the nucleus-to-cytoplasm ratio (N/C) of normal mucosa and that of cancer of the esophagus were calculated and statistically analyzed. The overall diagnostic accuracy for cancer was evaluated. Results: The average scanning time to obtain the LCM image of a specimen was 1.6 seconds. The LCM images acquired corresponded well to the conventional H&E light microscopic images in the esophagus, stomach, and colon. Cell wall, nucleus, cytoplasm, and tissue structural elements were simultaneously visualized by LCM scanning. A difference in N/C ratios between normal mucosa and cancer in the esophagus was statistically apparent when Welch's test (P =0.05) was applied. The overall diagnostic accuracy of the LCM study for cancer was 89.7 %. Conclusions: This novel method enables us to obtain an immediate serial virtual microscopic section through a fresh specimen, which has not actually been cut, although the resolution of the image obtained is still limited. These early results encourage us to develop imaging relevant to conventional histopathology alongside the development of LCM technology in the near future. We should aim at the in vivo application of LCM coupled to probes which can be introduced through the working channel of endoscopes.

Foram estudadas as medidas microcirúrgicas da região selar de 20 cadáveres com um máximo de 12 horas após o óbito. Utilizou-se fluoresceína como contraste para estudo do sistema vascular. O estudo em cadáveres com pouco tempo após o óbito e o uso de fluoresceína objetivaram evitar distorções na anatomia das estruturas causadas pela formolização ou pelo uso de contrastes a base de cola. A droga foi diluída em água ou, para se obter maior consistência, em gel de ultrassonografia. A injeção do contraste foi feita nas artérias carótidas comuns e o registro fotográfico realizado com três diferentes tipos de iluminação: 1) lâmpada de Wood ("luz negra"); 2) lâmpada de wood mais luz branca; 3) luz branca. Obteve-se melhor contraste quando se iluminou somente com a lâmpada de wood. As medidas realizadas mostraram, com frequência, resultados diferentes dos de outros autores. A técnica fotográfica com fluoresceína é simples e acessível, apresentando excelente qualidade como contraste vascular para estudos neuroanatômicos. The microsurgical anatomy of sellar regions in 20 cadavers with a 12 hours maximum post mortem was studied. Fluorescein was used as contrast for vascular anatomy examination. The use of fresh specimen and fluorescein had the objective of avoiding any distortion of normal anatomy. The drug was diluted with water or, in order to have more consistency, in gel for ultrasonography. Contrast injection was made into the internal carotid artery and photographs were taken with three different kinds of lighting: 1) only with Wood's lamp; 2) Wood's lamp and normal white light; 3) only with white light. Lighting only with Wood's lamp showed more contrast. After measurement, data often showed discrepancies with other authors' data. The photographic technique with fluorescein is simple and accessible; the drug showed high quality as a vascular contrast for neuroanatomic research.

A fresh specimen of an adult Atlantic Puffin, Fratercula arctica, was obtained from a local hunter at the Minarets (Akpait) on the east coast of Baffin Island on 3 August 2007. The measurements of this bird exceeded those of Atlantic Puffins from Newfoundland and Labrador (F. a. arctica) but fell within the limits of the High Arctic subspecies (F. a. naumanni). This specimen appears to be the first of the High Arctic subspecies to be collected on a potential breeding site in Canada. This subspecies is highly disjunct from the Low Arctic subspecies and is probably represented in Canada by only a few hundred individuals.

Single crystal MgO specimens having low load Vickers indentations were thinned in an ion milling machine employing a single ion gun, and their characteristics were investigated with optical microscopy and high voltage electron microscopy (HVEM). It was found that the state of cleanliness of the specimen chamber of the ion milling machine had a very marked influence on the quality of the thinned specimens. If the specimen chamber was not well cleaned before ion milling a fresh specimen, the latter tended to show amorphisation due to the deposition on the specimen of the debris left in the chamber from the previously ion-milled specimens. Such observations were made from MgO specimens ion milled in several different types of commercial ion milling machine employing a single gun. It is proposed that to obtain good-quality ion milled TEM specimens, it is important to clean the specimen chamber thoroughly prior to milling.

Background: Histological examination for gastorintestinal superficial lesions has been mainly based upon the light micorscopic examination of a thin-slice specimen under H-E staining. However, it takes a couple of days to create a slide grass for microscopic study. In order to obtain a simultaneous microscopic image for an untreated specimen, the authors introduced a laserscanning confocal microscopic study for a fresh sample of gastrointestinal mucosa. Methods: Fresh and untreated mucosal specimens in the esophagus, stomach and colon obtained by pinch biopsy or EMR are based in normal saline and examined by utilizing the laser-scanning confocal microscopy (LCM) with collecting the reflective light of 488- nm wave-length argon laser beam. Findings from the LCM image are compared with those of the conventional H-E staining in all specimens. Findings: Average scanning time of a specimen to obtain the LCM image was 1.6 seconds. The acquireed LCM images were corresponded well to the conventional H-E light microscopic imagnes in the esophagus, stomach and colon. Overall accuracy of LCM image for cancer diagnosis is 89.7%. Discussion: This novel method enables to obtain a simultaneous and a serial imaginary microscopic section on an actually non-cut fresh specimen, although the resolution of the obtained image is still limited. These early results encourage us to create an image relevant to conventional histology in accordance with the development of the LCM technology in the near future.

Figure 1. Adenoma of the Sigmoid Colon. A screening barium enema in a 78-year-old man with a positive test for fecal occult blood showed a flat round mass with a surrounding hump (Panel A, arrows). Panel B is an endoscopic view of the lesion. Sigmoidectomy was performed without complication. The resected fresh specimen, which measured 3.8 by 2.4 cm, is shown in Panel C. Panel D shows the longitudinal cut surface of the lesion (hematoxylin-eosin stain). No carcinomatous focus was found. The patient is well two years after surgery.

In an analytical laboratory, reference materials are used for several purposes, including the calibration of analytical instruments, periodic standardization of these instruments, and verification that the standardization is correct. Reference materials are also used to confirm that an analysis is performed with optimum precision and is not biased, and to determine the presence and effects of interferences. All of these applications require that proposed reference materials be evaluated for homogeneity. To this end, specimens are cut at regular intervals from bars selected for evaluation and identified accordingly. Across the entire cross section of each specimen, a flat surface is prepared as specified by the analytical method. The specimens are analyzed in random order by optical emission spectrometry, once at the center of the specimen and twice as close to the edge of the specimen as possible. If X-ray emission (fluorescent) spectrometry is used, two analyses are performed with a fresh specimen preparation before each analysis. Statistical analyses are performed on the values obtained for each element to determine whether there are significant differences in composition between center and edge or among specimens. This procedure has been used successfully for ten years to evaluate proposed reference materials prior to certification.

Roentgenologic studies by polytomography of the temporal bones of a 2,600-year-old Egyptian mummy (PUM-II) revealed an increase in radiographic density throughout the left temporal bone. The superior portions of the petrous bones were removed with an electric saw for study by undecalcified techniques. For histological control, a similar specimen was removed from a cadaver immediately after autopsy. Light microscopy of sections showed excellent preservation of the Haversian systems. The vascular channels were clearly seen, and they were similar in appearance to those of the fresh specimen. The lacunae contained osteocytes with good preservation of the nucleus. Osteoid seams were identified and the osteon activity was described for the first time in an Egyptian mummy. There was a low turnover type of bone, but there was no evidence of metabolic bone disease. We postulate that the increased radiological density of the left temporal bone was due to a greater amount of resin infiltration in this bone as compared to the right. It appeared that the resin acted as a fixative preserving the bone cells and related elements.

Low resistivity contacts to the bulk specimens of YBa2Cu3O7-y with Ag have been developed. In order to obtain homogeneous mixtures of YBa2Cu3O7-y and Ag we used Ag2O powder, The contact resistivity of 0.19μΩ·cm2 was obtained by annealing the specimen at 1, 100°C. The current versus voltage characteristic of the contact was found to be ohmic. The contact was made to the specimen which is exposed to air for 110 days; the contact resistivity after this period had elapsed was almost the same as the contact made to the fresh specimen. Using physical and chemical analysis it was found that all the Ag2O in YBa2Cu3O7-y was separated to Ag and that Ag existed in all voids.

An investigation into the relative elastic and non-elastic components of compressive deformity of the cervical spinal cord has been made. Sixteen patients were examined by computed myelography with the neck flexed and extended, and one fresh specimen of a human spinal cord was studied under reversible transverse compression with the specimen in a stretched and shortened state. In conditions of transverse compression cord tissue undergoes an alteration in viscoelastic properties, manifesting itself as reduced expansion of the compressed segment as the cord shortens, which in some cases is independent of whether the agent is seen to actually contact the cord. The presence of degenerative changes in cord substance probably accentuates this alteration in mechanical properties.

Validity of the compliance matching procedure has been examined for short fiber composites experimentally by studying the behavior of damaged specimens. Comparison between the behavior of damaged and fresh specimens shows that the load—COD (crack mouth opening displacement) curve and fracture load of a damaged specimen are different from that of a fresh specimen having a machined crack of length equal to the estimated crack length in the damaged specimen. However, it has been clearly established that the secondary compliance of the damaged specimen can be used to accurately estimate the extent of damage to the specimen by earlier loading. Therefore, the use of the compliance matching procedure is justified for estimating instantaneous crack length in short fiber composites.

A method for examining the viscera of rat and rabbit fetuses is described. Techniques used in detecting visceral alterations in rats and rabbits for routine teratogenicity screens have varied over the years. The method used should be quick and simple but at the same time must be accurate, reliable, and comprehensive. The procedure used in this lab is a complete and systematic necropsy of the fresh fetus requiring minimum equipment and time. The examination can be done immediately following cesarean section and yields an intact skeleton which can subsequently be processed for skeletal examination. The fresh specimen and the natural coloration of in situ organs makes color photography of visceral alterations clear and concise. Any lesion can be appropriately fixed for histopathic examination. This technique begins with the examination of the organs in the abdominal cavity and proceeds to the thorax. Of special interest is the procedure used to inspect the internal anatomy of the fresh fetal heart. A description of the internal examination of both rat and rabbit heads and eyes is also included.

When a fresh specimen of yellowfin tuna Neothunnus albacora was analyzed, the cross-sectional area of ordinary (white) muscle fiber was 3.5 times larger than that of dark muscle. As heating temperature was raised, the shear force of dark muscle became progressively larger than that of ordinary muscle. Simultaneously, the A band in the ultrastructure of both muscles was stained more densely. While the Z line in I band of dark muscle still remained, that of ordinary muscle disappeared completely by heat treatment at 60°C for 30 min. Therefore, the differences in toughness change during heat treatment correlated well with that fiber sizes and ultrastructures of the two types of muscle.

Glass rods for lamp and tube of about 4mm in diameter were provided with two electrodes at the distance of 30-40mm, and the apparent resistance, which is equivalent to the combination of volume and surface resistance connected pararell, were measured by the method of filament electrometer or direct deflection.In the first series of the experiment the specimens were washed by loundry soap and city water. In the season of high humidity (summer) the apparent resistance has changed after the preparation of the specimens showing a clear corelation with relative humidity. After half a year, and in the season of low humidity (winter) some glasses showed comparatively high surface resistance, keeping the apparent resistance at a constant level, being very near to the volume resestance. In the glasses of higher volume resistance, however, the change of surface resistance due to the change of humidity was clearly observed. After a year a lot of specimens gave the apparent resistance of as high as 10-100 times under the same atmospheric conditions.In the second series of the experiments the apparent resistance of the fresh specimen was compared with those aged a year and those treated with dilute hydrofluoric acid. It was observed that the apparent resistances of above three kinds of the specimens showed the similar trend of the change with the atmospheric conditions, and the initially quite different of numerical values came up to one and the same value with the laps of time.In the last series of experiments the apparent resistance of the specimens washed with, acid, alkali and salts was measured in the cell, to which, after evacuation, water vapour was introduced up to a certain saturation pressure. It was confirmed that the characteristic properties of apparent resistance vs. vapour pressure varied remarkably with the method of surface treatment.

Low resistivity contacts to bulk specimens of YBa 2 Cu 3 O 7− y with Ag have been developed 1 . In order to obtain homogeneous mixtures of YBa 2 Cu 3 O 7− ity and Ag we used Ag 2 O powder. The contact resistivity of 0.19 μΩ cm 2 was obtained by annealing the specimen at high temperature. The current versus voltage characteristic of the contact was found to be ohmic. The contact was made to the specimen which was then exposed to air for 110 days; the contact resistivity after this period had elapsed was almost the same as the contact made to the fresh specimen. Using physical and chemical analysis it was found that all the Ag 2 O in YBa 2 Cu 3 O 7− y was separated to Ag and that Ag existed in all voids.

Brilliant and striking preparations of the Vertebrate bony skeleton may be prepared for class-room demonstrations or study with a minimum of equipment and reagents. The method is applicable, with certain modifications, to fish, amphibia, reptiles, birds, and mammals and involves practically no dissection as the muscles are not removed but are rendered transparent. All the skeletal elements remain in position and every bone, no matter how minute, is clearly shown. Usually, large animals cannot be used as the treatment frequently results in maceration before the clearing process is complete, but full-grown rats, guinea pigs and mice are within the size range that can be readily prepared. The fresh specimen is eviscerated through an abdominal incision. It is especially important to remove all fragments of the heart, lungs and liver to prevent staining of the preparation. The brain may be left in situ. Fetal mammals and bird embryos require only evisceration. Older specimens with hair or feathers as well as adult amphibia and reptiles should be skinned. In larger fishes the scales should be removed as they also react with alizarin. The entire animal is then fixed in 95%o alcohol for several days or even weeks. The longer it is fixed the less danger there is of maceration in the subsequent treatment in potassium hydroxide. Formalin-fixed material is more difficult to clear and formalin is not recommended for routine work. Following fixation the specimen is placed in a 1% solution of potassiumn hydroxide and left there until the bones are clearly visible through the muscle. It is then transferred to a dilute solution of alizarin red S (alizarin mono-sodium sulphonate), one part of the dye to 10,000 to 40,000 parts of 1% potassium hydroxide and allowed to remain there until the bones are stained a deep red. If the

The application of histochemical techniques to skeletal muscle heralded a new era in muscle pathology. Whilst fibres appear uniform in formalin-fixed, paraffin-embedded tissue, in frozen sections enzyme histochemistry reveals a heterogeneous fibre population. Both physiological and pathological changes may be selective for one or other fibre type. The identification of different fibre types has therefore enhanced our understanding of skeletal muscle pathology and provided a valuable diagnostic tool. Electron microscopy, measurements of fibre size and biochemical analysis may give additional information. Muscle biopsy is a relatively simple procedure, but to obtain the maximum information it is essential firstly to select the most appropriate site for biopsy and secondly to handle the specimen correctly. Rough treatment of the fresh specimen, or misguided fixation, can destroy important diagnostic clues and introduce spectacular artifacts (Figures 1.1 and 1.2).

Validity of the compliance matching procedure has been examined for short fibre composites experimentally by studying the behaviour of damaged specimens. The load - COD curve and fracture load of a damaged specimen are different from that of a fresh specimen. However, it has been clearly established that the secondary compliance of the damaged specimen can be used to accurately estimate the extent of damage to the specimen by earlier loading.