How many lymph nodes are necessary to stage early and advanced adenocarcinoma of the sigmoid colon and upper rectum?

The lymph-node yields in specimens resected for colorectal adenocarcinoma show considerable variations, raising the question whether the minimum lymph-node number recommended by the UICC (International Union Against Cancer) for pN0 classification represents an appropriate quality standard for specimen work-up. The number of pericolic lymph nodes recovered from 568 archival surgical colorectal carcinoma specimens located in the sigmoid or upper rectum showed a highly statistically significant correlation with both the pT category and the presence of metastases (P<0.0005). The median lymph-node yield in standardized (i.e., resembling in size surgically removed cancer specimens) tumor-free specimens obtained during autopsies was 13 lymph nodes, compared with 20.5 when diverticula were present and more than 30 in specimens with chronic inflammation or from patients with systemic infections. In 48 pT2 and pT3 carcinoma specimens prospectively dissected in the same way, median numbers of 18 (pT2) and 23 (pT3) lymph nodes were detected (range between 8 and 39 nodes). The lymph-node numbers recommended in previous studies and by the UICC often seem to be too low to declare a specimen free of metastases. Although the great variation in lymph-node counts requires the recovery of all lymph nodes for pN0 classification, recommendations considering the pT status and additional factors like diverticula and inflammatory changes can be useful as a quality standard for specimen work up.     

Microsatellite analysis of breast carcinoma and corresponding local recurrences

Local recurrence is a serious complication of breast carcinoma that reduces quality of life and influences prognosis. The aim of this study was to determine whether local recurrences of breast carcinoma are genetically related to the primary tumours. Forty cases of locally recurrent breast carcinomas (median onset: 3.6 years after primary surgery) were analysed: 22 patients had undergone breast-conserving therapy and 18 mastectomy. Eighteen microsatellites on chromosomes 2p, 3p, 5q, 10q, 11p, 11q, 13q, 17q, 17p, 18p were amplified by PCR using fluorescent-labelled primers, automatically detected after polyacrylamide gel electrophoresis and analysed for loss of heterozygosity (LOH) or microsatellite instability (MSI). Follow-up data were available for 39 cases with a median value of 89 months. All LOH and MSI found in the primary tumours were also present in the corresponding recurrences, indicating that they are genetically related to the primary tumours and not secondary malignancies in the same breast. MSI was found in three cases, of which one harboured MSI at more than two loci. The median value of LOH per case was significantly higher in the recurrent (four per case) compared to the primary tumours (two per case; p < 0.001, Mann-Whitney test), reflecting the genotype of tumour progression. Early local recurrence was associated with specific LOH for TP53.15 (p = 0.018, log-rank test) in the primary tumours. LOH on D13S1699 or D17S855 was associated with lymph node metastases (p = 0.024 and p = 0.019, respectively; chi-square test). In addition, tumour grade, lack of oestrogen or progesterone receptor expression, young patient age and early appearance of local recurrence significantly correlated with poor survival. The development of local recurrence despite clear resection margins may result from residual DCIS distant from the invasive carcinoma, homing of circulating tumour cells, or genetically altered, histologically normal breast tissue not immediately adjacent to the invasive carcinoma.     

How to improve microscopic images obtained with consumer-type digital cameras

AIM: Digital imaging is useful in conventional photography because it immediately provides images, and the image quality can be improved afterwards by the use of computer programs. The major disadvantages of consumer-type digital cameras mounted on microscopes are (i) unequal illumination through the image, and (ii) a coloured background. A computer program was specifically adapted and refined to improve images obtained with consumer-type digital cameras mounted on microscopes. METHODS AND RESULTS: An approach using a division operation between the specimen image and a background image leads to homogeneous illumination throughout the image, with automatically corrected brightness and white background. The correct colour spectrum is preserved by correction of the histogram. This approach was obtained from the freeware computer program 'Image Arithmetic'. In a test, three different consumer-type digital cameras (Sony, Nikon, Olympus) on different microscopes were used to obtain images of different types of histological specimens (cervical smear, bone marrow biopsy, and colonic biopsy). The computer program dramatically improved the quality of images obtained with all tested cameras. CONCLUSION: Using this approach, even low-cost digital cameras mounted on microscopes produce brilliant images with homogeneous illumination and a white background, the image quality being comparable with expensive cameras especially designed for microscopes.     

An evaluation of a high‐pressure 11CO carbonylation apparatus

[¹¹C]Carbon monoxide (¹¹CO) is a versatile building block for the synthesis of Positron Emission Tomography (PET) radioligands. However, the difficulty of trapping ¹¹CO in a small solvent volume has limited its utility. We here report an evaluation of a simple, fully automated high‐pressure synthesizer prototype for the use in ¹¹C‐carbonylation reactions. [¹¹C]Carbon monoxide was easily prepared by online reduction of [¹¹C]carbon dioxide using either Mo(s) or Zn(s) as the reducing agent. The conversion yield of ¹¹CO was >99% when zinc was used as the reducing agent, and the corresponding value for Mo was approximately 71%. When the Zn or Mo column was constantly kept under inert atmosphere, no significant decrease in reducing properties was observed for more than 100 ¹¹CO productions. However, in our hands, Mo reductant was much easier to service. A total of nine functional groups were successfully radiolabeled using the ¹¹CO synthesizer prototype. All measured radiochemical yields exceeded 37%, and the ¹¹CO trapping efficiency was generally above 90%, except for the Suzuki coupling where the trapping efficiency was 80%. This high‐pressure synthesizer using [¹¹C]carbon monoxide as the labeling precursor is easy to operate allowing for ¹¹C‐carbonylation reactions to be performed in a high yield and in a routinely fashion.     

Insight into the active site nature of zeolite H-BEA for liquid phase etherification of isobutylene with ethanol

The nature of active acid sites of zeolite H-BEA with different Si/Al ratios (15–407) in liquid phase etherification of isobutylene with ethanol in a continuous flow reactor in the temperature range 80–180 °C has been explored. We describe and discuss data concerning the strength and concentration of acid sites of H-BEA obtained by techniques of stepwise (quasi-equilibrium) thermal desorption of ammonia, X-ray diffraction, low-temperature adsorption of nitrogen, FTIR spectroscopy of adsorbed pyridine and solid-state ²⁷Al MAS NMR. The average values of the adsorption energy of NH₃ on H-BEA were experimentally determined as 63.7; 91.3 and 121.9 mmol g⁻¹ (weak, medium, and strong, respectively). In agreement with this, a correlation between the rate of ethyl-tert-butyl ether synthesis and the concentration of weak acid sites (E_NH3 = 61.6–68.9 kJ mol⁻¹) has been observed. It was concluded that the active sites of H-BEA for this reaction are Brønsted hydroxyls representing internal silanol groups associated with octahedrally coordinated aluminum in the second coordination sphere.

The active sites of H-BEA zeolites for ETBE synthesis are the weak Brønsted acid sites representing internal silanol groups.     

The relationship between angle kappa and astigmatism after phacoemulsification with implanting of spherical and aspheric intraocular lens

Purpose:To determine the significance of any association between either change in angle kappa (K°) or the rectilinear displacement (L, mm) of the first Purkinje image relative to the pupil center and unexpected changes in astigmatism after phacoemulsification.Methods:Orbscan II (Bausch and Lomb) measurements were taken at 1, 2, and 3 months after unremarkable phacoemulsification in patients implanted with spherical (group 1, SA60AT, Alcon) or aspheric (group 2, SN60WF, Alcon) nontoric IOLs. The outputs were used to calculate L. Astigmatism, measured by autorefractometry and subjective refraction, was subjected to vector analysis (polar and cartesian formats) to determine the actual change induced over the periods 1–2 and 2–3 months postop.Results:Chief findings were that the mean (n, ±SD, 95%CI) values for L over each period were as follows: Group 1, 0.407 (38, ±0.340, 0.299–0.521), 0.315 (23, ±0.184, 0.335–0.485); Group 2, 0.442 (45, ±0.423, 0.308–0.577), 0.372 (26, ±0.244, 0.335–0.485). Differences between groups were not significant. There was a significant linear relationship between (A) the change in K (ΔK = value at 1 month-value at 2 months) and K at 1 month (x), where ΔK =0.668-3.794X (r = 0.812, n = 38, P = <0.001) in group 1 and ΔK = 0.263x -1.462 (r = 0.494, n = 45, P = 0.002) in group 2, (B) L and the J₄₅ vector describing the actual change in astigmatism between 1 and 2 months in group 2, where J₄₅ (by autorefractometry) =0.287L-0.160 (r = 0.487, n = 38, P = 0.001) and J₄₅ (by subjective refraction) =0.281L-0.102 (r = 0.490, n = 38, P = 0.002), and (C) J₄₅ and ΔK between 2 and 3 months in group 2, where J₄₅ (by subjective refraction) =0.086ΔK-0.063 (r = 0.378, n = 26, P = 0.020).Conclusion:Changes in the location of the first Purkinje image relative to the pupil center after phacoemulsification contributes to changes in refractive astigmatism. However, the relationship between the induced change in astigmatism resulting from a change in L is not straightforward.     

Ca2+ release-activated Ca2+ channel blockade as a potential tool in antipancreatitis therapy

Alcohol-related acute pancreatitis can be mediated by a combination of alcohol and fatty acids (fatty acid ethyl esters) and is initiated by a sustained elevation of the Ca²⁺ concentration inside pancreatic acinar cells ([Ca²⁺]_i), due to excessive release of Ca²⁺ stored inside the cells followed by Ca²⁺ entry from the interstitial fluid. The sustained [Ca²⁺]_i elevation activates intracellular digestive proenzymes resulting in necrosis and inflammation. We tested the hypothesis that pharmacological blockade of store-operated or Ca²⁺ release-activated Ca²⁺ channels (CRAC) would prevent sustained elevation of [Ca²⁺]_i and therefore protease activation and necrosis. In isolated mouse pancreatic acinar cells, CRAC channels were activated by blocking Ca²⁺ ATPase pumps in the endoplasmic reticulum with thapsigargin in the absence of external Ca²⁺. Ca²⁺ entry then occurred upon admission of Ca²⁺ to the extracellular solution. The CRAC channel blocker developed by GlaxoSmithKline, GSK-7975A, inhibited store-operated Ca²⁺ entry in a concentration-dependent manner within the range of 1 to 50 μM (IC₅₀ = 3.4 μM), but had little or no effect on the physiological Ca²⁺ spiking evoked by acetylcholine or cholecystokinin. Palmitoleic acid ethyl ester (100 μM), an important mediator of alcohol-related pancreatitis, evoked a sustained elevation of [Ca²⁺]_i, which was markedly reduced by CRAC blockade. Importantly, the palmitoleic acid ethyl ester-induced trypsin and protease activity as well as necrosis were almost abolished by blocking CRAC channels. There is currently no specific treatment of pancreatitis, but our data show that pharmacological CRAC blockade is highly effective against toxic [Ca²⁺]_i elevation, necrosis, and trypsin/protease activity and therefore has potential to effectively treat pancreatitis.     

Oncocytic versus mitochondrion‐rich follicular thyroid tumours: should we make a difference?

Aims: To separate true oncocytic neoplasms from mitochondrion‐rich non‐oncocytic lesions based on the intracellular relationship between major cell organelles, and to establish the diagnostic and clinical relevance of this distinction. Methods and results: Tissue samples from 276 follicular adenomas, 194 follicular carcinomas, 162 normal thyroids and 296 non‐neoplastic lesions were classified as conventional, mitochondrion‐rich or oncocytic based on the immunohistochemically assessed quantity and intracellular distribution of mitochondria and endoplasmic reticulum (ER) and nuclear position. Pathological and clinical features were compared among the groups. In oncocytes, densely packed mitochondria resulted in homogeneous immunolabelling of basal cytoplasmic regions, whereas ER and the nuclei were typically displaced to the apical position. This aberrant organelle distribution was not observed in non‐oncocytes, which allowed reliable distinction between oncocytic and mitochondrion‐rich lesions. Clinically, mitochondrial increase in non‐oncocytic lesions was associated with neoplasia, malignancy and higher cancer recurrence rates. Similar correlation, albeit less pronounced, was observed within the oncocytic tumour group. By contrast, oncocytic change per se was not associated with neoplasia, malignancy or cancer aggressiveness. Conclusions: True oncocytic neoplasms can be distinguished from mitochondrion‐rich non‐oncocytic tumours based on aberrant distribution of all major cell organelles. This distinction has immediate clinical relevance and should be implemented in practice.     

Finite element simulation of electrochemically reversible,quasi-reversible and irreversible linear sweep voltammetry at the wall tube electrode

A finite element approach for numerical simulation of linear sweep voltammetry at the wall tube electrode is presented. Working curves and surfaces are computed and reported which permit the analysis of reversible, quasi-reversible and irreversible voltammograms for all voltage scan rates.