Analysis of centroiding algorithms for non-diffracting structured and hollow structured laser beams.

This paper explores the potential of optical-based systems, specifically pseudo-non-diffractive beams, as an alternative for alignment. The study focuses on structured laser beams and hollow structured laser beams, which exhibit lower divergence and enhanced detection capabilities. The research objective is to analyze and compare centroiding algorithms in terms of accuracy and robustness to noise. The study compares the gamma-corrected and threshold-corrected center of gravity and correlation template matching. It also introduces a polarization-based algorithm.

Wavefront reconstruction of a non-diffracting structured laser beam.

The Structured Laser Beam (SLB) is a pseudo-non-diffracting laser beam that shares many characteristics with a Bessel beam. However, it can theoretically propagate over an unlimited distance while maintaining an extremely low inner core divergence of only 0.01 mrad. This makes it a promising candidate for precise long-distance alignment applications such as the alignment of particle accelerator components at CERN. In this work, a novel method to assess the symmetrical wavefront aberrations induced by an SLB generator is presented. Our approach is based on the analysis of a single-intensity distribution of an SLB. The coefficients of the Zernike polynomials are estimated using artificial intelligence before least-squares fitting is used to refine the result. This approach ensures that the fitting avoids local minima. This method provides a novel way to analyze the optical aberrations induced by the SLB generator.

Variability in the levels of fungicide residues in final beer as they are influenced by various sequences of agrochemicals used for treatment on hops.

The fate of during brewing of pesticides (organic compounds and copper) from hops was determined to reveal linkages between various pesticide sequences applied in hop yards and the level of pesticides in beer. For this purpose, laboratory-scale brewing trials were carried out with conventional hops from four localities in which pest control spray programs varied. Pesticide residue analysis in samples of hopped wort, young beer, and beer was carried out by liquid chromatography in tandem with mass spectrometry (LC-MS/MS). Cu concentrations were determined using inductively coupled plasma mass spectrometry (ICP-MS). The transfer rates (%) of individual pesticides were calculated to demonstrate their ability to be transferred from hops to the brewed solutions. The pesticides exhibited different transfer rates correlating well with their log p values; however, the obtained transfer data were not related to their concentration in hops. The average transfer rates calculated for ametoctradin (15% ± 5%), mandipropamid (38% ± 4%), boscalid (48% ± 5%), and azoxystrobin (47% ± 2%) increased in good correlation with their descending log P values. On the contrary, the transfer rates of copper residues were related to residual concentrations of copper in hops and exhibited logarithmic dependency. The carryover of the sum of all pesticides in the four samples ranged from 36% to 49%, averaging 42% ± 6%. The data showed no substantial influence of various pesticide spray sequences on the percentages of overall pesticide residues carried over into beer.

Variation of structured laser beam pattern and optimization for an alignment reference line creation.

The alignment of particle accelerators demands a dedicated measurement system based on a straight-line reference. This straight line can be provided by a laser beam. The alignment then involves accurately measuring the offset of accelerator components with respect to this light path. In order to be efficient, the laser beam needs to serve as a stable and straight reference for distances of several hundreds of meters. The attainable accuracy depends, among other parameters, on the laser spot size, which should ideally change very little over the distances at which the alignment system needs to operate. Due to the significant divergence of Gaussian laser beams, we propose using a structured laser beam (SLB) for alignment. Its transversal intensity profile is similar to a Bessel beam and consists of an intense inner core (IC) and concentric rings. The divergence of the IC, i.e., the growth of its size with distance, can be limited to 10µrad using a favorable generator configuration. Thus an SLB may be suitable as a straight-line reference for long-distance alignment applications. However, the SLB is distorted if obstructions cover parts of the outermost ring (OR) of the beam within, which should therefore also be small. In this paper, we investigate the relationship between the size of the IC and OR depending on the design parameters of the SLB generator. We use numerical simulations and experiments with different generators over distances up to 50 m to analyze the transversal intensity profile and wavefronts of different SLBs. The results indicate the general suitability of an SLB as a reference line for long-distance alignment but also expose tradeoffs between small IC and small OR. The findings outlined in the paper help to describe the optimal SLB parameters for given conditions.

Seasonal and geographical variations in insecticide and miticide residues in hop samples produced across three hop-growing regions in the Czech Republic during the years 2018-2020.

The aim of this study was to determine the presence of zoocide (insecticide and miticide) residues in hops collected in three hop-growing regions located in the Czech Republic, and to assess their zoocide profiles and residue levels in terms of variability in temperature and precipitation across the 2018-2020 seasons. Furthermore, the weather factors that influenced the occurrence of hop pests are described and discussed. During our 3-year survey, a total of 120 samples of whole-cone hops samples harvested in three hop-growing regions were analysed for the presence of 29 insecticides and miticides using the modified QuEChERS extraction method, followed by liquid chromatography-tandem mass spectrometry and gas chromatography-tandem mass spectrometry. A majority, 119 of 120 samples, contained a residue of at least one of the active substances surveyed in this study, and 34 analysed samples contained multiple residues with three to four zoocides presented. Concerning the most frequently detected zoocide residues, spirotetramat and/or its metabolites were found in 94.2% of the samples at levels ranging from 0.02 to 1.08 mg/kg. Of the other zoocides surveyed, residues of fenpyroximate, hexythiazox, bifenazate and lambda-cyhalothrin were routinely found in hop cone samples. Obtained data were then used for evaluating seasonal and geographical variations in the profile of zoocide residues among the hop-growing regions in the years 2018-2020, and the compliance with legal regulations concerning the use of zoocides on hops was ascertained. The results showed that (1) the profile and levels of zoocide residues found in the samples reflected seasonal prevalence of pest infestation on hop plants; (2) the strategy to control pests (especially aphids) used in most of hop yards was consistent across the seasons; and (3) a concentration of spirotetramat residues less than 1 mg/kg was typical for hops grown in the Czech Republic.

Fate and Behavior of Field-Applied Pesticides during Malting and Mashing Processes.

The present work aimed to study the fate of field-applied pesticides during malting and mashing processes. Twenty-four field-collected barley samples were subject to micromalting followed by lab-scale mashing to investigate the carryover of residual pesticides from barley to malt and then from malt to sweet wort. The citrate-buffered QuEChERS sample preparation method was adapted for simultaneous determination of 57 pesticide residues in grain, malt, spent grains, and sweet wort samples using ultra-performance liquid chromatography coupled with tandem mass spectroscopy (UPLC-MS/MS). Residues of four fungicides (fenpropimorph, pyraclostrobin, tebuconazole, and trifloxystrobin) and two insecticides (chlorpyrifos and pirimiphos-methyl), frequently found in the barley samples, were investigated in detail in this study. The carryover percentages of these pesticides to malt, against the concentration of residues in barley grain, ranged from 22% for pirimiphos-methyl up to 78% for fenpropimorph. The results confirm a general rule that residues of pesticides with log P values >2 remain on the malt, but it was found that their transfer potential is more related to its individual physical-chemical properties but does not much correlate to their log P values. In the second part of the study, a noticeable carryover from malt to sweet wort was observed for pyraclostrobin, fenpropimorph, and tebuconazole residues, and these values ranged from 2 to 15%. Moreover, the analysis of pesticide residues in spent grain after mashing revealed that the spent grain samples contain on average once as much pyraclostrobin and tebuconazole residues as the original malt. It was concluded that (1) pyraclostrobin and tebuconazole residues could be incorporated into or associated with macromolecules in barley grain to form "hidden" (bound) forms, and (2) the parent compounds are subsequently released from their hidden forms during mashing.

Targeted next generation sequencing of MLH1-deficient, MLH1 promoter hypermethylated, and BRAF/RAS-wild-type colorectal adenocarcinomas is effective in detecting tumors with actionable oncogenic gene fusions.

Oncogenic gene fusions represent attractive targets for therapy of cancer. However, the frequency of actionable genomic rearrangements in colorectal cancer (CRC) is very low, and universal screening for these alterations seems to be impractical and costly. To address this problem, several large scale studies retrospectivelly showed that CRC with gene fusions are highly enriched in groups of tumors defined by MLH1 DNA mismatch repair protein deficiency (MLH1d), and hypermethylation of MLH1 promoter (MLH1ph), and/or the presence of microsatellite instability, and BRAF/KRAS wild-type status (BRAFwt/KRASwt). In this study, we used targeted next generation sequencing (NGS) to explore the occurence of potentially therapeutically targetable gene fusions in an unselected series of BRAFwt/KRASwt CRC cases that displayed MLH1d/MLH1ph. From the initially identified group of 173 MLH1d CRC cases, 141 cases (81.5%) displayed MLH1ph. BRAFwt/RASwt genotype was confirmed in 23 of 141 (~16%) of MLH1d/MLH1ph cases. Targeted NGS of these 23 cases identified oncogenic gene fusions in nine patients (39.1%; CI95: 20.5%-61.2%). Detected fusions involved NTRK (four cases), ALK (two cases), and BRAF genes (three cases). As a secondary outcome of NGS testing, we identified PIK3K-AKT-mTOR pathway alterations in two CRC cases, which displayed PIK3CA mutation. Altogether, 11 of 23 (~48%) MLH1d/MLH1ph/BRAFwt/RASwt tumors showed genetic alterations that could induce resistance to anti-EGFR therapy. Our study confirms that targeted NGS of MLH1d/MLH1ph and BRAFwt/RASwt CRCs could be a cost-effective strategy in detecting patients with potentially druggable oncogenic kinase fusions.

Results of morphological screening for Lynch syndrome during the period 2013-2016.

The introduction of a screening system for Lynch syndrome in pathology laboratories in Plzen yielded 24 diagnoses of Lynch syndrome during the period of 2013-2016, 20 of them presenting with colorectal cancer. In 8 of those 24 cases germline mutations of MMR genes, previously not recognized as pathogenic with certainty, were detected. Although the frequency of Lynch syndrome in patients with colorectal cancer was only 0.34 % in total, following introduction of the universal immunohistochemical investigation of MMR (mismatch repair) proteins expression in all colorectal cancers examined in Sikl´s Institute of Pathology the frequency per year in this department reached 2.4 %. The results favor universal immunohistochemical screening for Lynch syndrome in colorectal and endometrial cancer cases over a selective approach based on a combination of clinical and morphological criteria. Increased effectiveness of the universal approach is not brought about only by higher sensitivity of the immunohistochemical examination per se, but also by the possibility of automation of the process leading to increased adherence even of pathologists not directly engaged in Lynch syndrome management. However, the introduction of a nation-wide universal screening system requires support from the government and health insurance companies. Keywords: colorectal cancer - endometrial cancer - immunohistochemistry - Lynch syndrome - MMR - screening.

Tracking, Behavior and Fate of 58 Pesticides Originated from Hops during Beer Brewing.

The study presents tracking of 58 pesticide residues associated with hops to estimate their carryover into brewed beer. The pesticides were spiked onto organic hops at a concentration of 15 mg/kg, and the wort was boiled with the artificially contaminated hops and fermented on a laboratory scale. Samples were collected during the whole brewing process and pesticide residues were extracted using a method known as QuEChERS (quick, easy, cheap, effective, rugged, and safe). An HPLC-HR-MS/MS method was developed and validated to identify and quantitate pesticide residues in treated hops, spent hops, hopped wort, green beer, and beer samples. Quantitation was achieved using standard addition with isotopically labeled standards. The carryover percentages into hopped wort and the percentages of decay reduction relative to the amount spiked on hops were calculated. The relationship between the partition coefficients n-octanol-water (log P values) and the residual ratios ( RW and RB) of a pesticide were evaluated to predict their behavior during hopping of wort and fermentation. Pesticides with a high log P values (>3.75) tended to remain in spent hops. The pesticides that have a low log P value up to approximately 3 could represent the demarcation lines of appreciable transfer rate of pesticides from hops to beer. Consequently, the pesticides were divided into three categories depending upon their fate during the brewing process. The most potential risk category represents a group involving the thermostable pesticides, such as azoxystrobin, boscalid, dimethomorph, flonicamid, imidacloprid, mandipropamid, myclobutanil, and thiamethoxam, which were transferred at high rates from the pesticide enriched hops into beer during the laboratory brewing trial. These results can be used as a guideline in the application of pesticides on hop plants that would reduce the level of pesticide residues in beer and their exposure in humans.

Inhibitory effect of hop fractions against Gram-positive multi-resistant bacteria. A pilot study.

AIM: Our research focused on the antimicrobial effects of purified hop (Humulus lupulus L.) fractions including α-bitter acids (humulones), ß-bitter acids (lupulones) and xanthohumol, and a commercial CO2 hop extract of bitter acids against reference and multi-resistant strains of Gram-positive and Gram-negative bacteria and against selected yeast strains. METHODS: In vitro testing of antimicrobial activity was performed according to standard testing protocols (EUCAST). The effects of hop extracts on bacterial/yeast strains at concentrations below MICs were also determined and the antimicrobial potential of hop extracts was compared with selected antibiotics using optical density measurement. RESULTS: The fractions were effective not only against reference strains of Gram-positive bacteria but, more importantly, against their methicillin- and vancomycin-resistant variants. No antimicrobial effect was detected against Gram-negative bacterial strains. Among the tested substances, xanthohumol was identified as the hop fraction with the most potent antimicrobial properties. It was also found that hop substances exerted their antimicrobial effects at concentrations considerably lower than the determined MICs, with the strongest effect in case of α-bitter acids in enterococci. CONCLUSION: The search for and research of new compounds with antimicrobial properties represents a possible solution to the current global problem of bacterial resistance. Our data suggest a desirable activity of hop fractions against some multi-resistant bacterial strains. Thus, hops might find use as a source of potential antimicrobial agents applicable in both human and veterinary medicine.

Antibiofilm activity of bioactive hop compounds humulone, lupulone and xanthohumol toward susceptible and resistant staphylococci.

Bacterial biofilms pose a serious medical problem due to their significant resistance to antimicrobials, and staphylococci are recognized as the most frequent cause of biofilm-associated infections. The hop plant (Humulus lupulus L.) contains substances that have been determined to act as anti-infective agents against bacteria, mainly in planktonic form. Therefore, we decided to investigate the antibiofilm properties of H. lupulus L.-derived compounds (humulone, lupulone and xanthohumol) against a selected group of Staphylococcus spp., including methicillin-susceptible and resistant strains. All tested hop compounds were shown to possess antimicrobial properties against all tested staphylococci, both planktonic and biofilm-dwelling, with no significant difference between resistant and susceptible strains. All compounds lowered the number of bacterial cells released from the biofilm, with the strongest effect seen for lupulone, followed by xanthohumol. Moreover, lupulone and xanthohumol were not only able to penetrate the biofilm and reduce the number of bacteria within it, but their higher concentrations (∼60 µg/mL for xanthohumol and ∼125 µg/mL for lupulone) reduced the number of surviving bacterial cells to zero.

Renal Cell Carcinoma With Leiomyomatous Stroma: A Group of Tumors With Indistinguishable Histopathologic Features, But 2 Distinct Genetic Profiles: Next-Generation Sequencing Analysis of 6 Cases Negative for Aberrations Related to the VHL gene.

We have studied a cohort of renal cell carcinomas (RCCs) with smooth-muscle stroma (N=6), which lacked any of following genetic aberrations: mutations in the VHL-gene-coding sequence, loss of heterozygosity of chromosome 3p, or hypermethylation of VHL. Using targeted next-generation sequencing, no intronic VHL mutations or mutations in selected genes involved in angiogenesis and genes frequently mutated in clear cell RCC were identified. Tumors were also tested for the presence of hotspot mutations in the TCEB1 gene with negative results in all cases. We conclude that there exists a group of RCCs with abundant leiomyomatous stroma, where the epithelial component is indistinguishable from conventional clear cell RCC and distinct from clear cell (tubulo-) papillary RCC and that these tumors lack aberrations related to the function of the VHL gene, mutations in genes involved in angiogenesis, and hotspot mutations in the TCEB1 gene.

High-grade renal cell carcinoma with emperipolesis: Clinicopathological, immunohistochemical and molecular-genetic analysis of 14 cases.

Emperipolesis has recently been described as a constant feature of "biphasic squamoid" papillary renal cell carcinoma (BPRCC). We also noticed this in some high-grade (HG) RCC, which promoted the present study to estimate the incidence of emperipolesis in RCCs and to describe them in further detail. 14 cases of HGRCC showing emperipolesis were retrieved from our registry. Microscopic examination of filed slides was supplemented with immunohistochemical and molecular-genetic analyses using paraffin embedded tissue. 12 of 14 patients were males with a mean age of 58.6 years (range 41-72 years). Tumor size ranged from 6-16.5 cm (mean of 8.8 cm). Follow up data were available for 8/14 patients (range 0.5-10 years). Metastases were documented in 6 cases. All tumors showed solid-alveolar growth patterns with focal pseudopapillary features, and were composed of large cells with bizarre nuclei and eosinophilic rhabdoid-like cytoplasm. Emperipolesis was a constant and prominent feature in large bizarre cells. All cases were positive for OSCAR, CANH 9, vimentin, cyclin D1, INI-1, and myoD1, while negative for melanocytic markers, CK 7, myoglobin, cathepsin K, and TFE3. VHL gene abnormalities were found in 6/9 analyzable cases, of which 2 demonstrated polysomy of chromosomes 7, 17. Emperipolesis is a rare histomorphologic feature which can be seen not only in BPRCCs but also in highgrade CCRCCs. All RCC cases with prominent emperipolesis fulfilled both morphologic and immunohistochemical diagnostic criteria of high-grade CCRCC. The majority of patients with available follow up information developed metastases.

Strong antimicrobial activity of xanthohumol and other derivatives from hops (Humulus lupulus L.) on gut anaerobic bacteria.

Anaerobic bacteria, such as Bacteroides fragilis or Clostridium perfringens, are part of indigenous human flora. However, Clostridium difficile represents also an important causative agent of nosocomial infectious antibiotic-associated diarrhoea. Treatment of C. difficile infection is problematic, making it imperative to search for new compounds with antimicrobial properties. Hops (Humulus lupulus L.) contain substances with antibacterial properties. We tested antimicrobial activity of purified hop constituents humulone, lupulone and xanthohumol against anaerobic bacteria. The antimicrobial activity was established against B. fragilis, C. perfringens and C. difficile strains according to standard testing protocols (CLSI, EUCAST), and the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) were calculated. All C. difficile strains were toxigenic and clinically relevant, as they were isolated from patients with diarrhoea. Strongest antimicrobial effects were observed with xanthohumol showing MIC and MBC values of 15-107 µg/mL, which are close to those of conventional antibiotics in the strains of bacteria with increased resistance. Slightly higher MIC and MBC values were obtained with lupulone followed by higher values of humulone. Our study, thus, shows a potential of purified hop compounds, especially xanthohumol, as alternatives for treatment of infections caused by select anaerobic bacteria, namely nosocomial diarrhoea caused by resistant strains.

Programmed death-1 (PD-1) receptor/PD-1 ligand (PD-L1) expression in fumarate hydratase-deficient renal cell carcinoma.

Fumarate hydratase-deficient renal cell carcinoma (FH-RCC) is a rare and aggressive tumor affecting mostly younger patients. This is the first study to assess the expression of programmed death-1 (PD-1) receptor/PD-1 ligand (PD-L1) in FH-RCC. Formalin-fixed paraffin-embedded samples from 13 FH-RCCs collected in an international multi-institutional study, were evaluated by immunohistochemistry (IHC) for PD-1/PD-L1 reactivity in tumor cells and tumor infiltrating lymphocytes (TILs). PD-1/PD-L1 expression was further evaluated by qPCR. By IHC, PD-1 was negative in tumor cells in all 13 cases. PD-L1 was positive in tumor cells in 2/13 cases, weak positive in 7/13, and negative in 4/13 cases, respectively. In TILs, PD-1 was positive in 1/13, weak positive in 3/13, and negative in 9/13 cases. In TILs, PD-L1 was weak positive by IHC in 5/13, and negative in 8/13 cases, respectively. qPCR confirmed the result for 2 of 3 IHC weak positive PD-1 samples. Of 7 IHC weak positive samples (in tumor cells), PD-L1 mRNA was detected in all 7 tumors. The majority of FH-RCCs did not express PD-1/PD-L1 by IHC, which was confirmed by molecular analysis. PD-1/PD-L1 expression in FH-RCC is restricted to a proportion of cases which may benefit from targeted therapies.

Analysis of 100-Year-Old Beer Originated from the Czech Republic.

Three bottles of different beers were found in 2015 during a reconstruction of the brewery of the Raven Trading s.r.o. company in Záhlinice, Czech Republic. Thanks to good storage conditions, it was possible to analyze their original characteristics. All three bottles contained most probably lager type beer. One beer had sulfuric and fecal off-flavors; it was bright with the original extract of 10.3° Plato. The second beer, with an original extract of 7.6° Plato, was dark and very acidic, resembling Lambic. DNA analysis proved the presence of Dekkera bruxellensis, which corresponded to its chemical profile (total acidity, FAN, ethyl acetate, total esters). The third beer contained traces of carbon dioxide bubbles, was light brown and slightly bitter, with an original extract 10.4° Plato. Because it obviously underwent a natural aging process, sweetness, honey, and fruity off-flavors were detected and transformation products of iso-α-acids were found.

Warthin-like papillary renal cell carcinoma: Clinicopathologic, morphologic, immunohistochemical and molecular genetic analysis of 11 cases.

Oncocytic papillary renal cell carcinoma (PRCC) is a distinct subtype of PRCC, listed as a possible new variant of PRCC in the 2016 WHO classification. It is composed of papillae aligned by large single-layered eosinophilic cells showing linearly arranged oncocytoma-like nuclei. We analyzed clinicopathologic, morphologic, immunohistochemical and molecular-genetic characteristics of 11 oncocytic PRCCs with prominent tumor lymphocytic infiltrate, morphologically resembling Warthin's tumor. The patients were predominantly males (8/11, 73%), with an average age of 59years (range 14-76), and a mean tumor size of 7cm (range 1-22cm). Tumors had the features of oncocytic PRCCs with focal pseudostratification in 8/11 cases and showed dense stromal inflammatory infiltration in all cases. Papillary growth pattern was predominant, comprising more than 60% of tumor volume. Tubular and solid components were present in 5 and 3 cases, respectively. Uniform immunohistochemical positivity was found for AMACR, PAX-8, MIA, vimentin, and OSCAR. Tumors were mostly negative for carboanhydrase 9, CD117, CK20, and TTF-1. Immunohistochemical stains for DNA mismatch repair proteins MLH1 and PMS2 were retained in all cases, while MSH2 and MSH6 were negative in 1 case. Tumor infiltrating lymphocytes (TILs) consisted of both B and T cells. Chromosomal copy number variation analysis showed great variability in 5 cases, ranging from a loss of one single chromosome to complex genome rearrangements. Only one case showed gains of chromosomes 7 and 17, among other aberrations. In 4 cases no numerical imbalance was found. Follow up data was available for 9 patients (median 47.6months, range 1-132). In 6 patients no lethal progression was noted, while 3 died of disease. In conclusion, Warthin-like PRCC is morphologically very close to oncocytic PRCC, from which it differs by the presence of dense lymphoid stroma. Chromosomal numerical aberration pattern of these tumors is variable; only one case showed gains of chromosomes 7 and 17. Warthin-like PRCC is a potentially aggressive tumor since a lethal outcome was recorded in 3/9 cases.

Study on the metabolism of 5,6-methylenedioxy-2-aminoindane (MDAI) in rats: identification of urinary metabolites.

1. 5,6-Methylenedioxy-2-aminoindane (MDAI) is a member of aminoindane drug family with serotoninergic effect, which appeared on illicit drug market as a substitute for banned stimulating and entactogenic drugs. 2. Metabolism of MDAI, which has been hitherto unexplored, was studied in rats dosed with a subcutaneous dose of 20 mg MDAI.HCl/kg body weight. The urine of rats was collected within 24 h after dosing for analyses by HPLC-ESI-HRMS and GC/MS. 3. The main metabolic pathways proceeding in parallel were found to be oxidative demethylenation followed by O-methylation and N-acetylation. These pathways gave rise to five metabolites, namely, 5,6-dihydroxy-2-aminoindane, 5-hydroxy-6-methoxy-2-aminoindane, N-acetyl-5,6-methylenedioxy-2-aminoindane, N-acetyl-5,6-dihydroxy-2-aminoindane and N-acetyl-5-hydroxy-6-methoxy-2-aminoindane, which were found predominantly in the form of corresponding glucuronides and sulphates. However, the main portion of administered MDAI was excreted unchanged. 4. Minor metabolites formed primarily by hydroxylation at various sites include cis- and trans-1-hydroxy-5,6-methylenedioxy-2-aminoindane, 5,6-methylenedioxyindan-2-ol and 4-hydroxy-5,6-methylenedioxy-2-aminoindane. 5. Identification of all metabolites except for glucuronides, sulphates and tentatively identified 4-hydroxy-5,6-methylenedioxy-2-aminoindane was supported by synthesised reference standards.

Morphological, immunohistochemical, and chromosomal analysis of multicystic chromophobe renal cell carcinoma, an architecturally unusual challenging variant.

Chromophobe renal cell carcinoma (ChRCC) is typically composed of large leaf-like cells and smaller eosinophilic cells arranged in a solid-alveolar pattern. Eosinophilic, adenomatoid/pigmented, or neuroendocrine variants have also been described. We collected 10 cases of ChRCC with a distinct multicystic pattern out of 733 ChRCCs from our registry, and subsequently analyzed these by morphology, immunohistochemistry, and array comparative genomic hybridization. Of the 10 patients, 6 were males with an age range of 50-89 years (mean 68, median 69). Tumor size ranged between 1.2 and 20 cm (mean 5.32, median 3). Clinical follow-up was available for seven patients, ranging 1-19 years (mean 7.2, median 2.5). No aggressive behavior was documented. We observed two growth patterns, which were similar in all tumors: (1) variable-sized cysts, resembling multilocular cystic neoplasm of low malignant potential and (2) compressed cystic and tubular pattern with slit-like spaces. Raisinoid nuclei were consistently present while necrosis was absent in all cases. Half of the cases showed eosinophilic/oncocytic cytology, deposits of pigment (lipochrome) and microcalcifications. The other half was composed of pale or mixed cell populations. Immunostains for epithelial membrane antigen (EMA), CK7, OSCAR, CD117, parvalbumin, MIA, and Pax 8 were positive in all tumors while negative for vimentin, TFE3, CANH 9, HMB45, cathepsin K, and AMACR. Ki67 immunostain was positive in up to 1 % of neoplastic cells. Molecular genetic examination revealed multiple chromosomal losses in two fifths analyzable tumors, while three cases showed no chromosomal numerical aberrations. ChRCC are rarely arranged in a prominent multicystic pattern, which is probably an extreme form of the microcystic adenomatoid pigmented variant of ChRCC. The spectrum of tumors entering the differential diagnosis of ChRCC is quite different from that of conventional ChRCC. The immunophenotype of ChRCC is identical with that of conventional ChRCC. Chromosomal numerical aberration pattern was variable; no chromosomal numerical aberrations were found in three cases. All the cases in this series have shown an indolent and non-aggressive behavior.