Familial non-syndromic clear cell renal cell carcinoma

The diagnosis of familial non-syndromic clear cell renal cell carcinoma is one of exclusion. In families presenting with clear cell RCC a germline VHL mutation and a constitutional translocation of chromosome 3 must be excluded before familial non-syndromic clear cell RCC can be diagnosed. Large familial non-syndromic clear cell RCC kindreds are uncommon and a predisposing gene has not been identified. However inheritance is autosomal dominant in most cases and age at onset is earlier than in sporadic cases. Recognition and appropriate screening of familial non-syndromic clear cell RCC cases will reduce morbidity and mortality. Large scale collaborative linkage studies may provide a basis for the identification of familial non-syndromic clear cell RCC susceptibility gene(s).     

Cytodiagnosis of clear cell hepatocellular carcinoma. A case report.

The cytologic features of clear cell hepatocellular carcinoma are described. This tumor may cause a diagnostic dilemma since it resembles other clear cell tumors originating in the adrenals, kidneys and ovaries. However, clear cell hepatocellular carcinoma possesses some characteristic features that permit a cytologic diagnosis to be made by fine needle aspiration, thus contributing to proper management.     

Identification of novel therapeutic targets in microdissected clear cell ovarian cancers

Clear cell ovarian cancer is an epithelial ovarian cancer histotype that is less responsive to chemotherapy and carries poorer prognosis than serous and endometrioid histotypes. Despite this, patients with these tumors are treated in a similar fashion as all other ovarian cancers. Previous genomic analysis has suggested that clear cell cancers represent a unique tumor subtype. Here we generated the first whole genomic expression profiling using epithelial component of clear cell ovarian cancers and normal ovarian surface specimens isolated by laser capture microdissection. All the arrays were analyzed using BRB ArrayTools and PathwayStudio software to identify the signaling pathways. Identified pathways validated using serous, clear cell cancer cell lines and RNAi technology. In vivo validations carried out using an orthotopic mouse model and liposomal encapsulated siRNA. Patient-derived clear cell and serous ovarian tumors were grafted under the renal capsule of NOD-SCID mice to evaluate the therapeutic potential of the identified pathway. We identified major activated pathways in clear cells involving in hypoxic cell growth, angiogenesis, and glucose metabolism not seen in other histotypes. Knockdown of key genes in these pathways sensitized clear cell ovarian cancer cell lines to hypoxia/glucose deprivation. In vivo experiments using patient derived tumors demonstrate that clear cell tumors are exquisitely sensitive to antiangiogenesis therapy (i.e. sunitinib) compared with serous tumors. We generated a histotype specific, gene signature associated with clear cell ovarian cancer which identifies important activated pathways critical for their clinicopathologic characteristics. These results provide a rational basis for a radically different treatment for ovarian clear cell patients.     

Three-dimensional distribution of the clear zone of migrating osteoclasts on dentin slices in vitro

Osteoclasts are cells that dynamically alternate resorption and migration on bone surfaces, and have the special structure called ruffled borders and clear zones by transmission electron microscopy (TEM). However, TEM features, especially the distribution of the clear zone of osteoclasts during migration, remains unclear. This study aimed to examine osteoclasts cultured on dentin slices by TEM and clarify the features of migrating osteoclasts, especially the three-dimensional distribution of clear zones. Osteoclasts obtained from mice were cultured with dentin slices for 72 h, and then cells were fixed and the tartrate-resistant acid phosphatase (TRAP) activity was detected. Specimens were embedded in Epon, then TRAP-positive cells were serially sectioned by alternating semithin and ultrathin sections. The cells were examined by TEM and the three-dimensional structures were reconstructed by computer. By TEM, most TRAP-positive cells were resorbing osteoclasts with ruffled borders and a clear zone. There were osteoclasts without ruffled borders, and these cells had clear zone-like structures and lamellipodia. The three-dimensional reconstruction showed that resorbing osteoclasts had rounded contours and ring-shaped clear zones encircling ruffled borders, and that osteoclasts without ruffled borders had irregular and flat shapes; the clear zone-like structures showed a dot or patch-like distribution. The presence of lamellipodia of the osteoclasts without ruffled borders shows that the cells are migrating osteoclasts. These results suggest that dot or patch-like distribution is the feature of the clear zone of osteoclasts during migration, and that these structures play the role of focal contacts and adhesion to the dentin surfaces during cell migration.     

Human gallbladder mucosa ultrastructure: evidence of intraepithelial nerve structures

The zinc iodide-osmium tetroxide (ZIO) fixative-staining method was used along with topographical ultrastructure to investigate cholecystectomized human gallbladders under light and electron microscopic techniques. This method delineated neuronal structures which may be involved in controlling the functions of the gallbladder epithelium. Three epithelial cell types were described in the surface epithelium: 1) Columnar clear cells; 2) dark, tuft osmiophilic cells; and 3) basal clear cells with electron-dense granules and showing intense ZIOphilic staining properties. While mucous granules were delineated in the first two cell types, the columnar epithelial clear cells are of uncertain function(s) and content but are probably absorptive cells. The small basal clear cells displaying intense ZIOphilia are associated with intraepithelial nerve endings. These nerve structures may have a sensory and/or motor function(s); they were detected throughout the gallbladder epithelial lining and mucosa.     

Cytomorphologic features of the rare epithelial-myoepithelial carcinoma of the salivary gland

The cytomorphologic features of an epithelial-myoepithelial carcinoma arising in the submandibular gland are reported. The aspiration smears were cellular, with two cell types present: dark cells showing attempted gland formation and clear cells with vacuolization. The clear cells frequently formed globular masses and had indistinct cytoplasmic membranes. These globular masses of clear cells were usually adjacent to the fragments of dark cells. A striking feature was the presence of numerous naked nuclei.     

The pollen tube clear zone:Clues to the mechanism of polarized growth

Pollen tubes usually exhibit a prominent region at their apex called the “clear zone” because it lacks light refracting amyloplasts. A robust, long clear zone often associates with fast growing pollen ...     

Different types of thermoconditional clear plaque-mutants and prophage induction by heat or cold of Serratia phage kappa.

Phage kappa of Seratia marcescens was treated with different mutagens to induce thermoconditional clear plaque-mutants. 176 mutants obtained were analysed by crosses and found to be located in clear plaque-region III. Two types resembling the mutants t2 and t1 of phage lambda were identified. Lysogens for the mutant 126 can be induced by heat and even by cold whereas they are scarcely inducible by uv. Nevertheless, a 126 prophage like a uv inducible ct 163 prophage can be sensitized to thermoinduction by short preirradiation if the cells are incubated for 30 to 45 min between uv exposure and heating. With ct 163 this time corresponds to the minimum extension of latent period after uv induction compared with infection at low moi. A mutant of clear plaque-region II, c154, shows an inverted thermoconditional behaviour forming clear plaques at 30 degrees C and turbid plaques due to lysogenization at 37 degrees C.     

Simple and inexpensive preparation of dried polyacrylamide gels for densitometry and/or storage

Polyacrylamide gels soaked in glycerol, dried at room temperature, and sealed with a clear sheet of inexpensive polypropylene are clear, flat, flexible, and stable. The technique is simpler and less expensive than others that have been described and does not involve the use of protein glues.     

The effect of orientation on the clarity of Hermann grid illusory lines.

To test the hypothesis that Hermann grid illusory lines are most clear when the grid is presented at a 45 deg, each of 20 participants underwent 10 trials in each of two conditions ('make the lines least clear' and 'make the lines most clear') which were run using a method of adjustment. A matched-pairs t-test applied to the means of the two conditions was significant beyond the 0.01 level. One-group t-tests supported the claim that the illusory lines are least clear for a vertical/horizontal orientation of the grid and most clear when it is rotated 45 deg. The results substantiate, in a systematic manner, 70 years of claims about the effects of grid orientation on illusory line clarity.     

A Simple Staining Procedure for Detecting the true Acrosome Reaction in Buffalo (Bubalus bubalis) Spermatozoa

A simple dual staining procedure for detecting the true acrosome reaction in dried smears of buffalo spermatozoa is described. Trypan blue is used first to differentiate live from dead spermatozoa and the dried smears which have been prepared are stained with Giemsa for acrosome evaluation. Four categories of spermatozoa were recognized: A) live, intact acrosome (acrosome pink, postnuclear cap clear); B) dead, intact acrosome (acrosome pink, postnuclear cap blue); C) live, detached acrosome (acrosome clear, postnuclear cap clear); and D) dead, detached acrosome (acrosome clear, postnuclear cap blue). The procedure is simple, rapid and convenient for assessing true acrosome reaction in buffalo spermatozoa. Simultaneous assessment of sperm viability and its acrosomal status in dried smears makes this procedure attractive because the true acrosome reaction can be studied thoroughly at a later state after the incubation period.     

Growth of a cell culture on porous capillaries

The growth of BHK-21 culture cells on the outer surface and in the clear space of capillaries made of phenylone or polyacrylnitrile was studied. The cultivation of cells in the clear space of the capillary extends their survival time and outlines possible ways of continuous cultivation of tissue cells which are cultivated in the attached state.     

High resolution clear native electrophoresis for in-gel functional assays and fluorescence studies of membrane protein complexes

Clear native electrophoresis and blue native electrophoresis are microscale techniques for the isolation of membrane protein complexes. The Coomassie Blue G-250 dye, used in blue native electrophoresis, interferes with in-gel fluorescence detection and in-gel catalytic activity assays. This problem can be overcome by omitting the dye in clear native electrophoresis. However, clear native electrophoresis suffers from enhanced protein aggregation and broadening of protein bands during electrophoresis and therefore has been used rarely. To preserve the advantages of both electrophoresis techniques we substituted Coomassie dye in the cathode buffer of blue native electrophoresis by non-colored mixtures of anionic and neutral detergents. Like Coomassie dye, these mixed micelles imposed a charge shift on the membrane proteins to enhance their anodic migration and improved membrane protein solubility during electrophoresis. This improved clear native electrophoresis offers a high resolution of membrane protein complexes comparable to that of blue native electrophoresis. We demonstrate the superiority of high resolution clear native electrophoresis for in-gel catalytic activity assays of mitochondrial complexes I-V. We present the first in-gel histochemical staining protocol for respiratory complex III. Moreover we demonstrate the special advantages of high resolution clear native electrophoresis for in-gel detection of fluorescent labeled proteins labeled by reactive fluorescent dyes and tagged by fluorescent proteins. The advantages of high resolution clear native electrophoresis make this technique superior for functional proteomics analyses.     

Long-term Patterns of Shifts between Clear and Turbid States in Lake Krankesjön and Lake Tåkern

During the past century, Lake Tåkern and Lake Krankesjön, southern Sweden, have shifted repeatedly between a state of clear water and abundant submerged vegetation, and a state of turbid water and sparse vegetation. Long-term empirical data on such apparently alternative stable state dynamics are valuable as complements to modeling and experiments, although the causal mechanisms behind shifts are often difficult to identify in hindsight. Here, we summarize previous studies and discuss possible mechanisms behind the shifts. The most detailed information comes from monitoring of two recent shifts, one in each lake. In the 1980s, L. Krankesjön shifted to clear water following an expansion of sago pondweed, Potamogeton pectinatus. Water clarity increased when the pondweed was replaced by characeans. Zooplankton biomass in summer declined and the concentration of total phosphorus (TP) was reduced to half the previous level. The fish community changed over several years, including an increasing recruitment of piscivorous perch (Perca fluviatilis). An opposite directed shift to turbid water occurred in Lake Tåkern in 1995, when biomass of phytoplankton increased in spring, at the expense of submerged vegetation. Consistent with the findings in L. Krankesjön, phyto- and zooplankton biomass increased and the average concentration of TP doubled. After the shift to clear water in L. Krankesjön, TP concentration has increased during the latest decade, supporting the idea that accumulation of nutrients may lead to a long-term destabilization of the clear water state. In L. Tåkern, data on TP are inconclusive, but organic nitrogen concentrations oscillated during a 25-year period of clear water. These observations indicate that intrinsic processes cause gradual or periodic changes in system stability, although we cannot exclude the possibility that external forces are also involved. During such phases of destabilization of the clear water state, even small disturbances could possibly trigger a shift, which may explain why causes behind shifts are hard to identify even when they occur during periods of extensive monitoring.     

Low clouds and cloud immersion enhance photosynthesis in understory species of a southern Appalachian spruce-fir forest (USA)

High-altitude forests of the southern Appalachian Mountains (USA) are frequently immersed in clouds, as are many mountain forests. They may be particularly sensitive to predicted increases in cloud base altitude with global warming. However, few studies have addressed the impacts of immersion on incident sunlight and photosynthesis. Understory sunlight (photosynthetically active radiation, PAR) was measured during clear, low cloud, and cloud-immersed conditions at Mount Mitchell and Roan Mountain, NC (USA) along with accompanying photosynthesis in four representative understory species. Understory PAR was substantially less variable on immersed vs. clear days. Photosynthesis became light-saturated between ～100 and 400 μmol · m(-2) · s(-1) PAR for all species measured, corresponding closely to the sunlight environment measured during immersion. Estimated daily carbon gain was 26% greater on clear days at a more open canopy site but was 22% greater on immersed/cloudy days at a more closed canopy site. F(v)/F(m) (maximum photosystem II efficiency) in Abies fraseri seedlings exposed to 2.5 min full sunlight was significantly reduced (10%), indicating potential reductions in photosynthesis on clear days. In addition, photosynthesis in microsites with canopy cover was nearly 3-fold greater under immersed (2.6 mmol · m(-2) · h(-1)) vs. clear conditions (0.9 mmol · m(-2) · h(-1)). Thus, cloud immersion provided more constant PAR regimes that enhanced photosynthesis, especially in shaded microsites. Future studies are needed to predict the survival of these refugial forests under potential changes in cloud regimes.     

Improving design and analysis of research: lessons from clinical research

There is a clear need to optimise the use of animals in research and to ensure that the studies that are done make a worthwhile contribution to scientific knowledge. Research that has an inappropriate study design, or is improperly analysed or interpreted, may mislead and is not defensible. Published reviews of the quality of design and analysis in clinical studies are used to see what lessons can be drawn in respect of animal research. The need for clear and full reporting of research is discussed.     

ALDH1-High Ovarian Cancer Stem-Like Cells Can Be Isolated from Serous and Clear Cell Adenocarcinoma Cells,and ALDH1 High Expression Is Associated with Poor Prognosis

Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are defined as a small population of cancer cells that have high tumorigenicity. Furthermore, CSCs/CICs are resistant to several cancer therapies, and CSCs/CICs are therefore thought to be responsible for cancer recurrence after treatment and distant metastasis. In epithelial ovarian cancer (EOC) cases, disease recurrence after chemotherapy is frequently observed, suggesting ovarian CSCs/CICs are involved. There are four major histological subtypes in EOC, and serous adenocarcinoma and clear cell adenocarcinoma are high-grade malignancies. We therefore analyzed ovarian CSCs/CICs from ovarian carcinoma cell lines (serous adenocarcinoma and clear cell adenocarcinoma) and primary ovarian cancer cells in this study. We isolated ovarian CSCs/CICs as an aldehyde dehydrogenase 1 high (ALDH1^high) population from 6 EOC cell lines (3 serous adenocarcinomas and 3 clear cell adenocarcinomas) by the ALDEFLUOR assay. ALDH1^high cells showed greater sphere-forming ability, higher tumorigenicity and greater invasive capability, indicating that ovarian CSCs/CICs are enriched in ALDH1^high cells. ALDH1^high cells could also be isolated from 8 of 11 primary ovarian carcinoma samples. Immunohistochemical staining revealed that higher ALDH1 expression levels in ovary cancer cases are related to poorer prognosis in both serous adenocarcinoma cases and clear cell adenocarcinoma cases. Taken together, the results indicate that ALDH1 is a marker for ovarian CSCs/CICs and that the expression level of ALDH1 might be a novel biomarker for prediction of poor prognosis.     

Histidine uptake in the epidermis of lizards and snakes in relation to the formation of the shedding complex

Mammalian epidermis utilizes histidine-rich proteins (filaggrins) to aggregate keratin filaments and form the stratum corneum. Little is known about the involvement of histidine-rich proteins during reptilian keratinization. The formation of the shedding complex in the epidermis of snakes and lizards, made of the clear and the oberhautchen layers, determines the cyclical epidermal sloughing. Differently from snakes, keratohyalin-like granules are present in the clear layer of lizards. The uptake of tritiated histidine into the epidermis of two lizards and one snake has been studied by autoradiography in sections at progressive post-injection periods. At 40 min and 1 hr post-injection keratohyalin-like granules were not or poorly labeled. At 3-22 hr post-injection most of the labeling was present over suprabasal cells destined to form the shedding complex, in keratohyalin-like granules of the clear layer, and in the forming a-layer but was low in the forming b-layer, and in superficial keratinized layers. The analysis of the shedding complex in the pad lamellae (a specialized scale used for climbing) of a gecko showed that the setae and the cytoplasm of clear cells among them are main sites of histidine uptake at 4 hr post-injection. In the snake most of the labeling at 4 hr post-injection was localized in the shedding complex along the boundary between the clear and oberhautchen layers. The present study suggests that, in the epidermis of lepidosaurian reptiles, the synthesis of a histidine-rich protein is involved in the formation of the shedding layer and, as in mammals, in a-keratinization.     

Cytoskeletal mechanisms of ooplasmic segregation in annelid eggs

Annelid embryos are comprised of yolk-deficient animal and yolk-filled vegetal blastomeres. This "unipolar" organization along the animal-vegetal axis (in terms of ooplasmic distribution) is generated via selective segregation of yolk-free, clear cytoplasm to the animal blastomeres. The pathway that leads to the unipolar organization is different between polychaetes and clitellates (i.e., oligochaetes and hirudinidans). In polychaetes, the clear cytoplasm domain, which is established through ooplasmic segregation at the animal side of the egg, is simply cut up by unequal equatorial cleavage. In clitellates, localization of clear cytoplasm to animal blastomeres is preceded by unification of the initially separated polar domains of clear cytoplasm, which result from bipolar ooplasmic segregation. In this article, I have reviewed recent studies on cytoskeletal mechanisms for ooplasmic localization during early annelid development. Annelid eggs accomplish ooplasmic rearrangements through various combinations of three cytoskeletal mechanisms, which are mediated by actin microfilaments, microtubules and mitotic asters, respectively. One of the unique features of annelid eggs isthat a homologous process is driven by distinct cytoskeletal elements. Annelid eggs may provide an intriguing system to investigate not only mechanical aspects of ooplasmic segregation but also evolutionary divergence of cytoskeletal mechanisms that operate in a homologous process.