Minna Kujala

Expression of growth factors and growth factor receptors in capillary hemangioblastoma.

To elucidate the mechanisms underlying the regulation of growth and differentiation of capillary hemangioblastoma we studied the expression of selected growth factors and growth factor receptors by immunocytochemistry. As stromal cells of capillary hemangioblastoma express high levels of vascular endothelial growth factor (VEGF) and placental growth factor (PIGF) mRNA, we studied the distribution of the corresponding VEOF and PIGF proteins. We also studied the expression of epidermal growth factor receptor (EGFR) and platelet-derived growth factor receptors (PDGFR) because their ligands have been reported to promote angiogenesis. The stromal cells expressed abundant EGFR and, in addition, some stromal cells expressed PDGFR-α but not PDGFR-β. In contrast, the endothelial cells co-expressed PDGFR-α and PDGFR-β. VEGF and PIGF were expressed by scattered stromal cells; however, more intense staining was observed in the endothelial cells of the intratumoral blood vessels, possibly indicating the secreted growth factors bound to their target receptors. We conclude that capillary hemangioblastomas express a variety of growth factor receptors and ligands, potentially involved in both autocrine and paracrine loops. The uniformly high EGFR expression is unique among brain tumors and may be associated with the typical morphology of capillary hemangioblastoma. The expression of highly angiogenic growth factors and their receptors may contribute to the rich vascularity of this enigmatic tumor.

Differential Regulation of Basolateral Cl−/HCO3− Exchangers SLC26A7 and AE1 in Kidney Outer Medullary Collecting Duct

SLC26A7 is a recently identified Cl(-)/HCO(3)(-) exchanger that co-localizes with AE1 on the basolateral membrane of Alpha intercalated cells (A-IC) in outer medullary collecting duct (OMCD). The purpose of these studies was to determine whether AE1 and SLC26A7 are differentially regulated in OMCD in pathophysiologic states. Toward this end, the expression and regulation of AE1 and SLC26A7 was examined in water deprivation, a condition known to increase the osmolality of the medulla. Rats were subjected to 3 d of water deprivation while having free access to food. Northern hybridizations demonstrated that in the outer medulla, the mRNA expression of SLC26A7 increased by approximately 300% (P < 0.01 versus control; n = 3), whereas the expression of AE1 decreased by approximately 50% (P < 0.05 versus control, n = 3) in water-deprived rats. Immunoblot analysis studies demonstrated that in the outer medulla, SLC26A7 abundance increased by approximately 3.5-fold (P < 0.02 versus control; n = 3), whereas the AE1 abundance decreased by approximately 55% (P < 0.05 versus control) in water deprivation. The expression of SLC26A7 remained unchanged in the kidney cortex and stomach in water deprivation, indicating the specificity of SLC26A7 upregulation in outer medulla. In situ hybridization indicated the exclusive expression of SLC26A7 in the outer medulla and double immunofluorescence labeling confirmed the co-localization of AE1 and SLC26A7 on the basolateral membrane of A-IC cells in OMCD. It is concluded that AE1 and SLC26A7 are differentially regulated in OMCD in water deprivation. On the basis of these results and previous functional studies indicating the activation of SLC26A7 activity by high osmolality, it is proposed that SLC26A7 may play an important role in bicarbonate reabsorption and or cell volume regulation in OMCD (specifically under hypertonic conditions).

Mutation screening of fumarate hydratase by multiplex ligation-dependent probe amplification: detection of exonic deletion in a patient with leiomyomatosis and renal cell cancer

Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a syndrome predisposing to cutaneous and uterine leiomyomatosis as well as renal cell cancer and uterine leiomyosarcoma. Heterozygous germline mutations in the fumarate hydratase (FH, fumarase) gene are known to cause HLRCC. On occasion, no FH mutation is detected by direct sequencing, despite the evident HLRCC phenotype in a family. In the present study, to investigate whole gene or exonic deletions and amplifications in FH mutation-negative patients, we used multiplex ligation-dependent probe amplification technology. The study material comprised 7 FH mutation-negative HLRCC patients and 12 patients affected with HLRCC-associated phenotypes, including papillary RCC, early-onset RCC, uterine leiomyomas, or uterine leiomyosarcoma. A novel FH mutation, a deletion of FH exon 1 that encodes the mitochondrial signal peptide, was detected in one of the HLRCC patients (1/7). The patient with the FH mutation displayed numerous painful cutaneous leiomyomas and papillary type renal cell cancer. Our finding, together with the two patients with whole FH gene deletion who had been detected previously, suggests that exonic or whole-gene FH deletions are not a frequent cause of HLRCC syndrome.

SLC26A6 and SLC26A7 Anion Exchangers Have a Distinct Distribution in Human Kidney

Background: The anion transporters SLC26A6 (PAT1) and SLC26A7, transporting at least chloride, oxalate, sulfate and bicarbonate, show a distinct expression and function in different mammalian species. They are expressed in kidney, but their exact localization in human kidney has not been studied. We therefore examined SLC26A6 and A7 expression in human kidneys. Methods: The localization of SLC26A6 and A7 in different segments of human nephrons was studied by RT-PCR and immunohistochemistry by comparing to the tubular markers PNRA, CD10, Tamm-Horsfall antigen, high molecular weight cytokeratin, CK7, AQP2 and H+V-ATPase. Results: In human kidney, SLC26A6 is expressed in distal segments of proximal tubules, parts of the thin and thick ascending limbs of Henle’s loops, macula densa, distal convoluted tubules and a subpopulation of intercalated cells of collecting ducts. SLC26A7 is expressed in extraglomerular mesangial cells and a subpopulation of intercalated cells of collecting ducts. Conclusion: Our results show that in human kidney SLC26A6 and A7 have a distinct, partially overlapping expression in distal segments of nephrons. The distribution partly differs from that found previously in rodent kidneys.

Sole ulcers in Finnish dairy cattle.

The Finnish Healthy Hooves project was set up to determine the frequency of, and risk factors for various hoof lesions in Finnish dairy herds. Data were collected in the years 2003 and 2004. A large dataset of over 74,000 cow-level observations recorded by hoof trimmers was merged with production data from the Finnish Agricultural Data Processing Centre Ltd. Ultimately, data from a single lactation from each of 16,792 cows in 703 herds were used for the analyses in this paper. Three-level hierarchical logistic models with hoof trimmer and farms (within hoof trimmer) as random effects were fit to data sets of tie stall (TS) and loose housing (LH) herds separately. The outcome of interest was the presence or absence of a sole ulcer in one or more legs of a cow during the lactation of interest. Cows examined once had a risk of sole ulcer 5.23% in tie stall herds and 7.58% in LH herds. As the number of examinations increased the odds of a diagnosis of sole ulcer increased substantially (2 and 3+ examinations had odds ratios (ORs) of 1.42 and 3.42 in TS herds and 2.77 and 6.89 in LH herds). Breed had a large effect on the risk of sole ulcer with Holsteins 2.89 times more likely to be affected than Ayrshires in TS herds and 2.94 times in LH herds. In TS herds, the presence of other hoof lesions such as haemorrhages (OR = 2.97), heel-horn erosions (OR = 2.10) and corkscrew claw (OR = 2.83) increased the risk of a sole ulcer developing. In LH herds, only haemorrhages (OR = 1.80) were a significant risk factor when parity was > or = 2. In TS herds, use of mats (compared to hard flooring) significantly reduced the risk of sole ulcers (OR = 0.49). The effect of parity on the risk of sole ulcer was greatest when parity > or = 4 but this effect was only significant in tie stalls (OR = 1.86). When analyses were restricted to cows with parity > or = 2, similar results were obtained for the risk factors identified above. In addition, parity became highly significant in TS and LH (OR 2.31 and 2.23, respectively when parity was 4+). In TS herds, herd average milk production was significantly associated with a decrease risk of sole ulcer (OR = 1.28 per 1000 kg decrease) but there was no effect of production at the cow level (measured as deviation from the herd mean). No significant effects of production were observed in LH herds.

Inherited forms of renal cell carcinoma.

It is estimated that up to 2% of renal cell cancer (RCC) clusters in families. Several forms of hereditary RCC have been characterized with specific clinical, histopathological, and genetic features. The most common of these is von Hippel-Lindau (VHL) disease caused by mutations in the VHL gene and predisposing to clear cell RCC. Predisposition to papillary RCC is present in hereditary leiomyomatosis and renal cell cancer (HLRCC) and hereditary papillary renal cell carcinoma (HPRC). Identification of the genetic defects causing these diseases has enlightened the molecular pathogenesis of RCC, and moreover, provided means to improve patient management. Genetic testing enables early diagnosis of the disease, after which individuals at-risk can be guided to regular surveillance. Screening facilitates detection of presymptomatic early tumors broadening treatment options and potentially improving prognosis. Thus, identification of individuals with inherited cancer susceptibility is important as special management of these patients improves disease outcome. The purpose of this review is to provide clues for identification and management of hereditary renal cancer patients in clinical practice.

Survey of interdigital phlegmon outbreaks and their risk factors in free stall dairy herds in Finland

BackgroundSevere outbreaks of interdigital phlegmon (IP) associated with a high morbidity and major economic losses have occurred in Finland in the past decade. A survey was performed to indicate the current occurrence of infectious hoof diseases and to identify herd level risk factors predisposing to an outbreak of IP.ResultsResponses to a questionnaire revealed that an outbreak of IP defined as morbidity ≥5% within the 1st month of the outbreak, had occurred in 18.0% of the respondent study farms. Risk factors for an outbreak included animal transport between herds, i.e. either animal purchase or contract heifer rearing, enlargement or renovation of the barn, and if the fields of the farm had been organically cultivated. Having any kind of mechanical ventilation in comparison to natural ventilation seemed to lower the risk of IP. Additionally, the farms that had experienced an outbreak of IP often had other infectious hoof diseases. However, it was unclear which disease appeared first.ConclusionsMore attention is needed before and during enlargement or renovation of the barn and substantial planning is crucial for every part of the enlargement process in dairy farms.