GaAs 904-nm laser irradiation improves myofiber mass recovery during regeneration of skeletal muscle previously damaged by crotoxin

This work investigated the effect of gallium arsenide (GaAs) irradiation (power: 5 mW; intensity: 77.14 mW/cm(2), spot: 0.07 cm(2)) on regenerating skeletal muscles damaged by crotoxin (CTX). Male C57Bl6 mice were divided into six groups (n = 5 each): control, treated only with laser at doses of 1.5 J or 3 J, CTX-injured and, CTX-injured and treated with laser at doses of 1.5 J or 3 J. The injured groups received a CTX injection into the tibialis anterior (TA) muscle. After 3 days, TA muscles were submitted to GaAs irradiation at doses of 1.5 or 3 J (once a day, during 5 days) and were killed on the eighth day. Muscle histological sections were stained with hematoxylin and eosin (H&E) in order to determine the myofiber cross-sectional area (CSA), the previously injured muscle area (PIMA) and the area density of connective tissue. The gene expression of MyoD and myogenin was detected by real-time PCR. GaAs laser at a dose of 3 J, but not 1.5 J, significantly increased the CSA of regenerating myofibers and reduced the PIMA and the area density of intramuscular connective tissue of CTX-injured muscles. MyoD gene expression increased in the injured group treated with GaAs laser at a dose of 1.5 J. The CTX-injured, 3-J GaAs laser-treated, and the CTX-injured and treated with 3-J laser groups showed an increase in myogenin gene expression when compared to the control group. Our results suggest that GaAs laser treatment at a dose of 3 J improves skeletal muscle regeneration by accelerating the recovery of myofiber mass.

Sponges as substrata and early life history of the tubulariid Zyzzyzus warreni (Cnidaria: Hydrozoa) in the Sao Sebastiao Channel, Brazil

The hydroid Zyzzyzus warreni is usually found in shallow coastal waters forming aggregations of solitary polyps embedded in demosponges. Early life history transformations and settlement responses by the actinulae of this hydroid were studied in the laboratory using 13 species of sponges and 2 species of algae collected in the Sao Sebastiao Channel (Brazil) as substrata. The absence of oral tentacles and mouth in the actinulae and early events of metamorphosis suggest that these larvae are unable to spend long periods in the plankton and attach quickly near conspecifics when a preferred substratum is encountered. The time required for settlement and the number of elicited settlings indicated four settlement responses: (a) frequent and short-time settlement, in actinulae exposed to Halichondria cebimarensis, Mycale angulosa, M. aff. americana, M. laxissima (skeleton) and Tedania ignis; (b) moderate and delayed settlement, in actinulae exposed to Aplysina caissara, A. fulva, Haliclona melana and M. microsigmatosa; (c) no settlement, in actinulae exposed to Suberites aurantiacus and to the algae Hypnea musciformis and Sargassum cymosum; and (d) lethal response, in actinulae exposed to Amphimedon viridis, Aplysilla rosea, Dragmacidon reticulatum and M. laxissima. These responses indicate a considerable degree of species discrimination by the actinulae and are consistent with substrata used by the hydroid in the natural environment.

Prediction and probability of neonatal outcome in isolated congenital diaphragmatic hernia using multiple ultrasound parameters

Objectives To evaluate the accuracy and probabilities of different fetal ultrasound parameters to predict neonatal outcome in isolated congenital diaphragmatic hernia (CDH). Methods Between January 2004 and December 2010, we evaluated prospectively 108 fetuses with isolated CDH (82 left-sided and 26 right-sided). The following parameters were evaluated: gestational age at diagnosis, side of the diaphragmatic defect, presence of polyhydramnios, presence of liver herniated into the fetal thorax (liver-up), lung-to-head ratio (LHR) and observed/expected LHR (o/e-LHR), observed/expected contralateral and total fetal lung volume (o/e-ContFLV and o/e-TotFLV) ratios, ultrasonographic fetal lung volume/fetal weight ratio (US-FLW), observed/expected contralateral and main pulmonary artery diameter (o/e-ContPA and o/eMPA) ratios and the contralateral vascularization index (Cont-VI). The outcomes were neonatal death and severe postnatal pulmonary arterial hypertension (PAH). Results Neonatal mortality was 64.8% (70/108). Severe PAH was diagnosed in 68 (63.0%) cases, of which 63 died neonatally (92.6%) (P < 0.001). Gestational age at diagnosis, side of the defect and polyhydramnios were not associated with poor outcome (P > 0.05). LHR, o/eLHR, liver-up, o/e-ContFLV, o/e-TotFLV, US-FLW, o/eContPA, o/e-MPA and Cont-VI were associated with both neonatal death and severe postnatal PAH (P < 0.001). Receiver-operating characteristics curves indicated that measuring total lung volumes (o/e-TotFLV and US-FLW) was more accurate than was considering only the contralateral lung sizes (LHR, o/e-LHR and o/e-ContFLV; P < 0.05), and Cont-VI was the most accurate ultrasound parameter to predict neonatal death and severe PAH (P < 0.001). Conclusions Evaluating total lung volumes is more accurate than is measuring only the contralateral lung size. Evaluating pulmonary vascularization (Cont-VI) is the most accurate predictor of neonatal outcome. Estimating the probability of survival and severe PAH allows classification of cases according to prognosis. Copyright (C) 2011 ISUOG. Published by John Wiley & Sons, Ltd.

Molluscicidal activity of some marine substances against the snail Biomphalaria glabrata (Mollusca, Planorbidae)

Freshwater snails of the genus Biomphalaria play a major role as intermediate hosts of Schistosoma mansoni, the etiologic agent of schistosomiasis. While Biomphalaria spp. control by molluscicides is one of the main strategies to reduce the snail population in infected areas, there are few effective molluscicides commercially available. Natural products may be considered as potentially useful and safe molluscicides. We have evaluated the molluscicidal activity of 12 extracts from ten marine organisms on adult and embryonic stages of Biomphalaria glabrata. Only extracts of the red algae Liagora farinosa and of the sponge Amphimedon viridis presented molluscicidal activity. Lethal concentration (LC)(50) values obtained were 120 mu g/mL for L. farinosa CH2Cl2 extract (apolar fraction) and 20 mu g/mL for A. viridis extract and halitoxin. The polar alga fraction and halitoxin had no effect on B. glabrata embryos. The algae apolar fraction was active on B. glabrata in all embryonic development stages, with LC50 values for blastulae at 42 mu g/mL, gastrulae at 124 mu g/mL, trochophore at 180 mu g/mL, and veliger at 222 mu g/mL. This is the first report of extracts from marine organisms which presented molluscicidal activity.

Assessment of the role of transcript for GATA-4 as a marker of unfavorable outcome in human adrenocortical neoplasms

Abstract Background Malignant neoplasia of the adrenal cortex is usually associated with very poor prognosis. When adrenocortical neoplasms are diagnosed in the early stages, distinction between carcinoma and adenoma can be very difficult to accomplish, since there is yet no reliable marker to predict tumor recurrence or dissemination. GATA transcription factors play an essential role in the developmental control of cell fate, cell proliferation and differentiation, organ morphogenesis, and tissue-specific gene expression. Normal mouse adrenal cortex expresses GATA-6 while its malignant counterpart only expresses GATA-4. The goal of the present study was to assess whether this reciprocal change in the expression of GATA factors might be relevant for predicting the prognosis of human adrenocortical neoplasms. Since human adrenal cortices express luteinizing hormone (LH/hCG) receptor and the gonadotropins are known to up-regulate GATA-4 in gonadal tumor cell lines, we also studied the expression of LH/hCG receptor. Methods We conducted a study on 13 non-metastasizing (NM) and 10 metastasizing/recurrent (MR) tumors obtained from a group of twenty-two adult and pediatric patients. The expression of GATA-4, GATA-6, and LH/hCG receptor (LHR) in normal and tumoral human adrenal cortices was analysed using reverse transcriptase-polymerase chain reaction (RT-PCR) complemented by dot blot hybridization. Results Messenger RNA for GATA-6 was detected in normal adrenal tissue, as well as in the totality of NM and MR tumors. GATA-4, by its turn, was detected in normal adrenal tissue, in 11 out of 13 NM tumors, and in 9 of the 10 MR tumors, with larger amounts of mRNA found among those presenting aggressive clinical behavior. Transcripts for LH receptor were observed both in normal tissue and neoplasms. A more intense LHR transcript accumulation was observed on those tumors with better clinical outcome. Conclusion Our data suggest that the expression of GATA-6 in human adrenal cortex is not affected by tumorigenesis. GATA-4 expression is more abundant in MR tumors, while NM tumors express more intensely LHR. Further studies with larger cohorts are needed to test whether relative expression levels of LHR or GATA-4 might be used as prognosis predictors.

Comparative analysis of the corps en cerise in several species of Laurencia (Ceramiales, Rhodophyta) from the Atlantic Ocean

Different species of Laurencia have proven to be a rich source of natural products yielding interesting bioactive halogenated secondary metabolites, such as terpenoids and acetogenins. It is shown that such compounds are accumulated in the spherical, reniform to claviform refractive inclusions called corps en cerise (CC), which are intensively osmiophilic and located mainly in the cortical cells of the thalli and also in trichoblast cells. Up to now, it was believed that CC were present only in these two kinds of cells. Recently, however, a species of Laurencia, L. marilzae, with CC in all cells of the thallus, i.e., cortical, medullary, including the pericentral and axial cells, as well as in the trichoblasts, was described from the Canary Islands, and subsequently also reported to Brazil and Mexico. Within the Laurencia complex, only Laurencia species produce CC. Since the species of Laurencia are targets of interest for the prospection of bioactive substances due to their potential antibacterial, antifungal, anticholinesterasic, antileishmanial, cytotoxic, and antioxidant activities, the present paper carries out a comparative analysis of the corps en cerise in several species of Laurencia from the Atlantic Ocean to obtain basic information that can support natural product bioprospection projects. Our results show that the number and size of the CC are constant within a species, independent of the geographical distribution, corroborating their use for taxonomical purposes to differentiate groups of species that present a lower number from those that have a higher number. In this regard, there was a tendency for the number of CC to be higher in some species of Laurencia from the Canary Islands. The presence of CC can also be used to distinguish species in which these organelles are present in all cells of the thallus from those in which CC are restricted to the cortical cells. Among the species analyzed, L. viridis displayed the most varied secondary metabolites composition, such as sesquiterpenes, diterpenes, triterpenes, all of which showed potent antiviral, cytotoxic, and antitumoral activities, including protein phosphatase type 2A (PP2A) inhibitory effects.

Lokalisation und Verteilung von Synapsen auf identifizierten Zellen im Zentralen Nervensystem von Drosophila melanogaster

Wir präsentieren zwei Techniken, mit deren Hilfe es erstmals möglich ist, zentralnervöse Synapsen im Zentralen Nervensystem (ZNS) von Drosophila melanogaster reproduzierbar darzustellen und experimentellen Methoden zugänglich zu machen. Die erste Technik beruht auf dem UAS/Gal4-System und ermöglicht die Expression markierter synaptischer Proteine in bekannten reproduzierbaren Neuronen. Die zweite Technik beruht auf der Einzel-Zelltransplantation und erlaubt die reproduzierbare Visualisierung von Synapsen in allen neuralen Zell-Linien des Drosophila Bauchmark.Mit Hilfe dieser Techniken konnte gezeigt werden, dass Neuriten im Bauchmark von Drosophila mindestens drei unterschiedliche Kompartimente aufweisen: 1. Primäre, häufig transversal verlaufende Neurite ohne Output-Synapsen, 2. Seitenneurite von vermutlich rein postsynaptischer Natur und 3. Seitenneurite, die präsynaptisch spezialisierte Regionen aufweisen. Des Weiteren konnten wir nachweisen, dass die Seitenneurite von Motroneuronen im ZNS vermutlich rein postsynaptisch sind. Weitere Beobachtungen veranlassen uns zu der Hypothese, dass motorneuronale Seitenneurite in Drosophila homolog oder analog zu Dendriten in Vertebraten sein könnten.Mit Hilfe der Transplantationstechnik untersuchten wir weiterhin die Funktion des Gens kakapo im ZNS. Obwohl kakapo für die Entwicklung von Synapsen an der Neuromuskulären Verbindung wichtig ist, konnten wir im ZNS keinen Phänotyp feststellen. Dies legt nahe, dass zwischen der Entwicklung von periphären und zentralnervösen Synapsen klare Unterschiede bestehen.

Biochar in perennial crops: nutritional, agronomical and environmental implications

Biochar is the solid C-rich matrix obtained by pyrolysis of biomasses, currently promoted as a soil amendment with the aim to offset anthropogenic C emissions, while ameliorating soil properties and growth conditions. Benefits from biochar seem promising, although scientific understandings are beginning to be explored. In this project, I performed a suite of experiments in controlled and in field conditions with the aims to investigate the effect of biochar on: a) the interaction with minerals; b) Fe nutrition in kiwifruit; c) soil leaching, soil fertility, soil CO2 emissions partitioning, soil bacterial profile and key gene expression of soil nitrification-involved bacteria; d) plant growth, nutritional status, yield, fruit quality and e) its physical-chemical changes as affected by long-term environmental exposure. Biochar released K, P and Mg but retained Fe, Mn, Cu and Zn on its surface which in turn hindered Fe nutrition of kiwifruit trees. A redox reaction on the biochar surface exposed to a Fe source was elucidated. Biochar reduced the amount of leached NH4+-N but increased that of Hg, K, P, Mo, Se and Sn. Furthermore, biochar synergistically interacted with compost increasing soil field capacity, fertility, leaching of DOC, TDN and RSOC, suggesting a priming effect. However, in field conditions, biochar did not affect yield, nutritional status and fruit quality. Actinomadura flavalba, Saccharomonospora viridis, Thermosporomyces composti and Enterobacter spp. were peculiar of the soil amended with biochar plus compost which exhibited the highest band richness and promoted gene expression levels of Nitrosomonas spp., Nitrobacter spp. and enzymatic-related activity. Environmental exposure reduced C, K, pH and water infiltration of biochar which instead resulted in a higher O, Si, N, Na, Al, Ca, Mn and Fe at%. Oxidation occurred on the aged biochar surface, it decreased progressively with depth and induced the development of O-containing functional groups, up to 75nm depth.

Mythos und Mathematik : Hermann Brochs Roman "Die Schuldlosen" und seine Essays

„Natürlich habe ich mich [...] unausgesetzt mit Mathematik beschäftigt, umso mehr als ich sie für meine erkenntnistheoretisch-philosophischen Studien brauchte, denn ohne Mathematik lässt sich kaum mehr philosophieren.“, schreibt Hermann Broch 1948, ein Schriftsteller, der ca. zehn Jahre zuvor von sich selbst sogar behauptete, das Mathematische sei eine seiner stärksten Begabungen.rnDiesem Hinweis, die Bedeutung der Mathematik für das Brochsche Werk näher zu untersuchen, wurde bis jetzt in der Forschung kaum Folge geleistet. Besonders in Bezug auf sein Spätwerk Die Schuldlosen fehlen solche Betrachtungen ganz, sie scheinen jedoch unentbehrlich für die Entschlüsselung dieses Romans zu sein, der oft zu Unrecht als Nebenarbeit abgewertet wurde, weil ihm „mit gängigen literaturwissenschaftlichen Kategorien […] nicht beizukommen ist“ (Koopmann, 1994). rnDa dieser Aspekt insbesondere mit Blick auf Die Schuldlosen ein Forschungsdesiderat darstellt, war das Ziel der vorliegenden Arbeit, Brochs mathematische Studien genauer nachzuvollziehen und vor diesem Hintergrund eine Neuperspektivierung der Schuldlosen zu leisten. Damit wird eine Grundlage geschaffen, die einen adäquaten Zugang zur Struktur dieses Romans eröffnet.rnDie vorliegende Arbeit ist in zwei Teile gegliedert. Nach einer Untersuchung von Brochs theoretischen Betrachtungen anhand ausgewählter Essays folgt die Interpretation der Schuldlosen aus diesem mathematischen Blickwinkel. Es wird deutlich, dass Brochs Poetik eng mit seinen mathematischen Anschauungen verquickt ist, und somit nachgewiesen, dass sich die spezielle Bauform des Romans wie auch seine besondere Form des Erzählens tatsächlich aus dem mathematischen Denken des Autors ableiten lassen. Broch nutzt insbesondere die mathematische Annäherung an das Unendliche für seine Versuche einer literarischen Erfassung der komplexen Wirklichkeit seiner Zeit. Dabei spielen nicht nur Elemente der fraktalen Geometrie eine zentrale Rolle, sondern auch Brochs eigener Hinweis, es handele sich „um eine Art Novellenroman“ (KW 13/1, 243). Denn tatsächlich ergibt sich aus den poetologischen Forderungen Brochs und ihren Umsetzungen im Roman die Gattung des Novellenromans, wie gezeigt wird. Dabei ist von besonderer Bedeutung, dass Broch dem Mythos eine ähnliche Rolle in der Literatur zuspricht wie der Mathematik in den Wissenschaften allgemein.rnMit seinem Roman Die Schuldlosen hat Hermann Broch Neuland betreten, indem er versuchte, durch seine mathematische Poetik die komplexe Wirklichkeit seiner Epoche abzubilden. Denn „die Ganzheit der Welt ist nicht erfaßbar, indem man deren Atome einzelweise einfängt, sondern nur, indem man deren Grundzüge und deren wesentliche – ja, man möchte sagen, deren mathematische Struktur aufzeigt“ (Broch).

Temperature effects on larval growth and survival in five species of Caribbean Echinoids

Under the global change scenario, the possible effects of ocean warming were investigated on the larvae of five species of Caribbean Echinoids: Echinometra lucunter, Echinometra viridis, Clypeaster rosaceus, Tripneustes ventricosus and Lytechinus williamsi. Their thermal tolerance was evaluated rearing them for six days under different temperature regimes (26, 28, 30, 32, 34, 36°C). The larval sensitivity to the treatments was evaluated on the base of survival and growth. The rearing at higher temperatures has revealed a great suffering state of the larvae by inducing both reduction of live larvae and abnormality in their development. The extent of impact of the treatments varied from species to species, evidencing different levels of thermal tolerance. Anyway, higher temperature treatments have shown a general lethal threshold at about 34°C for most of the species. As an exception, the lethal threshold of Echinometra species was 36°C, few larvae of which being still capable of survive at the temperature of 34°C. The studies have also analyzed the effect of water warming on the larvae growth in terms of size and symmetry. The results put in evidence the presence of a critical upper temperature (about 32°C) at which the larvae of all species reveal a great suffering state that translates in the reduction of size (i.e., of body, stomach and postero-dorsal arm) and abnormalities (i.e., strong difference in the lengths of the two postero-dorsal arms). As sea surface temperatures are predicted to increase of 4-5°C by 2100, the high percentage of abnormal larvae and their scarce survival observed at 32- 34°C treatments indicate that the early stages of these species could be affected by future global warming.

Effects of acute temperature increase on performance and survival of Caribbean echinoids

Climate change is occurring at a faster rate than in the past, with an expected increase of mean sea surface temperatures up to 4.8°C by the end of this century. The actual capabilities of marine invertebrates to adapt to these rapid changes has still to be understood. Adult echinoids play a crucial role in the tropical ecosystems where they live. Despite their role, few studies about the effect of temperature increase on their viability have been reported in literature. This thesis work reports a first systematic study on several Caribbean echinoids about their tolerance to temperature rise in the context of global warming. The research - carried out at the Bocas del Toro Station of the Smithsonian Tropical Research Institute, in Panama - focalized on the 6 sea urchins Lytechinus variegatus, L. williamsi, Echinometra lucunter, E. viridis, Tripneustes ventricosus and Eucidaris tribuloides, and the 2 sand dollars Clypeaster rosaceus and C. subdepressus. Mortality and neuromuscular well-being indicators - such as righting response, covering behaviour, adhesion to the substrate, spine and tube feet movements - have been analysed in the temperature range 28-38°C. The righting time RT (i.e., the time necessary for the animal to right itself completely after inversion) measured in the 6 sea urchin species, demonstrated a clearly dependence on the water temperature. The experiments allowed to determine the “thermal safety margin” (TSM) of each species. Echinometra lucunter and E. viridis resulted the most tolerant species to high temperatures with a TSM of 5.5°C, while T. ventricosus was the most vulnerable with a TSM of only 3°C. The study assessed that all the species already live at temperatures close to their upper thermal limit. Their TSMs are comparable to the predicted temperature increase by 2100. In absence of acclimatization to such temperature change, these species could experience severe die-offs, with important consequences for tropical marine ecosystems.

Common CFTR haplotypes and susceptibility to chronic pancreatitis and congenital bilateral absence of the vas deferens

CFTR mutations enhance susceptibility for idiopathic chronic pancreatitis (ICP) and congenital bilateral absence of the vas deferens (CBAVD); however, it is unknown why CFTR heterozygotes are at increased disease risk. We recently showed that common CFTR variants are associated with aberrantly spliced transcripts. Here, we genotyped for common CFTR variants and tested for associations in two ICP (ICP-A: 126 patients, 319 controls; ICP-B: 666 patients, 1,181 controls) and a CBAVD population (305 patients, 319 controls). Haplotype H10 (TG11-T7-470V) conferred protection (ICP-A: OR 0.19, P<0.0001; ICP-B: OR 0.78, P = 0.06; CBAVD OR 0.08, P<0.001), whereas haplotype H3 (TG10-T7-470M) increased disease risk (ICP-A: OR 8.34, P = 0.003; ICP-B: OR 1.88, P = 0.007; CBAVD: OR 5.67, P = 0.01). The risk of heterozygous CFTR mutations carriers for ICP (OR 2.44, P<0.001) and CBAVD (OR 14.73, P<0.001) was fully abrogated by the H10/H10 genotype. Similarly, ICP risk of heterozygous p.Asn34Ser SPINK1 mutation carriers (OR 10.34, P<0.001) was compensated by H10/H10. Thus, common CFTR haplotypes modulate ICP and CBAVD susceptibility alone and in heterozygous CFTR and p.Asn34Ser mutation carriers. Determination of these haplotypes helps to stratify carriers into high- and low-risk subjects, providing helpful information for genetic counseling.

Impact of Holocene climate changes on alpine and treeline vegetation at Sanetsch Pass, Bernese Alps, Switzerland

In order to infer reactions of treeline and alpine vegetation to climatic change, past vegetation changes are reconstructed on the basis of pollen, macrofossil and charcoal analysis. The sampled sediment cores originate from the small pond Emines, located at the Sanetsch Pass (connecting the Valais and Bern, Switzerland) at an altitude of 2288 m a.s.l. Today's treeline is at ca. 2200 m a.s.l. in the area, though due to special pass (saddle) conditions it is locally depressed to ca. 2060 m a.s.l. Our results reveal that the area around Emines was covered by treeless alpine vegetation during most of the past 12,000 years. Single individuals of Betula, Larix decidua and possibly Pinus cembra occurred during the Holocene. Major centennial to millennial-scale responses of treeline vegetation to climatic changes are evident. However, alpine vegetation composition remained rather stable between 11,500 and 6000 cal. BP, showing that Holocene climatic changes of +/− 1 °C hardly influenced the local vegetation at Emines. The rapid warming of 3–4 °C at the Late Glacial/Holocene transition (11,600 cal. BP) caused significant altitudinal displacements of alpine species that were additionally affected by the rapid upward movement of trees and shrubs. Since the beginning of the Neolithic, vegetation changes at Sanetsch Pass resulted from a combination of climate change and human impact. Anthropogenic fire increase and land-use change combined with a natural change from subcontinental to more oceanic climate during the second half of the Holocene led to the disappearance of P. cembra in the study area, but favoured the occurrence of Picea abies and Alnus viridis. The mid- to late-Holocene decline of Abies alba was primarily a consequence of human impact, since this mesic species should have benefitted from a shift to more oceanic conditions. Future alpine vegetation changes will be a function of the amplitude and rapidity of global warming as well as human land use. Our results imply that alpine vegetation at our treeline pass site was never replaced by forests since the last ice-age. This may change in the future if anticipated climate change will induce upslope migration of trees. The results of this study emphasise the necessity of climate change mitigation in order to prevent biodiversity losses as a consequence of unprecedented community and species displacement in response to climatic change.

Leydig-cell tumour in children: variable clinical presentation, diagnostic features, follow-up and genetic analysis of four cases

BACKGROUND: Testicular tumours are relatively uncommon in infants and children, accounting for only 1-2% of all paediatric solid tumours. Of these approximately 1.5% are Leydig-cell tumours. Further, activating mutations of the luteinizing hormone receptor gene (LHR), as well as of the G protein genes, such as Gsalpha (gsp) and Gialpha (gip2) subunits, and cyclin-dependent kinase gene 4(CDK4) have been associated with the development of several endocrine neoplasms. AIMS/METHODS: In this report, the clinical variability of Leydig-cell tumours in four children is described. The LHR-, gsp-, gip2- and CDK4 genes were investigated to establish the possible molecular pathogenesis of the variable phenotype of the Leydig-cell tumours. RESULTS: No activating mutations in these genes were found in the four Leydig-cell tumours studied. Therefore, the absence of activating mutations in LHR, as well as in both the 'hot spot' regions for activating mutations within the G-alpha subunits and in the regulatory 'hot spot' on the CDK4 genes in these tumours indicates molecular heterogeneity among Leydig-cell tumours. CONCLUSION: Four children with a variable phenotype caused by Leydig-cell tumours are described. A molecular analysis of all the 'activating' genes and mutational regions known so far was performed, but no abnormalities were found. The lessons learnt from these clinically variable cases are: perform ultrasound early and most importantly, consider discrepancies between testicular swelling, tumour size and androgen production.

A degradation-sensitive anionic trypsinogen (PRSS2) variant protects against chronic pancreatitis

Chronic pancreatitis is a common inflammatory disease of the pancreas. Mutations in the genes encoding cationic trypsinogen (PRSS1) and the pancreatic secretory trypsin inhibitor (SPINK1) are associated with chronic pancreatitis. Because increased proteolytic activity owing to mutated PRSS1 enhances the risk for chronic pancreatitis, mutations in the gene encoding anionic trypsinogen (PRSS2) may also predispose to disease. Here we analyzed PRSS2 in individuals with chronic pancreatitis and controls and found, to our surprise, that a variant of codon 191 (G191R) is overrepresented in control subjects: G191R was present in 220/6,459 (3.4%) controls but in only 32/2,466 (1.3%) affected individuals (odds ratio 0.37; P = 1.1 x 10(-8)). Upon activation by enterokinase or trypsin, purified recombinant G191R protein showed a complete loss of trypsin activity owing to the introduction of a new tryptic cleavage site that renders the enzyme hypersensitive to autocatalytic proteolysis. In conclusion, the G191R variant of PRSS2 mitigates intrapancreatic trypsin activity and thereby protects against chronic pancreatitis.