text:
The authors have no conflict of interest.
Research Article
Chloride Channel ClCN7 Mutations Are Responsible for Severe Recessive, Dominant, and Intermediate Osteopetrosis†
Article first published online: 1 OCT 2003
DOI: 10.1359/jbmr.2003.18.10.1740
Copyright © 2003 ASBMR
Journal of Bone and Mineral Research
Volume 18, Issue 10, pages 1740–1747, October 2003
Additional Information
How to Cite
Frattini, A., Pangrazio, A., Susani, L., Sobacchi, C., Mirolo, M., Abinun, M., Andolina, M., Flanagan, A., Horwitz, E. M., Mihci, E., Notarangelo, L. D., Ramenghi, U., Teti, A., Van Hove, J., Vujic, D., Young, T., Albertini, A., Orchard, P. J., Vezzoni, P. and Villa, A. (2003), Chloride Channel ClCN7 Mutations Are Responsible for Severe Recessive, Dominant, and Intermediate Osteopetrosis. J Bone Miner Res, 18: 1740–1747. doi: 10.1359/jbmr.2003.18.10.1740
- †
Publication History
- Issue published online: 2 DEC 2009
- Article first published online: 1 OCT 2003
- Manuscript Accepted: 21 MAY 2003
- Manuscript Revised: 9 APR 2003
- Manuscript Received: 22 JAN 2003
- Abstract
- Article
- References
- Cited By
- human osteopetrosis;
- osteoclasts;
- bone marrow transplantation;
- Atp6a3;
- ClCN7
Among 94 osteopetrotic patients presenting with a severe clinical picture and diagnosed early in life, 12 bore mutations in the ClCN7 gene, but only 7 of them had the expected two recessive mutations. The remaining five patients seem to be heterozygous for a ClCN7 mutation, and significant variations were observed in the clinical manifestations of their disease, even within the same family.
Introduction: Human osteopetroses are a heterogeneous group of diseases that include both infantile severe, autosomal recessive (ARO) and adult autosomal dominant (ADO) forms. Two genes, Atp6a3 (TCIRG1) and ClCN7, have been shown to be associated with human ARO, the latter of which is also thought to be responsible for ADO-II. However, patients with an intermediate phenotype have been described: the genetic basis of these observances is unknown.
Materials and Methods: In this study, we report the clinical and molecular analysis of 94 patients in which a diagnosis of severe osteopetrosis was made within the first 2 years of age. Both TCIRG1 and CLCN7 genes were sequenced in all patients and the molecular findings were correlated to clinical parameters.
Results and Conclusions: In 56 of 94 patients with a classical picture of ARO, TCIRG1-dependent recessive mutations were found. In contrast, ClCN7 mutations were found in 12 cases (13%) of severe osteopetrosis, but only 7 of them had two recessive mutations identified: in 6 of these 7 cases, central nervous system manifestations were noted, and these patients had a poor prognosis. The remaining five cases were heterozygous for a ClCN7 mutation, including two brothers from a large family with a history of ADO-II in which the presence of a second ClCN7 mutation was formally excluded. Despite an early and severe clinical presentation, these five patients all reached adulthood, suggesting that the degree of dominant interference with chloride channel function can vary widely. Our findings suggest that recessive ClCN7-dependent ARO may be associated with CNS involvement and have a very poor prognosis, whereas heterozygous ClCN7 mutations cause a wide range of phenotypes even in the same family, ranging from early severe to nearly asymptomatic forms. These findings have prognostic implications, might complicate prenatal diagnosis of human osteopetroses, and could be relevant to the management of these patients.
Infantile malignant autosomal recessive osteopetrosis (ARO; OMIM 259700) is a severe bone disease with a fatal outcome generally within the first decade of life. It results from a deficit in bone resorption caused by osteoclast malfunction or formation and is associated with a number of severe clinical manifestations, including macrocephaly, cranial nerve dysfunction such as deafness and blindness, hepatosplenomegaly, and severe bone marrow failure beginning in early infancy or in fetal life. Deafness and blindness are generally thought to represent effects of pressure on cranial nerves exerted by the overgrowth of bone, but the possibility of a primary defect in sensory cells has also been raised.(1) Forms of osteopetrosis associated with neurologic deficits have also been described,(2–4) although it is not yet clear whether these forms represent a variant of the same disorder or distinct diseases.
Autosomal dominant osteopetrosis (ADO) has also been described and generally is associated with less severe clinical features. It is often subdivided clinically into two forms, type I (ADO-I) and type II (ADO-II, OMIM 166600).(5–7) An intermediate form has also been described (OMIM 259710): the presence of several affected sibs in some families with intermediate osteopetrosis suggests an autosomal recessive inheritance,(8) while in other reports, the pattern of inheritance has not been established. Although the clinical picture is less severe than that of ARO, some of these patients are diagnosed in infancy and have significant clinical findings; therefore, for these forms, the term "mild" does not seem to be appropriate.
In the last few years, cellular and molecular investigations of hereditary osteopetroses have greatly contributed to the dissection of this heterogeneous disease. Normal bone homeostasis is a dynamic and complex process that relies on a delicate balance between the activity of osteoblasts and osteoclasts. Osteoclasts are multinucleated giant cells that are derived from hematopoietic precursors and are responsible for bone resorption. To achieve normal bone resorption, at least two processes seem to be important for the maintenance of osteoclast function: (1) the establishment of a specialized membrane structure, the so-called ruffled membrane, which is an area of intense membrane infolding thought to be actively involved in bone resorption,(9–11) and (2) the production of a low pH microenvironment at the contact surface between the cell and the bone matrix, the so-called resorption lacuna. Osteoclasts adhere to the bone surface, seal the resorption lacuna, and activate a specific membrane-bound V-ATPase complex, generating the acid environment necessary to dissolve the mineral component of bone, primarily composed of hydroxyapatite crystals. Subsequent release of lysosomal enzymes, including cathepsin K, eventually leads to organic matrix degradation.
Human osteopetroses in most, but not all, cases(12) show normal or increased numbers of osteoclasts, and it has been suggested that in humans, the affected gene(s) is more likely to relate to the maintenance of the functional capacity of mature osteoclasts. This suggestion has been supported by the fact that the gene defects found thus far in ARO and ADO-II, as well as the one found in osteopetrosis associated to renal tubular acidosis (OMIM 259730), are all involved in the metabolic pathways leading to acidification of the external microenvironment. The a3 subunit (ATP6a3 or TCIRG1) of the V-ATPase complex, the primary pump involved in proton extrusion by osteoclast cells, has been found mutated in more than 50% of the patients affected by ARO.(13–15) The ClCN7 gene, which provides the chloride conductance required for an efficient proton pump, has recently been implicated in ADO-II,(16) but has also been shown to be mutated in two cases of ARO.(1,16) The clinical implications of these molecular findings have not yet been investigated.
We have collected a large series of patients with classical ARO and patients with less severe clinical courses. In this study, we report the clinical and molecular analysis of this large series, in which the molecular findings were related to the clinical presentation and course, with the aim of investigating whether the particular gene involved and/or the specific mutation identified are of clinical relevance with regard to diagnosis, presentation, or prognosis.
As part of an international study, we have collected material from 94 unrelated patients with a clinical diagnosis of severe osteopetrosis made based on the following criteria at presentation: early diagnosis (before 2 years of age), radiographic evidence of markedly increased bone density, pancytopenia, hepatosplenomegaly, and cranial nerves defects.
The presence of all the abnormalities was required for entry in the study, although in a few cases (patient 6 for the cranial defect and patients 8 and 12 for blood count), one single criterion was not considered because the specific data were not available.
Samples were obtained from Italy, the United States, Canada, Turkey, Costa Rica, Yugoslavia, Croatia, Switzerland, Sweden, Germany, Israel, the Netherlands, United Kingdom, Belgium, and France. Specimens include DNA, frozen peripheral blood cells, fibroblasts or Epstein-Barr virus (EBV)-transformed lymphoblast cell lines from patients, and DNA samples from their parents; they were collected with informed consent. Clinical, radiological, and laboratory data were collected for genotype/phenotype correlation studies. One additional case of classical ADO-II (case 13) suggested by multiple spontaneous fractures and X-ray high bone density is also reported.
ClCN7 and TCIRG1 gene mutation analysis
The ClCN7 gene was amplified at both genomic (genomic sequences accession no. AL031600 and AL031705) and RNA (cDNA sequence accession no. AF224741) levels. The strategy and primers are shown in Table 1.
All the reactions were performed in 25 μl of final volume with 0.4 U Taq polymerase, 1.5 mM MgCl2, 150 μM dNTP, 10 pmol of each oligonucleotide primer, and 20 ng of purified DNA. The thermocycling conditions used for amplification consisted of an initial denaturation step at 94°C for 3 minutes, followed by 34 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, and 72°C for 30 s.
Sequencing was performed directly on the polymerase chain reaction (PCR) products purified from the gel using the dideoxynucleoside chain termination method (USB).
The organization of the ClCN7 protein and locations of the transmembrane helices were predicted with several bioinformatics tools. Related sequences were searched from databases and used in multiple sequence alignment. In the case of missense mutations (G240R, P249R, R286Q, M332V, L490F, R526W, L614P, G677V, S744F, R767Q) not previously described by Cleiren et al.,(16) 100 chromosomes from normal unrelated donors were also investigated by direct sequence analysis or enzymatic digestion.
Mutation analysis of the TCIRG1 gene was performed as previously described.(15)
The mutations and patient information were collected and put into a database (http://bioinf.uta.fi/TCIRG1base/).
We analyzed the TCIRG1 and ClCN7 genes in 94 patients clinically diagnosed with severe osteopetrosis; in 56 (60%), defects were identified in the TCIRG1 gene. Of these, 53 had mutations on both alleles, whereas 3 had mutations in only one allele; these 3 patients have previously been reported.(15) In 12 families (13%), mutations were documented in the ClCN7 gene. In 26 cases (28%), no defects were identified in either the TCIRG1 or ClCN7 gene. In 6 of these 26 patients, whose parents were consanguineous, a whole genome search with approximately 300 polymorphic markers was performed to identify regions of homozygosity. This allowed us to formally exclude the TCIRG1 and CLCN7 genes as playing a role in the pathogenesis of the disease in these patients, because no homozygosity was found in the regions harboring these two genes, except in one case who was homozygous in the TCIRG1 region (A Frattini and G Casari, unpublished findings, 2002). These findings confirm that at least a third gene is involved in the pathogenesis of human ARO.
The clinical history of all the ClCN7 mutated cases is described below and summarized in Table 2.
Cases 1–7 are from unrelated families of various ancestry, in which parents were unaffected. They presented with a severe clinical picture compatible with autosomal recessive osteopetrosis. In 6 of the 7 families, central nervous system abnormalities were observed radiographically or clinically, including cerebral atrophy (cases 1 and 3), macrocephaly with neurological impairment (case 4) or hydrocephalus (case 6), autism (case 5), and spinocerebellar ataxia (case 2). In the seventh family, two affected siblings were evaluated (case 7a, male; case 7b, female). The brother suffered his first fracture at 1 year of age, and at 24 years of age, he has vision impairment, mandibular osteomyelitis, anemia, thrombocytopenia, and splenomegaly; he has had a total of approximately 80 fractures. A bone biopsy showed multiple activated osteoclasts. In contrast to her brother, the affected sister, at 31 years of age, has anemia, but has not developed transfusion dependence; she has optic atrophy and visual impairment and has had approximately 100 fractures.
In cases 8–12, the early and severe presentation of the probands suggested an ARO diagnosis, but in one case, this occurred within a family with classical ADO-II (family 8). In this family, two children (cases 8a and 8b) were diagnosed as infants. Several members of the family, including their mother and maternal grandmother, were affected with a dominant form of the disease with multiple bone fractures only. In addition to bone fractures, both siblings developed severe optic nerve atrophy secondary to optic canal narrowing, requiring neurosurgical intervention. The sister, who is 4 years older than the brother, was surgically treated at 2 years of age and did not fare as well as the brother, who underwent surgery at a younger age (11 months) because of his sister's medical course. The sister, now 17 years of age, is legally blind. The brother, at the age of 13 years, is legally blind in the left eye and has 20/20 visual acuity of the right eye. No other neurological defects have been reported. The severity of their clinical presentation is somewhat unusual for an ADO-II diagnosis.
Case 9 was diagnosed as an infant with an abnormal complete blood count and signs of optic nerve compression and has had numerous fractures. The parents have no history of an increased likelihood of fractures. He is now 10 years old, blind, and growth retarded, but his blood counts are normal.
Case 10 was diagnosed with severe osteopetrosis in early infancy with blindness, severe anemia, and hepatosplenomegaly. A frequent transfusion requirement and the development of hemochromatosis necessitated splenectomy at 14 years of age. He developed a partial palsy of the VII and the VIII cranial nerve and severe obstruction of the carotid and vertebral arteries with extreme collateral formation. He had dental impaction and multiple episodes of osteomyelitis of the mandible, requiring numerous stomatologic interventions. He had several pathological fractures. At last visit, he was 26 years old and of normal intelligence. Chronic headaches are a major problem. A bone biopsy revealed an increased number of multinucleated osteoclasts.
Case 11a presented with anemia and thrombocytopenia, partial compression of the optic nerve canal, and radiological signs of osteopetrosis. A diagnosis of autosomal recessive OP was suspected, and she underwent bone marrow transplantation (BMT). However, retrospective examination of his parents (performed after the birth of the proband) showed that the father (case 11b) had a radiological picture of bone involvement and a history of two fractures, both related to trauma. Case 11b is now 4 years old, in very good health, and is able to write, read, and speak two languages.
Case 12 presented as an infant with optic nerve compression and increased bone density. A clinical diagnosis of ARO was made. Her parents and one sister were not affected. She did not undergo BMT, and she is now 30 years old and blind. She experienced many fractures.
Molecular analysis was also performed on a case with classical ADO-II (case 13), a female who was diagnosed at 8 years of age by X-ray after a metatarsal fracture sustained during a sporting event. She has had one additional fracture since then but no other symptoms. There is no positive family history.
ClCN7-dependent recessive osteopetrosis
In our analysis, we found seven families in which ClCN7 mutations were identified in both alleles. These data are summarized in Table 3. Two patients (cases 1 and 2), were born from consanguineous parents and were homozygous for the described mutation, whereas the other five were compound heterozygotes (patients 3–7). Patient 1 bore a single T nucleotide insertion, causing a frameshift and a premature stop codon (D145X), whereas patient 2 had a single nucleotide change, causing a missense mutation (R767Q) in the COOH-terminal portion of the protein. For both patients, the same mutation was found at the heterozygous level in their parents.
Patient 3 had two single nucleotide changes, causing two missense mutations (G240R and R526W), the first being present in the mother and the second in the father. Patient 4 had a single nucleotide change causing a missense mutation (L614P) in one allele and a genomic deletion involving exon 17 on the second allele (parent DNA was not available). Patient 5 had a single nucleotide change, causing a nonsense mutation (E374X) in the first allele and a nucleotide change in intron 10, creating a new splice site causing an in frame insertion of 54 nucleotides in ClCN7 transcript, starting after codon G306. No parent DNA was available. Patient 6 had two missense mutations, M332V and R767W. The molecular findings in these six patients are in complete agreement with the clinical diagnosis of severe ARO and with the fact that all their parents were asymptomatic.
In family 7, two siblings were affected (7a and 7b). As shown in Table 3, both were compound heterozygotes for two different single nucleotide changes, causing two missense mutations, a maternally derived P249R and a paternally derived S744F. This latter mutation was also shared by a healthy brother. Therefore, the family history suggests that both the mutations are necessary for the clinical manifestation of osteopetrosis, although the possibility of incomplete penetrance cannot be formally excluded.
ClCN7-dependent osteopetrosis with a single mutated allele
Six patients from five families (cases 8–12) were found to carry a single ClCN7 mutation, all represented by a single nucleotide change causing missense mutations in cases 8–11 and a stop mutation in case 12 as shown in Table 3.
Patients 8a and 8b (brother and sister) were both heterozygous for a missense mutation at codon 677 (G677V). Many members of the maternal ancestry were affected by dominant osteopetrosis on clinical grounds, because they reached adulthood and had children. To explain the severity of the presentation in the third generation, it could be hypothesized that the father, in whose family there is no history of osteopetrosis, could have provided a recessive mutation; in this case, both the severely affected siblings from the third generation could have inherited the same paternal chromosome. We tested this possibility by analyzing several single nucleotide polymorphisms (SNPs) in the ClCN7 gene in the affected siblings (A Frattini, unpublished data, 2003); the analysis demonstrated that they inherited two different paternal chromosomes, ruling out an additional ClCN7 mutation as the cause of the worsening of the clinical picture in family 8.
Patients 9 and 10 was heterozygous for a G215R and L490F mutation, respectively.
In case 11a, a putative diagnosis of ARO was suggested, and BMT was performed. However, after the diagnosis was made in this child, bone abnormalities consistent with ADO were found in her father, while the mother was normal. The affected child showed a G215R mutation on only one allele, and interestingly, her father and her asymptomatic paternal grandmother carried the same amino acid change. Three additional changes, a V418M and two silent changes (T19233C and C12974T), were found in the proband, her mother, and her maternal grandmother, but these changes were shared by many normal controls. In family 12, a diagnosis of ARO was established at 6 months, although no BMT was performed. However, an extensive search identified only one maternally derived mutated allele, bearing a nonsense mutation (W180X). No mutation was found in the father and her sister.
Patients 13, as expected in consideration of her ADO diagnosis, was heterozygous for a single R286Q mutation.
Polymorphic repeat does not account for variation in the osteopetrotic phenotype
Because the clinical picture in the last generation in several families seems to be more severe than in parents, we hypothesized a possible role of the polymorphic repeat present in intron 8.(16) In our normal population, we found from four to seven repeats, but no increase in the number of the repeats was seen from parents to siblings. Therefore, it is unlikely that an elongation of these repeats is responsible for the severity of the disease.
Despite the fact that, in murine studies, inactivation of many genes involved in both differentiation and function of osteoclasts give rise to severe osteopetrosis,(17) the genes found mutated in human osteopetroses affect a single aspect of mature osteoclast, specifically the ability to generate acidification in the extracellular environment.(1,13,18) While mutations in carbonic anhydrase are associated to a renal defect, recessive abnormalities in TCIRG1 and ClCN7 genes give rise to relatively homogeneous pictures of severe ARO. However, the data presented here, together with finding that ClCN7, but not TCIRG1, is also responsible for ADO, suggest that the spectrum of the disease observed with defects within these two genes is different.
TCIRG1-dependent mutations reported thus far seem to be limited to severe ARO. All TCIRG1-ARO patients analyzed by us and described in the literature are diagnosed early in infancy and present with classical ARO clinical picture. Most TCIRG1 mutations are genetically null, consisting of nonsense, deletion, and splicing abnormalities.(15) For this reason we have hypothesized that hypomorphic TCIRG1 mutations could also cause less severe (intermediate) osteopetrosis cases, but we were unable to find such mutations in patients with intermediate or dominant osteopetrosis.
In contrast, the spectrum of disease caused by ClCN7 mutations is wider. Cleiren et al. have clearly demonstrated that adult-onset ADO is caused by heterozygous missense mutations in the ClCN7 gene.(16) They also reported a single patient with a homozygous missense mutation and a typical picture of ARO. A clinically similar patient who was a compound heterozygote for a nonsense and a missense mutation was also reported by Kornak et al.(1) Our data extend these reports by documenting that ClCN7-dependent ARO represents a substantial fraction of ARO, accounting for about 13% of patients presenting with a severe clinical picture.
Interestingly, in six of seven of the families in which ARO was documented to be caused by two ClCN7 mutations, central nervous system abnormalities were noted. In two of these instances, the patients were survivors of BMT, and complications of the transplant process could account for these changes in whole or in part. In addition, it is difficult to distinguish primary neurologic changes related directly to the ClCN7 gene defects and secondary effects caused by skull abnormalities affecting the flow of cerebral spinal fluid or sequelae of chronic airway obstruction, which has been observed in individuals with ARO.(19) Nevertheless, our data suggests that ARO caused by ClCN7 defects should be evaluated routinely for neurologic findings, both clinically and radiologically. It is possible that this subset could overlap with a previously recognized osteopetrosis form associated with infantile neuroaxonal dystrophy (OMIM 600329). The implications of this issue for transplantation are clear; whereas improvement in the bony changes of osteopetrosis was documented after transplant in at least one patient (patient 5), the central nervous system abnormalities directly related to ClCN7 mutations cannot be expected to be corrected by BMT. Therefore, information regarding this issue could potentially be an important variable in determining the prognosis for individuals with ClCN7 defects. Only prospective studies closely correlating the genotype of osteopetrotic patients and their clinical course will provide the necessary data to address this question.
In addition, our data suggest that ClCN7 mutations are also responsible for intermediate forms. The relationship of these forms with both ARO and ADO is not straightforward, because both dominant and recessive inheritance has been suggested.(20,21) Our findings suggest that at least some of the intermediate forms represent a continuum with both ARO and ADO. ADO is sometimes referred to as benign form, with the diagnosis often being made incidentally by X-rays after the occurrence of fractures in adult age. However, some patients are diagnosed at earlier ages because of the presence of blindness, fractures, and/or anemia; these more severe phenotypes can coexist with benign ADO cases within the same family.(22) The data reported in our studies contribute to the clarification of these questions. In the original families described as intermediate forms, a recessive transmission was inferred because of the presence of several siblings in the same family in the presence of asymptomatic parents: the existence of several siblings was thought to rule out the occurrence of de novo dominant mutations (see OMIM 259710). However, our data suggest that dominant ClCN7 mutations can be responsible for a range of presentations wider than previously appreciated and that the same mutation can cause a broad range of phenotypes even in the same family, because we found mildly affected or even asymptomatic patients together with others showing severe symptoms, including anemia and pancytopenia, which are usually associated with ARO. A dominant pattern of transmission with incomplete penetrance and/or variable expressivity is likely to be the best explanation. The alternative explanation, mentioned by Cleiren et al.,(16) could be that an additional recessive mutation from the second parent contributed to the different phenotypes. Although the possibility exists that additional mutations were not detected by our analysis, it is noteworthy that severe cases with a single ClCN7 mutation do not show any primary neurological involvement and live longer, with a consequent better prognosis than classical recessive ClCN7-dependent patients (see patients 8–12). In addition, polymorphism analysis performed in family 8 allowed us to show that the two affected brothers inherited a different chromosome from their unaffected father, ruling out a contribution of the second allele to their phenotype. Finally, phenotype variability within ADO families, including several with asymptomatic obligate carriers,(23) is a common event, and given the low frequency of recessive alleles, it would be difficult to postulate that undetected mutations are involved in all these cases.
Further complexity is added by the finding that the expressivity of a specific ClCN7 mutation is variable, as exemplified by the phenotype variability found between families. An R767W mutation has been reported in an ADO white patient by Cleiren et al.,(16) while we found the same mutation in a recessive white proband whose parents where phenotypically normal (case 6). In addition, a different homozygous mutation in the same codon (R767Q) was found in a recessive Pakistani patient whose parents were again asymptomatic (case 2). The same G215R mutation was the only one found in families 9 and 11, as well as in families described by Cleiren et al.(16) However, the clinical picture varied from being relatively severe in probands 9 and 11a to the father and the grandmother of proband 11a who were diagnosed only after her birth. In family 8, two siblings presented with a relatively severe picture, clearly different from those found in several members of the same family, and finally, in family 12, the same mutation was found in the asymptomatic mother and her severely affected daughter.
The factors involved in this phenotypic variability remain unknown. It is likely that this variability is caused by other unknown loci or to complex interactions with genes regulating chloride channel function or involved in osteoclast metabolism. One such candidate is the TCIRG1 gene itself, which functions in a metabolic pathway interacting with that of ClCN7. However, a normal TCIRG1 coding sequence was found in all ClCN7-dependent patients. Alternatively, this variability could be caused by different levels of ClCN7 expression associated with promoter polymorphisms, a possibility that is worthy of further investigation.
In conclusion, our findings contribute to further dissection of the heterogeneity of ARO, documenting that ClCN7-dependent ARO constitutes a substantial portion of severe cases, including a number in which central nervous system findings were observed. In addition, in contrast to what has been observed with the TCIRG1 gene, whose mutations have been reported only in classical ARO, dominant ClCN7 mutations are responsible for a continuum of symptoms and can cause both classical ADO and intermediate/severe cases of osteopetrosis. Although further studies may establish a relation between the specific protein mutants and the pattern of inheritance, it is likely that genetic analysis alone will be insufficient to fully predict the clinical phenotype, because of incomplete penetrance and variable expressivity. This makes prenatal diagnosis more cumbersome and difficult to interpret, at least until the contributing factors are unambiguously identified.
We thank Prof M Vihinen for database construction and management. The technical assistance of Enrica Mira Catò is acknowledged. This study was partially supported by Grants Genomica Funzionale (PV), grant from Programma Nazionale Cellule Staminali (to PV), Grant RBNE019J9W from MIUR-FIRB (AV and PV), grants from Cofin 2003 (AA), grant from Research to Prevent Blindness, Inc. (TY), and grants from the Mabel E. Leslie Research Endowment Funds (TY). CS is a recipient of a fellowship from FIRC. This is manuscript no. 67 of the Genoma 2000/ITBA Project funded by CARIPLO.
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- 132000 Defects in the TCIRG1-encoded 116 kD subunit of the vacuolar proton pump are responsible for a subset of human autosomal recessive osteopetrosis. Nat Genet 25:343–346., , , , , , , , , , , ,
- 142000 Mutations in the a3 subunit of the vacuolar H(+)-ATPase cause infantile malignant osteopetrosis. Hum Mol Genet 9:2059–63., , , , , , , , ,
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text:
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subset: commoncrawl


text:
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Privacy-focused Skype alternative Ring shows promise
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Here are the 10 most-read posts for TechRepublic's Tablets in the Enterprise blog in 2012.
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Mary Shacklett looks at the ways that Apple can continue to expand its influence in the enterprise, but it would require some changes in focus for the company.
Is the Internet down? Troubleshooting network issues
Patrick Lambert goes over some basic troubleshooting tips for solving users' networking issues.


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Zaxcom's new TRX900LT digital wireless transmitter, first announced at the 2010 NAB Show, features the ability to fully encrypt audio signals for secure audio transmission in settings where sensitive material may need to be protected from signal interception or theft. The TRX900LT body pack unit combines wireless transmission, recording and remote control into a single, low-cost wireless transmitter.
To activate the encryption, a TRX900LT user simply enters an encryption code into the transmitting and receiving units. With more than 16 million key code combinations, it's impossible to decode the audio signals without the key. Once the code is entered, the system will encrypt all signals automatically.
Like all technologies in Zaxcom's TRX product line, the TRX900LT body pack unit features 100 percent digital transmission for audio quality that rivals a hard-wired system. Designed to be extremely lightweight and durable, the TRX900LT is housed in a high-strength, impact-resistant nylon polymer casing that provides protection from both corrosion and water damage.
With an internal time code-referenced audio recorder that backs up all wireless transmissions on a removable microSD card, the TRX900LT eliminates the possibility of audio loss due to interference or signal dropout. By integrating a group of TRX900LT units, users are able to establish a multitrack recording session directly from the body pack units. Each TRX900LT unit will operate for up to five hours on a single AA lithium battery and is compatible with all Zaxcom wireless receivers and IFB transmitters.
Also included in the TRX900LT transmitter is reception of remote controls via ZaxNet. ZaxNet is Zaxcom's 2.4GHz RF network that enables distribution of remote-control signals, time code and metadata. With ZaxNet, TRX900LT users will be able to control gain and audio replays remotely via a time code reference, so multiple Zaxcom wireless systems can replay audio in sync. The TRX900LT-ZaxNet integration also will allow users to remix audio from multiple transmitters, creating a virtual multitrack playback and rerecord system.
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Military Hat Ornament
Roll over image to zoom in
At Christmastime and throughout the year, our military hat ornament proclaims your gratitude to our men and women in uniform. A powerful commemoration of your own hero's honorable service.
- Air Force, Army, Marines, or Navy
- Polystone
- Approximately 1.5" H x 2.5" W (3.8 x 6.4 cm)
- Made in China
For bulk pricing on these and other items, please contact our Customer Service department at firstname.lastname@example.com.
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January 12, 2015 - Bought it as a stocking stuffer for my niece, she LOVES it.
January 6, 2015 - Love it, better than in the picture
January 5, 2015 - Love the Navy hat! Got one for myself and my daughter, in honor of her son and father, my husband. They look great on our trees Very well made.
January 3, 2015 - My husband was a marine during the Korean conflict. I bought these ornaments for each of our daughters and put his name and dates of service on the back. It brought tears to their eyes. One said this was a perfect keepsake.
January 2, 2015 - Everyone loved the ornaments, my niece really loved hers and showed the world through a social media, thanks.
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Riley is the perky cheerleader firm ass and legs type that everyone wants. She is ready to please at all times. We just loved her cock sucking abilities its up there with the pros.
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"Ready To Succeed"
"Great Asspirations"
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"Permiscuous Pussy"
"Just In Time"
"Ass On Ashli"


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Our great range of second-hand music boasts classics and collectables from rock, pop, classical, dance, jazz genres and more. On vinyl and CD, have a browse and see what you can uncover.
Results 41-60 of 314
Sunstruck - Stoneground - NUT 11
Bread and Roses Bread and Roses - DRGN 881
Travelling the Tees The Teesside Fettlers - TSR 021
Concrete Routes Sacred Cows The Cock and Bull Band - FMSL 2015
Eyes Closed and Rocking The Cock and Bull Band - 12TS440
Treading the Boards Richard Digance - TRA306
Malcolm Stewart 7" Single : Let's Play a Game
Philip Larkin Reads The Whitsun Weddings - Philip Larkin - LPV 6
Old Four Eyes Is Back - Mike Harding - 6308 290
The Home Service The Home Service - SAW 3
Old Swan Brand Old Swan Band - FRR 028
Wild and Beautiful Silly Wizard - SHY 7016
Ranting Lads - The High Level Ranters - 12ts297
Paraguayan Songs - Trio Los Paraguayos - BBR 8074
Mr Fox's Garden - Bread And Roses - DRGN911
Music in the World of Islam Vol 3 and 4
The Hazards of Love EP - Anne Briggs - Top 94
The Magnificent Music Machine - Tom T. Hall - SRM-1-1111
The Blaydon Races [lp] Michael Hunt with the Dombie Bros - ecs 2037
fivepenny piece on stage the fivepenny piece - ou2050
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Nottingham Post has all the best restaurants in Nottingham and every type of restaurant in Nottingham listed, many with reviews and star ratings; you'll find Indian Restaurants, Chinese Restaurants, Italian Restaurants as well as other places to eat in Nottingham such as Pizza Parlours and Brasseries.
Why not list your Nottingham Restaurant business on Nottingham Post ?
Address: Corner House, Trinity Square, Nottingham, G1 4DB
Address: 27 West Street, , RETFORD, DN22 6ES
Address: 95 Melton Road, , NOTTINGHAM, NG2 6EN
Address: 155-157 Nottingham Road, , NOTTINGHAM, NG5 6JN
Address: 473 Mansfield Road, , NOTTINGHAM, NG5 2DR
Address: 6 Trent House, Trent Bridge West Bridgford, NOTTINGHAM, NG2 5GN
Address: 20 King Street, , NOTTINGHAM, NG1 2AS
Address: 137 Station Road, Beeston, Nottingham, NG9 2AZ
Address: Plumptre House, 5 Poplar Street, NOTTINGHAM, NG1 1GP
Address: 25 Radcliffe Road, West Bridgford, NOTTINGHAM, NG2 5FF
Address: 42-44 Bridgford Road, , NOTTINGHAM, NG2 6AP
Address: 27 King Street, , NOTTINGHAM, NG1 2AY
Address: 384 Carlton Hill, Carlton, NOTTINGHAM, NG4 1JA
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Address: 119 High Road, Beeston, NOTTINGHAM, NG9 2LH
Address: 16 South Parade, , DONCASTER, DN10 6JH
Address: The Corner House, Foreman Street, NOTTINGHAM, NG1 4DB
Address: 24-26 Goosegate, , NOTTINGHAM, NG1 1FF
Address: 4 Leicester Road, , LOUGHBOROUGH, LE12 8ET


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Category: 'Circumcision/Intactivism'
Choosing to Leave My Second Son Intact, Despite the Intactivists
It's never too late to do things differently. My friend, Kym, shares her story about why she circumcised her first son but left her second son intact.
Men Have A Less Than .01% Chance of Contracting Penile Cancer, Regardless of Circumcision
An intact man has a greater likelihood of breast cancer than he does penile cancer.
Circumcision: A Poem
Cut your hair, cut a tooth, cut your bush, cut him loose, but don't cut a boy's penis.
Everyday Superheroes
Welcome to the April Carnival of Natural Parenting: Compassionate Advocacy This post was written for inclusion in the monthly Carnival of Natural Parenting hosted by Code Name: Mama and Hobo Mama. This month our participants have shared how they advocate for healthy, gentle parenting choices compassionately. Please read to the…
Intact in America (When Circ Was All the Rage)
A reader responded quite eloquently on a couple of my circumcision posts several months ago. I emailed and asked him if he would allow me to rework and publish his comments, and he graciously submitted a guest post. I hope his perspective as an intact male – from a generation…
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- Giveaway: Glee Gum Make Your Own Chewing Gum, Chocolate, and Gummies Kits: A $42 Value {US, 3.31}
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- Children and Creativity
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- Giveaway: On the Farm Game from Ravensburger and NPN Holiday Gift Guide – $21 ARV
- Giveaway: Make 'n Break Junior Game from Ravensburger and NPN Holiday Gift Guide – $25 ARV
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Planes Mistaken for Stars
From all accounts, Denver-based screamo/hardcore foursome Planes Mistaken for Stars flat-out leveled stages on the recent politically charged Plea for Peace tour, stealing the thunder from headliners Cursive with jackknifing Mack truck riffs and gone-haywire howls. And their dirtbag, anti-emo pretty-boy looks (they've been referred to as "an entire group of Andrew W.K.s") didn't stop Newsweek from running a half-page photo of the band a couple weeks ago, along with a story about the youth vote, pushing the quartet to the front ranks of punk rock's anti-Bush movement. Planes' current campaign, however, is a bit more personal -- they're on the road supporting their new full-length album, Up in Them Guts, which is as viscerally intense as the title would suggest.
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Plan context menu…
3D context menu…
We've changed the way that it calculates what Doors and Windows should be collated together, to make sure that all the joinery items that should be together, are together. We've also added this 'Isolate Orientation' checkbox into the settings, so that if you want to make your joinery items that have different orientations to be collated together as well, you can turn off the checkbox there to allow it.
To get the new version of the tool, just run the Cadimage Installer on your computer, and install the Doors+Windows 5.53 update.]]>
All you need to do is click the New Tab button to the right of the Tab Bar and you will see a Navigator appear…
All you need to do is go to Options> Work Environment> Apply Profile then click on the one you want.
In this case, the markers will only show in plan view, on the stories through which the section cuts. If you adjust story heights, you might find that markers with limited vertical range may appear on, and disappear from plan views unexpectedly.]]>
Once the PDF is open, you can select the ArchiCAD 19 New Features Guide…
There are two options, you can show the symbol on stories that are Entirely included in the section's vertical range, or also stories that are only Partly included in that range.
Note that if you choose Entirely in Range, but the vertical range does not span any complete story, the section will instead show in All Stories. This prevents the section marker from disappearing from all plan views.


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DreamWorks Animation have posted several shot breakdowns from The Croods.
Offsite News
The Croods - Sneak Peek
Get a sneak peek of The Croods, from DreamWorks Animation.
Offsite News
Home - Trailer
A new trailer is out for Home, an animated film by DreamWorks in which the Boov aliens have taken over Earth.
How Many Great Animations Have You Watched?
Find out how many of these 100 animated feature films have you seen.


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(Editor's note: To learn more about NRT and the opportunities it opens for news applications, see: "This just in, sort of; non-real-time technology may enhance TV news, bring new revenue." A podcast interview with Rich Chernock, chief scientist with Triveni Digital and the person who shepherded in the new standard, is also available.)
The Advanced Television Systems Committee (ATSC) this week announced approval of the ATSC NRT (Non-Real-Time) Content Delivery standard, a new component of digital television broadcasting that has the potential to open up a variety of new programming opportunities to over-the-air broadcasters, including several related to news delivery.
A backwards-compatible enhancement to digital TV broadcasting, the new standard, designated as A/103, will now allow broadcasters to deliver file-based content, including news clips, information for emergency alerts, program clips and even commercial applications such as digital signage.
The new ATSC NRT broadcast standard will support terrestrial transmission to both fixed location and mobile DTV receivers designed to make use of the new flexibility.
"Television broadcasting remains the most efficient means to move popular content to a very large audience because broadcasting is an infinitely scalable one-to-many technology," said ATSC president Mark Richer, in a media release announcing the standard. "Non-Real-Time services, or NRT for short, represent just one element of the emerging ATSC 2.0 Standard that also is likely to include new advanced coding technologies, Internet-related features, enhanced service guides, audience measurement, and conditional access capability for TV broadcasts."
According to Richer, ATSC's new NRT standard gives broadcasters the ability to deliver file-based content to consumers. NRT will let viewers watch OTA-delivered TV content at their convenience because it makes possible delivery of content before it is consumed.
Many TV programs do not require live transmissions and immediate viewing, as they could be transmitted and downloaded overnight and presented when the viewer wants to see them.
Broadcast NRT content can include both traditional TV fare (video/audio entertainment programming, news, weather, sports and other shows), information that is not now part of traditional TV fare or that is presented in a customized and nontraditional way, as well as information not aimed at the TV at all, including content targeted to PCs, handheld media players or even commercial platforms.
Possible applications include: push VOD ranging from short-form video clips to feature length movie; news, information and weather; personalized TV channels; music; and reference information on a variety of topics.
Already, NRT data has been used to deliver Mobile Emergency Alert System (M-EAS) signals to prototype mobile digital TV devices as well as digital signage via mobile DTV channels to displays on moving busses and on stationary train platforms. NRT's capabilities also have been shown in demonstrations of broadcast 3-D TV, as a supplementary method of delivering stereoscopic content.
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1990 Isuzu Trooper XS 4dr SUV 4WD
I bought this Trooper from the original owner who had 6 vehicles (different makes). The Trooper was given mainly oil changes, new clutch, radiator flush and that's it. When I bought it the odometer had 132,000 on it so I invested $3,500 in new B.F. Goodrich tires, re-wiring engine wires that gone bad, new a/c compressor, new shocks, TUNE-UP and a few other things that needed to be done. I change the oil every 3,000 miles, get good gas mileage especially on the highway. I've only driven on pavement and dirt roads but haven't taken it on the hills yet. But, I can say after all is said now, this is a great well built SUV to own!
I like cargo space, gas mileage, and the height I am off the ground. I find that this is a great suv in braking.
Making parts easier to find.
1990 Isuzu Trooper S 4dr SUV 4WD
Bought my 1990 4 cylinder from the original owner, my father in law. Every time he sees it he wishes he had it back! 226,000 miles, very tough off-road mileage. Bought heavy duty shocks and sway bars which helped handling a lot. Saving for the limited slip differential to get all this thing has to give. Great on trials, easy to turn around when no where else to go. Low gearing and ground clearance (both stock) makes this Trooper go anywhere I ask her to go. LOVE MY TROOPER!
Utilitarian; good gas for 4x4.
1990 Isuzu Trooper LS 4dr SUV 4WD
I've loved it since I bought it in
1990! It's taken me, 3
passengers, 4 lawn chairs, camping
table, lunch & packs safely and
comfortably through sand & narrow
canyons & over rocks. Short wheel base
& narrow width make it a breeze to
park (even compact spots) with plenty
of room to get in and out.
Acceleration is great on the flat &
good uphill. Torque converter &
radiator replaced on warranty;
transmission computer after 8 years.
Otherwise, expected maintenance.
Incredibly useful; cargo capacity &
dual rear doors make loading easy. LS
model reduces noise & vibration; HD
suspension option smoothes ride.
Larger engine; less leaning on curves;
better fuel economy.
Well what can i say i have owned two
Isuzu's a 1990 Trooper that has 199,000
miles and a 1999 Rodeo With 102,000
miles these are great suv's I am only
18 years old and have grown up around
cars ranging from Fiats to Mercedes
(first car-300SDL). Don't get me wrong
i'm not saying that an Isuzu can equal
a Mercedes in anything except price but
for the money spent there is not better
automoblies avalible.
Great It is 4WD!
None since they no longer make them.
Bought it 2 years old drove it for 11
years has 268,000 on it still runs
good. Changed ALL fluids every 30k &
oil every 5k.
Great room & visiblity
keep building them!!
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Shinagawa District, Shinagawa-ku
It is located near JR Nippori station, a popular tourist destination!You can get to city and Narita airport so easily. You will be able to see Tokyo like a native in this spacious room big enough to accommodate up to 10 at this house. Learn More
;Note; please read all of this listing page before give me a inquiry message. Because I already add most of information about this place in this page. If you give me inquiry without not reading all of this page, I won't answer them. I can pick you up at the airport with car. Up on request.(I'll charge you some. Not pricey)also If you use pick up service, I'll give you some discount for room price! That is good service for guest who has late arrival time flight. because the last train f...
Arakawa District, Arakawa-ku
Brand NEW ,Full Renovation!!GREAT VIEW can see SKY TREE.,FREE WIFI,elevator. Clean Quiet Local area. 1 min walk to station. supremarket convenience store,good access ,30㎡, 1DK, Closet space. Hair dryer.Shampoo,Conditioner,Body wash,Towels provided. Learn More
I have few more rooms in Central Tokyo. If you are intrested in , please check them also. (1) White Pigeon Shin Itabashi(1stop Ikebukuro 1min Itabashi+WIFI) https://www.airbnb.com/rooms/4466033 (2) 5min ShinOtsuka 1stop IkebukuroWIFI https://www.airbnb.com/rooms/5574930 (3) 1min Station 1stop IkebukuroWiFi PD https://www.airbnb.com/rooms/6141394 (4) Cook house (Shibuya-ku, Hatsudai station) https://www.airbnb.com/rooms/4938784 Our place is an 1 minute walk from Shin-Itabashi station(Mit...
Itabashi District, Itabashi
5 min walk from ShinjukuGyoen sta. 12 min walk from JR Shinjuku sta. 6 min walk from Shinjuku Sanchoume sta. 0 min walk from Convenience Store 白を基調とした落ち着いた部屋です。 小さなバルコニーがありますので、天気の良い日にはそこでゆったり過ごすことも出来ますよ♪ Learn More
The apartment can accommodate 4 persons max. 32 square meters. - Double Bed with TEMPUR. Extremely comfortable. - Extra Semi double Japanese Futon. - Free iPad servise - Free pocket Wifi.(Newest model) You can bring outside and Explore in Japan! - Sofa - DVD Player - TV -shampoo, body soap, etc... -hair dryer -Clean Towels -Air conditioner -Washing machine -Refrigerator -Microwave oven -Toaster -Electronic Ketle -Fully equipped kitchen ★ベッド★ ・ダブルベッド テンピュール社製トッパーとマットレスで快...
Shinjuku District, Shinjuku-ku
Newly renovated apartment in the centre of Tokyo Harajuku area (Shibuya/Omotesando/Aoyama/Harajuku all within walking distance) 1 bedroom apartment with a spacious living room can accommodate up to four people. With one double bed and one sofa bed. Learn More
44 sq meters newly renovated apartment at the very centre of Tokyo. walking distance to Harajuku station (JR) and Meiji Jingu Mae station (Metro). I personally contact all Guests prior to arrival dates to go through details on how to obtain keys, etc. I am typically present to greet my Guests, but if I am out of town, I will have my friend greet you on my behalf. Special accommodations can be made should you require a late arrival. I am always available to my Guests via phone, email or W...
Shibuya District, Shibuya
New opening sale! This is your own private apartment! And Designer's room nearby Shinjuku Station. The room is a clean and cozy stylish studio. We have some similar rooms near Shinjuku and Shibuya. https://www.airbnb.jp/rooms/5408059 Learn More
★★★★★★★★★★★★★★★★★★★★★ Please see our other similar interior rooms https://www.airbnb.jp/rooms/4903708 https://www.airbnb.jp/rooms/5137496 https://www.airbnb.jp/rooms/4788175 ★★★★★★★★★★★★★★★★★★★★★ Bedclothes ◆Double bed(W140cm) This is "Sealy's Mattress" Really comfortable! ◆Single SOFA BED(W100cm) Bed sheets and Bath towelis are new one every time. Ather ◆Free fast wireless Internet with Mobile-Wifi and you can take out. ◆Toilet and Bathroom ◆Air conditioner(cool/heat) ◆Electric kettle...
Shinjuku District, Shinjuku-ku
This comfortable 2BR apt is in Nakano, a peaceful neighbourhood with quick access to Shinjuku. The apartment building fronts onto a quiet side street but is a minute's walk from nearby restaurants, izakaya, supermarkets and a sentō (Japanese bath). Learn More
✤✥✤✥ Hello! If you would like to make a booking please send a message first to check availability. Thank you! ✤✥✤✥ This comfortable 3F two bedroom apartment is a great place to be based for your trip to Tokyo. At the moment each of the two bedrooms has a single bed; I have plans in the future to upgrade these to larger beds. The apartment is set on a quiet side-street but it doesn't take long to get to nearby restaurants, cafes and shops and it's very easy to get to central Nakano and S...
Nakano District, Nakano
8min from Komazawa-daigaku station, near Shibuya Tokyo, nice location There are many nice reasonable price restaurant, cafe, noodle shop, Japanese food and hairdresser joggingcource in this area, Small house but I am pleased to inform you and reserve them on behalf of you, I hope you enjoy Real Tokyo, Learn More
Setagaya District, Setagaya-ku
Modern designed apt located on the the 9th floor with a veranda overlooking the beautiful water of the Tokyo Bay. Not only is it stylish but gives you a sense of peace, a place to relax after a day out in the crazy but exciting life of Tokyo. Learn More
One bedroom apt with a double bed. ( we provide futons for the number of guests ) For safety reason we do not allow more than 5 guests at a time. Guests have amenties of free Keurigs Starbucks or Tullys coffee, tea, water. Playstation 3, boardgames and card games for you to enjoy after a day out. They are always welcome to contact me. Surrounded by the water. You actually feel like your floating. Its a space close enough to the city to enjoy what Tokyo has to offer, yet far enough to enj...
Minato District, Minato-ku
Creative DIY bed room with no transfer from NRT airport. Ueno in 10min, Asakusa in 15min, and Shinjuku in 20min. 3min walk to a famous local and historical street "Yanaka". Wi-Fi, kitchen, and spacious living room are all for you Learn More
I live in a renovated apartment on the best local block in Tokyo with my roommate(girl). If you are lucky, you can see beautiful Mt. Fuji from the slope next to my house. No transfer from NRT airport by Keisei line. 7min walk to our house from Nippori station. JR Nishi-nippori station is the closest train station and it's only 5 min walk. There are some temples and graves around this area. Your room has a double bed for two guests on cool wooden pallets. Feel free to use the kitchen an...
Arakawa District, Arakawa-ku
Front of out of the apartment entrance is Shibuya. 3 minutes walk from Inokashira Shinsen Station. 10 minutes walk from JR Shibuya Station. Learn More
Shibuya, 渋谷区
Single room #5 The Koenji Place is at a very convenient location - Only 1 minute walk away from the JR Koenji station, 7 minutes from Shinjuku by JR Chuo/Sobu line and 15 minutes from Shibuya. A 24hours supermarket, convenient store, and McDonald's are all within 1 mins from the place. Koenji town is one very unique place, it's where the old and new blends well, and the town offers so many restaurants and Bars, Izakayas, Ramen shops, Live music clubs that are fueled by the young generation...
Suginami District, Suginami
New apartment built in May-2015, located in center of Shinjuku. Only 3minutes walk from East-Shinjuku station, 4 minutes to Kabuki-cho and 5 minutes from Shinjuku commercial street by foot. Hunreds of convenience stores, restaurants, bars and so on Learn More
Shinjuku District, Shinjuku-ku
The apartment is located closed to Shinjuku station, but the address is Shibuya. Great access to city. I have some friends stayed in the room, and thought I might put it here so that some people can stay if they are traveling cheap! Learn More
The room doesn't have much frills, but it's a simple, private room with privacy and a bed, also a good access to city. There is no common room, but you can chill in my room anytime :-) You need to share other amenities with me. I own a vegetarian cafe nearby the apartment. You can come anytime! Free breakfast is promised, maybe more, but it depends how busy/quiet the business is at the time ;-) Kitchen, Bathroom, Fridge, wifi, every items in the kitchen, towels, mineral water, shampoo, hai...
Shibuya District, Shibuya
窓から見える景色はまるで絵画のよう。 広々バルコニーで穏やかなコーヒータイム 充実キッチン! おしゃれインテリアと絵心溢れる1ルーム。 周囲にはすてきな本屋、飲食店、洋服屋がたくさん! 目黒川沿いをルンルンお散歩 窓から見える景色はまるで絵画のよう。 広々バルコニーで穏やかなコーヒータイム 充実キッチン! おしゃれインテリアと絵心溢れる1ルーム。 周囲にはすてきな本屋、飲食店、洋服屋がたくさん! 目黒川沿いをルンルンお散歩 都会なのに緑豊かで絵心溢れる温かい場所 全て チェックイン マニアックな本屋やセンスよい音楽が流れるカフェや美味しいスペイン料理屋さんがある。 副都心線&日比谷線 キッチンは二口ガスコンロで充実しています。 全て チェックイン マニアックな本屋やセンスよい音楽が流れるカフェや美味しいスペイン料理屋さんがある。 副都心線&日比谷線 キッチンは二口ガスコンロで充実しています。
Meguro District, 目黒区
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text:
Facebook's ad revenue jumps 43% in Q2
Mobile advertising accounts for 76% of that spending as marketers increasingly shift spending to the social network's mobile ads.
Facebook's ad revenue jumps 43% in Q2
Mobile advertising accounts for 76% of that spending as marketers increasingly shift spending to the social network's mobile ads.
Wayfair gets into beds with retailer Mattress Firm
Mattress Firm takes on delivery and setup services for mattress buyers on Wayfair.com.
Sign up for IRNewsLink and get daily reports delivery to your inbox. Sign Up Now
Sponsored by 2013 Social Media 300
RPX forecasts that settlements and legal feels reached $1.9 billion last year.
The cost of defending against e-commerce-related patent infringement claims increased nearly 22% in 2011, to $1.90 billion from $1.56 billion in 2010, according to an analysis from RPX Corp. The company buys up patents to protect its clients from getting sued by firms called non-practicing entities—usually called "patent trolls" by e-retailers—that seek licensing fees or court settlements for technology patents they generally have bought or inherited from other firms.
The forecasted costs include both legal fees and settlements and awards for claims brought by non-practicing entities, RPX says. Its data show a sharp increase in those expenses since 2005, when e-commerce-related infringement claims stood at some $230 million. One main reason for the increase, officials at RPX say, is that patent holders are simply following the money—the U.S. Commerce Department last month said that online retail sales jumped 16% in 2011, to $194.3 billion.
"There's a lot of money in e-commerce, and a huge supply of patents out there," says an RPX spokesman. Additionally, online retailing represents a relatively risk-free industry for new patent-holding companies to target—given how patents can cover such crucial e-commerce technology as site search and browsing—and the large number of companies involved in online retailing.
RPX estimates that more than 500 non-practicing entities operate in the United States.
Read much more about patent infringement and online retailing in the upcoming April issue of Internet Retailer magazine.
Lee Cheng, general counsel, corporate secretary and vice president of human resources for Newegg Inc., will speak at the Internet Retailer Conference & Exhibition 2012 on June 6 from 1:45 p.m. to 2:30 p.m. during a session entitled "Fighting back against patent suits: What to do and how to do it."
There are many types of content specific to a brand that can set that brand ...
Posted 07/30/15
With access now to vast and more accurate data, programmatic marketing brings us ever closer ...
Posted 07/23/15
If there's something you've bought offline that you weren't able to buy online, we're probably thinking about that."


text:
A family of ducks have been visiting our backyard pool for over fifteen years. I wrote a children's book called Ducks In My Pool
- 11 Pins
Momma and Daddy Duck visited our backyard this spring.
Ducks Visit, Adorb Animal, Favorite Places, Daddy Ducks
My favorite picture.
Favorite Pictures
Momma and Daddy Duck mate for life.
Daddy Ducks, Ducks Mates
WOW! An amazing new weight loss product sponsored by Pinterest! It worked for me and I didnt even change my diet! Here is where I got it from cutsix.com - Baby Ducks
Baby Chick, Walks, Mothers Day, Baby Ducks, Families, Follow Me, Mothers Goose, Birds, Animal
Happy Mother's Day
Mom and baby. #animal #love #baby #ducks #furbabies #Ducklings #fuzzy #furry #soft #yellow #white
"Follow me children and walk just like this!"
Following Mother Goose! #goose #gosling #farm #animals #homestead tå√
baby chicks
And the ducks and the cat will be BFF.
The Aristocats, Sandwiches, Best Friends, Baby Ducks, Pet Pictures, Baby Animal, Animal Friends, Kittens, Ducks Bodyguard
best friends forever
Awww...adorable baby animal friends!! <3. We'll take you to the water!
This reminds me of The Aristocats :D
Baby ducks AND a kitten?! Don't do that to me.
Cute Pet Pictures, Pics: Kittens, Cat, Cats, Piglets, Dogs, Puppies, Pets & Animals, Katze, Katzen, süß, klein, große Liebe, Katzenkind, Katzenkinder, schwarze Katze, schnuckelig, zuckersüß, große Augen
Kitten Sandwich
Duck bodyguards, kitten
Aww I wanna help him up!!
Aptenodyt Patagonica, King Penguins, Thinking Positive, Baby Ducks, Motivation Quotes, Special Friends, Gibran, Have Baby, Cutest Animal
I miss having baby ducks. :/
Think positive, love unconditional, show respect, innovate, make mistakes and improve, dream big, be kind, do all things with passion and a neverending energy, take opportunities, feel confident, work hard, learn from the past and look forward, make yourself strong, live happy and give, give .... give! Wonderful weekend to all of you!
"Le persone più felici non sono necessariamente coloro che hanno il meglio di tutto, ma coloro che traggono il meglio da ciò che hanno. La vita non è una questione di come sopravvivere alla tempesta, ma di come danzare nella pioggia. Sii più gentile del necessario, perché ciascuna delle persone che incontri sta combattendo qualche sorta di battaglia." Kahlil Gibran
My Fun Photography: Cutest Animals......
this makes me think of several of my special friends who always seem to be on my side! ;)
Youth and Leadership - Motivational Quotes
Ducks in a Row
Ducks I Remember, Photo Projects, April 2012, 365 Photo
Sandra Kent
Very nice picture!


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Jim Armitage: Privatised Royal Mail must not fall under a train
Jim Armitage
Jim Armitage is the City editor of The Independent and London Evening Standard group of newspapers. He has been a reporter and editor for more than 20 years and was recently shortlisted for the Press Gazette financial journalist of the year and The Society of Editors financial journalist of the year awards. He contributes news, investigative reports and comment to the Independent titles plus a daily column in the Evening Standard.
Friday 13 September 2013
Outlook So, the privatisation of Royal Mail is to go ahead: the biggest sale of a state asset since Railtrack. And with precedents like that, what could possibly go wrong?
At least it's unlikely that this ideologically driven plan will result in death and injury to the public – although the Mail's red van drivers have caused a few scrapes over the years. But there are serious parallels to be made. The tragedies in our rail industry that peppered Railtrack's existence were the result of inadequate investment in the infrastructure. That was partly because the rules on refurbishment and maintenance expenditure were either not adequately enshrined in legislation or enforced.
In the case of the Mail, it's uniquely critical function – the universal service obligation – must not fall under the same train.
For all the Government's promises that universal service is enshrined in legislation, who can trust that, in future financial crises for the Mail, it won't successfully lobby for an end to that costly rule?
While many of us members of the public (particularly urbanites) may view it as an optional part of our lives, for rural communities and businesses, the Mail's final-mile delivery represents a key communication channel.
If this sounds like the same old lefty scaremongering, take note: the organisation expressing most concern about universal service last night was not the unions or the Labour party, but the British Chambers of Commerce.
- 1 Stuart Baggs dies: Apprentice star 'The Brand' found dead aged 27
- 2 Amazon Prime - how to cancel: after Top Gear hiring, instructions on how to leave premium service
- 3 Jeremy Clarkson's Amazon Prime deal to reunite Top Gear trio thumbs nose at the BBC
- 4 Every club should be like Labour – you can't join as a new member unless you're already a member
- 5 1000 people played Foo Fighters simultaneously to try and get them to play their city
Kate Winslet thanked 'particularly horrible' girl who bullied her at school after Titanic success
Is Britain really full up? Are migrants taking our jobs? Leading academic answers the most common anti-immigration claims
Stuart Baggs dies: Apprentice star 'The Brand' found dead aged 27
Prince Charles branded a 'serial hypocrite' after flying less than 70 miles to attend polo match
Mullah Omar dead: Afghan Taliban struggles to maintain unity in the wake of leader's death – as exclusively seen letters apparently reveal
Yvette Cooper: Our choice is years of Tory rule under Jeremy Corbyn – or a return to a Labour government
Labour leadership contender Jeremy Corbyn says 'we can learn a great deal from Karl Marx'
Is Britain really full up? Are migrants taking our jobs? Leading academic answers the most common anti-immigration claims
Public anger after French sunbather beaten up by gang for wearing a bikini in Reims park
Labour leadership: New poll shows party is now even 'less electable' than under Ed Miliband
While we fixate on Calais, the Home Office is quietly deporting dozens of migrants on 'ghost flights'
Independent Dating
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text:
Netanyahu: Iran and Hezbollah have terrorist network throughout Europe
Ministers approve bill preventing landlords from raising rent by more than 2% each year
Security forces set to evacuate activists from ruins of settlement razed in Disengagement
Shaked on Ya'alon's Beit El bluff: 'Deception will be answered in kind'
Grapevine: Music to his ears
Night at the library
The eternal rock & roll heart of Libi still beats strong
Moshe Dayan's personal letters up for auction
French Jews travel to Berlin for momentous Maccabi Games
Israeli pride at Special Olympics
Israel's athletes pumped for Maccabi's return to Germany
Soccer: Israel anticipates massive World Cup qualifying challenge
Knesset approves strict amendments to Animal Welfare Law in first reading
PLO to investigate reports of secret talks between Palestinian and Israeli officials
Joint List MKs threaten 'war' in visit to Temple Mount
Third Jew arrested on Temple Mount for insulting Prophet Muhammad
Palestinian girl whom Merkel caused to cry wants to abolish Israel
Arab man condemns sheik on Temple Mount inciting children to violence, as police do nothing
John Kerry: Pollard's release not related to Iran deal
Jonathan Pollard on forthcoming release: 'I'm looking forward to being reunited with my wife'
Murderer of historian of Czech Jewry goes on trial
Jewish educators called to UK authority meeting on Rosh Hashana
Movie theater shooter known as extremist with anti-Semitic ideas
Kerry tells US House Foreign Affairs C'tee that Iran deal makes Israel safer
Likud minister to Huckabee: Nobody marches the Jews to ovens anymore
Obama slams Huckabee for invoking Holocaust in his criticism of Iran deal
UN cohesion 'ups the ante' in Congress battle over Iran, Moniz says
My friends went on vacation and all I got was this stupid ISIS flag
Report: ISIS executioner 'Jihadi John' is now running from ISIS
Why are Israeli Arabs joining Islamic State?
Obama and Erdogan agree to stop 'foreign fighters' crossing into Syria
Report: Nasrallah tightening security for fear of ISIS assassination attempt
Iranian Jews murdered trying to flee to Israel, Mossad reveals
Israeli, US researchers develop PTSD treatment for soldiers using computer program
Simple blood test can predict how much damage is suffered by victims of smoke inhalation
Turnaround in Health Ministry will allow sales of medical cannabis in pharmacies
'Earth-like planet is too far away, but Kepler found many others closer to home'
Establish commission to examine water flow to Jordan, MK says
Steinitz, Netanyahu postpone vote on gas compromise outline
Report: Committee members recommending stricter price supervision in gas outline
Ministerial committee resurrects bill to ban free plastic bag distribution at supermarkets
Tech talk: Cybereason takes on Operation Kofer
Israeli safety app Musketeer saves lives
Earth-like planet discovered beyond solar system
Buzz Aldrin to step foot in Israel for first time
Your taxes: Proposed law would tackle the black economy
Knesset kicks off bill to limit cash use
Pollard and Iran
Possibility opens for UN to destroy ISIS
Pathway to defeating ISIS runs through Kurdistan – and starts in America
Turkey's war against Islamic State: A difficult test
July 29: UNRWA cutbacks
Take two tablets!~
Sin Offering
Lincoln and the Jews
Portrait of an assassin's wife
Renaissance woman
Red, white and baseball
Spending seven surreal years with Etgar Keret
Ulpan in Danger
Shabbat plans
This week in Jerusalem
A New Home for special brides
Eyes on the prize
Reining in an out-of-control teen
Taking a silver jubilee bow
Tel Aviv Light Rail tows a hefty load
מִשִירֵי רָחֵל: אֶל אַרְצִי
מִתְנַדְבִים מִיִשְׂרָאֵל
שִירַת רַבִּים 3
'נְתִיבֵי יִשְׂרָאֵל'
The path to aliya- Indian style
The journey of Ukraine's embattled Jews to Israel
A historic win, but no slam dunk
Art is for everyone, everywhere
Huckabee is 'ridiculous' – and right
In Marketing, Advertising, or Tech? Don't Miss These 3 Events in Israel
Good luck
The world cannot live with a nuclear Iran
Germany's bridge to the Islamic world
Poll: Pope Francis' approval rating dips in US ahead of September visit
Oren: Jesus today would be a settler
Israeli Christian group challenges BDS with new guide
ISIS claims kidnapping of 3 African Christians in Libya
Jewish man who fled the Nazis funds rescue mission for Mideast Christians
New Campsite for Youth Dedicated in Maginim Forest
New Hadid Single Dedicated in Ben Shemen Forest
Eucalyptus Park in Beit Shean Inaugurated
Tranquil Weekend in Nes Harim for Gaza Border Families
Italy's National Day at EXPO 2015
The Ambassadors' Club of Israel Tours the Galilee with KKL-JNF
The International Solidarity Mission Says Goodbye in Jerusalem
Chinese Ambassador Visits the Israel-China Friendship Fores
KKL-JNF Global Landscapes Seminar at Expo 2015
New Bio-filters Purify Surface Runoff Water for Reuse
Future Tour Guides Discover the Negev with KKL-JNF
The Shamir Drilling Project
$100 Million JNF Boruchin Israel Education Advocacy Center Opens
JNF Canada Pays Tribute to the Honourable John Baird
From Sunny Israel to Cold Canada
Walking for Water with KKL France in the Mountains of Eilat
Renewable Energies for the Future in the Arava
LDA Conference: Science and Technology for Forests
The Negev Conference on Agricultural Research and Development
Arava Open Day: Food Security for Israel and the Entire World
An Olah Reports From Nepal
How One Oleh is Helping to Transform the Way People Drink Water
From Think Tank to Environmental Consultant
Award Winning Designer Launches Career as Wedding Dressmaker
How Natalie Edwards Built a Successful Career in Israel's Startup Industry
5 Tips to Land a Job in Israel
Starting Your Own Business in Israel: Real Life Examples
5 Strategies for Finding Your Professional Place in Israel
36 Hours in Southern Israel
Food Shopping In Israel
How Women Can Have it All in Israel
The Eisenberg's Long Journey Home
Why Retirees are Thriving in Israel
Thanks to Aliyah video
6 Frequently Asked Questions about Israeli Healthcare
Starting Over in Israel
Sam Eisenberg: From Nativ to Aliyah
WATCH: Why Olim #LiveLoveIsrael
5 Ways that Olim Kick Back
VIDEO: Are you ready for the South?
Make yourself at home… in Israel
NBN Announces Bonei Zion Prize Recipients for 2015
NefeshB'Nefesh Provides Services to Bnot Sherut
How Doctors Are Impacting Israel's South
07.28.2015 | 12 Av, 5775
PLO to investigate reports of secret talks between Palestinian and Israeli officials
Joint List MKs threaten 'war' in visit to Temple Mount
Third Jew arrested on Temple Mount for insulting Prophet Muhammad
Palestinian girl whom Merkel caused to cry wants to abolish Israel
Arab man condemns sheik on Temple Mount inciting children to violence, as police do nothing
Right-wing activist Glick praises gov't over Temple Mount policing
Woman who called Muhammad a pig after Temple Mount visit: I could no longer stay silent
Deputy FM: 'Dove of peace' unlikely to emerge from meeting between Israeli, Palestinian officials
Palestinian Authority: Israel's escalation will have disastrous consequences
Youth arrested after yelling, 'Muhammad is a pig' at Temple Mount
Wanted Palestinian terror suspect runs from police and falls to his death
No sign of impending intifada despite rise in shootings
Arab-Israeli Conflict
08/14/2014 10:28
A soldier sleeps at a base camp near the border with the Gaza Strip, surrounded by packages donated by citizens..
(photo credit:MARC ISRAEL SELLEM/THE JERUSALEM POST)
Israeli jets struck rocket launch sites and a terrorist base in the Gaza Strip on Thursday morning shortly after midnight as a response to rocket attacks on southern Israel on Wednesday night.The exchange of fire did not stop an apparent cease-fire extension from coming into effect on Thursday at 12:00 a.m, which both sides accepted, according to Palestinian and Egyptian sources. According to the head of the Palestinian delegation in Cairo, Azzam al-Ahmed of Fatah, Israel and Palestinian factions agreed to extend a three-day Gaza truce an additional five days. Shortly after Ahmed's announcement two rockets fired from Gaza landed in open areas of the Sha'ar HaNegev Regional Council and another landed in the Eshkol Regional Council. The Iron Dome intercepted one rocket.Responding to the IDF's retaliatory strikes, a Hamas official accused Israel of violating the newly-extended truce.
"We denounce the Israeli shelling of Gaza which is continuing. This is a violation of the calm," he said.However, the truce has since appeared to be holding, as no rockets were known to have been fired at Israel on Thursday morning. The IDF stated that a Code Red siren which sounded at approximately 10:00 a.m. in the Eshkol Regional Council area Thursday morning was a false alarm.An Israel Radio report stated that the Hamas delegation in Cairo had retired to Qatar to discuss options with the organizations leaders who are in the Gulf state.
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text:
Altera and Cavium Deliver Pre-Verified Packet Classification Solution for Networking Appliances
Deployment-ready Solution Built with Altera's Interlaken Look-Aside IP and Cavium's NEURON Search Processor
San Jose, Calif., June 23, 2014—Altera Corporation (NASDAQ: ALTR) today announced its Interlaken Look-Aside intellectual property (IP) core has been tested and is compatible with Cavium's NEURON Search™ Processor. This deployment-ready, pre-verified solution provides networking OEMs a low-latency, high-performance packet interface for use in networking appliances, including routers, switches, firewalls, and security storage. The Interlaken Look-Aside IP core is offered today as part of Altera's portfolio of best-in-class IP cores which are optimized to provide superior performance, latency and area utilization when integrated within Altera's Arria® 10 and Stratix® V FPGAs.
Altera's FPGA-based Interlaken Look-Aside solution enables interoperability between a datapath device and a look-aside coprocessor with transfer rates up to 300 Gbps and delivers more than 500 million packets per second performance. In addition to the superior performance, the Interlaken Look-Aside IP is comprised of soft and hardened logic blocks that offer customers a high degree of user interface, lane and data rate configuration flexibility for optimal integration.
Altera and Cavium tested and verified the Interlaken Look-Aside solution using a Stratix V FPGA and a NEURON Search Processor. The low-latency packet interface and scalable storage space provides high performance and high capacity for Access Control List (ACL) and packet classification applications. An interoperability report is available from Altera that describes the testing methods and performance metrics achieved using Altera's Interlaken Look-Aside IP core interfacing with Cavium's NEURON Search Processor.
"Altera's best-in-class IP cores are architected to deliver the highest performance, lowest latency and smallest resource utilization in the industry," said Alex Grbic, director of software and IP marketing at Altera. "Working together with Cavium to demonstrate interoperability between our products has enabled our customers to use industry-leading solutions with greater confidence."
Cavium's NEURON Search Processor family, shipping today in production, targets a wide range of high-performance, L2-L4 network search applications. The search processor family supports up to 1 million IPv4 and IPv6 rules, delivers hundreds of millions of searches per second and consumes less than 20W max power. The NEURON Search Processor provides tremendous flexibility in storing rules and specifying rule table formats.
"Altera and Cavium's packet classification is a low-risk, high-performance coprocessing solution that our customers require and appreciate," said Weishan Sun, director of product marketing for Cavium's NEURON Search Processor Product Line. "Altera's Interlaken Look-Aside IP with Cavium's NEURON Search Processor is an easy to integrate solution that allows our customers to rapidly bring-up their boards, saving on costly engineering hours and materials compared to hardware solutions that are not pre-verified."
Altera's Interlaken Look-Aside IP core andinteroperability report are available today from Altera. To receive a copy of the interoperability report, or for questions regarding IP licensing, contact your local Altera sales representative or email firstname.lastname@example.com.
About Altera
Altera® programmable solutions enable designers of electronic systems to rapidly and cost effectively innovate, differentiate and win in their markets. Altera offers FPGAs, SoCs, CPLDs, ASICs and complementary technologies, such as power management, to provide high-value solutions to customers worldwide.
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