bluuebunny/crossref_metadata_2025 huggingface dataset:
Mosaicism of isochromosome 18p.
Authors: Göcke H. , Muradow I. , Zerres K. , Hansmann M.
Published: March 2007

The first case of isochromosome 18p as a mosaic in a male fetus diagnosed by amniocentesis is reported. After termination of the pregnancy at 21 weeks gestation biopsies from different fetal organs as well as from the placenta were taken and set up for long term cell cultures. The distribution of the normal and abnormal cell‐line in fetal organs was unequal. Unexpectedly the metaphases in the placenta and the lymphocyte culture all showed a normal karyotype.

The tetrasomy 18p is associated with a uniform phenotype, even in fetal life, characterized by a small head with protuberant occiput, low‐set ears with posterior rotation, epicanthic fold, sharp pinched nose, convex and poorly formed philtrum, high‐arched palate, retrognathia, flexion contractures of fingers, short and broad hallux, hypoplastic penis, angulation of clavicles and mild scoliosis.

The propositus showed no growth retardation.
Prospective risk in reciprocal translocation heterozygotes at amniocentesis as determined by potential chromosome imbalance sizes. Data of the european collaborative prenatal diagnosis centres
Authors: Daniel A. , Boué A. , Gallano P.
Published: March 2007

The Data of the European Cooperative Prenatal Diagnosis Laboratories (Boué and Gallano, 1984) of 596 prenatal (amniocyte) diagnoses of familial rep was examined as to relationships between balanced/unbalanced result and ascertainment, carrier parent and chromosome imbalance size (percentage haploid autosome length). Each rearrangement was graphed once with actual (unbalanced result) or potential (normal or balanced result) imbalances plotted with trisomy as the ordinate and monosomy as the abscissa. The graphed data was divided into 15 regions, each of 2·0 per cent trisomy and 0·75 per cent monosomy and the rate of unbalanced pregnancies determined for each region. The highest rates of chromosomally unbalanced progeny (excluding regions with inadequate data) were found closest to the origin (i.e. associated with the smallest imbalances) and these were for ascertainment category 1 (previous rep unbalanced child) 22·3 per cent for maternal carriers and 39 per cent for paternal carriers. Overall in pooled data for this ascertainment category (without reference to the imbalance graphs) there were for paternal carriers 28·6 per cent unbalanced pregnancies and for maternal carriers 18·1 per cent. The graphed data, therefore, revealed the higher rates associated with some of the rep with small potential (combined duplication/ deficiency) imbalances. Lesser rates were observed for ascertainment category 2 (carrier parent with a history of recurrent miscarriage) with overall percentages of imbalanced progeny ranging from 2·7 (paternal carriers) to 4·7 (maternal carriers). Again, higher rates were revealed in graphed data for small potential imbalances. All unbalanced results for this group (ascertainment category 2) plotted in the region closest to the origin with rates of 16 per cent (maternal carriers) and 9·5 per cent (paternal carriers) in this region. Remarkably in both ascertainment groups 1 and 2 there was no significant difference in the size of the imbalanced segments for unbalanced progeny. In ascertainment group 1 this was (dup/def; mean ±S.D.): 1·09±0·77/0·47 ± 0·45 and in ascertainment group 2: 1·09 ±0·80/0·66±0·71. From the graphed data which arguably denote viability relationships, a trisomy was approximately 2·7 times as likely to survive until amniocentesis as a monosomy of equivalent size.

It is proposed that given further data, risk estimates could be determined for rep heterozygotes using the present approach where empiric data (from the family history or an analysed series of similar rep) is not available.
Prenatal detection of cystic fibrosis; comparative study of maltase and alkaline phosphatase activities in amniotic fluid
Authors: Claass Andreas H. W. , Kleijer Wim J. , Van Diggelen Otto P. , Van Der Veer Eveline , Sips Herman J.
Published: March 2007

The potential value of microvillar enzymes in the prenatal diagnosis of cystic fibrosis (CF) has previously been demonstrated and is corroborated in the present comparative study. Maltase and alkaline phosphatase (ALP) activities were studied in the amniotic fluids of 57 pregnancies with a 1 in 4 risk for CF or with a known CF outcome and in 489 controls.

A simple assay for maltase activity (MU‐maltase) with the fluorogenic substate 4‐methylumbelliferyl α‐glucoside, offers great technical advantages and an at least equal detection rate of CF, when compared to the previously used test with maltose as substrate.

Intestinal ALP was estimated either as phenylalanine inhibitable activity (PI‐ALP) or as the proportions of residual activity in the presence of the inhibitors phenylalanine or homoarginine. MU‐maltase and PI‐ALP appeared the most successful methods: both tests were able to detect 14 of the 16 (88 per cent) pregnancies with fetal CF. Each of the two tests alone also allowed a correct prediction in 24 of the 25 pregnancies at risk but with normal outcome; however all 25 cases could be correctly predicted by a combined evaluation.

It is suggested that more than one intestinal enzyme activity should be evaluated to allow optimal results in the prenatal monitoring of pregnancies at high risk for CF.
The in utero diagnosis and management of alloimmune thrombocytopenia
Authors: Lynch Lauren , Bussel James , Goldberg James D. , Chitkara Usha , Wilkins Isabelle , Macfarland Jan , Berkowitz Richard L.
Published: March 2007

A case is presented in which percutaneous umbilical sampling (PUBS) was utilized in the second and third trimesters for the diagnosis and management of a pregnancy at risk for neonatal alloimmune thrombocytopenia (NAIT).
Easurements of placental, decidual, and fetal proteins before and after chorionic villus sampling
Authors: Stabile I. , Warren R. , Rodeck C. , Grudzinskas J. G.
Published: March 2007

Circulating placental [human chorionic gonadotrophin (hCG), Schwangerschafts protein 1 (SP1), pregnancy‐associated plasma protein A (PAPP‐A), decidual (pregnancy protein) 12 (PP12), and fetal alphafetoprotein (AFP)] proteins were measured immediately before and within 1 h in 18 women undergoing diagnostic chorionic villus sampling (CVS) in the first trimester. An elevation of serum AFP levels was consistently seen, while fluctuations in excess of 10 per cent of the pre‐CVS levels of SP1 and PP12 were seen in the majority of patients. Fluctuations in hCG and PAPP‐A were consistently less than 10 per cent of pre‐CVS values. Post‐CVS changes in levels were not apparently associated with any feature of the technique, the pregnancy, or its outcome (one missed abortion). As feto‐maternal haemorrhage is a common event, anti‐D should be offered to rhesus‐negative women undergoing CVS. In the prediction of subsequent miscarriage, only hCG and PAPP‐A measurements should be considered.
First trimester diagnosis of Wolman's disease
Authors: Van Diggelen O. P. , Von Koskull H. , Ämmälä P. , Vredeveldt G. T. M. , Janse H. C. , Kleijer W. J.
Published: March 2007

Wolman's disease was diagnosed in the first trimester of pregnancy by the direct demonstration of acid lipase deficiency in chorionic villi. The diagnosis was confirmed by studies on cultured chorionic villus cells and fetal skin fibroblasts. Acid lipase activity was assayed with both 4‐methylumbelliferyl‐palmitate and radiolabelled cholesterol oleate as substrates. The higher specificity of the enzyme for the latter, natural, substrate makes it superior in prenatal diagnosis.
The fetal intrahepatic umbilical vein as an alternative to cord needling for prenatal diagnosis and therapy
Authors: Nicolini U. , Santolaya J. , Ojo O. E. , Fisk N. M. , Hubinont C. , Tonge M. , Rodeck C. H.
Published: March 2007

Seventy‐one fetal blood samplings (FBS) were attempted from the intrahepatic portion of the umbilical vein (IHV) at 18–34 weeks; 54 were attempted primarily and 17 secondarily after a failed attempt at the placental cord insertion. Fetal blood was obtained in 89 per cent of the cases. Intravascular transfusion (IVT) was attempted on 31 occasions and successful in 24 (77 per cent). In all cases of failed sampling or transfusion via the IHV, prenatal diagnosis and/or therapy was accomplished using alternative procedures. On only one occasion was the procedure postponed. There were no losses or neonatal morbidity attributable to the procedure.

FBS from the IHV may be considered as an alternative approach to sampling the placental cord insertion. It is recommended in cases where the approach to the placental cord insertion is difficult or hazardous.
Prenatal diagnosis and management of fetal ovarian cysts
Authors: Rizzo Nicola , Gabrielli Sandro , Perolo Antonella , Pilu Gianluigi , Cacciari Alfredo , Domini Remigio , Bovicelli Luciano
Published: March 2007

Congenital ovarian cysts are a pathological condition which can be diagnosed in utero by ultrasound. We report 14 consecutive diagnoses of fetal ovarian cysts, obtained in the second and third trimesters of pregnancy. Congenital ovarian cysts have almost invariably a good prognosis, and no change in the standard obstetrical management is required. However, the evolution of the disease in utero is extremely variable. The cyst may increase in size, decrease or even disappear, or undergo complications such as torsion and rupture, which may carry some risks to the fetus. When a fetal ovarian cyst is detected, serial ultrasound examinations should be performed. If one of the above complications is suspected, the option of prompt caesarean section should be considered.
Prenatal detection of trisomy 9 mosaicism
Authors: Schwartz S. , Ashai S. , Meijboom E. J. , Schwartz M. F. , Sun C. C.‐J. , Cohen M. M.
Published: March 2007

Chromosomal mosaicism in amniotic fluid cells poses a serious dilemma in prenatal diagnosis since the observation may represent: (1) pseudomosaicism—an inconsequential tissue culture artefact; or (2) true mosaicism—occurring in approximately 0.0 per cent of amniocenteses with a significant impact on pregnancy outcome. Mosaicism for trisomy 9 was observed in an amniotic fluid specimen obtained for advanced maternal age with two cell lines [46,XX (46 per cent)/47,XX, + 9 (54 per cent)] present in each of four culture flasks. Since more than 75 per cent of newborns with trisomy 9 mosaicism have complex cardiac malformations, a fetal echocardiogram was obtained at 20 weeks' gestation and interpreted as normal. A fetal blood sample (22 weeks' gestation) disclosed only a single trisomy 9 cell among the 100 metaphases analysed. However, a second fetal echocardiogram performed at the time of blood sampling suggested a non‐specific cardiac anomaly. Fetal autopsy following elective pregnancy termination revealed several malformations including severe micrognathia, persistence of the left superior vena cava, and skeletal anomalies. Cytogenetic studies of cell cultures derived from several fetal tissues demonstrated trisomy 9 ranging from 12 to 24 per cent.
A 1 and 6 month follow‐up of prenatal diagnosis patients who lost pregnancies
Authors: Black Rita Beck
Published: March 2007

This paper reports results of an exploratory study of prenatal diagnosis patients who experienced voluntary terminations of pregnancy following the detection of an abnormality or spontaneous miscarriages. The 121 participants were part of the national collaborative Chorionic Villus Sampling and Amniocentesis Study. They completed semi‐structured telephone interviews and mailed questionnaires at 1 month and 6 months after the pregnancy losses. Scores on the Profile of Mood States showed that mood levels improved significantly over time. However, there were some declines in loss‐related support from partners and others. The persisting distress and difficulties of a minority highlight the variability in women's responses to pregnancy losses. Women who lost pregnancies later in gestation, showed the greatest mood disturbances at initial assessments, used professional mental health assistance after the loss, or reported less satisfactory loss‐related support from significant others showed the greatest levels of mood disturbance at the six‐month assessment. Follow‐up contacts with patients who lose pregnancies should be used to inform women about the variation in possible grief reactions, to assess the extent of support the women are receiving from their partners and significant others, and to provide additional follow‐up or referral of those experiencing the greatest distress.
Alpha‐fetoprotein and acetylcholinesterase in twins discordant for neural tube defect
Authors: Johnson Virginia P. , Vidgoff Jaclyn , Wilson Nancy , Madison Dean
Published: March 2007

Twins concordant for elevated alpha‐fetoprotein (AFP) and acetylcholinesterase (AChE) and discordant for neural tube defect (NTD) and sex are reported. A literature review reveals instances of termination of twin pregnancies with one normal and one abnormal fetus, partly based on concordant high AFP and positive AChE (although discordant on ultrasound examination). The levels of AFP and AChE in twin pregnancies are probably a function of the number of layers of fetal membranes separating twin sacs: dichorionic, diamniotic membranes allow transfer of AFP; monochorionic, diamniotic membranes allow transfer of both AFP and AChE. Cautious interpretation of biochemical findings and reliance on high resolution ultrasonography are suggested.
Prenatal karyotyping in malformed fetuses
Authors: Rizzo Nicola , Pittalis Maria Carla , Pilu Gianluigi , Orsini Luigi Filippo , Perolo Antonella , Bovicelli Luciano
Published: March 2007

Experience with prenatal karyotyping of 237 fetuses with sonographic evidence of malformation is reported. Abnormal karyotype was found in 40 cases (16‐8 per cent): chromosomal aberrations were found in 19 of the 178 fetuses with an isolated structural anomaly (10‐6 per cent) and in 21 of the 59 fetuses with multiple malformations (35‐6 per cent). Detailed cytogenetic and morphological information concerning fetuses affected by omphalocele, duodenal atresia, hydrocephalus, multicystic kidney, unilateral hydronephrosis and cystic hygroma is reported. The need for a very careful ultrasound evaluation of fetal anatomy in these pregnancies is stressed, as the risk of a chromosomal anomaly depends mainly on the existence of more than one ultrasonically diagnosed structural defect.
Diagnosis and significance of cystic hygroma in the first trimester
Authors: Cullen Mark T. , Gabrielli Sandro , Green Jacqueline J. , Rizzo Nicola , Mahoney Maurice J. , Salafia Carolyn , Bovicelli Luciano , Hobbins John C.
Published: March 2007

Thirty cases of cervical cystic hygroma were diagnosed in the first trimester of pregnancy. Karyotype analysis was available in 29 (97 per cent). Fifteen (52 per cent) had a chromosomal abnormality. The ultrasound appearance was described as posterior cervical, lateral cervical, or cervical hygroma with hydrops. Of the 14 euploid embryos, six were electively aborted, two are undelivered, and six have been delivered as phenotypically normal infants. In cases in which the chromosomes were normal and the pregnancy continued, all lesions resolved by 18 weeks.
Duplication of the long arm of chromosome 13 secondary to a recombination in a maternal intrachromosomal insertion (shift)
Authors: Vekemans M. , Morichon‐Delvallez N.
Published: March 2007

A rare chromosomal aberration consisting of a chromosomal shift was found in a woman who had prenatal diagnosis because she had previously had a malformed girl with phenotypic features compatible with the diagnosis of Patau syndrome. Chromosome analysis using G, C, and NOR banding showed a direct intrachromosomal insertion of bands 13q12 to 13q14 onto the short arm of chromosome 13 at band 13p13. We discuss this observation and compare it with other published reports of chromosomal shifts.
Blood chemistry of human fetuses in the second and third trimesters
Authors: Legras B. , Clerc C. , Ruelland A. , Vialard J. , Millon J. , Cloarec L.
Published: March 2007

Six biochemical parameters and four enzyme activities were determined from the serum of 76 healthy and 56 pathological human fetuses between the 20th and 38th week of pregnancy. In the normal fetuses studied within that period, creatinine, immunoglobulin M, lactate dehydrogenase, and gamma‐glutamyltransferase increased; haemoglobin F and glucose progressively decreased; and alkaline phosphatase was at a peak around the 26th week; cholesterol and triglycerides were always low. The same parameters were also measured in some of the pathological fetuses and compared with their normal counterparts.
Catamenial hemoptysis from endobronchial endometriosis in a child with type 1 von Willebrand disease
Authors: Martire Baldassarre , Loizzi Michele , Cimmino Antonietta , Peruzzi Sonia , De Mattia Delia , Giordano Paola
Published: March 2007

Catamenial hemoptysis is a rare condition characterized by cyclic pulmonary hemorrhage, synchronous with menses and associated with the presence of intrapulmonary or endobronchial endometrial tissue. Because of the paucity of cases reported in the literature, information regarding the natural history is limited and also the optimal diagnostic workup and management of these patients are not well defined. In this report, we present a case of endobronchial endometriosis in a 12‐year‐old female diagnosed by bronchoscopy and immunocytochemical assay, associated with type 1 von Willebrand disease. Pediatr Pulmonol. 2007; 42:386–388. © 2007 Wiley‐Liss, Inc.
Internet and written respiratory questionnaires yield equivalent results for adolescents
Authors: Raat H. , Mangunkusumo R.T. , Mohangoo A.D. , Juniper E.F. , Van Der Lei J.
Published: March 2007

This study compared results from Internet and written questionnaires about respiratory symptoms in order to find out if both forms of the survey yielded the same answers. One thousand seventy‐one students, ages 13 to 17, were asked to complete either an Internet or a written questionnaire. The demographic characteristics of the participants equalled those of the general Dutch adolescent population. Participants were randomly assigned to fill out an electronic or written questionnaire. In addition to eight items from the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire, two items on doctor visits (medical attention) regarding asthma or allergic disease during the past 12 months were included. The participation rate was 87%. The Internet version of the questionnaire showed fewer missing answers than the written version, but this was not statistically significant. The respiratory items did not show statistically significant score differences between the Internet and written modes of administration, and there was no visible trend for higher respectively lower scores by either mode of questionnaire administration. From these results, we conclude that respiratory questionnaires may be provided to adolescents electronically rather than on paper, since both approaches yielded equal results. To generalize these findings, we recommend repeated studies in other settings. Pediatr Pulmonol. 2007; 42:357–361. © 2007 Wiley‐Liss, Inc.
Binding of antifusion peptides with HIVgp41 from molecular dynamics simulations: Quantitative correlation with experiment
Authors: Strockbine Bentley , Rizzo Robert C.
Published: March 2007

Peptides based on C‐terminal regions of the human immunodeficiency virus (HIV) viral protein gp41 represent an important new class of antiviral therapeutics called peptide fusion inhibitors. In this study, computational methods were used to model the binding of six peptides that contain residues that pack into a conserved hydrophobic pocket on HIVgp41, an attractive target site for the development of small molecule inhibitors. Free energies of binding were computed using molecular mechanics Generalized Born surface area (MM‐GBSA) methods from molecular dynamics (MD) simulations, which employed either explicit (TIP3P) or continuum Generalized Born (GB) water models and strong correlations between experimental and computational affinities were obtained in both cases. Energy decomposition of the TIP3P‐MD results (r2 = 0.75) reveals that variation in experimental affinity is highly correlated with changes in intermolecular van der Waals energies (ΔEvdw) on both a local (residue‐based, r2 = 0.94) and global (peptide‐based, r2 = 0.84) scale. The results show that differential association of C‐peptides with HIVgp41 is driven solely by changes within the conserved pocket supporting the hypothesis that this region is an important drug target site. Such strong agreement with experiment is notable given the large size of the ligands (34 amino‐acids) relative to the small range of experimental affinities (2 kcal/mol) and demonstrates good sensitivity of this computational method for simulating peptide fusion inhibitors. Finally, inspection of simulation trajectories identified a highly populated π‐type hydrogen bond, which formed between Gln575 on the receptor and the aromatic ring of peptide ligand Phe631, which could have important implications for drug design. Proteins 2007. © 2007 Wiley‐Liss, Inc.
Protein kinase D enzymes are dispensable for proliferation, survival and antigen receptor‐regulated NFκB activity in vertebrate B‐cells
Authors: Liu Ping , Scharenberg Andrew M. , Cantrell Doreen A. , Matthews Sharon A.
Published: March 2007

To investigate the importance of protein kinase D (PKD) enzymes we generated a PKD‐null DT40 B‐lymphocyte cell line. Previously we have shown that PKDs have an essential role in regulating class II histone deacetylases in DT40 B‐cells [Matthews, S.A., Liu, P., Spitaler, M., Olson, E.N., McKinsey, T.A., Cantrell, D.A. and Scharenberg, A.M. (2006) Essential role for protein kinase D family kinases in the regulation of class II histone deacetylases in B lymphocytes. Mol. Cell Biol. 26, 1569–1577]. We now show that PKDs are also required to regulate HSP27 phosphorylation in DT40 B‐cells. However, in contrast to previous observations in other cell types, PKD enzymes do not regulate basic cellular processes such as proliferation or survival responses, nor NFκB transcriptional activity downstream of the B cell antigen receptor. Thus, PKDs have a selective role in DT40 B‐cell biology.
Recent Applications of Biotechnology in Wine Production
Authors: Colagrande O. , Silva A. , Fumi M. D.
Published: March 2007

Over the last 20 years wine‐making technology has seen remarkable developments, which have improved the quality of the wines produced and have also made it possible to prepare wines with a range of characteristics. In this field, some biotechnological techniques have been of fundamental importance. Among these technological innovations, enzymic treatments, selected yeasts, improvement of microbial starters, and microorganism immobilization are reviewed here. A concise report is presented on the possibilities offered by enzymes that catalyze the transformation of polysaccharides, proteins, and polyphenols. We illustrate the importance of the characteristics of wine yeast and of a genetic improvement program for the wine industry, as well as the complexity of the genetic modifications of commercial wine yeast strains. We summarize the research on the optimization of new processes such as the use of immobilized microorganisms and cell‐recycle batch fermentation in the preparation of wines and sparkling wines.
Measurement of Unequal DNA Partitioning in Tetrahymena pyriformis Using Slit‐Scanning Flow Cytometry
Authors: Sweeney Pamela J. , Srienc Friedrich , Fredrickson A. G.
Published: March 2007

Slit‐scanning flow cytometry allows analysis of the locations of components along the long axes of cells. This is useful in determining how the components of a dividing cell are partitioned into the two daughter cells. Ciliated protozoa, unlike most eukaryotic cells, do not partition their DNA equally between daughter cells. We have used slit‐scanning flow cytometry to measure the distribution of this unequal DNA partitioning in the ciliate Tetrahymena pyriformis. Our results show that the difference in the amounts of DNA alloted to sister nuclei varies from cell to cell but, on the average, increases with the DNA content of the mother cell. However, the average difference in the fraction of the mother cell DNA partitioned to each daughter cell is more or less independent of the DNA content of the mother cell and is about 8.5% of that DNA content. Slit‐scanning flow cytometry also allows determination of the DNA distributions of dividing and newborn cells, determinations that are difficult to make with good statistical precision by other means. The measured newborn and dividing cell DNA distributions are broad, and this must be the result of repeated rounds of unequal partitioning of DNA.
Biotreatment of a Point‐Source Industrial Wastewater Arising in 3, 4‐Dichloroaniline Manufacture Using an Extractive Membrane Bioreactor
Authors: Brookes P. R. , Livingston A. G.
Published: March 2007

A novel membrane bioreactor has been used to detoxify an industrial “point‐source” wastewater containing aniline, 4‐chloroaniline (4CA), 2, 3‐dichloroaniline (23DCA), and 3, 4‐dichloroaniline (34DCA). This wastewater arises during the process of manufacturing 34DCA. Direct biological treatment of this wastewater is difficult without some form of pretreatment or dilution due to its inorganic composition. The reactor system described overcomes this problem by selectively extracting the anilines from the wastewater by means of a membrane separation step and transferring them into a biological growth compartment (biomedium) where they are biodegraded by specifically enriched microorganisms. This article reports the use of a membrane bioreactor configuration that uses shell and tube membrane modules linked to a CSTR bioreactor via a recirculating biomedium flow. The reactor system has successfully extracted and biodegraded the targeted pollutants from industrial wastewater samples over an operating period of 5 months with removal efficiencies of over 99% at wastewater residence times of less than 2 h.
Binding of Lysozyme onto a Cation‐Exchange Microporous Membrane Containing Tentacle‐Type Grafted Polymer Branches
Authors: Tsuneda Satoshi , Shinano Hironori , Saito Kyoichi , Furusaki Shintaro , Sugo Takanobu
Published: March 2007

Ion‐exchange adsorption of lysozyme to the sulfonic acid (SO3H) group on polymer chains grafted onto microporous polyethylene hollow‐fiber membranes was examined. The lysozyme solution was forced to permeate across the hollow fiber. Diversely anchored SO3H groups, i.e., SP and SS groups, were introduced into the membrane by reaction of the glycidyl methacrylate‐grafted membrane with propanesultone and sodium sulfite, respectively. The resulting SP and SS group‐containing membranes, designated as SP‐T and SS‐T fibers, respectively, had 95 and 77 % water flux of the original membrane, respectively. The binding capacity of lysozyme as a function of the SO3H group density was compared between the SP‐T and SS‐T fibers from measurement of the ion‐exchange breakthrough curves during the permeation of lysozyme solution across the SP‐T and SS‐T fibers. The binding capacity of lysozyme to the SP‐T fiber remained constant, independent of the SP group density, whereas that to the SS‐T fiber increased linearly with increasing SS group density. This difference was explained by means of a model whereby lysozyme adheres onto the SP group‐containing grafted polymer branches, while the SS group‐containing grafted polymer branches hold lysozyme in a tentacle‐like manner.
Monoclonal Antibody Process Development Using Medium Concentrates
Authors: Bibila Theodora A. , Ranucci Colette S. , Glazomitsky Konstantin , Buckland Barry C. , Aunins John G.
Published: March 2007

A fed‐batch process using concentrated medium was evaluated for its ability to improve cell culture longevity and final monoclonal antibody (MAb) titers for two monoclonal antibody producing cell lines. It was found to result in up to 7‐fold increases in final antibody titers compared to batch culture controls. Although the development of cell line specific fed‐batch protocols is critical to the development of cost‐efficient largescale production processes, the use of complete medium concentrates provided us with a quick and simple method for producing large quantities of antibodies in the early stages of process development, thus accelerating early work on purification process development, analytical development, biochemical characterization, and safety studies. Insights gained from the concentrated medium fed‐batch approach were valuable for the development of refined, cell line specific feeding strategies yielding final MAb titers on the order of 1–2 g/L. Process development data on the effects of inhibitory growth byproducts, medium osmolarity, and the mode of nutrient feed addition on culture longevity and MAb production and information on culture metabolic behavior were successfully incorporated in the development of the optimized fed‐batch protocols.
Reduction of Nonselective Adsorption of Proteins by Hydrophilization of Microfiltration Membranes by Radiation‐Induced Grafting
Authors: Kim Min , Kojima Jun , Saito Kyoichi , Furusaki Shintaro , Sugo Takanobu
Published: March 2007

Alcoholic hydroxyl groups were introduced into a polyethylene microfiltration (MF) membrane by radiation‐induced graft polymerization of 2‐hydroxyethyl methacrylate (HEMA), vinyl acetate (VAc), and glycidyl methacrylate (GMA). Subsequently, the VAc‐ and GMA‐grafted membranes were quantitatively hydrolyzed into monool (single hydroxyl) and diol groups, respectively. The pure water flux of the modified membrane and the saturation capacity of bovine γ‐globulin onto the membrane were determined as a function of alcoholic hydroxyl group density. The threshold value for hydrophilization ranged between 5 and 7 mol of alcoholic hydroxyl group per kilogram of original MF membrane. Masking of the polyethylene surface with grafted polymer branches containing the diol groups was effective because approximately 70% of the pure water flux of the original MF membrane was maintained. Irrespective of the pore diameter of the original MF membrane, saturation capacities on the modified membrane correlated well with the diol group density. Saturation capacities of bovine γ‐globulin and bovine serum albumin were reduced to 1 mg/m2 of the membrane. In addition, the binding interaction changed from irreversible to reversible.
Enhanced Productivity through Gratuitous Induction in Recombinant Yeast Fermentations
Authors: Napp Scott J. , Da Silva Nancy A.
Published: March 2007

The advantages of gratuitous induction for GAL‐regulated cloned gene (lacZ) product synthesis were evaluated for the yeast Saccharomyces cerevisiae. The growth, yield, and productivity of a gratuitous (gal1) strain were compared with those of an otherwise isogenic, nongratuitous (GAL1) strain. Batch studies clearly demonstrated the improvements possible in product synthesis when the inducer is not metabolized by the yeast cells; both β‐galactosidase specific and volumetric activities were superior for the gal1 strain. At equivalent metabolizable sugar concentrations, the productivity of the gratuitous strain exceeded that of the nongratuitous strain by 180%. The effects of initial inducer concentration and induction time were also examined. For the gratuitous strain, galactose:glucose ratios as low as 0.1 still gave maximum β‐galactosidase volumetric activity. A 5‐fold higher ratio was necessary for full induction with the nongratuitous strain, and productivity was substantially lower relative to the gal1 strain. A comparison of various times for galactose addition indicated that productivity is highest when the gratuitous culture is induced for the entire batch fermentation.
Nanobiotechnology: Protein‐Nanomaterial Interactions
Authors: Kane Ravi S. , Stroock Abraham D.
Published: March 2007

We review recent research that involves the interaction of nanomaterials such as nanoparticles, nanowires, and carbon nanotubes with proteins. We begin with a focus on the fundamentals of the structure and function of proteins on nanomaterials. We then review work in three areas that exploit these interactions: (1) sensing, (2) assembly of nanomaterials by proteins and proteins by nanomaterials, and (3) interactions with cells. We conclude with the identification of challenges and opportunities for the future.
Unintended consequences of increasing block tariffs pricing policy in urban water
Authors: Dahan Momi , Nisan Udi
Published: March 2007

We exploit a unique data set to estimate the degree of economies of scale in water consumption, controlling for the standard demand factors. We found a linear Engel curve in water consumption: each additional household member consumes the same water quantity regardless of household size, except for a single‐person household. Our evidence suggests that the increasing block tariffs (IBT) structure, which is indifferent to household size, has unintended consequences. Large households, which are also likely to be poor given the negative correlation between income and household size, are charged a higher price for water. The degree of economies of scale found here erodes the effectiveness of IBT price structure as a way to introduce an equity consideration. This implication is important in view of the global trend toward the use of IBT.
Tide‐induced head fluctuations in a confined aquifer with sediment covering its outlet at the sea floor
Authors: Li Hailong , Li Guanyi , Cheng Jianmei , Boufadel Michel C.
Published: March 2007

A confined coastal aquifer usually extends under the sea for some distance with its submarine outlet covered by a thin layer of sediment with properties dissimilar from the aquifer. Previous theoretical studies neglected this outlet capping. In this paper an analytical solution is derived for a confined aquifer subject to tidal fluctuations with a leaky boundary condition at the outlet capping. For the cases that offshore length of the aquifer and/or the leakance of the outlet capping are either zero or infinity, existing solutions in literature are obtained. It is shown that our analytical solution can also be used to describe the tidal wave propagation in a confined aquifer extending infinitely under a tidal river. The full solution agreed well with the observations in a piezometer in the Jahe River Basin in China, 200 m inland from the coastline where the tide‐induced head fluctuations without delay were observed but cannot be explained using previous analytical and numerical solutions ignoring the effect of the outlet capping. The sensitivity analysis showed that the combined actions of the tidal loading and outlet capping lead to complex dependency of the head fluctuation on the outlet capping's leakance. When the offshore confined aquifer is short, the inland head fluctuation increases with the outlet capping's leakance. In this case significant negative phase shift occurs if the leakance is small. For offshore aquifer length greater than a threshold value the inland head fluctuations are independent of this length and the outlet capping's leakance.
Magnetic properties and potential field modeling of the Peculiar Knob metamorphosed iron formation, South Australia: An analog for the source of the intense Martian magnetic anomalies?
Authors: Schmidt Phillip W. , McEnroe Suzanne A. , Clark David A. , Robinson Peter
Published: March 2007

Magnetic property measurements show that the strongly metamorphosed Peculiar Knob iron formation (IF), South Australia, is coarse‐grained, high‐grade hematite with variable amounts of magnetite and maghemite. This body exhibits a relatively low magnetic susceptibility (<0.3 SI) that cannot explain the associated intense magnetic anomaly, 30,000 nT, in terms of induced magnetization alone. Peculiar Knob IF possesses an extremely intense (∼120 A m−1) remanence, directed steeply upward. This ancient remanence reinforces the local Earth's field (inclination −63°). A simple geological model, constrained by drilling and physical property measurements, explains both the observed magnetic and gravity anomalies, consistent with the Poisson theorem. Koenigsberger ratios (Qs) of 10 and greater, as found here, are rare in nature. We postulate that acquisition of a thermoremanent magnetization (TRM) by the ore during postmetamorphic cooling from above the Curie/Néel temperature accounts for the intense remanence and high Qs. Although the hematite is in the multidomain size range, the coercivity is higher than expected. Also, the natural remanent magnetization (NRM) values are less than 10% of the expected value for a saturated TRM of hematite. On the basis of reflected light, scanning electron microscope observations, and rock magnetism, we propose that the common fine intergrowths of a very small amount of magnetite and/or maghemite within the hematite host are responsible for the relatively high coercivity and contribute to the NRM. These intergrowths are not normal exsolution lamellae and were likely present at high temperature. This study suggests that coarse‐grained hematite‐rich bodies that carry TRM and have been subjected to high‐grade (>680°C) metamorphism may be possible sources for some of the prominent Martian anomalies.
IUPAC-NIST Solubility Data Series. 82. Alcohols with Water—Revised and Updated: Part 1. C4 Alcohols with Water
Authors: Goral Marian , Wisniewska-Goclowska Barbara
Published: March 2007

The mutual solubility and related liquid–liquid equilibria of C4 alcohols with water are exhaustively and critically reviewed. Reports of experimental determination of solubility in three chemically distinct binary systems that appeared in the primary literature prior to end of 2004 are compiled. For all the systems sufficient data are available to allow critical evaluation. All data are expressed as mass percent and mole fraction as well as the originally reported units. In addition to the standard evaluation criteria used throughout the Solubility Data Series, a new method based on the evaluation of all experimental data for a given homologous series of saturated alcohols was used.
Genomic identification and
in vitro
reconstitution of a complete biosynthetic pathway for the osmolyte di-
myo
-inositol-phosphate
Authors: Rodionov Dmitry A. , Kurnasov Oleg V. , Stec Boguslaw , Wang Yan , Roberts Mary F. , Osterman Andrei L.
Published: March 2007

Di-
myo
-inositol 1,1′-phosphate (DIP) is a major osmoprotecting metabolite in a number of hyperthermophilic species of archaea and bacteria. Although the DIP biosynthesis pathway was previously proposed, genes encoding only two of the four required enzymes, inositol-1-phosphate synthase and inositol monophosphatase, were identified. In this study we used a comparative genomic analysis to predict two additional genes of this pathway (termed
dipA
and
dipB
) that remained missing. In
Thermotoga maritima
both candidate genes (in an originally misannotated locus TM1418) form an operon with the inositol-1-phosphate synthase encoding gene (TM1419). A predicted inositol-mono-phosphate cytidylyltransferase activity was directly confirmed for the purified product of
T. maritima
gene
dipA
cloned and expressed in
Escherichia coli
. The entire DIP pathway was reconstituted in
E. coli
by cloning of the TM1418–TM1419 operon in pBAD expression vector and confirmed to function in the crude lysate.
31
P NMR and MS analysis revealed that DIP synthesis proceeds via a phosphorylated DIP intermediate, P-DIP, which is generated by the
dipB
-encoded enzyme, now termed P-DIP synthase. This previously unknown intermediate is apparently converted to the final product, DIP, by an inositol monophosphatase-like phosphatase. These findings allowed us to revise the previously proposed DIP pathway. The genomic survey confirmed its presence in the species known to use DIP for osmoprotection. Among several newly identified species with a postulated DIP pathway,
Aeropyrum pernix
was directly proven to produce this osmolyte.
Assisted reproductive technologies do not alter mutation frequency or spectrum
Authors: Caperton Lee , Murphey Patricia , Yamazaki Yukiko , McMahan C. Alex , Walter Christi A. , Yanagimachi Ryuzo , McCarrey John R.
Published: March 2007

Assisted reproductive technologies (ARTs) have now contributed to the birth of >3 million babies worldwide, but concerns remain regarding the safety of these methods. We have used a transgenic mouse model to examine the effects of ARTs on the frequency and spectrum of point mutations in midgestation mouse fetuses produced by either natural reproduction or various methods of ART, including preimplantation culture, embryo transfer,
in vitro
fertilization, intracytoplasmic sperm injection, and round spermatid injection. Our results show that there is no significant difference in the frequency or spectrum of
de novo
point mutations found in naturally conceived fetuses and fetuses produced by
in vitro
fertilization, intracytoplasmic sperm injection, or round spermatid injection. These results, based on analyses of a transgenic mouse system, indicate that with respect to maintenance of genetic integrity, ARTs appear to be safe.
Actin Dynamics in Papilla Cells of Brassica rapa during Self- and Cross-Pollination
Authors: Iwano Megumi , Shiba Hiroshi , Matoba Kyoko , Miwa Teruhiko , Funato Miyuki , Entani Tetsuyuki , Nakayama Pulla , Shimosato Hiroko , Takaoka Akio , Isogai Akira , Takayama Seiji
Published: March 2007

The self-incompatibility system of the plant species Brassica is controlled by the S-locus, which contains S-RECEPTOR KINASE (SRK) and S-LOCUS PROTEIN11 (SP11). SP11 binding to SRK induces SRK autophosphorylation and initiates a signaling cascade leading to the rejection of self pollen. However, the mechanism controlling hydration and germination arrest during self-pollination is unclear. In this study, we examined the role of actin, a key cytoskeletal component regulating the transport system for hydration and germination in the papilla cell during pollination. Using rhodamine-phalloidin staining, we showed that cross-pollination induced actin polymerization, whereas self-pollination induced actin reorganization and likely depolymerization. By monitoring transiently expressed green fluorescent protein fused to the actin-binding domain of mouse talin, we observed the concentration of actin bundles at the cross-pollen attachment site and actin reorganization and likely depolymerization at the self-pollen attachment site; the results correspond to those obtained by rhodamine-phalloidin staining. We further showed that the coat of self pollen is sufficient to mediate this response. The actin-depolymerizing drug cytochalasin D significantly inhibited pollen hydration and germination during cross-pollination, further emphasizing a role for actin in these processes. Additionally, three-dimensional electron microscopic tomography revealed the close association of the actin cytoskeleton with an apical vacuole network. Self-pollination disrupted the vacuole network, whereas cross-pollination led to vacuolar rearrangements toward the site of pollen attachment. Taken together, our data suggest that self- and cross-pollination differentially affect the dynamics of the actin cytoskeleton, leading to changes in vacuolar structure associated with hydration and germination.
The Mismatch Repair Protein MLH1 Marks a Subset of Strongly Interfering Crossovers in Tomato
Authors: Lhuissier Franck G.P. , Offenberg Hildo H. , Wittich Peter E. , Vischer Norbert O.E. , Heyting Christa
Published: March 2007

In most eukaryotes, the prospective chromosomal positions of meiotic crossovers are marked during meiotic prophase by protein complexes called late recombination nodules (LNs). In tomato (Solanum lycopersicum), a cytological recombination map has been constructed based on LN positions. We demonstrate that the mismatch repair protein MLH1 occurs in LNs. We determined the positions of MLH1 foci along the 12 tomato chromosome pairs (bivalents) during meiotic prophase and compared the map of MLH1 focus positions with that of LN positions. On all 12 bivalents, the number of MLH1 foci was ∼70% of the number of LNs. Bivalents with zero MLH1 foci were rare, which argues against random failure of detecting MLH1 in the LNs. We inferred that there are two types of LNs, MLH1-positive and MLH1-negative LNs, and that each bivalent gets an obligate MLH1-positive LN. The two LN types are differently distributed along the bivalents. Furthermore, cytological interference among MLH1 foci was much stronger than interference among LNs, implying that MLH1 marks the positions of a subset of strongly interfering crossovers. Based on the distances between MLH1 foci or LNs, we propose that MLH1-positive and MLH1-negative LNs stem from the same population of weakly interfering precursors.
Two Host-Induced
Ralstonia solanacearum
Genes,
acrA
and
dinF
, Encode Multidrug Efflux Pumps and Contribute to Bacterial Wilt Virulence
Authors: Brown Darby G. , Swanson Jill K. , Allen Caitilyn
Published: March 2007

Multidrug efflux pumps (MDRs) are hypothesized to protect pathogenic bacteria from toxic host defense compounds. We created mutations in the
Ralstonia solanacearum acrA
and
dinF
genes, which encode putative MDRs in the broad-host-range plant pathogen. Both mutations reduced the ability of
R. solanacearum
to grow in the presence of various toxic compounds, including antibiotics, phytoalexins, and detergents. Both
acrAB
and
dinF
mutants were significantly less virulent on the tomato plant than the wild-type strain. Complementation restored near-wild-type levels of virulence to both mutants. Addition of either
dinF
or
acrAB
to
Escherichia coli
MDR mutants KAM3 and KAM32 restored the resistance of these strains to several toxins, demonstrating that the
R. solanacearum
genes can function heterologously to complement known MDR mutations. Toxic and DNA-damaging compounds induced expression of
acrA
and
dinF
, as did growth in both susceptible and resistant tomato plants. Carbon limitation also increased expression of
acrA
and
dinF
, while the stress-related sigma factor RpoS was required at a high cell density (>10
7
CFU/ml) to obtain wild-type levels of
acrA
expression both in minimal medium and in planta. The type III secretion system regulator HrpB negatively regulated
dinF
expression in culture at high cell densities. Together, these results show that
acrAB
and
dinF
encode MDRs in
R. solanacearum
and that they contribute to the overall aggressiveness of this phytopathogen, probably by protecting the bacterium from the toxic effects of host antimicrobial compounds.
Acidobacteria
Phylum Sequences in Uranium-Contaminated Subsurface Sediments Greatly Expand the Known Diversity within the Phylum
Authors: Barns Susan M. , Cain Elizabeth C. , Sommerville Leslie , Kuske Cheryl R.
Published: March 2007

The abundance and composition of bacteria of the phylum
Acidobacteria
were surveyed in subsurface sediments from uranium-contaminated sites using amplification of 16S rRNA genes followed by clone/sequence analysis. Analysis of sequences from this study and public databases produced a revised and greatly expanded phylogeny of the
Acidobacteria
phylum consisting of 26 subgroups.