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PubMed1991

The spindle is required for the process of sister chromatid separation in Drosophila neuroblasts.

We have studied two aspects of the process of sister chromatid separation in the Drosophila melanogaster neuroblasts. First, we analyzed the requirement of a functional spindle for sister chromatid separation to take place using microtubule depolymerizing drugs such as colchicine or a reversible analogue (MTC). Incubation of this tissue in colchicine causes the cells to block irreversibly at metaphase and no significant levels of sister chromatid separation were observed even after long periods of incubation. Exposure of neuroblasts to MTC also causes cells to block at metaphase, but after reversion most of the cells enter anaphase and are thus able to complete sister chromatid separation. These results imply that a functional spindle is required for sister chromatid separation. Second, we studied the role of heterochromatin during chromatid pairing and subsequent separation in chromosomes which carry either one or two extra pieces of heterochromatin. The results indicate that sister chromatids establish strong pairing along the translocated heterochromatin. During the early stages of anaphase, these chromosomes separate first the centromeric region and later the regions bearing extra heterochromatin. These results indicate that constitutive heterochromatin plays an important role for sister chromatid pairing and might be involved in the process of separation.

Animals↗
PubMed1992

Energetics of the strand separation transition in superhelical DNA.

In this paper the values of three free energy parameters governing the superhelical strand separation transition are determined by analysis of available experimental data. These are the free energy, a, needed to initiate a run of separation, the torsional stiffness, C, associated with interstrand winding of the two single strands comprising a separated site and the coefficient, K, of the quadratic free energy associated to residual linking. The experimental data used in this analysis are the locations and relative amounts of strand separation occurring in the pBR322 DNA molecule and the measured residual linking, both evaluated over a range of negative linking differences. The analytic method used treats strand separation as a heteropolymeric, co-operative, two-state transition to a torsionally deformable alternative conformation, which takes place in a circular DNA molecule constrained by the constancy of its linking number. The values determined for these parameters under the experimental conditions (T = 310 K, pH = 7.0, monovalent cation concentration = 0.01 M) are a = 10.84(+/- 0.2) kcal/mol, C = 2.5(+/- 0.3) x 10(-13) erg/rad2 and K = 2350(+/- 80) RT/N, where N is the molecular length in base-pairs. In order to assess the accuracy of the author's theoretical methods, these free energy parameters are incorporated into the analysis of superhelical strand separation in different molecules and under other conditions than those used in their evaluation. First, the temperature dependence of transition is treated, then superhelical strand separation is analyzed in a series of DNA molecules having systematic sequence modifications, and the results of these theoretical analyses are compared with those from experiments. In all molecules, transition is predicted in the range of linking differences where it is seen experimentally. Moreover, it occurs at the specific sequence locations that the analysis predicts, and with approximately the predicted relative amounts of transition at each location. The known sensitivities of this transition to changes of temperature and to small sequence modifications are predicted in a quantitatively precise manner by the theoretical results. The demonstrated high-level precision of these theoretical methods provides a tool for the screening of DNA sequences for sites susceptible to superhelical strand separation, some of which may have regulatory or other biological significance.

Base Composition↗
PubMed1984

Effects of mother-litter separation and reunion on maternal aggression and pup mortality in lactating hamsters.

This study examined the effects of a 24 hr mother-litter separation on maternal aggression and pup mortality in hamsters. There were four lactating groups tested for aggression in their home cages on days 5 and 15 postpartum: a group that was separated from their litters for 24 hr, a group that was not separated from their litters and two 24 hr separation groups in which the litters were returned 30 and 120 min prior to the aggression test. Nonreceptive, estrous cycling animals were used as controls (tested twice, 10 days apart) and as intruders. Measures of aggression recorded during the 10 min tests included the number of attacks, fights, chases and intruder retreats. Pup mortality was examined in the 4 lactating groups and in two additional non-tested groups, one of which was separated from their litters for 24 hr. Lactating hamsters initiated significantly higher levels of aggression than control animals. Mother-litter separation (24 hr) significantly decreased levels of aggression and these levels were restored when litters were replaced 30 and 120 min prior to aggression tests. There were few differences in levels of aggression between days 5 and 15 of lactation. Pup mortality was significantly greater between days 5-6 and 15-16 of lactation among those groups that were separated from their litters compared to those groups that were not separated. Significantly higher mortality levels were found between days 7 and 15 of lactation among the 2 groups in which pups were returned prior to the aggression test compared to all other groups.

Aggression↗
PubMed1989

The effect of a distribution of separations upon intramolecular distances in biopolymers, as determined by radiationless energy transfer.

In a fluorescent donor group and a nonfluorescent acceptor group are incorporated into a biopolymer, so that radiationless energy transfer occurs between the two groups, the apparent separation of the groups, as determined by energy transfer, will be influenced by the existence of a distribution of separations. This might arise from the presence of significant localized flexibility at the sites of attachment of the two groups, or from internal flexibility involving the biopolymer itself. If a Gaussian form is assumed for the distribution of separations of the donor and acceptor groups, the efficiency of transfer is dependent upon the width of the distribution, as well as the average distance between the groups. Significant differences may thereby arise between the true average separation and the separation computed from transfer efficiencies by the usual procedures. The deviations are different for transfer efficiencies computed from quantum yields and from decay times. They become more important with increasing width of the distribution of separations and increasing efficiency of transfer. In general, if a distribution of separations is present, the average separation is most reliably computed by procedures which take into account the effects of this distribution.

Biopolymers↗
PubMed1988

Enantiomeric separations and their application in pharmaceutical analysis using chiral eluents.

The most important problems of enantiomeric separations using chiral eluents are discussed. The methods have been compared with respect to enantiomeric purity of reagents, reagent selection and separation variables. The most important considerations for methods based on inclusion-complexation with different CDs and on using chiral counter ions are summarised and compared. A new possibility for the separation of enantiomeric compounds with the aid of a combination of ion-pair and inclusion-complex formation has been introduced. As a consequence of this investigation, the selectivity of the separation can be significantly improved; those ionic isomers and enantiomers which cannot be separated or are unsatisfactorily separated by ion-pair chromatography or by inclusion-complex formation, can be separated by the combined technique. Comparison of methods applicable for enantiomeric separations is also given with respect to solving specific analytical tasks in the field of pharmaceutical analysis.

Journal Article↗
PubMed2006

Early deprivation, but not maternal separation, attenuates rise in corticosterone levels after exposure to a novel environment in both juvenile and adult female rats.

Separation from the maternal nest alters the hypothalamic-pituitary-adrenal (HPA) axis stress response in adult male rats, but little research has addressed how separation affects female rats. The following experiments investigated how early maternal separation from postnatal day (PND) 2 to 14 affected stress-induced corticosterone and ACTH after exposure to an open field in juvenile and adult female rats. Female rats were separated for 5 h daily from mother and littermates (early deprivation: ED), separated from mother but not littermates (maternal separation: MS), or animal facility reared (AFR). Male siblings were left with the mother rat during separation. Female rats were exposed to an open field arena either during the juvenile period (PND 30) or during adulthood (PND 80-100). Results show that ED juvenile female rats showed a lower corticosterone stress response than MS and AFR female rats when measured at 5 min post-stress, but no difference at 20 or 60 min post-stress. In adulthood, ED female rats showed comparable elevations of corticosterone as MS and AFR rats at 5 min post-stress but lower elevations at 20 min. In terms of behavior, there were no significant effects of early experience. However, in adulthood, ED and MS rats tended to show a decreased proportion of inner grid crossings of the open field compared to AFR rats, suggesting a tendency for increased anxiety in these two separation groups.

Adrenocorticotropic Hormone↗
PubMed2004

Generic, highly selective and robust capillary electrophoresis method for separation of a racemic mixture of glitazone compounds.

A generic, highly selective, and robust capillary electrophoresis (CE) method was developed for separation of a racemic mixture of three available glitazone compounds (also known as thiazolidinediones) in active pharmaceutical ingredients (API) and tablets. The method separated the R and S enantiomers of balaglitazone, pioglitazone and rosiglitazone, and showed that the samples contained an equal (50:50) quantity of the enantiomers as a mixture. After a simple extraction of samples with acetonitrile:water (80:20), separation was performed using a combination of two cyclodextrins: sulfobuthylether-beta-cyclodextrin (SB-beta-CD) and dimethyl-beta-cyclodextrin (DM-beta-CD) in the electrolyte at pH 8.0. The method showed a very good specificity, and all separations were achieved with a resolution (Rs) over 3.0. The developed CE method was then validated. The Rs for the separations were 3.5 for balaglitazone enantiomers, 3.5 for pioglitazone enantiomers, and 3.7 for rosiglitazone. The squared correlation coefficients (r2) were found to be 0.999 for all three compounds. The range of the CE method (injection volume was approximately 4 nl) was demonstrated to be from 1.0 to 2.4 ng. The R.S.D. in the repeatability study was found to be less than 0.5 for area/area ratio (and 3.0% for area) for all three compounds. The R.S.D. in the intermediate precision study was found to be less than 0.7 for area/area ratio (and 4.5% for area) for all three compounds. Generally, the method showed good robustness. Resolution between the enantiomers peak was maintained acceptable throughout the small variations around the pH value of the buffer, different capillary, CE instrument and electrolytes ion strength capacity, but changes in concentration of cyclodextrins and acetonitrile showed significant effects on separations and affected the resolution. The validation results showed that the CE method was suitable for separation of the racemic mixtures of the three glitazone drugs. The CE method was then applied for routine test during the drug and formulation development work of balaglitazone. Due to the achieved results from this work, it is the authors' belief that this method can easily separate other glitazone racemic mixtures.

Cyclodextrins↗
PubMed2005

Comparison of the performance of conventional microparticulates and monolithic reversed-phase columns for liquid chromatography separation of eleven pollutant phenols.

The performance of isocratic separations of 11 pollutant phenols (PP) using monolithic (Chromolith RP-18e) and conventional reversed-phase 5 microm (Luna and Purospher C18) and 4 microm (Synergi C12) particulate size columns, selected from high purity silica materials, has been compared. The separations have been optimized based on a previously optimized separation in which a reversed-phase C18 Luna column and acetonitrile as organic modifier were used, allowing the separation of all phenols tested in 23 min. The optimization process was carried out for each column by studying the effect of the mobile phase (acetonitrile as organic modifier, pH, flow-rate) on phenols separation. Under the optimized separation conditions, all phenols were separated in less than 23 min for all columns tested. Asymmetry factors were further evaluated and used to estimate column efficiency using the Dorsey-Foley equation. The efficiency and asymmetry factors were lower for Chromolith than for Purospher and Luna columns respectively. The Chromolith column was finally selected, due to its lower flow resistance, analysis time and good efficiency and asymmetry factors. The PPs separation was achieved in 3 min. The asymmetry factors were in the range 0.9-1.5 using 50mM acetate buffer (pH = 5.25)-ACN (64:36, v/v) as mobile phase, T=45 degrees C and 4.0 ml min(-1) flow-rate.

Environmental Pollutants↗
PubMed2005

Phase system selectivity and two-dimensional separations in liquid column chromatography.

Correlations between the separation selectivity in aqueous and non-aqueous reversed-phase systems and in normal-phase LC systems were investigated for samples containing different numbers of two repeat structural elements. Such samples are best separated in "orthogonal" two-dimensional chromatographic systems, showing selectivity for one type of the repeat structural element only in the first dimension and for the other structural element only in the second dimension. The number of resolved compounds improves as the degree of orthogonality of the separation systems increases with decreasing correlation between the selectivities for the sample structural distribution in the two dimensions. Orthogonal systems with non-correlated selectivities for each repeat structural element provide the highest number of separated peaks and regular arrangement of the peaks over the two-dimensional retention space according to the individual structural element distribution and the best use of the available peak capacity. Fully orthogonal systems are difficult to find in practice. Partially orthogonal system with correlated selectivities for one structural type distribution, but with one system non-distinguishing the distribution for the other structural element are still useful for the two-dimensional separations. The correlations between the selectivities for repeat regular structural increments were employed to evaluate the suitability of phase systems for two-dimensional HPLC separations. The selectivity correlation in various reversed-phase and normal-phase systems was evaluated for two sample types: (1) Various RP columns show significantly inversely correlated selectivities for acyl lengths and numbers of double bonds distribution, but the differences in the double bond selectivity can be used for practical separations of triacylglycerols with the same equivalent carbon numbers. (2) Synthetic EO-PO block (co)oligomers with two-dimensional distribution of oxyethylene and oxypropylene monomer units were separated according to the two distribution types using on-line two-dimensional reversed-phase-normal-phase LC with a C18 column in the first dimension and an aminopropyl silica column in the second dimension.

Chromatography, Liquid↗
PubMed2004

Dynamics of nonionic surfactant-rich phase separation and recovery of dyes.

Cloud point separation of selected dyes was studied. The use of dyes made possible observation of the dynamics of surfactant-rich phase separation by color video. The pictures were interpreted by means of ImageC software and degrees of whiteness were calculated. It was found that separation was slow and equilibrium was not achieved even after a period of more than 10 h. The separated surfactant-rich phase had a heterogeneous structure. The globules of the surfactant-rich phase were also observed in the micellar aqueous phase. The surfactant concentration could be as high as one or two orders of magnitude above the expected cmc values and was not decreased to critical micelle concentration by centrifugation. The presence of sodium chloride was important and improved separation. Separation of dyes was in the range 73-98% and depended upon the surfactant, the temperature, the electrolyte content, and the dye. In each system considered, appropriate conditions had to be selected to obtain high recovery of the dye. The presence of the electrolyte was the most important parameter and it improved the separation of dyes. However, addition of the electrolyte could also account for precipitation of the dyes, as observed for the systems containing Direct Pink. The best recoveries were observed for Direct Yellow and oxyethylated nonylphenol (98% at 55 degrees C in the presence of NaCl). Centrifugation gave recoveries similar to those for prolonged heating but it shortened the time of phase separation.

Coloring Agents↗
PubMed2005

MEKC-LIF of gamma-amino butyric acid in microdialysate: systematic optimization of the separation conditions by factorial analysis.

Micellar electrokinetic chromatography allows the efficient separation of biogenic amines and amino acids in biological samples. Analytes of interest, sample composition, and sample matrix may vary between studies, which necessitates optimization of separations to meet the requirements and conditions of an experiment. Factorial analysis is an efficient tool to accomplish this type of optimization involving multiple interacting factors. The present study describes an optimization procedure for separation of the inhibitory neurotransmitter GABA utilizing capillary electrophoresis with laser induced fluorescence detection. Standards labeled with the flourogenic reagent 3-(2-furoyl)quinoline-2 carboxaldehyde were separated with varying concentrations of borate buffer, beta-cyclodextrin, sodium dodecyl sulfate and pH. The optimized separation method had a correlation coefficient between concentration of GABA and fluorescent signal of 0.98, and was linear in the desired concentration range of 25 nM-10 microM. Glutamic acid, aspartic acid and taurine were also quantified using this separation. When applied to microdialysate collected from the region of the suprachiasmatic nucleus, this separation was able to measure daily variations in GABA levels. The factorial design experiment has proven to be a useful tool, allowing adjustments in the separation of neurotransmitters based on individual requirements.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2006

Newly synthesized tetraoxa-diaza crown ether derivatives versus commercialized crown ethers in the separation of positional isomers with capillary electrophoresis.

Three new tetraoxa-diaza derivatives of 1,4,10,13-tetraoxa-7,16-diazacyclo-octadecane (R-1, R-2 and R-3) and three commercially available crown ethers, 18-crown-6 (18C6), (+)-18-crown-6-tetracarboxilic acid (18C6H4) and 1,4,10,13-tetraoxa-7,16-diazacyclo-octadecane, were investigated to separate the positional isomers of aminophenol, aminobenzoic acid and aminocresol. The running electrolyte, in which the crown ethers were dissolved, was a 50 mM Tris solution adjusted to pH 2.0 with hydrochloric acid. Using 50 mM H3PO4 buffer, whose pH was adjusted to 2.0 with Tris, or only hydrochloric acid solution with the same pH, did not allow good separations for the tested components. The effect of the crown ether concentration on the separation of the 11 positional isomers was studied in the concentration range of 10-50 mM. The best separations were achieved using the 18C6 and the 18C6H4 crown ethers: 9 isomers out of 11 could be separated within one run. The m- and p-aminophenol isomers could not be separated under the investigated experimental conditions. The newly synthesized tetraoxa-diaza crown ether derivatives were only found suitable for the separation of aminobenzoic acid positional isomers. The macrocyclic ring of the tetraoxa-diaza crown ethers was not able to form a stable inclusion complex with the tested positional isomers. Consequently, the aminophenol and aminocresol isomers were not separated, the isomers migrated with the same or very similar velocities.

Crown Ethers↗
PubMed1977

Aortic stenosis: echocardiographic cusp separation and surgical description of aortic valve in 22 patients.

Diminished echocardiographic aortic cusp separation is used as one indicator of the severity of aortic stenosis. To test the validity of this index, 22 patients--12 (55 percent) with isolated aortic valve disease and 10 (45 percent) with aortic stenosis associated with mitral or coronary artery disease--underwent M mode echocardiographic examination before aortic valve replacement. Tracings of diagnostic quality were obtained without difficulty from all 22 patients. Cardiac catheterization was performed in 21 patients. Echocardiographic cusp separation was measured from the apparent mid-systolic orifice and from the outer periphery of the anterior cusp to the outer periphery of the posterior cusp (maximal peripheral cusp separation). Mid-systolic cusp separation varied in nearly every patient, depending on the angle of leaflet presentation to the ultrasonic beam. Maximal peripheral cusp separation measured 16 mm in 18 of 22 patients (82 percent); it indicated neither the severity of the aortic stenosis as documented with cardiac catheterization nor the mobility of the cusps seen at operation. The surgical and echocardiographic descriptions of leaflet and aortic root calcification were similar. In situ examination of aortic cusp separation indicated that diseased aortic valves are not comparable with normal valves or valves with uncomplicated congenital obstruction. The aortic leaflets were curled, fused, calcified and deformed from their natural state of coaptation. Abnormal thickening and limited or eccentric mobility of the aortic leaflets were useful indicators of the cause of valve disease, but cusp deformity secondary to aortic stenosis invalidated mid-systolic cusp separation and maximal peripheral cusp separation as indicators of the severity of aortic stenosis.

Adult↗
PubMed2001

Separation of hydrophobic polymer additives by microemulsion electrokinetic chromatography.

Microemulsion electrokinetic chromatography (MEEKC) has been applied to the separation of some phenolic antioxidants [Irganox 1024, Irganox 1035, Irganox 1076, Irganox 1010, Irganox 1330, Irgafos 138, Irganox 168 and 2,6-di-tert.-butyl-4-methylphenol (BHT)]. Due to the extremely hydrophobic nature of these analytes, they could not be separated using standard MEEKC conditions and two alternative approaches were investigated. Using an acidic buffer (phosphate, pH 2.5) to effectively suppress the electroosmotic flow, the addition of 2-propanol to the aqueous phase of the microemulsion buffer to improve partitioning of the analytes, and a negative separation voltage, separation of five of the analytes in under 10 min was possible. The second approach, using a basic buffer (borate, pH 9.2) and a positive separation voltage resulted in complete resolution of all eight analytes. A mixed surfactant system comprising the anionic sodium dodecyl sulfate (SDS) and neutral Brij 35 was used to reduce the overall charge and with it the mobility of the droplets, and hence the separation time. Using an optimised MEEKC buffer consisting of 2.25% (w/w) SDS, 0.75% (w/w) Brij 35, 0.8% (w/w) n-octane, 6.6% (w/w) 1-butanol, 25% (w/w) 2-propanol and 64.6% (w/w) 10 mM borate buffer (pH 9.2) the eight target analytes were baseline separated in under 25 min. For these analytes, MEEKC was found to be superior to micellar electrokinetic chromatography in every respect. Specifically, the solubility of the analytes was better, the selectivity was more favourable, the analysis time was shorter and the separation efficiency was up to 72% higher when using the MEEKC method. Detection limits from 5.4 to 26 microg/ml were obtained and the calibration plot was linear over more than one order of magnitude. The optimised method could be applied to the determination of Irganox 1330 and Irganox 1010 in polypropylene.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2001

Metrics of separation in chromatography.

A new metric, separation measure, S, for chromatographic separation is proposed. Unlike other metrics such as resolution, separation number, and some versions of peak capacity, the new metric provides a consistent, additive measure of the separation of pairs of peaks as well as the separation capacities of arbitrary intervals within the analysis time. The attribute of additivity means that the separation measure of any separation interval is equal to the sum of the separation measures of its subintervals. Practical aspects of the measurement of S are also addressed. In addition to definition of S, a definition of peak capacity, n, that is consistent with S, and includes useful features of other known definitions of n is proposed for an arbitrary time interval.

Chromatography↗
PubMed1990

Isotachophoretic separation of organic acids in biological fluids.

The operating conditions for the isotachophoretic separation of organic acids were evaluated. At pH values ranging from 2.90 to 4.25 both relative step heights and molar flow-rates were determined experimentally for 26 anions. Comparing the observed values with simulated data, highly significant (p = 0.0001) correlation coefficients of 0.993 and 0.920, respectively, were found at pH 3.50. Whereas the concentration of the leading electrolyte did not affect the relative step heights, it increased the molar flow-rates significantly. The same applied to the detection current. The time of analysis was observed to be a function of the concentration of the leading electrolyte. However, the time elapsed between injection of the analyte and its detection depended solely on the volume and not on the amount of analyte injected. In isotachophoresis, incomplete separation of two compounds is indicated by the occurrence of a mixed zone which can hardly be distinguished from a pure zone. Thus, knowledge of the separation capacity is a prime prerequisite in optimizing the system for the analysis of biological fluids. The separability of nine equimolar pairs of anions was determined at pH values ranging from 2.90 to 4.25. Although two ionogenic constituents would separate only when their migration rates in the mixed state were different, no clear correlation was observed between separation capacity and difference in relative mobility. Separability, however, was found to increase with increasing concentration of the leading electrolyte. While the separation capacity was not influenced by the electric current, it was significantly affected by the volume injected. In subsequent analyses of serum, cerebrospinal fluid, seminal plasma and prostatic fluid, a variety of organic acids could be detected. Calibration graphs for the detected anions revealed a detection limit of 1 nmol and linearity over their biological concentration ranges. Further, the isotachophoretic results correlated well with high-performance liquid chromatographic and enzymatic analyses of citric acid and lactic acid in human seminal plasma and cerebrospinal fluid, respectively.

Acids↗
PubMed2003

Separation of mixtures of acidic and basic peptides at neutral pH.

Mixtures of acidic and basic peptides composed of the phosphorylated and nonphosphorylated forms of peptide substrates for kinases and a phosphatase were separated by capillary electrophoresis (CE) in buffer conditions compatible with live mammalian cells. The separation of such mixtures was especially challenging given the high salt and neutral pH of the requisite physiologic buffers. Due to poor peak reproducibility in bare capillaries, several strategies were implemented to improve the electrophoretic separation of the peptide mixtures. Covalent coating of the capillary with the neutral polymer poly(dimethylacrylamide) (PDMA) resulted in a 2-fold improvement in the migration time RSD, but required the use of hydrodynamic flow to overcome the differing electrophoretic mobilities (microeo) of the peptides at neutral pH. This parabolic fluid flow diminished separation efficiency almost 5-fold. Polarity switching during the CE run was used to overcome the opposed microeo, but required the retention of hydrodynamic flow and consequent reduction in separation efficiency. The most efficient separations were seen with the use of covalently-linked, charged polymer coatings to maintain electroosmotic flow and to reduce wall interactions. Two such coatings were tested in the current study. Relative to the PDMA coating, an anionic poly(acrylate) improved the average migration time RSD of six peptides from 1.3 to 0.85% and average separation efficiency from 4.8 to 18.0 (x 10(4) plates/m). Likewise, cationic poly([3-(methacryloylamino)propyl]-trimethylammonium) improved the average migration time RSD of eight peptides from 1.2 to 1.1% and average separation efficiency from 4.8 to 33.9 (x 10(4) plates/m). These findings will be of value to the growing number of applications for analytical techniques utilizing CE for cellular analysis and biochemical studies.

Amino Acid Sequence↗
PubMed1998

Separation of omphalopagus twins: unique reconstruction using syngeneic cryopreserved tissue.

Omphalopagus twin girls were admitted for evaluation of possible separation and repair at age 7 days. Prenatal sonographic diagnosis occurred late in the third trimester and was followed by cesarean section delivery shortly thereafter. Results of extensive evaluation over the next 7 days including x-rays, computed tomography and ultrasound scan of the head and torso, and cardiac catheterization showed: the gastrointestinal tracts were separate and normal, the livers were joined but had separate biliary and vascular systems, and the hearts were separate with vastly different anatomy and function. One twin (twin A) had a normal heart with a small insignificant VSD. Twin B had a single ventricle, an incompetent A-V valve, stenotic pulmonic valve, ASD, PDA, and congenital heart block. Hemodynamic support of twin B was almost entirely from twin A. The vascular communications between the two consisted of a major connection between the internal mammary arteries and large arterial and venous connections traversing the joined livers. Because of continued deterioration of twin B, separation was undertaken at age 15 days. The separation included dividing the liver and the multiple large vascular connections. Two teams then reconstructed each twin separately. Twin B began showing signs of cardiac decompensation shortly after separation in spite of placement of a pacemaker, pulmonary artery banding, and ligation of the PDA. Cardiac function rapidly deteriorated and she died. Tissue from her chest wall was cryopreserved and placed in the tissue bank. Twin A underwent closure of her abdomen, and received a temporary bovine pericardial patch over the chest defect. She subsequently underwent placement of a graft of twin B's rib cage to bridge the bony chest defect and skin flap closure. She is presently taking a normal diet and thriving at home at age 18 months. The use of cryopreserved tissue from a syngeneic source provides a unique method of reconstruction in this situation.

Abdomen↗