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Is mechanical bowel preparation really necessary in colorectal surgery?

OBJECTIVE: To determine the outcome of colorectal surgery without mechanical bowel preparation. DESIGN: A descriptive, analytical and observational study. PLACE AND DURATION OF STUDY: Combined Military Hospital, Kharian and Pano Aqil, from September 1998 to April 2003. SUBJECTS AND METHODS: Forty-seven patients underwent debridement/resection and repair/primary anastomosis of colon and upper rectum without bowel preparation. Of these, 16 patients were operated in emergency. The anastomosis was carried out with polyglactin (vicryl) interrupted, full thickness single layer and no patient had defunctioning colostomy. Third generation cephalosporin, cefotaxime or ceftazidime and metronidazole were given perioperatively, repeated during surgery if lasted for more than 2 hours and continued for 3-5 days postoperatively. RESULTS: Anastomoses were ileocolic in 29.7%, colicocolic in 61.7% and colorectal in 14.8% cases. Anastomotic failure was seen in 4.2% and wound infection in 8.5% cases. There was one mortality (2.1%) due to unrelated cause. CONCLUSION: Mechanical bowel preparation is not necessary for safe colorectal surgery.

Adolescent↗

The central Z-disk region of titin is assembled from a novel repeat in variable copy numbers.

The giant sarcomeric protein titin (also described as connectin) is composed mainly of immunoglobulin (Ig)-like and fibronectin type III (fn3)-like domains arranged consecutively. At both ends of the molecule, these domains are interrupted by sequence insertions. The amino terminus of titin is localized in the Z-disk, a structure of great variability in different muscle types. We have determined the ultrastructural position of sequences in this region of the molecule in skeletal and cardiac muscle by immunoelectron microscopy using antibodies directed against unique epitopes. Titin molecules entering the Z-disk from two half sarcomeres do not significantly overlap, showing that the amino terminus is at the centre of the Z-disk. A serine/proline rich site, which can be phosphorylated by kinases in developing muscle tissues, was identified near the amino terminus of titin. Sequence analysis revealed the presence of a novel 45 residue repeat ('Z-repeats') in this region of the molecule. The number of titin Z-repeats varies due to differential splicing. We propose that this mechanism is a means of assembling Z-disks of variable thickness and mechanical strength.

Amino Acid Sequence↗

Structural comparison of tektins and evidence for their determination of complex spacings in flagellar microtubules.

Recent structural studies indicate that a tektin heteropolymer forms a unique protofilament of flagellar microtubules. We report here the sequence of tektin C (approximately 47 kDa), predicted from its cDNA (GenBank U38523), compared to tektins A (approximately 53 kDa) and B (approximately 51 kDa) from sea urchin (Strongylocentrotus purpuratus) sperm flagellar microtubules, and compared to partial sequences reported from mouse and human. We are now able to make several observations concerning the tektin family: (1) their common structural features, (2) a comparison of their structure to intermediate filament proteins, and (3) their possible organization in the tektin filament polymer. The predicted amino acid sequence identities/similarities are: for tektins A and C, 42/54%, for tektins A and B, 34/51%; for tektins B and C, 29/42%; for tektin C and a partial cDNA clone from mouse testis, 55/65%; and for tektin B and a partial cDNA clone from the human brain, 45/47%. The three tektins (and the human clone) possess the exact sequence repeat RPNVELCRD. The structural pattern of all three tektin polypeptides is similar to intermediate filament proteins. Tektins are predicted to form extended rods composed of two alpha-helical segments (approximately 180 residues long) capable of forming coiled coils, which are interrupted by short non-helical linkers. The two segments are homologous in sequence and secondary structure, indicating a gene duplication event prior to the divergence of the three tektins. Along each tektin rod cysteine residues occur with a periodicity of approximately 8 nm, coincident with the axial repeat of tubulin dimers in microtubules. From EM data and calculations of secondary structure, the segment length of tektin AB heterodimers is likely to be 16 nm. Both segments of tektin C may be 24 nm long, but one may be 16 nm. On the basis of the available evidence, we propose that coassembly of tektin AB heterodimers with tektin C dimers produces filaments with overall repeats of 8, 16, 24, 32, 40, 48 and 96 nm, generating the basis for the complex spatial arrangements of axonemal components.

Amino Acid Sequence↗

Effects of intermittent androgen suppression on androgen-dependent tumors. Apoptosis and serum prostate-specific antigen.

BACKGROUND: Since postcastration progression of tumors to an androgen-independent state appears to be linked to the cessation of androgen-induced differentiation of tumorigenic stem cells, the authors hypothesized that the replacement of androgens at the end of a period of apoptotic regression might result in the regeneration of differentiated tumor cells with further apoptotic potential. METHODS AND RESULTS: To determine the effect of intermittent exposure of androgens on the androgen-dependent Shionogi carcinoma, the tumor was transplanted into a succession of male mice, each of which was castrated when the estimated tumor weight became about 3 g. After the tumor had regressed to 30% of the original weight, it was transplanted into the next noncastrated male. This cycle of transplantation and castration-induced apoptosis was repeated successfully four times before growth became androgen-independent during the fifth cycle. In four of Stage C and three of Stage D patients with prostate cancer, androgen withdrawal was initiated with cyproterone acetate (100 mg/d) and diethylstilbestrol (0.1 mg/d) and then maintained with cyproterone acetate in combination with the luteinizing hormone-releasing hormone agonist, goserelin acetate (3.6 mg/month). After 6 or more months of suppression of serum prostate-specific antigen (PSA) into the normal range, treatment was interrupted for 2 to 11 months. After recovery of testicular function, androgen-withdrawal therapy was resumed when serum PSA increased to a level of about 20 micrograms/l. This cycle was repeated sequentially to a total of two to four times over treatment periods of 21 to 47 months with no loss of androgen dependence. CONCLUSIONS: These results demonstrate that intermittent androgen suppression can be used to induce multiple apoptotic regressions of a tumor; they also suggest that the cyclic effects of such treatment on prostate cancer can be followed by the sequential measurement of serum PSA levels.

Adenocarcinoma↗

Cross-species amplification of soybean (Glycine max) simple sequence repeats (SSRs) within the genus and other legume genera: implications for the transferability of SSRs in plants.

We investigated the transferability of 31 soybean (Glycine max) simple sequence repeat (SSR) loci to wild congeners and to other legume genera. Up to 65% of the soybean primer pairs amplified SSRs within Glycine, but frequently, the SSRs were short and interrupted compared with those of soybeans. Nevertheless, 85% of the loci were polymorphic within G. clandestina. Cross-species amplification outside of the genus was much lower (3%-13%), with polymorphism restricted to one primer pair, AG81. AG81 amplified loci in Glycine, Kennedia, and Vigna (Phaseoleae), Vicia (Vicieae), Trifolium (Trifolieae), and Lupinus (Genisteae) within the Papilionoideae, and in Albizia within the Mimosoideae. The primer conservation at AG81 may be explained by its apparent proximity to the seryl-tRNA synthetase gene. Interspecific differences in allele size at AG81 loci reflected repeat length variation within the SSR region and indels in the flanking region. Alleles of identical size with different underlying sequences (size homoplasy) were observed. Our findings and the emerging patterns in other plant studies suggest that in contrast to animals, successful cross-species amplification of SSRs in plants is largely restricted to congeners or closely related genera. Because mutations in both the SSR region and the flanking region contribute to variation in allele size among species, knowledge of DNA sequence is essential before SSR loci can be meaningfully used to address applied and evolutionary questions.

Base Sequence↗

The effect of phenylpropanolamine on the urethral pressure and heart rate is retained after repeated short-term administration in the unanaesthetized, conscious dog.

The present study was performed to test whether high dose agonist (phenylpropanolamine) administration twice a day causes desensitization of urethral alpha-adrenoceptors in vivo. Urethral pressure was measured on five consecutive days of phenylpropanolamine treatment of the unanaesthetized, conscious dog, and the method is described in detail. Phenylpropanolamine hydrochloride proper (75 mg, 5.1-6.1 mg kg-1), and a sustained-release formulation, both significantly increased urethral pressure and decreased heart rate. Interruption of administration for two to three days did not alter the effect. The present results suggest that the effect of phenylpropanolamine was retained, and that the alpha-adrenoceptors of dog urethra did not desensitize after repeated administration of the alpha-adrenoceptor agonist phenylpropanolamine twice a day.

Adrenergic Agents↗

Patent side branches do not affect coronary blood flow in internal thoracic artery-left anterior descending anastomosis: an experimental study.

BACKGROUND: It has been reported that large side branches of internal thoracic artery grafts may steal flow from the coronary circulation. Material an. METHODS: To assess the importance of the side branches, we measured the proximal and distal flow and pressures (mean subclavian artery pressure and mean arterial anastomotic pressure) at baseline and during infusion of adenosine (0.5 mg/kg/min) in 10 Landrace pigs in which an internal thoracic artery-left anterior descending anastomosis was constructed without interruption of the side branches. The difference between proximal and distal flow was considered to represent the blood flow of the internal thoracic artery side branches. Measurements were then repeated after surgical occlusion of all the side branches. RESULTS: At baseline, blood flow of the side branches represented 18% of the total flow in the proximal internal thoracic artery, and this percentage remained constant under the infusion of adenosine, which caused a 220% increase of the cardiac index and a 368% increase of the proximal flow. The infusion reduced the gradient along the left internal thoracic artery (mean subclavian artery pressure-mean arterial anastomotic pressure) from 15 to 10 mm Hg (P =.02) as the result of a lower mean subclavian artery pressure, although the mean arterial anastomotic pressure remained constant. Interruption of all the side branches resulted in a small and not significant increase in distal flow even after adenosine infusion. CONCLUSION: These observations suggest that blood flow in the side branches is minimal either at baseline and under combined systemic and coronary vasodilation. Clinically significant flow steal from the coronary circulation to the internal thoracic artery side branches seems then unlikely.

Anastomosis, Surgical↗

Pattern of Eimeria oocyst output and repeatability in naturally infected suckling Rhön lambs.

The faeces of 14 Rhön lambs were examined every second day between 7 and 99 days of age for the presence of Eimeria oocysts. Eimeria absata, E. bakuensis, E. faurei, E. granulosa, E. intricata, E. ovinoidalis, E. pallida, E. parva and E. crandallis/weybridgensis were identified. The predominant species were E. ovinoidalis, E. parva, E. crandallis/weybridgensis and E. bakuensis. Using a statistical model, the oocyst excretion rate was described as a sequence of periods with decreasing levels and varying length ('excretion periods') interrupted by intervals with no or very low oocyst counts. Several variables could be deduced from these two parameters, including the length of an excretion period and the maximum output during an excretion period. Thc estimated repeatability for oocyst counts for the different species ranged from 0.05 to 0.41. This result provides a starting point for possible genetic selection based on faecal oocyst counts of Rhön sheep for resistance to Eimeria infections.

Animals↗

Molecular basis for expression of common and rare fragile sites.

Fragile sites are specific loci that form gaps, constrictions, and breaks on chromosomes exposed to partial replication stress and are rearranged in tumors. Fragile sites are classified as rare or common, depending on their induction and frequency within the population. The molecular basis of rare fragile sites is associated with expanded repeats capable of adopting unusual non-B DNA structures that can perturb DNA replication. The molecular basis of common fragile sites was unknown. Fragile sites from R-bands are enriched in flexible sequences relative to nonfragile regions from the same chromosomal bands. Here we cloned FRA7E, a common fragile site mapped to a G-band, and revealed a significant difference between its flexibility and that of nonfragile regions mapped to G-bands, similar to the pattern found in R-bands. Thus, in the entire genome, flexible sequences might play a role in the mechanism of fragility. The flexible sequences are composed of interrupted runs of AT-dinucleotides, which have the potential to form secondary structures and hence can affect replication. These sequences show similarity to the AT-rich minisatellite repeats that underlie the fragility of the rare fragile sites FRA16B and FRA10B. We further demonstrate that the normal alleles of FRA16B and FRA10B span the same genomic regions as the common fragile sites FRA16C and FRA10E. Our results suggest that a shared molecular basis, conferred by sequences with a potential to form secondary structures that can perturb replication, may underlie the fragility of rare fragile sites harboring AT-rich minisatellite repeats and aphidicolin-induced common fragile sites.

Alleles↗

Spinocerebellar ataxia type 1 in China: molecular analysis and genotype-phenotype correlation in 5 families.

BACKGROUND: Twelve genetic types of autosomal dominant hereditary ataxia have been recently identified and the genes responsible for most of them cloned. Molecular identification of the type of ataxia is important to determine the disease prevalence and its natural history in various populations. OBJECTIVES: To perform molecular analysis of 75 Chinese families affected with spinocerebellar ataxia (SCA) and to evaluate the spectrum of mutations in these genes and the correlation between genotypes and phenotypes in Chinese patients. SETTING: Neurogenetics Unit, China-Japan Friendship Hospital, Beijing, China. METHODS: One hundred nine patients from 75 kindreds diagnosed as having autosomal dominant SCA, 16 patients with sporadic SCA or spastic paraplegia, 280 control chromosomes of the Chinese population, and 120 control chromosomes of the Sakha population were selected for this study. We conducted detailed mutational analysis by direct sequencing of polymerase chain reaction products amplified from genomic DNA. RESULTS: Spinocerebellar ataxia type 1 (SCA1) was identified in 5 families with 12 studied patients. All affected family members were heterozygous for a CAG repeat expansion in the SCA1 gene containing 51 to 64 trinucleotide repeats. Normal alleles had 26 to 35 repeats. Spinocerebellar ataxia type 1 accounted for 7% of the studied Chinese families with ataxia. In addition, we determined the frequency of a single vs double CAT interruption in 120 control chromosomes of the Siberian Sakha population, which has the highest known prevalence of SCA1, and compared this with 280 control chromosomes from the Chinese populations. The results show that 64.7% of the Siberian normal alleles contain a single CAT interruption, whereas 92% of the Chinese had more than 1 interruption. CONCLUSIONS: Spinocerebellar ataxia type 1 is responsible for 7% of affected families in the Chinese population. A correlation between the prevalence of SCA1 and the number of CAT interruptions in the trinucleotide chain suggests that a CAT-to-CAG substitution may have been the initial event contributing to the generation of expanded alleles and influencing relative prevalence of SCA1.

Adult↗

R loop mapping of the 18S and 28S sequences in the long and short repeating units of Drosophila melanogaster rDNA.

Cleavage of D. melanogaster rDNA with the Eco R1 restriction endonuclease reveals two major classes of repeating units: a long class of 17 kilobases (kb) and a short class of 11.4 kb. R loop mapping has been used to determine the topography of the sequences corresponding to the 18S and 28S rRNAs in both classes, including a cloned member (Dm103) of the long class. This mapping procedure derives from a novel reaction that we discovered between single-stranded RNA and homologous regions in duplex DNA molecules. In high formamide and at an elevated temperature, the RNA pairs with one of the two DNA strands in the region of homology to form an R loop in which one element is an RNA/DNA duplex and the other is single-stranded DNA (Figure 1). Mapping is accomplished by visualization of R loops in the electron microscope. The R loop map of Dm103 parallels that determined independently by Glover and Hogness (1977) from an analysis of its restriction fragments. Both maps indicate that the 28S rDNA in this cloned unit is divided into two blocks by a 5 kb insertion segment. R loop mapping of a population of long units obtained directly from the rDNA (that is, without cloning) has demonstrated that this interruption of the sequence coding for the 28S rRNA is a characteristic of the long class. By contrast, the 28S rDNA in the short units that we examined is not interrupted by an insertion segment. Otherwise, the R loop maps of the long and short units do not differ significantly. The two classes therefore correspond to repeating units that do (IN+) or do not (IN-) contain an insertion segment. Models for the transcription and function of these two classes of repeating units are discussed.

Animals↗

Sequence analysis of the CAG triplet repeats region in the Huntington disease gene (IT15) in several mammalian species.

Huntington's disease (HD) is a progressive neurodegenerative disease linked to abnormally expanded CAG repeats in the first exon of the IT15 gene. Neither HD or other glutamine expansion triplet disease has not yet been described in other species as human. In this study, we sequenced the region containing the CAG repeat expansion region of the corresponding IT15 gene in several mammalian species. In all investigated species, the number of CAG found is equal or inferior to the normal range of human IT15 alleles. The longer uninterrupted CAG stretch was found in the pig and consists of 18 CAG. In the other species, the CAG stretch was shorter and/or interrupted by other triplets. The proline encoding region following the CAG stretch is conserved in the mammalian species studied here and the number of proline is few or not reduced compared to the number of the glutamine codon repeats region.

Amino Acid Sequence↗

Excision-amplification of mitochondrial DNA during senescence in Podospora anserina. DNA sequence analysis of three unique "plasmids".

During senescence in the filamentous fungus Podospora anserina, specific regions of the mitochondrial genome, termed senDNA are excised, ligated and amplified. We have cloned in their entirety three such autonomously replicating plasmids, alpha, beta and epsilon senDNA. None of these plasmids displayed cross-hybridization nor did we detect any significant DNA homology by computer analysis. The complete DNA sequence of the 2.5 kb alpha, the 5.5 kb epsilon and about 3.4 kb of the 9.8 kb beta senDNA is presented (kb = 10(3) base-pairs). These sequences were analyzed for the presence of consensus sequences common to introns, and it was found that alpha senDNA has the characteristics of a group II intron, epsilon senDNA contains three group I introns, and beta senDNA did not show relevant sequences in the 3.4 kb examined. Comparison of the 5' and 3'-flanking sequences of alpha senDNA with oxi 3 (Co I) amino acid sequences from Neurospora crassa and Saccharomyces cerevisiae revealed significant homology and provided strong support that the excised alpha senDNA itself consists entirely of an intron. Upstream from the oxi 3 gene a transfer RNA cysteine sequence was detected. beta senDNA contained four tRNA sequences, aspartic acid, serine, valine and tryptophan, and sequences homologous to URFC (untranslated reading frame C) as well as two new URFs. epsilon senDNA contained sequences homologous to ATPase 8 and URFl; URFl was interrupted by three group I introns. The excision site sequences, as located by S1 nuclease mapping were unique for each senDNA. Analysis for repeated units showed that each plasmid contained elements which could be involved in secondary structure required for the alignment of distal ends preparatory to excision. These results are interpreted in terms of the structural requirements of mobile elements including the possible involvement of reverse transcriptase in the excision-ligation-amplification process.

Amino Acid Sequence↗

New compound curved needle TG6-C PLUS.

The benefits of a compound curve needle design will be seen readily when used as suggested. The surgeon will be able to position the needle repeatedly and accurately between the wound edges from needle entry through tissue, to exit to the opposite side of the wound. Continuous or interrupted corneal wound closure can be done consecutively without varying more than a few tenths of a millimeter in the positioning of the needle from bite to bite. During retrieval the compound curve needle has yet another advantage. The flat curve of the distal needle body minimizes the tissue distortion as the needle body is pulled through. The wound remains in close apposition. In wound closure, though an important and essential attribute of a surgical needle is its "sharpness", sharpness eventually reaches a maximum. The very sharp needles such as the new EthiconTM TG PLUS needles, have such little tactile feedback that shape and needle strength as exemplified by the compound curvature design become of primary importance. These combinations provide the consistent and accurate needle passage necessary for corneal wound apposition, with all of its optical as well as anatomic consequences, in the final outcome of contemporary keratoplasty and cataract procedures.

Cornea↗

Establishment and characterization of a highly tumorigenic human diploid endometrial cancer cell line.

A new cell line designated CUME-1 has been established from a poorly differentiated endometrial adenocarcinoma of the uterus. This cell line grew well without interruption for more than 88 months and 110 serial passages were successively carried out. The cells were highly tumorigenic in nude mice (85%). Repeated karyotype analyses from early (4th) to late (55th) passages of this cell line revealed a diploid stable clone in each passages without any noticeable structural or numerical aberrations. But from the 80th passage, a subpopulation with reciprocal translocation between chromosomes 1q and 9q consistently appeared and was observed in about 30% of the cells. This cell line is one of the rare examples of experimentally proved tumorigenic cells of human solid tumor origin that retains the diploid karyotype in vitro. HLA typing indicated the presence of DR4, DR13, DQ3 and DQ6. Cytosol estrogen and progesterone receptors were found both in fresh primary tumor and in this cell line. Gonadotropin-releasing hormone (Gn-RH) receptor mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) in cultured cells. Using the single-strand conformation polymorphism (SSCP) technique, we have screened CUME-1 cells for p53 mutation in exons 4 to 9. No mobility shift was observed. This cell line may be useful in studying the in vitro chromosomal evolution of the cell line and the in vivo properties of human endometrial adenocarcinoma.

Adenocarcinoma↗

Biochemical treatment of precancerous oral lesions: the effectiveness of various analogues of retinoic acid.

Leukoplakia is neither a clinical, aetiological nor histopathological entity. Therefore treatment is difficult particularly in multifocal and advanced lesions. Since 1970, we have tested the therapeutic effect of different derivatives of all-trans-retinoic acid. The study includes 75 cases with homogeneous leukoplakia without or with minimal epithelial dysplasia. Over 60% of the cases treated showed positive early results. In follow-ups from 1 to 6 years about 45% of cases showed complete or partial remission. The rest showed relapses or even progression. In cases with recurrence without changes in the morphological characteristics, positive effects were achieved with 1 to 4 repeated courses of therapy. All derivatives of retinoic acid tested so far have shown undesirable side effects with systematic manifestations or local symptoms: interruption of treatment (4) or reduction of dosage (9) were unavoidable for that reason. Regarding the side effects, retinoic acid should only be given under clinical supervision. Of the derivates tested to date, aromatic retinoid seems to have the best curative potential in homogenic leukoplakia.

Humans↗

The "second stalk" of Escherichia coli ATP synthase: structure of the isolated dimerization domain.

The b subunit of E. coli F(0)F(1)-ATPase links the peripheral F(1) subunits to the membrane-integral F(0) portion and functions as a "stator", preventing rotation of F(1). The b subunit is present as a dimer in ATP synthase, and residues 62-122 are required to mediate dimerization. To understand how the b subunit dimer is formed, we have studied the structure of the isolated dimerization domain, b(62-122). Analytical ultracentrifugation and solution small-angle X-ray scattering (SAXS) indicate that the b(62-122) dimer is extremely elongated, with a frictional ratio of 1.60, a maximal dimension of 95 A, and a radius of gyration of 27 A, values that are consistent with an alpha-helical coiled-coil structure. The crystal structure of b(62-122) has been solved and refined to 1.55 A. The protein crystallized as an isolated, monomeric alpha helix with a length of 90 A. Combining the crystal structure of monomeric b(62-122) with SAXS data from the dimer in solution, we have constructed a model for the b(62-122) dimer in which the two helices form a coiled coil with a right-handed superhelical twist. Analysis of b sequences from E. coli and other prokaryotes indicates conservation of an undecad repeat, which is characteristic of a right-handed coiled coil and consistent with our structural model. Mutation of residue Arg-83, which interrupts the undecad pattern, to alanine markedly stabilized the dimer, as expected for the proposed two-stranded, right-handed coiled-coil structure.

Amino Acid Sequence↗

Transient EEG patterns during sleep in healthy newborns.

24 healthy full-term newborns underwent polygraphic recordings of EEG, EMG, EOG, ECG, abdominal and thoracic respiration during day-time-sleep. Transient EEG patterns (rhythmic alpha and beta activity, spikes/sharp waves and frontal sharp transients) were visually evaluated and quantified. Rhythmic alpha activity is not very prominent. It is found in all states of sleep and appears only as scattered waves and interrupted sequences. Their duration varies from 1 to 5 sec. Rhythmic beta activity is a feature of quiet and active-REM sleep, repeatedly in spindle-shaped formations. "Typical" sleep spindle activity, however, is very rare. In a few cases rhythmic beta activity also appears in the form of the delta brush pattern. Spikes/sharp waves are sporadic in appearance, non-repetitive in nature and occur mostly multifocal with parietal predominance. They are more frequent in quiet sleep (1 per 2.3 min on average) than in active-REM sleep (1 per 18 min), and were rare in transitional states of sleep (1 per hour). "Frontal sharp transients" show great variations of configuration. They occur most frequently in quiet sleep. Often they appear only unilaterally. Their voltage maximum is frontal, fronto-temporal or praecentral. They are more frequent in REM sleep after wakefulness than after quiet sleep. The individual as well as the sleep-state related frequencies of spike/sharp waves and "frontal sharp transients" tend to differ widely with a range of a factor 10. In some of the newborns these graphoelements are not observed at all during certain sleep phases.

Electroencephalography↗