The long-term effects of early oral feeding following minimal invasive esophagectomy.

A nil-by-mouth regime with enteral nutrition via an artificial route is frequently applied following esophagectomy. However, early initiation of oral feeding could potentially improve recovery and has shown to be beneficial in many types of abdominal surgery. Although short-term nutritional safety of oral intake after an esophagectomy has been documented, long-term effects of this feeding regimen are unknown. In this cohort study, data from patients undergoing minimal invasive Ivor-Lewis esophagectomy between 04-2012 and 09-2015 in three centers in Netherlands were collected. Patients in the oral feeding group were retrieved from a previous prospective study and compared with a cohort of patients with early enteral jejunostomy feeding but delayed oral intake. Body mass index (BMI) measurements, complications, and nutritional re-interventions (re- or start of artificial feeding, start of total parenteral nutrition) were gathered over the course of one year after surgery. One year after surgery the median BMI was 22.8 kg/m2 and weight loss was 7.0 kg (9.5%) in 114 patients. Patients in the early oral feeding group lost more weight during the first postoperative month (P = 0.004). However, in the months thereafter this difference was not observed anymore. In the early oral feeding group, 28 patients (56%) required a nutritional re-intervention, compared to 46 patients (72%) in the delayed oral feeding group (P = 0.078). During admission, more re-interventions were performed in the delayed oral feeding group (17 vs. 46 patients P < 0.001). Esophagectomy reduces BMI in the first year after surgery regardless of the feeding regimen. Direct start of oral intake following esophagectomy has no impact on early nutritional re-interventions and long-term weight loss.

[Local wound care: evidence-based treatments and dressings]. / Lokale wondzorg: evidence-based behandelingen en verbandmaterialen.

Patients with open wounds require specific local-wound care. There is huge variety in methods of local-wound care. This is due not only to the many different types of wounds but also to the widely varying preferences of doctors and nurses, and to the lack of strong evidence and relevant guidelines regarding the most appropriate form of local-wound care. In 19 systematic literature reviews from the Cochrane Collaboration on surgical and traumatic wounds and ulcers (venous, arterial and diabetic) information can be found on local-wound treatment. Eleven of these reviews were limited to strong evidence for the use of: tissue adhesives for surgical and traumatic wounds; foam dressings in surgical wounds healing by secondary intention; (intermittent) compression, bilayer artificial skin and lidocaine-prilocaine cream for venous ulcers; hydrogel and in-shoe orthotics for diabetic-foot ulcers; negative pressure for chronic wounds, and cleansing of acute wounds with clean tap water.

Activation of the urease of Schizosaccharomyces pombe by the UreF accessory protein from soybean.

Plant orthologs of the bacterial urease accessory genes ureD and ureF, which are required for the insertion of the nickel ion at the active site, have been isolated from soybean ( Glycine max L. Merr.), tomato ( Lycopersicon esculentum) and Arabidopsis thaliana. The functionality of soybean UreD and UreF was tested by measuring their ability to complement urease-negative mutants of Schizosaccharomyces pombe, a eukaryote which produces a "plant-like" urease of ~90 kDa. The S. pombe ure4 mutant was complemented by a 12-kb fragment of S. pombe genomic DNA, which was shown by PCR to contain a putative ureD gene. However, ure4 was not complemented by a UreD cDNA soybean, expressed under the control of a strong promoter. In contrast, an S. pombe ure3 mutation was complemented by both a 10-kb fragment of S. pombe DNA containing ureF and the UreF cDNA from soybean. Soybean Eu2 is a candidate urease accessory gene; its product cooperates with the Eu3 protein in activating apourease in vitro. However, the sequences of UreD and UreF transcripts from two eu2/eu2 mutants, recovered as RT-PCR products, revealed no mutational alteration, suggesting that Eu2 encodes neither UreD nor UreF.

Examination of lactococcal bacteriophage c2 DNA replication using two-dimensional agarose gel electrophoresis.

The ori locus of the prolate-headed lactococcal bacteriophage c2 supports plasmid replication in Lactococcus lactis in the absence of phage infection. To determine whether phage c2 DNA replication is initiated at the ori locus in vivo and to investigate the mechanism of phage DNA replication, replicating intermediates of phage c2 were analyzed using neutral/neutral two-dimensional agarose gel electrophoresis (2D). The 2D data revealed that c2 replicates via a theta mechanism and localized the initiation of theta replication to the ori region of the c2 genome.

Prevalence, risk factors and prevention of pressure ulcers in Dutch intensive care units. Results of a cross-sectional survey.

OBJECTIVE: Evaluating the prevalence, risk factors and prevention of pressure ulcers in Dutch intensive care units (ICUs). DESIGN: Cross-sectional design. SETTING: ICUs of acute care hospitals that participated in the 1998 and 1999 national prevalence surveys. Data were collected on 1 day in each year. PATIENTS: Eight hundred fifty patients admitted to Dutch ICUs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Six categories of data were collected: (1) characteristics of the institution, (2) characteristics of the ward, (3) characteristics of the patients (age, sex, date of admission, reason for admission), (4) risk assessment using the Braden scale and two additional risk factors (malnutrition and incontinence), (5) severity of the pressure ulcers and (6) supportive surface used. The prevalence of pressure ulcers was 28.7%. In a forward logistic regression analysis, four risk factors were significantly associated with the presence of pressure ulcers: infection, age, length of stay and total Braden score. Of the patients at high risk of developing pressure ulcers but without actual pressure ulcers, 60.5% were positioned on a support system. Only 36.8% of the patients who were determined to need repositioning were actually being turned. CONCLUSIONS: The prevalence of pressure ulcers in Dutch ICUs is high and their prevention is flawed, especially as regards the use of support systems. Patients for whom turning is indicated are not being turned. Predicting pressure ulcers in ICU patients is difficult and needs further investigation.

Hemodynamic effects of intermittent manual lung hyperinflation in patients with septic shock.

OBJECTIVE: The purposes of this study were to investigate the hemodynamic changes induced by intermittent manual lung hyperinflation (MHI) and to assess if these changes are adverse enough to warrant prohibition of MHI as a routine procedure in the care of patients with septic shock. DESIGN: The study's design was experimental prospective. SETTING: The settings were university hospital intensive care units. PATIENTS: Subjects included 13 consecutive mechanically ventilated patients with septic shock who met the inclusion criteria. MEASUREMENTS AND RESULTS: Phasic MHI-related increments in mean inspiratory airway pressure were concordant to changes in mean pulmonary artery pressure (MPAP) (r(2) = 0.67) with a 0.6 mm Hg rise in MPAP per cm H(2)O airway pressure. The magnitude of MPAP changes was not reflected in magnitude of stroke volume index (SVI) (r(2) = 0.06). On average, MHI did not induce statistically significant hemodynamic changes and mean values returned to baseline level within 15 minutes. SVI during MHI increased slightly in 9 patients, from 37 +/- 15 (mean +/- SD) to 41 +/- 17 mL/m(2) (P <.05), and decreased in 4, from 60 +/- 10 to 50 +/- 14 mL/m(2) (not significant). Patients with an increase in SVI had lower baseline values for SVI, cardiac index, and left ventricular stroke work index (P <.05) and higher values for systemic vascular resistance index compared with patients with a decrease in SVI (P <.05). Left ventricular stroke work index was higher in patients with a decrease in SVI than in patients with an increase in SVI (52 +/- 9 vs 34 +/- 8; P <.05). Tidal volume increased from 499 +/- 176 mL before MHI to 587 +/- 82 mL, 5 minutes after MHI (P <.05) with a return to baseline values within 15 minutes after the procedure. CONCLUSION: The hemodynamic effects of intermittent MHI in patients with septic shock are relatively small and insignificant and seem to be related to the cardiovascular state before the procedure. The risk of inducing hemodynamic changes with MHI should not be considered as a contraindication in patients with septic shock who are mechanically ventilated.

Enhanced susceptibility to cytotoxic T lymphocytes without increase of MHC class I antigen expression after conditional overexpression of heat shock protein 70 in target cells.

Antigenic peptides have been found associated with heat shock proteins (HSP) including cytoplasmic HSP70 and heat shock cognate protein 70 as well as the endoplasmic reticulum-resident glucose-regulated protein 94. Recently, HSP70 transfection has been reported to increase MHC class I cell surface expression and antigen presentation on mouse melanoma B16 cells (Wells et al., Int. Immunol. 1998. 10: 609). To analyze the effect of HSP70 on MHC class I cell surface expression and lysability of target cells we transfected a human melanoma cell line with the rat Hsp70-1 gene using the Tet-On system for conditional overexpression of HSP70. Induction of HSP70 did not increase cell surface expression of HLA class I molecules in general or individual HLA-A and B antigens in particular. Nonetheless, induction of HSP70 enhanced susceptibility of these cells to lysis by allospecific CTL. The same effect was observed using an HLA-A2-restricted tyrosinase-specific CTL clone after pulsing the tyrosinase-negative target cells with the specific peptide. Thus, HSP70 induction can increase killing by CTL without affecting MHC class I cell surface expression or antigen processing. This effect of HSP70 appears to be different from the commonly found protection exerted by HSP70 against stress like heat shock, and might be mediated by improving CTL-induced apoptosis.

[Obstetric problems followed by stroke]. / Obstetrische problemen: soms een voorbode van naderend onheil uit onverwachte hoek.

Obstetrical problems sometimes portend manifestations of atherosclerosis, as illustrated by two case reports. The first patient had the combination of hyperhomocysteinaemia due to chronic vitamin deficiencies in the diet, and smoking. The second was also a smoker and had a genetically determined mild hyperhomocysteinaemia, aggravated by chronic vitamin deficiencies resulting from poor dietary habits; she also had an increased folic acid requirement because of use of anti-epileptic drugs in combination with a familial predisposition for premature atherosclerotic manifestations. The first patient had four pregnancies, two of which ended in intrauterine foetal death due to placental infarction, and one in the birth of a dysmature boy. The second patient's four pregnancies ended twice in abortion and twice in the birth of a dysmature child; in one of the latter cases placental infarction was observed. Both women subsequently suffered cerebrovascular accidents while in addition, older cerebral infarctions were found to be present. Women with recurrent abortion, pre-eclampsia, placental infarction, placental detachment and foetal growth retardation should be examined, even if other risk factors are also present, for (mild) hyperhomocysteinaemia, and treated for it with vitamin suppletion (folic acid, vitamins B6 and B12), even although admittedly more research is necessary to make certain that such treatment has a preventive effect on the manifestations of this disorder.

The development of a national registration form to measure the prevalence of pressure ulcers in The Netherlands.

To gain insight into the prevalence of pressure ulcers in Dutch healthcare institutions, a national registration form to measure the prevalence of pressure ulcers annually in different healthcare settings was developed based on a literature study and responses from a Delphi panel. The reliability and the feasibility of the form devised were tested in a pilot study conducted in a university hospital, a nursing home, and in a home healthcare setting. Interrater reliability of the grading system varied between the institutions from 0.49 to 0.97 (Cohen's Kappa). In the home healthcare setting, interrater reliability was 0.80 (Pearson correlation coefficient) for the total score on the Braden scale. The prevalence rates were 10.1% (n = 368) in the university hospital, 12.7% (n = 1,541) in the home healthcare setting, and 83.6% (n = 122) in the nursing home, although the latter figure seemed to be somewhat exaggerated. The most common lesions were found on the sacrum and below the knee (heel and malleolus). The pilot study concluded that it is possible to collect accurate and reliable data on the scope and severity of pressure ulcers with a uniform instrument in different healthcare settings.

Transcription analysis of the prolate-headed lactococcal bacteriophage c2.

A detailed transcription map of the prolate-headed lactococcal phage c2 has been constructed. Transcription of about one-third of the genome, encoding 22 open reading frames, began within the first 2 min of infection and produced at least 12 overlapping transcripts that persisted until lysis occurred at 30 min after initiation of infection. The remaining two-thirds of the genome, encoding 17 open reading frames, was divergently transcribed, beginning between 4 and 6 min after initiation of infection, and resulted in at least 18 overlapping transcripts that persisted until lysis. Five very strong, simultaneously active, and probably unregulated early promoters and a single positively regulated late promoter were identified. The late promoter had an extended -10 sequence, had a significant basal level of activity in the uninduced state, and was induced to high activity by a phage gene product. The complex overlapping pattern of transcripts resulted from the action of the multiple early promoters, inefficient termination of transcription, and (possibly) processing of a late precursor transcript(s). Phage proteins were not required for these processes, and the host RNA polymerase was probably used for both early and late transcription.

Mapping of ure1, ure2 and ure3 markers in fission yeast.

The following urease genes of the fission yeast Schizosaccharomyces pombe have been mapped by induced haploidization and tetrad analysis--ure1: chromosome are III-L; ure2 and ure3: chromosome are I-R. The previously determined tps19-rad1 interval (11-12 cM) has been increased to 18 cM. A convenient medium for rapidly scoring the ure gene markers of fission yeast was developed.

An origin of DNA replication from Lactococcus lactis bacteriophage c2.

An origin of DNA relication was identified in the intergenic region between the early and late gene regions of prolate lactococcal phage c2. A DNA fragment containing this origin, designated ori, was shown to direct DNA replication in Lactococcus lactis but not in Escherichia coli. A comparison of ori with the corresponding regions of other prolate phages revealed strict conservation of the nucleotide sequence in one half of this intergenic region. This conserved region alone would not support DNA replication. No open reading frames were identified in the ori fragment, suggesting that host factors alone are sufficient to initiate DNA replication at ori. A novel class of lactococcal vectors and E. coli-L. lactis shuttle vectors based on ori have been constructed.

Purification and characterization of urease from schizosaccharomyces pombe.

The urease from the ascomycetous fission yeast Schizosaccharomyces pombe was purified about 4000-fold (34% yield) to homogeneity by acetone precipitation, ammonium sulfate precipitation, DEAE-Sepharose ion-exchange column chromatography, and if required, Mono-Q ion-exchange fast protein liquid chromatography. The enzyme was intracellular and only one species of urease was detected by nondenaturing polyacrylamide gel electrophoresis (PAGE). The native enzyme had a M(r) of 212 kDa (Sepharose CL6B-200 gel filtration) and a single subunit was detected with a M(r) of 102 kDa (PAGE with sodium dodecyl sulfate). The subunit stoichiometry was not specifically determined, but the molecular mass estimations indicate that the undissociated enzyme may be a dimer of identical subunits. The specific activity was 700-800 micromols urea.min-1.mg protein-1, the optimum pH for activity was 8.0, and the Km for urea was 1.03 mM. The sequence of the amino terminus was Met-Gln-Pro-Arg-Glu-Leu-His-Lys-Leu-Thr-Leu-His-Gln-Leu-Gly-Ser-Leu-Ala and the sequence of two tryptic peptides of the enzyme were Phe-Ile-Glu-Thr-Asn-Glu-Lys and Leu-Tyr-Ala-Pro-Glu-Asn-Ser-Pro-Gly-Phe-Val-Glu-Val-Leu-Glu-Gly-Glu-Ile- Glu- Leu-Leu-Pro-Asn-Leu-Pro. The N-terminal sequence and physical and kinetic properties indicated that S. pombe urease was more like the plant enzymes than the bacterial ureases.

Sequencing and analysis of the prolate-headed lactococcal bacteriophage c2 genome and identification of the structural genes.

The 22,163-bp genome of the lactococcal prolate-headed phage c2 was sequenced. Thirty-nine open reading frames (ORFs), early and late promoters, and a putative transcription terminator were identified. Twenty-two ORFs were in the early gene region, and 17 were in the late gene region. Putative genes for a DNA polymerase, a recombination protein, a sigma factor protein, a transcription regulatory protein, holin proteins, and a terminase were identified. Transcription of the early and late genes proceeded divergently from a noncoding 611-bp region. A 521-bp fragment contained within the 611-bp intergenic region could act as an origin of replication in Lactococcus lactis. Three major structural proteins, with sizes of 175, 90, and 29 kDa, and eight minor proteins, with sizes of 143, 82, 66, 60, 44, 42, 32, and 28 kDa, were identified. Several of these proteins appeared to be posttranslationally modified by proteolytic cleavage. The 175- and 90-kDa proteins were identified as the major phage head proteins, and the 29- and 60-kDa proteins were identified as the major tail protein and (possibly) the tail adsorption protein, respectively. The head proteins appeared to be covalently linked multimers of the same 30-kDa gene product. Phage c2 and prolate-headed lactococcal phage bIL67 (C. Schouler, S. D. Ehrlich, and M.-C. Chopin, Microbiology 140:3061-3069, 1994) shared 80% nucleotide sequence identity. However, several DNA deletions or insertions which corresponded to the loss or acquisition of specific ORFs, respectively, were noted. The identification of direct nucleotide repeats flanking these sequences indicated that recombination may be important in the evolution of these phages.(ABSTRACT TRUNCATED AT 250 WORDS)

Molecular biology of lactococcal bacteriophage c2.

The 22163 bp genome of the lytic prolate-headed lactococcal phage c2 was fully sequenced. Mapping of restriction sites and RNA transcripts demonstrated the presence of early and late genes. Early and late promoters were identified. The early region contained 21 ORFs, with predicted protein products of 4.4-32.9 kDA, all reading right to left. Significant similarity was found between a putative protein encoded by an early region ORF and the erf (essential recombination function) gene product of Salmonella phage P22. The late genes for which a function has been identified, all of which read from left to right, included a possible holin gene, and genes encoding three major and six minor phage structural proteins. Analysis of the cohesive termini revealed complementary, non-symmetrical, 9-base single-stranded 3' extended DNAs. The exploitation of phage sequence data and analysis of phage genomes to find ways of inhibiting phage replication is discussed.

Sequencing and analysis of the cos region of the lactococcal bacteriophage c2.

The cohesive termini of the DNA genome of the lactococcal bacteriophage c2 were directly sequenced and appeared to be complementary, non-symmetrical, 9-nucleotide single-stranded, 3' extended DNAs, with the following sequence: 5'-GTTAGGCTT-3' 3'-CAATCCGAA-5'. DNA located on either side of the cohesive ends was sequenced and several repeats and a region with the potential for a DNA bend were found. Previously sequenced cos regions of 13 other bacteriophages were also examined for similar sequence features. All of the bacteriophages from gram-positive hosts had 3' extended DNA termini, in contrast to the bacteriophages from gram-negative hosts, which had 5' extended DNA termini. All bacteriophages had a region of dyad symmetry close to the cohesive termini. A 7.3 kb DNA fragment of the c2 genome containing the cos sequences was cloned; transduction experiments demonstrated that these cloned sequences could act as a substrate for packaging enzymes of phage c2.

Components of glycine reductase from Eubacterium acidaminophilum. Cloning, sequencing and identification of the genes for thioredoxin reductase, thioredoxin and selenoprotein PA.

The genes encoding thioredoxin reductase (trxB), thioredoxin (trxA), protein PA of glycine reductase (grdA) and the first 23 amino acids of the large subunit of protein PC of glycine reductase (grdC) belonging to the reductive deamination systems present in Eubacterium acidaminophilum were cloned and sequenced. The proteins were products of closely linked genes with 314 codons (thioredoxin reductase), 110 codons (thioredoxin), and 158 codons (protein PA). The protein previously called 'atypically small lipoamide dehydrogenase' or 'electron transferring flavoprotein' could now conclusively be identified as a thioredoxin reductase (subunit mass of 34781 Da) by the alignment with the enzyme of Escherichia coli showing the same typical order of the corresponding domains. The thioredoxin (molecular mass of 11742 Da) deviated considerably from the known consensus sequence, even in the most strongly conserved redox-active segment WCGPC that was now GCVPC. The selenocysteine of protein PA (molecular mass of 16609 Da) was encoded by TGA. The protein was highly similar to those of Clostridium purinolyticum and Clostridium sticklandii involved in glycine reductase. Thioredoxin reductase and thioredoxin of E. acidaminophilum could be successfully expressed in E. coli.

Sequence analysis of the lysin gene region of the prolate lactococcal bacteriophage c2.

Approximately 80% of the genome of the prolate-headed lactococcal bacteriophage c2 was cloned into shuttle vectors pSA3 and pFX3 in Escherichia coli and transferred to Lactococcus lactis. A 1.67-kilobase EcoRV fragment containing the gene for the phage lysin was identified and the position and orientation of the phage lysin gene in the physical map of the phage were determined. The phage lysin was expressed in E. coli and its sequence was determined and compared with the sequences of other bacteriophage lytic genes. The sequence was similar, but not identical, to that of the related lactococcal phage m13, having a number of silent substitutions and an apparent deletion that altered the carboxy terminus of the protein. Possible alternative translation initiation codons for the lysin gene and two possible alternative mechanisms for access of the lysin enzyme to the cell wall are discussed. An open reading frame upstream of the putative lysin gene was found to be 177 base pairs longer than that reported for phage m13. A codon usage table for the lysin genes of several phages as well as for reported gene sequences from L. lactis and lactococcal bacteriophages is presented.