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Biomedical subjects

M Ochs

Publications and source records attributed to M Ochs.

At least 37 records · Page 2Linked to original sources

Combined use of prostacyclin and higher perfusate temperatures further enhances the superior lung preservation by Celsior solution in the isolated rat lung.

BACKGROUND: The poor tolerance of the lung to ischemia and reperfusion (IR) still represents one of the limitations in clinically successful lung transplantation. Modified Euro-Collins (EC) is routinely used in lung preservation, but alternative solutions have been developed for improvement of pulmonary preservation. Celsior is an extracellular solution that has significantly reduced the IR-induced pulmonary damage in animal studies. So far, no extensive experimental studies exist concerning the influence of Celsior on pulmonary gas exchange following IR. METHODS: In an extracorporeal rat lung model 10 lungs, each, were preserved with Celsior (CE) and Celsior/prostacyclin (CEPC, 6 microg/100 ml) at 4 degrees and 15 degrees C, each, and compared to low-potassium Euro-Collins (EC-40, 40 mmol/liter potassium). After 2 hours of ischemia lungs were reventilated and reperfused using a roller pump. Oxygenation in terms of oxygen partial tension in the left atrial effluent, pulmonary vascular resistance (PVR), peak inspiratory pressure, and wet/dry ratio were monitored for 50 minutes. Furthermore, edema formation was evaluated by light microscopy. Statistical analysis was performed using ANOVA models. RESULTS: Compared to the EC-40 group, oxygenation was increased and amount of edema was reduced in most Celsior-preserved organs (p<0.032) with exception of the CEPC group at 4 degrees C (p = 0.06). Additional application of prostacyclin did not have any significant effect on oxygenation in the Celsior group. However, after temperature elevation of the CEPC perfusate to 15 degrees C, a superior partial tension of oxygen was observed (p<0.023) in contrast to the 4 degrees C groups CE and CEPC. The lowest PVR was found in the CE 4 degrees C group (p<0.02). CONCLUSIONS: Celsior provides better lung preservation than EC-40 solution. Application of prostacyclin at higher perfusate temperatures results in additional functional improvement. In vivo experiments and ultrastructural analysis are warranted for further evaluation of Celsior in lung preservation.

Animals↗

Stereological estimation of the volume weighted mean volumes of alveoli and acinar pathways in the rat lung to characterise alterations after ischaemia/reperfusion.

The aim of this study was to characterise pulmonary reimplantation injury in isolated, perfused rat lungs following 2 h of cold ischaemia, and 50 min. of in vitro reperfusion. The effects of 2 differently composed lung preservation solutions (low potassium Euro-Collins and Celsior; each n = 5) were examined in comparison with untreated, nonischaemic control lungs (n = 3). After fixation by vascular perfusion and tissue collection by systematic random sampling, the volume weighted mean volume (Vv) of alveoli and acinar pathways was estimated by light microscopic stereology using the method of point sampled intercepts in plastic embedded, Azan-stained material. Significantly higher Vv of alveoli and acinar paths was found in the Celsior group than in Euro-Collins preserved lungs. However, in the controls the size of acinar pathways was similar to Celsior preserved lungs whereas alveolar size was comparable to preservation with Euro-Collins. The between-animal coefficient of variation of alveoli was very low in controls and Celsior preserved but higher in the Euro-Collins group. Size distribution of alveoli and acinar paths in 15 size classes was largely homogeneous in all groups tested. In the Euro-Collins group the fractions of both class 1-alveoli and class 1-acinar paths significantly exceeded those of the other groups. Widely expanded alveoli (size classes 13-15) only occurred after preservation with Celsior whereas wider acinar paths (size class 15) were found in the Celsior group and in the controls. It is concluded that lung preservation with low-potassium Euro-Collins and Celsior solutions may act differently on distinct spaces in the distal gas-exchange regions of lungs. This may be due to selective effects on pulmonary surfactant activity and on elastic tissue elements in the alveolar ducts, respectively. Additionally, the method of point sampled intercepts is considered to be an efficient tool to evaluate the effects of different preservation solutions on lung parenchyma.

Animals↗

Ultrastructural alterations in intraalveolar surfactant subtypes after experimental ischemia and reperfusion.

Ischemia and reperfusion (I/R) result in surfactant dysfunction. Whether the impairment of surfactant is a consequence or a cause of intraalveolar edema formation is still unknown. The cumulative effects of lung perfusion, ischemic storage, and subsequent reperfusion on surfactant ultrastructure and pulmonary function were studied in a rat isolated perfused lung model. The left lungs were fixed for electron microscopy by vascular perfusion either immediately after excision (control; n = 5) or after perfusion with modified Euro-Collins solution (EC), storage for 2 h at 4 degrees C in EC, and reperfusion for 40 min (n = 5). A stereological approach was chosen to discriminate between intraalveolar surfactant subtypes of edematous regions and regions free of edema. Intraalveolar edema seen after I/R in the EC group occupied 36 +/- 6% (mean +/- SEM) of the gas exchange region as compared with control lungs (1 +/- 1%; p = 0.008). Relative intraalveolar surfactant composition showed a decrease in surface active tubular myelin (3 +/- 1 versus 12 +/- 0%; p = 0.008) and an increase in inactive unilamellar forms (83 +/- 2 versus 64 +/- 5%; p = 0.008) in the EC group. These changes occurred both in edematous (tubular myelin, 3 +/- 1%; unilamellar forms, 88 +/- 6%) and in nonedematous regions (tubular myelin, 4 +/- 3%; unilamellar forms, 77 +/- 5%). The ultrastructural changes in surfactant were associated with an increase in peak inspiratory pressure during reperfusion. In conclusion, surfactant alterations seen after I/R are not directly related to the presence of edema fluid in the alveoli. Disturbances in intraalveolar surfactant after I/R are not merely the result of inactivation due to plasma protein leakage but may instead be responsible for an increased permeability of the blood-air barrier, resulting in a vicious cycle of intraalveolar edema formation and progressing surfactant impairment.

Animals↗

Ultrastructural pathology of the alveolar type II pneumocytes of human donor lungs. Electron microscopy, stereology, and microanalysis.

Alveolar type II pneumocytes (PII) were studied in 12 human donor lungs perfused with modified Euro-Collins solution during single-lung transplantation (SLTx). While one lung was transplanted, the contralateral donor lung (cDL) was fixed at the time of SLTx for examination by electron microscopy, stereology, and microanalysis. Three groups were then formed: group A (n = 7), cDL without contusions, uneventful early postoperative course; group B (n = 3), cDL with conclusions, uneventful early postoperative course; group C (n = 2), cDL without contusions, early postoperative respiratory dysfunction. The major findings were that the presence of contusions had no effect on PII ultrastructure and that intracellular surfactant-storing lamellar bodies of cDL in group C were characterized by a higher volume-to-surface ratio (VsR) and larger area per cell profile than group A. Correlation analysis based on pooled data (groups A and C) showed that ischaemic time had little effect on PII ultrastructure and bore no relationship to postoperative clinical variables. The duration of preoperative donor intubation had a pronounced influence on ultrastructure and postoperative clinical variables. The stereologically estimated amount of intracellular surfactant and mitochondrial VsR were the only ultrastructural parameters that were significantly associated with early postoperative oxygenation. Lamellar bodies were the only ultrastructural components found to have a significant relationship to postoperative intubation time. The ultrastructural integrity of type II pneumocytes of human donor lungs is an important determinant of early respiratory function following clinical lung transplantation.

Adolescent↗

The past, present, and future of lung transplantation.

BACKGROUND: The history of lung transplantation from the first human transplant performed in 1963 to the present is reviewed with particular focus on the added challenges because of the contaminated bronchus, exposure of the graft to airborne organisms, the poor blood supply to the bronchus, and the problem of reperfusion pulmonary edema. METHODS: The technical aspects of single and double sequential lung transplantation are reviewed, as are the current indications for single, double sequential, and heart/lung transplantation. Criteria for lung transplant recipients, in addition to their primary disease are noted, as are absolute and relative contraindications. The standard criteria for donor selection are also reviewed. RESULTS: The results of single, double sequential, and heart-lung transplantation over the past 10 years as reported by the International Society for Heart and Lung Transplantation Database are reviewed. In addition, the statistics of the lung and heart-lung transplantation program at the University of Colorado Health Sciences Center are reviewed, including the current immunosuppressive regimens and early and late monitoring for infection and rejection. This experience includes 3 early deaths in the first 53 patients for an operative mortality of 5.6%, with a 1-year actuarial survival of 90%. CONCLUSIONS: During the past decade remarkable improvement in the result of single and double sequential lung transplantation have occurred. As 1-year, actuarial survival is now approaching 90% at some institutions. Living related lobar transplantation, new antirejection agents, chimerism, and xenograft transplantation are areas for continuing and future investigation. The shortage in donor organ supply continues to be a very significant factor in limiting human lung transplantation.

Forecasting↗

Surface signaling in transcriptional regulation of the ferric citrate transport system of Escherichia coli: mutational analysis of the alternative sigma factor FecI supports its essential role in fec transport gene transcription.

Ferric citrate induces transcription of the ferric citrate transport genes (fec) in escherichia coli by binding to the outer membrane receptor protein FecA without entering the cell. The signal elicited by ferric citrate crosses the outer membrane via TonB, ExbB, and ExbD. FecR transmits the signal across the cytoplasmic membrane and activates FecI located in the cytoplasm. FecI belongs to a subgroup of sigma factors that respond to extracytoplasmic stimuli. Chromosomal insertion and deletion mutations were generated in fecI; the resulting mutants were totally devoid of FecA production and fecB-lacZ expression. Iron starvation did not derepress fec transport gene transcription in fecI mutants. Missense point mutations were generated in the predicted helix-turn-helix motif of FecI to examine its role in transcription initiation. Replacement of glutamate by alanine (E141A) at the third position in the first helix reduced the residual activity of FecI in the absence of ferric citrate to 30% of the wild-type level, but induced fec transcription almost normally n the presence of ferric citrate. Mutant FecI(K145E) displayed 156% of the activity of wild-type FecI in the absence of ferric citrate and conferred full induction by ferric citrate. Mutant FecI(K155E), which has a mutation in the second helix, showed 9% of the wild-type activity in the presence of ferric citrate and 78% in the absence of ferric citrate. The reduced activity of FecI(K155E) was also shown in vitro by DNA binding assays with cell lysates; in gel retardation experiments FecI(K155E) reduced the electrophoretic mobility of fecA promoter-containing DNA less than did wild-type FecI. fecI is not autoregulated, as demonstrated by the lack of FecI-induced fecI-lacZ expression in vivo and by the lack of specific fecI transcription in vitro. Instead, formation of fecI mRNA requires sigma 70. We conclude that transcription of the fec transport genes is regulated by FecI, which responds to ferric citrate via FecR. fecI and fecR co-transcription is inhibited by the iron-loaded Fur repressor, which then results in a low level of transcription of the fec transport genes.

Amino Acid Sequence↗

Effects of ischaemia and preservation on the ultrastructure of the bronchiolar epithelium. A quantitative electron microscopic study of human and canine lungs.

In ten cases of clinical human single-lung transplantation, the nontransplanted Euro-Collins-preserved contralateral lungs were examined using electron microscopy to determine the effects of ischaemia on the bronchiolar epithelium. Existing structural damage at the time of transplantation was characterized using this approach, and nine nonpreserved canine single lungs were also investigated to identify the impact of ischaemia. The study revealed a significant correlation between the duration of ischaemia and the mitochondrial surface-to-volume ratio, which can serve as a morphometric criterion for mitochondrial damage, in canine lungs. However, this correlation was not found in the human donor lungs. Further examination of human donor lungs showed slight to moderate damage to the endoplasmic reticulum and nuclear chromatin. In addition, various degrees of damage to mitochondrial structure, ranging from inconspicuous to severe, were found. The mitochondrial surface-to-volume ratio can be considered to be a suitable criterion for the quantification of ischaemic damage of the bronchiolar epithelium under experimental conditions. Ultrastructural analysis of human donor lungs revealed intact bronchiolar epithelial cell structures at the time of transplantation, reflecting adequate organ preservation with Euro-Collins solution.

Adolescent↗

Transcriptional regulation of ferric citrate transport in Escherichia coli K-12. Fecl belongs to a new subfamily of sigma 70-type factors that respond to extracytoplasmic stimuli.

Transcription of the ferric citrate transport system of Escherichia coli K-12 is repressed by Fe(2+)-Fur and activated by ferric citrate. Ferric citrate does not have to enter the cytoplasm; it initiates a signal transduction mechanism by binding to the outer membrane receptor FecA. Presumably, a conformational change is transmitted in a TonB-dependent manner to the FecR protein. FecR activates FecI, and FecI activates transcription of the fecABCDE transport genes. In this communication, FecI was isolated after cloning fecI downstream of an ideal ribosome-binding site. Overexpressed FecI formed inclusion bodies which were solubilized and purified in active form using a mild detergent. FecI, in conjunction with RNA polymerase core enzyme, directed transcription from the fecA promoter in an in vitro run-off transcription assay. Furthermore, FecI retarded the electrophoretic mobility of a specific 75 bp DNA fragment located upstream of fecA. An in vivo competition experiment between the fecA promoters of wild-type and mutant strains identified the nucleotide positions 2747, 2749, 2751 and 2753, located within the 75 bp fragment, as important for FecI-induced transcription. Mobility band shift of fecA promoter DNA caused by cell lysates required growth of cells in the presence of ferric citrate and expression of FecA, FecI and FecR. These data support the previous assignment of FecI, based on sequence homologies, to a new subfamily of eubacterial RNA polymerase sigma 70 factors that respond to extra-cytoplasmic stimuli and regulate extracytoplasmic functions.

Bacterial Proteins↗

Regulation of citrate-dependent iron transport of Escherichia coli: fecR is required for transcription activation by FecI.

Citrate-dependent Fe3+ transport into Escherichia coli K-12 is induced by iron and citrate. The inducer is probably ferric dicitrate which does not have to be taken up into the cytoplasm to induce transcription of the fec transport genes. Two regulatory genes, fecI and fecR, located upstream of the fecABCDE transport genes, are required for induction. We report that in vivo the chromosomally encoded FecI protein activates transcription of the fecA and fecB transport genes in response to ferric citrate and the FecR protein. Cells expressing chromosomally and plasmid-encoded truncated FecR derivatives no longer responded to ferric citrate and expressed the fec transport genes constitutively. The smallest active FecR derivative contained 59 amino acid residues as compared to the 317 residues of wild-type FecR. Constitutive induction was lower than induction of the FecR wild-type strain by ferric citrate. It is concluded that the N-terminal portion of FecR activates FecI and that the C-terminal portion of FecR responds to ferric citrate. Transcription of the fec transport genes is positively regulated by FecI and FecR and negatively regulated by the Fe2(+)-Fur repressor. Transcription activation and repression may occur independently of each other.

Amino Acid Sequence↗

Differential diagnosis of jaw pain in the elderly.

While many diseases are marked by pain in the mandible or maxilla, a number of these conditions appear to be more prevalent in people 65 years and older. People in this age group often have a number of medical problems and take a variety of medications, so clear-cut diagnosis of jaw pain can be difficult. Memory deficits or concomitant somatic complaints can further complicate the diagnosis. This article presents a differential for jaw pain in the elderly and reports on a pertinent case.

Aged↗

Electron spectroscopic imaging (ESI) and electron energy loss spectroscopy (EELS) of multilamellar bodies and multilamellar body-like structures in tannic acid-treated alveolar septal cells.

We used electron spectroscopic imaging (ESI) and electron energy loss spectroscopy (EELS) to compare multilamellar bodies (MLB) of Type II alveolar epithelial cells with MLB-like structures that are present in various alveolar septal cells after fixation with tannic acid. Despite their structural similarity in conventional transmission electron microscopy, the phosphorus signal recorded by both ESI and EELS was considerably higher in multilamellar bodies than in MLB-like structures. This indicates that they are different in chemical composition.

Animals↗

Selecting routine outpatient tests for older patients.

In the routine management of elderly patients, the usual objective of diagnostic testing is to prevent the loss of independence when chronic disorders interfere with activities of daily living. Simple in-office tests and other outpatient procedures are the mainstay for diagnosing and monitoring dementia, incontinence, impairment of vision and hearing, alterations in nutrition, musculoskeletal disease, pulmonary and cardiovascular disease, malignancy, and endocrine disorders. In selecting diagnostic tests, physicians must weigh the alternatives based on patient needs, cost-effectiveness, and the availability of nearby medical services.

Aged↗

Surgical management of the hip in the elderly patient.

Hip surgery is the most common major orthopedic procedure performed in the elderly. The indications are fracture and pain secondary to degenerative arthritis. Patients undergoing hip replacement for arthritis have excellent outcomes with decreased pain, increased mobility, and a low mortality. Age should not be a contraindication to hip replacement, with patient selection being made on the basis of symptomatology and overall health. In hip fracture, the prognosis is more guarded. Poor functional outcome results from complications of the fracture, such as avascular necrosis of the femoral head and fracture nonunion in femoral neck fractures and instability with delayed weight bearing in intertrochanteric fractures. In addition, patients sustaining hip fracture are more likely to have significant comorbidity and subsequent perioperative complications. Pressure ulcers, delirium, deep venous thrombosis, urinary retention and urinary tract infection, and cardiac events are the most frequent complications seen. These complications can be anticipated and prevented with careful preoperative assessment and post-operative prophylactic management. A team approach including the orthopedic surgeon, primary care physician, nursing staff, and physical therapists is essential for optimal outcome.

Age Factors↗

Successful prehospital airway management by EMT-Ds using the combitube.

OBJECTIVE: To evaluate the ability to train emergency medical technicians-defibrillation (EMT-Ds) to effectively use the Combitube for intubations in the prehospital environment. METHODS: This was an 18-month prospective field study in which EMT-Ds were trained how and in what situations to use the Combitube. Data were then obtained for all patients in whom Combitube insertion was attempted. Indications for use of the Combitube included: unconsciousness without a purposeful response, absence of the gag reflex, apnea or respiratory rate less than 6 breaths/min, age more than 16 years, and height at least 5 feet tall. Contraindications were: obvious signs of death, intact gag reflex, inability to advance the device due to resistance, or known esophageal pathology. Data were entered prospectively from the San Diego County EMS QANet database for prehospital providers. RESULTS: Twenty-two EMT-D provider agencies, involving approximately 500 EMT-Ds, were included as study participants. Combitube insertions were attempted in 195 prehospital patients in cardiorespiratory arrest, with appropriate indication for Combitube use. An overall successful intubation rate (defined as the ability to successfully ventilate) of 79% was observed. Identical success rates for medical and trauma patients were noted. The device was placed in the esophagus 91% of the time. Resistance during insertion was the major reason for unsuccessful Combitube intubations. An overall hospital admission rate of 19% was observed. No complications were reported. CONCLUSION: EMT-Ds can be trained to use the Combitube as a means of establishing an airway in the prehospital setting. Future studies will need to further evaluate its effect on patient outcome.

California↗

Evaluation of pulmonary edema: stereological versus gravimetrical analysis.

Assessment of lung edema by gravimetrical analysis is a standard method to evaluate the severity of experimentally induced ischemia/reperfusion (IR) injury. The aim of this study was to compare gravimetrical assessment of pulmonary edema with a stereological approach which allows for qualitative and quantitative distinction between intravascular and edematous fluids by light microscopy. Eight experimental groups which differed in mode of preservation, ischemic storage and pharmacological treatments were studied in an extracorporeal rat lung model. Analysis of the pooled data showed that the wet/dry ratio values mainly reflected the amount of intra-alveolar edema (r(s) = 0.442; p = 0.0057) but only stereological assessment of edema formation revealed differences depending on the treatment used. Only stereological data correlated significantly with oxygen tension measured at the end of reperfusion (r(s) = -0.530; p = 0.0009). We conclude that gravimetry is of minor functional importance compared to assessment by stereological methods which prove to be a reliable and efficient tool for the evaluation of IR injury in the different experimental settings.

Animals↗