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Whole body arginine metabolism and nitric oxide synthesis in newborns with persistent pulmonary hypertension.

Despite the potential relevance of the L-arginine-nitric oxide (NO) pathway in the pathophysiology of pulmonary hypertension, no in vivo studies of the kinetics of arginine and NO have been conducted previously in this population. The terminal guanidino N-atom of L-arginine is the precursor for NO, which is oxidized to the stable inorganic nitrogen oxides, nitrite (NO2-) and nitrate (NO3-). Thus, synthesized NO is detected in serum or urine as NO2- and NO3-. The purpose of this investigation was to compare studies of whole body arginine metabolism twice in nine patients with persistent pulmonary hypertension of the newborn (PPHN), using a primed constant i.v. infusion of L-[guanidino-15N2,5,5(2)H2]arginine and L-[5,5,5(2)H3]leucine, first during acute pulmonary vasoconstriction and again during convalescence, and thereby to characterize quantitative aspects of whole body arginine kinetics and NO production, as estimated from the rate of transfer of the 15N-guanidino-label of arginine to urinary nitrate (15NO3-). Arginine flux rates were 84.1 +/- 8.6 mumol.kg-1 h-1 (mean +/- SEM) during acute pulmonary hypertension and increased to 125 +/- 13.2 (p < 0.05) during convalescence, whereas leucine fluxes were unchanged (168.5 +/- 15 versus 178.8 +/- 10.2 mumol.kg-1 h-1), and comparable to those reported in healthy newborns. During convalescence total urinary nitrate excreted increased by 66% (p < 0.05), urinary 15NO3- increased from 0.29 +/- 0.07 to 0.74 +/- 0.15 mumol.d-1 (p < 0.05), and the rate of plasma arginine conversion to NO increased from 10.3 +/- 2.2 to 45.6 +/- 13 mumol.d-1 (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Arginine↗

Role of IgA versus IgG in the control of influenza viral infection in the murine respiratory tract.

The roles of IgG and secretory IgA in the protection of the respiratory tract (RT) against influenza infection remain unclear. Passive immunization with Ab doses resulting in serum IgG anti-influenza virus Ab titers far in excess of those observed in immune mice has compounded the problem. We compared the effects of i.v. anti-influenza virus IgG and i.v. anti-influenza virus polymeric IgA (pIgA) mAb administered in amounts designed to replicate murine convalescent serum or nasal Ab titers, respectively. A serum anti-influenza virus IgG titer 2.5 times the normal convalescent serum anti-influenza virus IgG titer was required for detectible Ab transudation into nasal secretions, and a serum IgG titer 7 times normal was needed to lower nasal viral shedding by 98%. Anti-influenza virus pIgA at a nasal Ab titer comparable to that seen in convalescent mice eliminated nasal viral shedding. The RT of influenza-infected pIgA- or IgG-protected mice were studied by scanning electron microscopy. Only pIgA was found to prevent virally induced pathology in the upper RT, suggesting that IgG did not prevent viral infection of the nose, but neutralized newly replicated virus after infection had been initiated. In contrast, IgG, but not pIgA, was found to prevent viral pathology in the murine lung. Our results help to resolve the controversy of IgA- vs IgG-mediated protection of the RT; both Abs are important, with plasma IgG Ab serving as the back-up for secretory IgA-mediated protection in the nasal compartment, and IgG being the dominant Ab in protection of the lung.

Animals↗

Immunoglobulin M-capture biotin-streptavidin enzyme-linked immunosorbent assay for detection of antibodies to dengue viruses.

A biotin-streptavidin system was adapted to an IgM-capture ELISA for detection of dengue antibodies in human sera. To develop this assay, high titers of antibodies to flavivirus were purified by ion-exchange chromatography (DEAE-cellulose) and labeled with biotin. Heavy chain-specific goat anti-human IgM was first bound to the wells of a polystyrene microtiter plate, followed by binding of IgM in test specimens, and the use of tetravalent dengue antigens (dengue 1-4), biotin-labeled anti-flavivirus IgG, and streptavidin-peroxidase conjugate. The sensitivity and specificity of the IgM-capture biotin-streptavidin ELISA (IgM-BS-ELISA) in acute sera were 83.3% of patients with dengue infection and 95.3% of nondengue-infected cases, respectively. The positive predictive value was 92.4% and the negative predictive value was 89.2%. The efficiency of test was 90.4%. In convalescent sera, the sensitivity and specificity of IgM-BS-ELISA were 100% and 92.6%, respectively. The predictive values of positive and negative results were 90.3% and 100%, respectively. The efficiency of test was 95.6%. The agreement rate of IgM-BS-ELISA and standard hemagglutination inhibition test was good: kappa (kappa) values were 0.79 for acute sera and 0.91 for convalescent sera. The correlation between two methods was quite good, with correlation coefficients (r) of 0.76 for acute sera and 0.85 for convalescent sera (P < 0.001). The results indicate that the IgM-BS-ELISA is highly sensitive, specific, simple to perform, and rapid.

Acute Disease↗

Back to work more quickly after an inguinal hernia repair.

Tension-free hernia repair plus recovery expectancy statements return personnel to work more quickly. On the day of primary inguinal hernia repair, patients were given statements about their likelihood of returning from convalescent leave after 7 days and performing nonstrenuous work. Similar statements were given to them by telephone at 72 hours postoperatively and at a 1-week follow-up appointments. Seventy-four percent of the 73 patients returned to nonstrenuous work within 7 days, and 90% returned to strenuous work within 30 days. In this small sample, 385 work days were saved from the Navy's recommended 14 days of convalescent leave. By combining recovery expectancy statements with an effective surgical procedure, it is possible to avoid prolonged convalescence, thereby enhancing military readiness.

Absenteeism↗

Levels of serum vascular cell adhesion molecule-1 in measles.

While the levels of serum vascular cell adhesion molecule-1 (VCAM-1) have been investigated in many diseases, they have not, to our knowledge, been studied in patients with measles. Serum VCAM-1 and creatinine levels were determined in six adolescent and adult Japanese patients with measles in the acute febrile phase and defervescent afebrile phase. Serum VCAM-1 levels were significantly higher in the acute febrile phase than in the convalescent afebrile phase, but no significant difference between the serum creatinine levels was shown in the two phases. Our study revealed that the high serum VCAM-1 level in the acute febrile phase later decreased in the afebrile convalescent phase. The measurement of circulating VCAM-1 may be useful for the assessment of convalescence in patients with measles.

Acute Disease↗

[Dynamics of changes in magnesium (Mg) serum levels in patients suffering from infectious mononucleosis].

The aim of the study was to assess the dynamics of changes in magnesium (Mg) serum levels in the acute, symptomatic period of infectious mononucleosis and during the convalescence period, when clinical symptoms were no longer present. 50 subjects were included, among them 26 patients (14 women and 12 men) at the age of 16 to 27 years in the acute, symptomatic stage of the disease. The diagnosis of infectious mononucleosis was based on the clinical, hematological, biochemical and serologic criteria. Mg concentration was measured 3 times: on the 1st day and in the 2nd week of hospitalization, and also in the convalescence period--3 weeks after the regression of clinical symptoms. The control group consisted of 24 healthy individuals (5 men and 5 women) at the age of 17 to 26 years. Mg concentration was measured once in this group. All measurements of Mg serum concentration were performed using the method of atomic absorption spectrophotometry (AAS) at the wavelength of 285.2 nm. We observed significantly higher Mg serum concentrations in patients with acute, symptomatic infectious mononucleosis than in healthy individuals. During the convalescence period Mg serum concentrations were lower but still statistically higher than in the control group.

Acute Disease↗

[Ciliary activity of the ciliated epithelium of the airways in patients with acute pneumonia].

A TV unit was employed to study ciliary activity of bronchial ciliated epithelium in 53 patients with acute pneumonia (AP) at its height and in convalescence. Ciliary impairment of the epithelium is independent of AP clinical form (lobular or croupous) or of dissemination of the inflammation though is related to endobronchitis activity. The ciliary dysfunction emerged in all the patients at the height of the disease and in 65.3% of convalescents. Latent dysfunction was revealed by thermal and euphylline tests in 34.7% of the convalescents. Diagnostic criteria of ciliary dysfunction are proposed.

Acute Disease↗

[Valvular regurgitation in patients with Kawasaki disease and in healthy children: a pulsed Doppler echocardiographic study].

Valvular lesions in the acute stage of Kawasaki disease were observed using pulsed Doppler echocardiography. The subjects consisted of 65 patients with Kawasaki disease (2 months-6 2/12 years) who had been followed from the acute stage. The age-matched 113 controls were selected from 661 healthy children (2 months-14 years of age). In the acute stage of Kawasaki disease, tricuspid regurgitation (TR) was detected in 31 (48%), pulmonary regurgitation (PR) in 22 (34%) and mitral regurgitation (MR) in 17 (26%). There was no aortic regurgitation (AR). In the convalescent stage, TR was found in 26 (40%), PR in 20 (31%), and MR in 11 (17%), but no AR was detected. The incidence of each valvular regurgitation between the acute and convalescent stages in patients with Kawasaki disease did not differ significantly. Furthermore, there was no significant difference in the incidence of valvular regurgitation between patients with Kawasaki disease and the normal controls. In nine patients, however, valvular regurgitation in the acute stage had disappeared by the convalescent stage, and two patients had developed a new pansystolic murmur in the acute stage. We estimated the incidence of pathologic valvular involvement in Kawasaki disease to be 11/65 (17%). The incidence of valvular involvement in patients with coronary artery aneurysms was significantly higher than that of patients without coronary artery aneurysms (p less than 0.01). It was concluded that mild and transient valvular regurgitation, which cannot be detected by auscultation, may occur in some patients in the acute stage of Kawasaki disease. These may be caused by acute inflammation of the valve related to coronary artery lesions. In view of the Doppler echocardiographic findings in normal controls, these regurgitations should be distinguished from "physiological" ones.

Acute Disease↗

Analysis of immune function in herpes zoster patients: demonstration and characterization of suppressor cells.

We sought to identify imbalances of immune regulatory cells that might contribute to the depression of cell-mediated immunity that occurs during an episode of herpes zoster. Peripheral blood mononuclear cells (PBMC) were obtained from patients with herpes zoster during the acute (less than 7 days after disease onset) and convalescent (more than 10 days after disease onset) phases of illness and from healthy seropositive donors. The PBMC were analyzed for: lymphoproliferative responses to varicella-zoster virus (VZV) antigens, Leu-3 (helper/inducer):Leu-2 (cytotoxic/suppressor) ratios, and percentages of suppressor cells as defined by coexpression of the Leu-2 and OKM1 antigens. Significantly depressed proliferative responses of VZV antigens and Leu-3:Leu-2 ratios, and increased percentages of Leu-2+ OKM1+ suppressor cells were observed in PBMC of acute phase herpes zoster patients as compared with the PBMC of convalescent patients or healthy donors. These differences were also observed in individual patients sequentially studied during both phases of disease. Cryopreserved acute phase PBMC suppressed the proliferative response of autologous convalescent phase PBMC to VZV antigens, but not to herpes simplex virus (HSV) antigens. The acute phase PBMC suppressor cell was radiation sensitive and was identified as a Leu-2+ cell by fluorescence-activated cell sorting. Thus, depression of cell-mediated immunity during the acute phase of herpes zoster was associated with a relative increase of lymphocytes expressing a suppressor cell phenotype and the activation of a radiosensitive Leu-2+ suppressor cell with some degree of antigen specificity.

Acute Disease↗

Western blot analysis of the human serum response to Mycoplasma hominis.

Mycoplasma hominis is a human genital pathogen with importance in postpartum pregnancy complications (postpartum fever/endometritis). Previous research has suggested that serum antibody levels to M. hominis are important in predicting which groups of women are at risk. M. hominis strain PG21 was examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blots to determine which protein antigens would be good candidates for use in serologic tests. Western blots of strain PG21 were probed with acute and convalescent human sera from patients with culture-confirmed M. hominis infections, animal sera directed against strain PG21 and other M. hominis strains, and sera from patients with confirmed infections from other sexually transmitted diseases (STD). Western blot analysis showed that the prenatal (Groups I, II, III) and convalescent (Group IV) M. hominis human sera reacted with proteins with apparent MWs of 106, 67, 46, and 40 kilodaltons (kDa). Only the convalescent sera (Group IV) and the prenatal sera (Group III) reacted with proteins having apparent MWs of 58 and 50 kDa. Animal antisera directed against all strains of M. hominis examined showed that these proteins were reactive in all strains, and other STD human sera did not react with proteins in strain PG21 corresponding to the apparent MWs of 50 and 58 kDa. Preliminary evidence suggested that proteins with the apparent MWs of 50 and 58 kDa may be viable candidates for use in serologic tests for the detection of human anti-M. hominis antibodies and help to eliminate cross-reactivity observed with whole-cell lysates.

Adult↗

[Changes in the blood plasma lipoprotein system of patients in the acute and recovery periods of myocardial infarct].

Repeated assessments of changes in plasma lipoprotein spectrum in 160 patients with large-focal myocardial infarction, and the aggregation of high-density lipoproteins (HDLP) in 26 of those demonstrated specific quantitative and qualitative shifts that differed in acute infarction and convalescence. Total cholesterol, HDLP cholesterol and triglyceride levels on day 60 after the attack evidenced that atherogenic disturbances of lipid metabolism were virtually similar to those seen in chronic coronary patients. Lipoprotein disorders are more marked in patients with anginal episodes punctuating their convalescence as compared to those whose convalescence is uneventful.

Adult↗

Studies of lymphocytotoxins in infectious mononucleosis: reduced lymphocyte killing in the acute phase.

Fifty sera from twenty-five patients with uncomplicated infectious mononucleosis (IM) were tested for lymphocytotoxic activity (LCA) against pools of lymphocytes collected from (a) normal donors (b) ten patients during the acute phase of IM and (c) four of these patients during the convalescent phase. The LCA of twenty-four sera from patients with systemic lupus erythematosus (SLE) was also tested against the same panels of cells. The percentage of acute phase lymphocytes killed by sera from patients with IM or SLE was significantly less (P less than 0.001 and P less than 0.05 respectively) than the percentage of normal or convalescent cells killed by the same sera. This reduction could not be explained by lymphocyte donor variation as the same donors were used in the acute and convalescent phases of IM. It is suggested that changes in the surface characteristics of acute IM lymphocytes cause a temporary failure of interaction with circulating lymphocytotoxins. Such a phenomenon could have important influences on the self-limiting nature of the disease.

Acute Disease↗

Efficacy and immunogenicity of acellular pertussis vaccine by manufacturer and patient age.

Acellular pertussis vaccines, which have been used in Japan since 1981, vary in antigenic constituents among manufacturers. First, to assess the immunogenicity by manufacturer and patient age, 161 children aged 3 months to 2 years were immunized by acellular pertussis vaccine from one of three Japanese manufacturers, Biken, Takeda, or Kitasato. Anti-pertussis toxin antibody responses for children immunized with Takeda and Kitasato vaccines were comparable with patients with pertussis in the convalescent stage, and anti-pertussis toxin antibody response for Biken vaccine was far higher than those of convalescing patients. Anti-filamentous hemagglutinin antibody responses for the children given the three vaccines were far higher than those of the patients. Weak serotype 1.3 agglutinin responses were observed only in children administered the Takeda vaccine. Comparing these antibody responses among various age groups, the immunogenicity of acellular vaccines in children aged 3 to 6 months was comparable with children aged 2 years. Second, to assess the manufacturer-specific efficacy, 495 households of patients with pertussis were surveyed from 1981 to 1988. The estimated efficacy of the acellular pertussis vaccines in children aged 2 to 8 years was 82%, and there were no major differences in the secondary attack rates among children immunized with acellular pertussis vaccine from each manufacturer, ie, 12.5% (1/8) for Biken, 11.1% (2/18) for Takeda, and 5.9% (1/17) for Kitasato. We conclude from these two studies that similar efficacy was observed in children aged 2 years or older for acellular pertussis vaccines from the three manufacturers, which produced anti-pertussis toxin antibody responses comparable with patients with pertussis and far higher antifilamentous hemagglutinin antibody responses than in the convalescing patients, and that age did not affect the immunogenicity of acellular vaccines.

Age Factors↗

Laparoscopic cholecystectomy in acute cholecystitis. What is the optimal timing for operation?

OBJECTIVE: To review the results of laparoscopic cholecystectomy (LC) in patients with acute cholecystitis with attention to cost and clinical outcome. DESIGN: Retrospective study. SETTING: Large private metropolitan teaching hospital. PATIENTS: Four hundred forty-six patients had LCs at our institution between January 1993 and February 1995. Acute cholecystitis, confirmed by clinical, laboratory, operative, and histopathological findings, was present in 60 patients. MAIN OUTCOME MEASURES: The medical history, laboratory findings, gallbladder ultrasounds, timing of operation from the onset of symptoms, conversion rates to open procedures, operative times, intraoperative findings, complications, postoperative length of stay, cost of operative procedures and hospitalizations, and convalescence times were collected. RESULTS: Laparoscopic cholecystectomy was attempted in 16 patients within 72 hours of the onset of symptoms of acute cholecystitis (group 1), in 19 patients with symptoms between 4 and 7 days (group 2), and in 25 patients with symptoms lasting more than 7 days (group 3). The only factor (eg, preoperative laboratory and ultrasound findings) that affected the outcome of the operation was duration of symptoms prior to operation. Patients who had LC done within 72 hours of the onset of symptoms had lower rates of conversion to open procedures, less difficult operations, shorter operative times, less costly procedures, and a shorter convalescence than those with symptoms for longer than 72 hours prior to operation. The conversion rates in patients operated within and after 72 hours were 12% and 30%, respectively. There were no bile duct injuries and no mortalities. CONCLUSIONS: Laparoscopic cholecystectomy can be performed safely in most patients with acute cholelithiasis. However, we found that the duration of symptoms prior to LC affected the outcome; the conversion rates, hospital costs, and convalescence times increased in operated-on patients with symptoms for more than 72 hours. In our opinion, interval cholecystectomy may be a superior option in this latter group of patients.

Acute Disease↗

Viral proteins VP2, VP6, and NSP2 are strongly precipitated by serum and fecal antibodies from children with rotavirus symptomatic infection.

Rotavirus-specific IgA has been correlated with immune protection against rotavirus reinfection and symptomatic disease. Systemic and mucosal antibody responses were determined by an enzyme-linked immunosorbent assay in 11 infants with severe rotavirus gastroenteritis. Geometric mean titers of antirotavirus serum IgG and IgA antibodies were significantly higher during the convalescence of the disease (P < 0.001 vs. acute-phase titers). Rotavirus-specific fecal sIgA antibodies increased 4 times during the convalescence in 9 (81.8%) children (P < 0.001). The serum IgG and IgA antibody and fecal sIgA antibody responses to individual rotavirus polypeptides were characterized by radioimmunoprecipitation assay (RIPA) using Staphylococcus aureus protein A and the lectin jacalin to precipitate IgG- and IgA-immune complexes, respectively. The main IgG response was directed toward the structural viral proteins VP2, VP4, and VP6 and toward the nonstructural protein NSP2. Serum IgA reactivity was detected by RIPA in all serum samples, with major responses to VP2, VP6, and NSP2. Interestingly, fecal sIgA in convalescent samples reacted strongly toward NSP2 and VP6. These data reinforce the antigenic importance of rotaviral proteins other than VP4 and VP7, such as VP2, VP6, and NSP2, as main targets in the immune response to rotavirus.

Antibodies, Viral↗

Resistance of recent measles virus wild-type isolates to antibody-mediated neutralization by vaccinees with antibody.

The neutralization capacity of sera from Luxembourgian adolescent vaccinees and from Nigerian women with measles-induced immunity to a number of measles virus strains was compared. Although both cohorts were matched for their hemagglutination inhibition and standard neutralization titers, 12 of the 22 late convalescent sera, and only 6 of 24 vaccinees neutralized all viruses. Similarly, only 2 of 20 viruses were not neutralized by at least 75% of late convalescent sera, in comparison to 10 of 20 viruses that resisted neutralization by at least 75% of the vaccinees. The more resistant viruses were not limited to a certain clade. One Nigerian virus was resistant to neutralization by 30% of the late convalescent women and by 75% of vaccinees. These results suggest that qualitative differences in neutralizing antibodies may reduce further protection of infants by passively acquired immunity against wild-type viruses when vaccinated girls become mothers.

Adolescent↗

Day care after operations for hernia or varicose veins: a controlled trial.

Alternative systems of care after operations for varicose veins or hernia were compared in a total of 360 selected patients, of whom 121 were allocated to be managed in an acute ward for 48 h, 122 in a convalescent hospital for 48 h and 117 to be discharged directly home to the care of the district nursing sister and general practitioner. There were no deaths or major complications. Anaesthetic or surgical problems caused 5 patients (3 convalescent and 2 day care) to be retained in hospital on the day of operation. Minor complications were recorded in approximately one-third of the patients. The majority of these were effectively dealt with by the district nursing sister and only one-third of the complications needed the attention of the general practitioner. Two of the ward patients and 1 of the convalescent patients required readmission to hospital (1 per cent in all). No significant difference was demonstrated in the medical outcome between the three groups after operation. Day care was the most economical of the three systems of care. Inquiry into the patients' opinions elicited the highest proportion of favourable responses in the day care group.

Clinical Trials as Topic↗

Evaluation of active human herpesvirus 6 infection by reverse transcription-PCR.

Monitoring of active human herpesvirus 6 (HHV-6) infection is important for distinguishing between reactivation and latency of the virus. The reverse transcription polymerase chain reaction (RT-PCR) may be a useful tool in order to distinguish active and latent HHV-6 infection. An RT-PCR assay detecting 4 different HHV-6 gene transcripts was established. Samples of peripheral blood mononuclear cells (PBMCs) were collected from patients with exanthem subitum and used to evaluate the reliability of the assay. After confirming the reliability of the assay, RT-PCR was used to determine whether HHV-6 reactivation occurs in children with hypercytokinemia. Three gene transcripts (U31, U39, and U94) were detected in 90-100% of the PBMC samples collected from febrile period of exanthem subitum patients, from which HHV-6 was isolated. The two gene transcripts encoding the late proteins U31 and U39, however, were not detected in samples collected during the convalescent period that contained no infectious virus. The putative latency associated gene transcript, U94, was detected in 2 (10%) of the 20 convalescent samples, and another immediate early gene transcript, U90, was also detected in 3 (15%) of the 20 convalescent samples. The frequency of HHV-6 reactivation in patients with hypercytokinemia, suggesting monocyte/macrophage activation, was studied. Only 9 of 17 patients diagnosed with Kawasaki disease and 1 patient diagnosed with juvenile rheumatoid arthritis were positive for HHV-6 DNA in their PBMCs samples. Neither the U31 gene nor the U94 gene transcript was detected in any of the 10 samples. An RT-PCR assay screening for both immediate early and late genes may be useful for monitoring active HHV-6 infection. No HHV-6 reactivation was found in patients with hypercytokinemia using the RT-PCR assay.

Cytokines↗