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PubMed2006

Public sector nurses in Swaziland: can the downturn be reversed?

BACKGROUND: The lack of human resources for health (HRH) is increasingly being recognized as a major bottleneck to scaling up antiretroviral treatment (ART), particularly in sub-Saharan Africa, whose societies and health systems are hardest hit by HIV/AIDS. In this case study of Swaziland, we describe the current HRH situation in the public sector. We identify major factors that contribute to the crisis, describe policy initiatives to tackle it and base on these a number of projections for the future. Finally, we suggest some areas for further research that may contribute to tackling the HRH crisis in Swaziland. METHODS: We visited Swaziland twice within 18 months in order to capture the HRH situation as well as the responses to it in 2004 and in 2005. Using semi-structured interviews with key informants and group interviews, we obtained qualitative and quantitative data on the HRH situation in the public and mission health sectors. We complemented this with an analysis of primary documents and a review of the available relevant reports and studies. RESULTS: The public health sector in Swaziland faces a serious shortage of health workers: 44% of posts for physicians, 19% of posts for nurses and 17% of nursing assistant posts were unfilled in 2004. We identified emigration and attrition due to HIV/AIDS as major factors depleting the health workforce. The annual training output of only 80 new nurses is not sufficient to compensate for these losses, and based on the situation in 2004 we estimated that the nursing workforce in the public sector would have been reduced by more than 40% by 2010. In 2005 we found that new initiatives by the Swazi government, such as the scale-up of ART, the introduction of retention measures to decrease emigration and the influx of foreign nurses could have the potential to improve the situation. A combination of such measures, together with the planned increase in the training capacity of the country's nursing schools, could even reverse the trend of a diminishing health workforce. CONCLUSION: Emigration and attrition due to HIV/AIDS are undermining the health workforce in the public sector of Swaziland. Short-term and long-term measures for overcoming this HRH crisis have been initiated by the Swazi government and must be further supported and increased. Scaling up antiretroviral treatment (ART) and making it accessible and acceptable for the health workforce is of paramount importance for halting the attrition due to HIV/AIDS. To this end, we also recommend exploring ways to make ART delivery less labour-intensive. The production of nurses and nursing assistants must be urgently increased. Although the migration of HRH is a global issue requiring solutions at various levels, innovative in-country strategies for retaining staff must be further explored in order to stem as much as possible the emigration from Swaziland.

Journal Article↗
PubMed2000

[Comparative studies of the adsorption behavior of plasma proteins to heparin-coated surfaces].

Over the past 10 years, investigations have shown that heparin coating optimizes haemocompatibility in extracorporeal circulation systems. To date, however, the mechanisms involved have not been identified. 20 ml of fresh human blood were circulated in an in vitro closed-loop model with and without heparin coating. Using a newly developed ELISA, a quantitative analysis of the adsorbed plasma proteins was done. In addition, changes in coagulation, complement and blood cell releasing factors were measured by ELISA methods: prothrombin fragment 1 + 2 (F1 + 2), thrombin-antithrombin-III complex (TAT), PMN-elastase, beta-thromboglobulin (beta-TG) and terminal complement-complex (TCC). In uncoated tubing, high concentrations of fibrinogen, fibronectin, prothrombin, vitronectin, alpha 2-macroglobulin, von Willebrand factor and complement factor C3 were found. Large amounts of antithrombin-III, HMWK and C1-esterase inhibitor were found on the heparinized surfaces. In addition, the concentrations of the soluble plasma protein markers F1 + 2, FPA, PMN-elastase, TCC and beta-TG were significantly lower (p < 0.05) in the presence of heparin coating. The haemocompatibility of "foreign" surfaces depends largely on the extent to which the processes of activation and inhibition of humoral and cellular mediators permanently occurring at the natural endothelium can be simulated. Owing to the low adsorption of procoagulatory and proinflammatory enzymes, heparin-coated surfaces demonstrate significantly improved haemocompatibility.

Adsorption↗
PubMed2004

The LSM 510 META - ConfoCor 2 system: an integrated imaging and spectroscopic platform for single-molecule detection.

Fluorescence Correlation Spectroscopy (FCS) has developed into a routine method to quantitatively study diffusion of molecules and kinetic processes. FCS has recently become popular to complement live cell imaging with biophysical information. The enabling technology has been commercially realised by combining laser scanning microscopes and fluorescence correlation spectrometers in one integrated platform. This article provides an overview of the Zeiss solution of such a combined instrument, the LSM 510 META / ConfoCor 2 system. We focus on the instrumental set up as well as technical advances and improvements in the software that controls FCS data acquisition and evaluation. In addition, we outline the calibration of the instrument and the work flow for data analysis with emphasis on in vivo pharmacological applications.

Calibration↗
PubMed1989

Antigens, antibodies and immune complexes in cerebrospinal fluid of patients with cerebral gnathostomiasis.

Methods for the detection of antigens, antibodies and immune complexes in the cerebrospinal fluid (CSF) of patients with neurological manifestations suggestive of cerebral gnathostomiasis were developed, in the hope that they may be useful in confirming the diagnosis of Gnathostoma spinigerum infection. Gnathostoma antigens were determined by a sandwich enzyme linked immunosorbent assay (ELISA) using antibodies from rabbits immunized with the excretory/secretory (ES) antigens obtained from the in vitro supernatant fluid in which the third-stage G. spinigerum larvae were maintained. With a biotin streptavidin procedure, the presence of G. spinigerum antigens as low as 2 ng in one ml of CSF could be detected. An indirect ELISA was used for the quantitation of IgG antibodies in the paired serum and CSF of these patients. A complement consumption method was used for the detection of immune complexes in the concentrated CSF specimens. Of the 11 patients with clinical signs and symptoms suggestive of having G. spinigerum infection involving the central nervous system, only one patient had antigens detected in the CSF and in this one patient no antibody could be demonstrated. One other patient had immune complexes in her CSF. All remaining patients had IgG antibodies demonstrable in the CSF specimens. These data suggest that the detection of IgG antibodies in CSF is more reliable than the other two methods in confirming the diagnosis of cerebral gnathostomiasis.

Animals↗
PubMed1986

Integrated cardiopulmonary approach to exercise testing in pediatrics.

In the pediatric age groups exercise evaluation of cardiac and pulmonary disease has had a lesser impact on clinical care than in adults. Exercise evaluation complements clinical information, echocardiography, hemodynamics and lung function studies at rest to provide reproducible, quantitative information to answer questions of capacity for physical work, employment, insurability and participation in recreation and sports. To ensure serial studies in children measurements should be noninvasive and minimize demands of patient cooperation. Exercise evaluation should include evaluation of lung function at rest to exclude pulmonary limitations of exercise. Because of the close link of cardiovascular and pulmonary responses to exercise, assessment of ventilation and pulmonary gas exchange should be an integral part of exercise evaluation. Computerized systems can enhance the accuracy and precision of such measurements and can provide a data base for evaluation of individual patients and the natural history of treated congenital cardiac defects and pulmonary disease.

Adolescent↗
PubMed1980

Properties of the general acyl-CoA dehydrogenase from pig liver.

The properties of the general acyl coenzyme A dehydrogenase from pig liver and the stable reduced dehydrogenase . product and oxidized dehydrogenase . acetoacetyl-CoA complexes of the dehydrogenase were investigated. The enzyme has a molecular weight of 178,000 to 183,000 determined by gel filtration chromatography and by gel electrophoresis at different acrylamide concentrations. The subunit molecular weight is 45,000 based on acrylamide gel electrophoresis in the presence of dodecyl sulfate which agrees with the minimum molecular weight calculated from the flavin:protein ratio and the amino acid analysis. The subunits are identical, or very similar, as judged by quantitative NH2-terminal analysis and mapping of tryptic peptides. Immunochemical analyses by the complement fixation technique show that the structure of the oxidized enzyme is different from the structure of enzyme . acyl-CoA complexes whether the flavin in these complexes is in the oxidized or reduced state. The amino acid analysis, isoelectric point, and a procedure for crystallizing the dehydrogenase are also reported.

Acyl-CoA Dehydrogenases↗
PubMed1980

Immune-complexes in malignant skin melanoma: quantitation by three methods, purification, and characterization of their non-tumor related constituents.

Circulating immune-complexes (ICs) and complement system (C) were studied in sera from 10 patients with malignant skin melanoma. The presence of ICs was demonstrated in all of them by three distinct methods. Moreover, ICs were purified in all the patient sera. The fact that ICs were detected in each serum and that the results of the analysis obtained by the different methods were not overlapping is discussed. It is stressed that the differences among the intrinsic characteristic of each technique are responsible for these discrepancies. The C profile was altered if compared with normal values, but it was not strictly characteristic for an ICs disease. In the purified ICs C3, C4, IgM, IgC1, and IgG3 were demonstrated; C1q, C1s, IgA, IgG2, and IgG4 never. It is noteworthy that a rheumatoid activity was found in the purified material and that this activity was significantly correlated with serum immuno-conglutinins (IKs) titre.

Adult↗
PubMed1996

The use of monoclonal antibodies and flow cytometry in the diagnosis of paroxysmal nocturnal hemoglobinuria.

We have characterized the erythrocytes, granulocytes, and platelets of 54 patients with paroxysmal nocturnal hemoglobinuria (PNH) with antibodies to glycosylphosphatidylinositol-anchored proteins (anti-CD55, anti-CD59, and anti-CD16) and flow cytometry to establish the usefulness of this technique in the diagnosis of this disorder. All patients demonstrated either completely (PNH III) or partially (PNH II) deficient red cells and granulocytes. Anti-CD59 best demonstrated PNH II red cells, which were present in 50% of the patients. The proportion of abnormal granulocytes was usually greater than the proportion of abnormal red cells; 37% of the patients had >80% abnormal granulocytes. Anti-CD55 did not delineate the erythrocyte populations as well as did anti-CD59. Either anti-CD55 or anti-CD59 could be used equally well to analyze granulocytes; anti-CD16 did not demonstrate cells of partial deficiency. Platelets could not be used for detailed analysis as the normal and abnormal populations were not well distinguished. Flow cytometry of erythrocytes using anti-CD59 or of granulocytes using either anti-CD55 or anti-CD59 provides the most accurate technique for the diagnosis of paroxysmal nocturnal hemoglobinuria; it is clearly more specific, more quantitative, and more sensitive than the tests for PNH that depend upon hemolysis by complement (the acidified serum lysis [Ham] test, the sucrose lysis test, and the complement lysis sensitivity [CLS] test).

Adolescent↗
PubMed1971

Specific inactivation of the fourth complement component. I. In vivo studies.

Intravascular injection of guinea pigs with the fourth complement component (C4) inactivator from shark serum resulted in severe serum C4 depletion that lasted for several hours. Active and direct passive Arthus reactions did not develop or were extremely mild in such C4 inactivator-treated animals as judged by gross and histological observation. In consideration of the specificity of the C4 inactivator, its independence of cofactors, and its failure to generate biologically active materials in the process of C4 inactivation, it is concluded that complement-dependent immune injury can be suppressed or avoided by interruption of the complement cascade at a single early step. The depression of the Arthus reaction beyond the time of C4 recovery prompted quantitative speculations concerning (i) the in vivo rate of C5a generation under conditions of limited C4 supply and (ii) the "saturation" of immune complexes. This view is supported by in vitro demonstration of functional inactivation or "saturation" of immune precipitates by repeated incubation with fresh guinea pig complement. It seems therefore conceivable that even temporary complement inactivation may have a profound beneficial effect on complement-dependent immune injuries.

Animals↗
PubMed2006

Multiple innate inflammatory responses induced after systemic adenovirus vector delivery depend on a functional complement system.

Excessive complement activation can result in extreme tissue damage and systemic inflammatory responses, similar to innate immune responses rapidly elicited after systemic adenovirus (Ad) injections. To determine if Ad interactions with the complement system impact upon Ad-induced innate immune responses, we injected Ad into complement-deficient, C3-knockout mice (C3-KO) or wild-type mice (WT) and quantitatively compared multiple anti-Ad innate immune responses in both strains of mice. In Ad-treated WT mice, we noted rapid increases in plasma KC levels (1 h post injection), followed by increases in IL-6, IFN-gamma, RANTES, IL-12(p40), IL-5, G-CSF, and GM-CSF and subsequently thrombocytopenia. Conversely, in Ad-treated C3-KO mice, many of these inflammatory responses were significantly blunted, including the avoidance of Ad-induced thrombocytopenia. Global liver transcriptome responses in Ad-treated WT mice were assessed by RT-PCR-validated gene array analysis and were found to be also significantly affected by the lack of complement activity in Ad-treated C3-KO mice. Finally, our results confirmed the ability of high dose Ads to transduce hepatocytes despite a lack of complement activity. In summary, Ad interactions with the mammalian complement system are significant and likely initiate and/or exacerbate many of the inflammatory responses noted after systemic Ad injections.

Adenoviridae↗
PubMed1989

Effect of long-term normalization of serum complement levels on the course of lupus nephritis.

PURPOSE: We compared the long-term outcome of patients with lupus nephritis in whom normalization of complement levels (CH50) was sustained by adjustment of immunosuppressive therapy to those patients with persistently low complement levels despite similar immunosuppression in whom therapy was adjusted solely on the basis of clinical disease activity. PATIENTS AND METHODS: Thirty-nine female patients with lupus nephritis recruited from 1972 to 1979 were prospectively studied (mean follow-up, 116.7 +/- 11 months). Entry criteria included initial renal biopsy, low CH50, and elevated anti-DNA antibody levels. A second biopsy was performed in 24 patients after an interval of 40.6 +/- 5 months. Treatment was started with prednisone (1 mg/kg/day). Azathioprine at a dose of 1.5 to 2.0 mg/kg/day was added if complement was not normalized by prednisone alone. Twenty-five of 39 patients had normal complement levels within six months (Group 1), and immunosuppressive therapy was tapered but continuously readjusted to the lowest dosage that preserved normal CH50 and maintained clinical remission. Eight of these 25 patients subsequently became persistently hypocomplementemic due to inadequate drug intake (Group 1B), whereas the complement levels continued to be controlled in the other 17 patients (Group 1A). Despite similar therapy, the remaining 14 patients did not achieve normalization of complement within the initial six months of therapy, and therefore future treatment decisions were based solely on clinical symptoms (Group 2). Renal pathologic lesions were classified according to World Health Organization criteria and a semi-quantitative chronicity index. RESULTS: During the first six months, there were no significant differences in clinical or histologic features between patients in whom complement levels were controlled and patients in whom complement levels were not controlled. After a mean observation period of 10 years, however, patients with consistent normalization of complement (Group 1A) did much better than patients with only short-term complement control (Group 1B) or persistent hypocomplementemia (Group 2). Both groups with low complement levels had a similar outcome with significantly worse kidney and patient survival. Life-table analysis demonstrated that the differences in outcome between complement-controlled and complement-uncontrolled groups became apparent only after five or more years of follow-up. Patients with a low chronicity score on initial biopsy whose complement level was controlled did uniformly well with no renal failure or death. (ABSTRACT TRUNCATED AT 400 WORDS)

Azathioprine↗
PubMed1982

Immunochemical determination of Forssman and blood group A-active glycolipids in human gastric mucosa by inhibition assay of liposome lysis.

A simple liposome immunoassay, liposome immune-lysis inhibition (LILI) assay, is described for quantitative determination of individual glycolipid antigens. Liposomes containing fluorogenic marker, 4-methylumbelliferyl phosphate, were prepared from sphingomyelin, cholesterol, dicetylphosphate and standard glycolipid. Release of trapped markers from these liposomes by antibody and complement (liposome lysis) was inhibited by preincubating the antibody with test glycolipid incorporated into inhibitor liposomes. Based on the competitive inhibition, it was possible to quantitate each glycolipid antigen in less than picomolar amounts. The sensitivity and specificity of the assay were examined with purified glycolipid standards. LILI assay has been applied for the determination of Forssman glycolipid and blood group A-active glycolipid in human gastric mucosa and cancer tissues.

ABO Blood-Group System↗
PubMed2006

Cutaneous immune responses in the common carp detected using transcript analysis.

In order to detect new immune-related genes in common carp (Cyprinus carpio L.) challenged by an ectoparasitic infection, two cDNA libraries were constructed from carp skin sampled at 3 and 72h after infection with Ichthyophthirius multifiliis. In a total of 3500 expressed sequence tags (ESTs) we identified 82 orthologues of genes of immune relevance previously described in other organisms. Of these, 61 have never been described before in C. carpio, thus shedding light on some key components of the defence mechanisms of this species. Among the newly described genes, full-length molecules of prostaglandin D2 synthase (PGDS), the CC chemokine molecule SCYA103, and a second gene for the carp beta(2)-microglobulin (beta(2)m), beta(2)m-2, were described. Transcript amounts of the genes PGDS, interferon (IFN), SCYA103, complement factor 7 (C7), complement factor P (FP), complement factor D (FD) and beta(2)m-2 were evaluated by real-time quantitative PCR (RQ-PCR). Samples from skin, blood and liver from fish challenged with I. multifiliis were taken at 3, 12, 24, 36 and 48h post infection. Higher expression levels of most of these transcripts were observed in skin from uninfected fish, compared to the transcript levels detected in blood and liver from the same animals. Also, there was significant down-regulation of the genes PGDS and beta(2)m-2 in skin, whilst significant up-regulation was observed for the C7 and SCYA103 genes in liver of fish infected with the parasite. These results confirm the active role of fish skin in the immune response against infections, acting as an important site of expression of immune-related molecules.

Amino Acid Sequence↗
PubMed1992

[The correlation between measurements performed by intravascular echography and quantitative digital angiography in large- and small-caliber arteries].

Intravascular ultrasound is a new promising technique that appears as a very useful complement to standard angiography in the study of atherosclerosis. Specifically, intravascular ultrasound provides by itself, information of great value concerning vessel wall characteristics. Nevertheless, before taking any decision with the results provided by this new technique, we must validate this information with that provided by other more conventional techniques like standard angiography. Accordingly, we have analyzed in 25 patients, 50 images of the abdominal aorta and 77 images of the coronary arteries taken from arterial segments free of atherosclerotic involvement. With the use of a calibrated small grid located over the patient's thorax, we have analyzed "in the same points" the images obtained with intravascular ultrasound and those provided by contrast angiography using a quantitative angiographic analysis. In the abdominal aorta the vessel diameter measured by intravascular ultrasound and angiography was 17.8 +/- 0.39 mm vs 18.6 +/- 0.42 mm (NS), whereas in the coronary arteries it was 2.76 +/- 0.39 mm vs 2.98 +/- 0.37 mm (NS), respectively. The correlation between both techniques was good for both types of arteries (r = 0.93 in the abdominal aorta and r = 0.87 in the coronary arteries). We conclude, that in our experience and with the methodology used, there is a good correlation between the measurements of the vessel diameters obtained by both intravascular ultrasound and digital angiography in arteries of different sizes, being the correlation even better in large arteries.

Adult↗
PubMed1983

Juvenile periodontitis as a model for neutrophil function: reduced binding of the complement chemotactic fragment, C5a.

The binding of the chemotactic complement fragment, C5a, to peripheral blood neutrophils of Localized Juvenile Periodontitis (LJP) patients and normal controls was quantitated using iodinated human C5a and a rapid centrifugation assay. There was a significant reduction in the number of binding sites per cell on neutrophils from the patient group, whereas the binding affinity remained the same as control values.

Binding Sites↗
PubMed2006

The state of rehabilitation research: art or science?

Rehabilitation research has been criticized as not standing up enough to the rigors of scientific method to be called "science." The field has been portrayed as slow to promote its scientific achievements and to include them under the rubric of evidence-based rehabilitation. Following in the footsteps of psychology, rehabilitation as a broad-based discipline has faced many similar obstacles in achieving scientific status. Controversy exists about what exactly constitutes rehabilitation science versus its art and its respective multidisciplinary domains. The conception of these domains is directly related to current methods available to assess the state of the discipline and its research accomplishments. I used quantitative methods, such as randomized clinical and/or controlled trials (RCTs) and systematic reviews, to assess the status of rehabilitation research. Findings suggest that, as a field, rehabilitation makes significant contributions to science, measurable by the number and quality of RCTs and systematic reviews conducted so far on topics of critical importance for clinical care. In "artful" complement, qualitative approaches can be used as research tools to aid investigators in seeking knowledge beyond that obtained by quantitative methods, assessing many complexities associated with the various contexts of rehabilitation research. Other requirements to develop a common vision of rehabilitation science are also discussed.

Art↗
PubMed2001

Complementation of a speA negative Streptococcus pyogenes with speA: effects on virulence and production of streptococcal pyrogenic exotoxin A.

We have shown previously that an isogenic SPEA-negative Streptococcus pyogenes strain did not attenuate virulence in a murine model of necrotizing fasciitis. The aim of this study was to confirm that streptococcal pyrogenic exotoxin A (SPEA) is not crucial for streptococcal invasiveness in murine invasive infection. The SPEA-negative S. pyogenes (H326) was complemented with speA extra-chromosomally to create strain H361 which produced 2.2-fold more SPEA compared with the parental speA(+)wild-type (H305). The growth phase-regulated expression of SPEA in vitro was unaffected in this strain. Complementation with speA resulted in reduced virulence and bacterial counts in invasive murine infection. SPEA production was quantitated from muscle tissue of infected mice. However, H361 did not produce more SPEA than H305 in vivo. We conclude that SPEA does not play a key role in invasive murine streptococcal infection.

Animals↗
PubMed1997

Quantitation of ERCC-2 gene expression in human tumor cell lines by reverse transcription-polymerase chain reaction in comparison to northern blot analysis.

Excision repair cross-complementing rodent repair deficiency genes (ERCC) are human genes implicated in nucleotide excision repair. ERCC-2 has been implicated in the repair of DNA damaged by chemotherapeutic agents, and may thus play an important role in anticancer drug resistance. ERCC-2 gene expression is low in primary tumor samples rendering it difficult to quantitate. We have developed a semiquantitative method to measure ERCC-2 gene expression utilizing reverse transcription-polymerase chain reaction (RT-PCR). Total RNA extracted from established human tumor cell lines was reverse-transcribed to obtain cDNA. Serially diluted reference ERCC-2 DNA fragment was amplified by PCR to obtain a 617-bp fragment. A standard curve was then created using densitometry readings of the 617-bp bands on agarose gel. A fixed amount of sample cDNA from each cell line was amplified at the same time and the resultant PCR product was read by densitometer. Using the standard curve, ERCC-2 gene expression in a given amount of total RNA was quantitated and normalized to beta-actin expression. There was minimal variation in three repeated experiments with PCR amplification. ERCC-2 gene expression determined by this semiquantitative PCR was also correlated to ERCC-2 quantitation by Northern blot analysis, with a significant concordance (r = 0.912, P = 0.0002). We also successfully applied this sensitive method to quantify five clinical glioma samples.

Actins↗