Mechanical Anisotropy Assessment in Kidney Cortex Using ARFI Peak Displacement: Preclinical Validation and Pilot In Vivo Clinical Results in Kidney Allografts.

The kidney is an anisotropic organ, with higher elasticity along versus across nephrons. The degree of mechanical anisotropy in the kidney may be diagnostically relevant if properly exploited; however, if improperly controlled, anisotropy may confound stiffness measurements. The purpose of this study is to demonstrate the clinical feasibility of acoustic radiation force (ARF)-induced peak displacement (PD) measures for both exploiting and obviating mechanical anisotropy in the cortex of human kidney allografts, in vivo. Validation of the imaging methods is provided by preclinical studies in pig kidneys, in which ARF-induced PD values were significantly higher ( , Wilcoxon) when the transducer executing asymmetric ARF was oriented across versus along the nephrons. The ratio of these PD values obtained with the transducer oriented across versus along the nephrons strongly linearly correlated ( R2 = 0.95 ) to the ratio of shear moduli measured by shear wave elasticity imaging. On the contrary, when a symmetric ARF was implemented, no significant difference in PD was observed ( p > 0.01 ). Similar results were demonstrated in vivo in the kidney allografts of 14 patients. The symmetric ARF produced PD measures with no significant difference ( p > 0.01 ) between along versus across alignments, but the asymmetric ARF yielded PD ratios that remained constant over a six-month observation period post-transplantation, consistent with stable serum creatinine level and urine protein-to-creatinine ratio in the same patient population ( p > 0.01 ). The results of this pilot in vivo clinical study suggest the feasibility of 1) implementing symmetrical ARF to obviate mechanical anisotropy in the kidney cortex when anisotropy is a confounding factor and 2) implementing asymmetric ARF to exploit mechanical anisotropy when mechanical anisotropy is a potentially relevant biomarker.

Evaluating Renal Transplant Status Using Viscoelastic Response (VisR) Ultrasound.

Chronic kidney disease is most desirably and cost-effectively treated by renal transplantation, but graft survival is a major challenge. Although irreversible graft damage can be averted by timely treatment, intervention is delayed when early graft dysfunction goes undetected by standard clinical metrics. A more sensitive and specific parameter for delineating graft health could be the viscoelastic properties of the renal parenchyma, which are interrogated non-invasively by Viscoelastic Response (VisR) ultrasound, a new acoustic radiation force (ARF)-based imaging method. Assessing the performance of VisR imaging in delineating histologically confirmed renal transplant pathologies in vivo is the purpose of the study described here. VisR imaging was performed in patients with (n = 19) and without (n = 25) clinical indication for renal allograft biopsy. The median values of VisR outcome metrics (τ, relative elasticity [RE] and relative viscosity [RV]) were calculated in five regions of interest that were manually delineated in the parenchyma (outer, center and inner) and in the pelvis (outer and inner). The ratios of a given VisR metric for all possible region-of-interest combinations were calculated, and the corresponding ratios were statistically compared between biopsied patients subdivided by diagnostic categories versus non-biopsied, control allografts using the two-sample Wilcoxon test (p <0.05). Although τ ratios non-specifically differentiated allografts with vascular disease, tubular/interstitial scarring, chronic allograft nephropathy and glomerulonephritis from non-biopsied control allografts, RE distinguished only allografts with vascular disease and tubular/interstitial scarring, and RV distinguished only vascular disease. These results suggest that allografts with scarring and vascular disease can be identified using non-invasive VisR RE and RV metrics.

Evaluation of the Og34C filter paper technique in lymphatic filariasis prevalence studies.

Currently there are several tests which can be used for monitoring and evaluating lymphatic filariasis prevalence in mass drug administration (MDA) programs with the ICT as most commonly accepted field test. However, other techniques to determine circulating antigen are available that also could be suitable in these programs. Our aim was to determine the suitability of the Og4C3 filter paper technique to determine antigen prevalence for lymphatic filariasis. We compared multiple antigen methods during the course of baseline clinical prevalence studies in Papua, New Guinea in over 800 subjects. We found that the ICT and Og4C3 filter paper techniques and using blood from the ICT card in the Og4C3 ELISA were equivalent in performance and that the serum Og4C3 ELISA test detected significantly more antigenic individuals. In addition, we found that modification of the Og4C3 assay by removing the boiling step did not affect its performance. Our results indicate that the Og4C3 filter paper technique is suitable for use in elimination of transmission of lymphatic filariasis monitoring and evaluation programs, quality control of ICT testing could be accomplished by using blood from the ICT test card, and with appropriate logistics in place, venous sampling and testing by the serum Og4C3 ELISA is achievable in monitoring and evaluation and would better identify areas with low level antigenaemia prevalence and possible ongoing transmission.

Epidemiological assessment of continuing transmission of lymphatic filariasis in Samoa.

Ongoing transmission of lymphatic filariasis (LF) was assessed in five Samoan villages by measuring microfilaraemia (Mf), circulating filarial antigen (CFA) and antibody prevalence. Compared to the other villages, Fasitoo-Tai had a significantly higher Mf prevalence (3·2%), CFA prevalence (14·6%) and antibody prevalence in children (62·0%) (P<0·05). Puapua had a significantly lower CFA prevalence (2·5%), no detectable Mf-positive individuals and significantly low antibody prevalence in children (7·9%) (P<0·05). Siufaga, previously believed to be LF-free, recorded >1% CFA prevalence and a high antibody prevalence in children (46·6%). Overall, antibody prevalence in children appeared to reflect the transmission dynamics in the villages and, in Siufaga, identified an area of ongoing transmission. The Filariasis Cellabs Enzyme-Linked Immunosorbent Assay (CELISA), based on recombinant antigen Bm14, to detect antibodies, could potentially be a promising diagnostic tool for inclusion in future surveillance in the South Pacific.

ARFI ultrasound for in vivo hemostasis assessment postcardiac catheterization, part II: pilot clinical results.

In this second of a two part series, we present pilot clinical data demonstrating Acoustic Radiation Force Impulse (ARFI) ultrasound for monitoring the onset of subcutaneous hemostasis at femoral artery puncture sites (arteriotomies), in vivo. We conducted a randomized, reader-blinded investigation of 20 patient volunteers who underwent diagnostic percutaneous coronary catheterization. After sheath removal (6 French), patients were randomized to treatment with either standard of care manual compression alone or, to expedite hemostasis, manual compression augmented with a p-GlcNAc fiber-based hemostatic dressing (Marine Polymer Technologies, Danvers MA). Concurrent with manual compression, serial ARFI imaging began at the time of sheath removal and continued every minute for 15 min. Serial data sets were processed with custom software to (1) estimate the time of hemostasis onset, and (2) render hybrid ARFI/B-Mode images to highlight displacements considered to correspond to extravasted blood. Images were read by an observer blinded to the treatment groups. Average estimated times to hemostasis in patient volunteers treated with manual compression alone (n = 10) and manual compression augmented by hemostatic dressing (n = 9) were, respectively, 13.00 +/- 1.56 and 9.44 +/- 3.09 min, which are statistically significantly different (p = 0.0065, Wilcoxon two-sample test). Example images are shown for three selected patient volunteers. These pilot data suggest that ARFI ultrasound is relevant to monitoring subcutaneous bleeding from femoral arteriotomies clinically and that time to hemostasis was significantly reduced by use of the hemostatic dressing.

The argument for integrating vector control with multiple drug administration campaigns to ensure elimination of lymphatic filariasis.

BACKGROUND: There is a danger that mass drug administration campaigns may fail to maintain adequate treatment coverage to achieve lymphatic filariasis elimination. Hence, additional measures to suppress transmission might be needed to ensure the success of the Global Program for the Elimination of Lymphatic Filariasis. DISCUSSION: Vector control successfully eliminated lymphatic filariasis when implemented alone or with mass drug administration. Challenges to lymphatic filariasis elimination include uncertainty of the exact level and duration of microfilarial suppression required for elimination, the mobility of infected individuals, consistent non-participation of some infected individuals with mass drug administration, the possible development of anti-filarial drug resistance and treatment strategies in areas co-endemic with loasis. Integration of vector control with mass drug administration can address some of these challenges. The potential benefits of vector control would include: (1) the ability to suppress filariasis transmission without the need to identify all individual 'foci of infection'; (2) minimizing the risk of reestablishment of transmission from imported microfilaria positive individuals; and (3) decreasing the risk of dengue or malaria transmission where, respectively, Aedes or Anopheles are lymphatic filariasis vectors. SUMMARY: With adequate sustained treatment coverage, mass drug administration should meet the criteria for elimination of lymphatic filariasis. However, it may be difficult to sustain sufficiently high mass drug administration coverage to achieve lymphatic filariasis elimination in some areas, particularly, where Aedes species are the vectors. Since vector control was effective in controlling and even eliminating lymphatic filariasis transmission, integration of vector control with mass drug administration will ensure the sustainability of transmission suppression and thereby better ensure the success of national filariasis elimination programs. Although trials of some vector control interventions are needed, proven vector control strategies are ready for immediate integration with mass drug administration for many important vectors. Vector control is the only presently available additional lymphatic filariasis control measure with the potential for immediate implementation.

Use of Brugia Rapid dipstick and ICT test to map distribution of lymphatic filariasis in the Democratic Republic of Timor-Leste.

The newly-introduced Brugia Rapid dipstick for filarial antibodies and ICT filarial antigen card test were used to confirm historical data on the distribution of lymphatic filariasis in the Republic of Timor-Leste. Twelve out of thirteen districts were confirmed as being endemic. Brugian filariasis predominates, with an average prevalence of 11.6%. The average prevalence of Bancroftian filariasis was 1.1%. The study demonstrated that the Brugia Rapid test can provide useful information about the distribution of Brugian filariasis in circumstances where it is difficult or impossible to obtain night blood samples for microfilariae.

Increase in cellular immune responses in Onchocerca-infected cattle after treatment with the microfilaricide, milbemycin.

Lymphocytes from Onchocerca-infected steers treated with the microfilaricide, milbemycin showed increased proliferation when challenged with antigen from Dirofilaria immitis, concanavalin A, tuberculin and tetanus toxoid, compared with untreated animals. This paper confirms that Onchocerca infection induces immunosuppression to filarial and non-filarial antigens. It raises the possibility that filarial-induced immunosuppression may increase the susceptibility to mycobacterial infections and reduce the efficacy of vaccinations and strongly indicates that further research is required.

Adaptive immunity in melioidosis: a possible role for T cells in determining outcome of infection with Burkholderia pseudomallei.

Melioidosis is a potentially fatal disease caused by the bacterium Burkholderia pseudomallei. Individuals with subclinical melioidosis have no apparent clinical signs or symptoms, and are identified only by positive serology. The present study is the first to investigate cell-mediated immune (CMI) responses following in vitro stimulation with B. pseudomallei antigens in peripheral blood mononuclear cells (PBMC), collected under field conditions in Papua New Guinea (PNG) from individuals with exposure to B. pseudomallei (n = 13). While five had a clinical history of melioidosis (C(+)), the remaining individuals (n = 8) were seropositive, yet healthy with no clinical history of melioidosis (S(+)/C(-)). Proliferation and IFN-gamma production were significantly greater in lymphocyte cultures from S(+)/C(-) individuals compared to C(+) individuals (P < 0.001 and P < 0.05, respectively). These findings demonstrate that compared to C(+) patients, individuals with subclinical melioidosis have a stronger CMI response to B. pseudomallei antigens in vitro. Such a response may be essential for protection against disease progression.

Community empowerment in the control of lymphatic filariasis in Misima, Milne Bay Province using diethylcarbamazine in combination with albendazole.

We report the successful implementation of a community-based lymphatic filariasis control program using annual single-dose treatment with diethylcarbamazine (DEC) in combination with albendazole. The target population included over 28,000 people in the Samarai Murua District, Milne Bay Province, Papua New Guinea. A community-based delivery model was as effective as the standard health services delivery model. The number of people tested in 1998 before mass drug administration (MDA) and in 1999, one year after treatment, were 1644 and 942 respectively; the number who received mass treatment was 29,883 in 1998 and 28,965 in 1999. The prevalence of antigenaemia decreased significantly from 19% to 12%. The cost of running the program also decreased by 50%. The total number of trained health staff required to conduct the MDA program declined from 62 in 1998 to 12 in 1999, a reduction of 81%, with a cost saving in salary and allowances. A salient organizational initiative that surfaced was the use of local expertise in the private sector as a catalyst for obtaining funds from external sources to manage and facilitate the program which was conducted with locally available resources.

Prevalence of filarial antigenaemia in Papua Nnew Guinea: results of surveys by the School of Public Health and Tropical Medicine, James Cook University, Townsville, Australia.

During the period from 1991 to 1997 the School of Public Health and Tropical Medicine, James Cook University carried out filariasis surveys in several parts of Papua New Guinea using the newly introduced Onchocerca gibsoni monoclonal (Og4C3) and immunochromatographic test (ICT) antibody-based assays for filarial antigen and, in some cases, a Knott's test for microfilariae. The average prevalence of filarial antigenaemia and microfilaraemia was 56% and 35% respectively confirming earlier survey results that filariasis is hyperendemic in many parts of the country. The antigen tests detected 25% more cases than the Knott's test and the simplicity of the ICT and its capacity to produce almost instant results make it an ideal tool for surveys.

Antibodies to Japanese encephalitis virus in human sera collected from Irian Jaya. Follow-up of a previously reported case of Japanese encephalitis in that region.

Japanese encephalitis virus (JEV) and other arboviruses are demonstrating an emergence in the southern part of New Guinea Island. JE was previously unknown in this part of the world until 1995 when it was found in the Torres Strait, northern Australia. In this study 96 sera collected from residents of the Timika region of Irian Jaya were tested for antibodies to JEV and related arboviruses by epitope-specific blocking ELISA. Of the 9 sera deemed to be positive for JEV antibodies by ELISA, 5 were collected from persons indigenous to Timika, and who had not travelled to regions where JE is known to be active. This indicates that these individuals were infected with JEV in the Timika area and supports a recent report of a clinical case of JE in this region. Non-immune expatriates visiting or working in the Lowland areas of Irian Jaya and/or Papua New Guinea should consider immunization against JE. Precautions should always be taken to avoid being bitten by any mosquito both in the daytime and at night.

Characterization of 13 novel band 3 gene defects in hereditary spherocytosis with band 3 deficiency.

Hereditary spherocytosis (HS) is a common hemolytic anemia of variable clinical expression. Pathogenesis of HS has been associated with defects of several red cell membrane proteins including erythroid band 3. We have studied erythrocyte membrane proteins in 166 families with autosomal dominant HS. We have detected relative deficiency of band 3 in 38 kindred (23%). Band 3 deficiency was invariably associated with mild autosomal dominant spherocytosis and with the presence of pincered red cells in the peripheral blood smears of unsplenectomized patients. We hypothesized that this phenotype is caused by band 3 gene defects. Therefore, we screened band 3 DNA from these 38 kindred for single strand conformational polymorphisms (SSCP). In addition to five mutations detected previously by SSCP screening of cDNA, we detected 13 new band 3 gene mutations in 14 kindred coinherited with HS. These novel mutations consisted of two distinct subsets. The first subset included seven nonsense and frameshift mutations that were all associated with the absence of the mutant mRNA allele from reticulocyte RNA, implicating decreased production and/or stability of mutant mRNA as the cause of decreased band 3 synthesis. The second group included five substitutions of highly conserved amino acids and one in-frame deletion. These six mutations were associated with the presence of comparable levels of normal and mutant band 3 mRNA. We suggest that these mutations interfere with band 3 biosynthesis leading thus to the decreased accumulation of the mutant band 3 allele in the plasma membrane.

Opsonic human antibodies from an endemic population specific for a conserved epitope on the M protein of group A streptococci.

This study demonstrates the presence of epitope-specific opsonic human antibodies in a population living in an area endemic for group A streptococci (GAS) infection. Antibodies recognizing a conserved C-terminal region epitope (p145, sequence in single letter amino acids: LRRDLDASREAKKQVEKALE) of the M protein of GAS were isolated from human patients by affinity chromatography and were shown to be of the immunoglobulin G1 (IgG1) and IgG3 subclasses. These antibodies could reduce the number of colonies of serotype 5 GAS in an in vitro opsonization assay by 71-92%, compared with an equal amount of IgG from control adult donors living in non-endemic areas and without antibodies to p145. Addition of the peptide, p145, completely inhibited this opsonization. Indirect immunofluorescence showed that p145-specific antibodies were capable of binding to the surface of M5 GAS whereas control IgG did not. Using chimeric peptides, which contain overlapping segments of p145, each 12 amino acids in length, inserted into a known helical peptide derived from the DNA binding protein of yeast, GCN4, we have been able to further define two minimal regions within p145, referred to as pJ2 and pJ7. These peptides, pJ2 and pJ7, were able to inhibit opsonization by p145 specific antibodies. Finally, we have observed an association between the age-related development of immunity to GAS and the acquisition of antibodies to the conserved epitope, p145, raising the possibility of using this epitope as a target in a prophylactic vaccine administered during early childhood.

Maternal selenium levels and sudden infant death syndrome (SIDS).

BACKGROUND: The possibility is tested that low anti-oxidant status and/or low levels of selenium (Se) might predispose to Sudden Infant Death Syndrome (SIDS). AIM: This study was undertaken to collect evidence on the Se status of pregnant and non-pregnant women and newborn babies and to establish whether babies who later died of cot death had significantly divergent levels of blood Se at birth. METHODOLOGY: Aliquots of blood were collected from all newly pregnant mothers in Tasmania and from the cords of all newborn babies. These were analysed for Se and glutathione peroxidase (GPx) content and compared by season and with non-pregnant, age standardised blood donors in three areas of Tasmania and three mainland Australian States. RESULTS: Cot death babies' cordbloods were not significantly different in Se or GPx-status, in this small sample, from those of other babies, nor was a seasonal variation in these parameters demonstrated among 390 randomly selected Tasmanian mothers. Mothers-to-be showed a decrease in enzyme levels during pregnancy and Tasmanian blood donors had significantly lower levels than donors from other States. CONCLUSION: While no evidence can safely be drawn about a relationship between Se or GPx-status and SIDS, this study provides base level measures for populations showing that Tasmanian residents have low levels of these anti-oxidants.

Haematology, red cell metabolism and blood chemistry of the black-faced cormorant Leucocarbo fuscescens.

1. Haematology, red cell metabolism and blood chemistry of five fledgeling black-faced cormorants Leucocarbo fuscescens were studied and the results were compared with previously reported data on several other sea-birds. 2. The mean erythrocyte count of the cormorant is similar to that of penguins but lower than that of flying, non-diving sea-birds. The cormorant's red cell mean cell volume (MCV) is lower than that of penguins but higher than that of non-diving sea-birds. 3. Leucocyte numbers are within expected limits for avian species. 4. Red cell enzymes: glucose phosphate isomerase, phosphofructokinase, aldolase and enolase are higher in the cormorant than in the little penguin; glyceraldehyde phosphate dehydrogenase, monophosphoglyceromutase, pyruvate kinase, lactate dehydrogenase and glutathione reductase are lower. 5. Haemoglobin electrophoresis showed a typical avian haemoglobin pattern.

Haematology of the Australian eastern quoll, Dasyurus viverrinus--II. Red cell enzymes and metabolic intermediates.

1. The activity of 21 red cell enzymes and three red cell metabolic intermediates were measured in adult Dasyurus viverrinus and compared with published data on other marsupials. 2. Phosphofructokinase (PFK), glyceraldehyde dehydrogenase (GAPD) and phosphoglycerate kinase (PGK) were elevated in comparison to other marsupials. 3. Enolase (ENO) and 2,3-diphosphoglycerate (2,3 DPG) were lower than in other marsupials.