Haematologic complications from human babesiosis: a case report.

Formerly known as Nantucket fever, babesiosis is increasing in incidence across the Northeastern United States. Because of its emerging health risk globally, it is important to be aware of its various presenting manifestations. We present the case of a middle-aged man with haemolytic anaemia from Babesia microti infection.

Neurological impairment in a surviving twin following intrauterine fetal demise of the co-twin: a case study.

It has been established that twin pregnancies are at an increased risk for complications, including the risk of morbidity or mortality for one or both of the infants. Cerebral palsy and other associated neurological deficits also occur at higher rates in twin pregnancies. This report examines two cases of intrauterine demise of one twin with subsequent survival of the co-twin. In both cases, the surviving infant suffered significant neurological sequelae. Impairments observed in these two cases include multicystic encephalomalacia and periventricular leukomalacia as well as the subsequent development of cerebral palsy. This case study explores the predisposing factors, incidence, pathophysiology, consequences, and future research implications of these findings.

Neurovirulence of Cryptococcus neoformans determined by time course of capsule accumulation and total volume of capsule in the brain.

Cryptococcal meningitis remains a significant opportunistic infection in HIV-infected individuals worldwide, despite availability of antiretroviral therapies in developed nations. Current therapy with amphotericin B is difficult to administer and only partially effective. Mechanisms of cryptococcal neuropathogenesis are still not clearly defined. In the present study, we used a C57Bl/6 mouse model with intravenous inoculation of three isogenic strains of Cryptococcus neoformans: H99, Cap59, and Pkr1-33. These strains differ in their capsule production and are normocapsular, hypocapsular, and hypercapsular, respectively. We studied the role of capsule in the morbidity and mortality of our host animal. Surprisingly, we found that the hypercapsular strain was least virulent while the strains that produced less capsule were more virulent and had higher concentrations of organism in the brain. These results suggest that neurovirulence is related to total capsule volume and rate of capsule accumulation in the brain, rather than the amount of capsule produced per organism. Therapies which decrease central nervous system dissemination and inhibit replication rates in the brain may be more effective than therapies which target capsule production.

A transmissive laser speckle imaging technique for measuring deep tissue blood flow: an example application in finger joints.

BACKGROUND AND OBJECTIVE: Laser speckle perfusion imaging (LSPI) is a minimally invasive optical measure of relative changes in blood flow, providing real-time, high resolution, two-dimensional maps of vascular structure. Standard LSI imaging uses a light-reflective geometry that limits the measurement to a thin surface layer of 0.2-1 mm. The objective of this study was to test a new LSI instrument geometry with the laser source opposed to the image capture plane (light transmissive). Captured light then travels the entire tissue thickness (10-15 mm), sampling much deeper regions of interest than conventional optical imaging techniques. STUDY DESIGN: Reflective-light (conventional) and transmissive-light LSI modes were used to measure finger joint blood flow during a timed tourniquet occlusion of the brachial artery in volunteer participants. RESULTS: There was greatly increased visibility of vessels underlying the skin in the light-transmissive mode LSI mode. Established LSI algorithms were shown to still work in the light-transmissive mode, despite decorrelation due to finite laser coherence length and the light passing through a tissue thickness of 10-15 mm. CONCLUSION: Transmissive LSI can be used to measure blood flow deep (10-15 mm) into tissues. This could be useful for non-invasive measurements of finger joint synovial blood flow in diagnosing and treating peripheral vascular disorders, such as rheumatoid arthritis.

Kinetics of blood flow during healing of excisional full-thickness skin wounds in pigs as monitored by laser speckle perfusion imaging.

BACKGROUND/PURPOSE: The laser speckle perfusion imaging (LSPI) system is a new, non-invasive technique for rapidly and reproducibly measuring tissue perfusion. The high resolution and frame rate of the LSPI overcome many of the limitations of traditional laser Doppler imaging techniques. Therefore, LSPI is a useful means for evaluating blood flow in a variety of situations. The present study investigates the ability of the LSPI system to detect temporal changes in blood flow during the healing of cutaneous wounds in a well-characterized animal model. METHODS: Full-thickness excisional skin wounds (2 x 2 cm) were created on the backs of juvenile female red Duroc pigs. Every week post-injury, the wounds were measured and photographed, and normalized blood flow values were determined using the LSPI system. RESULTS: Tissue perfusion values were available after complete re-epithelialization and removal of the eschar, at day 21. At this point, wound blood flow was significantly elevated as compared with the surrounding, uninvolved skin. Wound blood flow declined steadily during healing, and approached normal values by day 35 post-injury. CONCLUSION: The kinetics of blood flow during excisional wound healing in the red Duroc model are comparable with that previously observed in laser Doppler imaging of healing human skin wounds and hypertrophic scars. These results therefore confirm that the red Duroc is a good model of human wound healing, and further indicates that the LSPI is an excellent technique for evaluating angiogenesis and neovascularization during healing in this and other models.

Endoscopic laser speckle imaging of tissue blood flow: applications in the human knee.

This work represents the first clinical data acquired with the endoscopic laser speckle imaging (eLSPI) system, a new diagnostic tool developed for real-time imaging of tissue blood flow during endoscopic surgical procedures. eLSPI was used to image tissue perfusion in the medial compartment of the knee of five patients requiring arthroscopic knee surgery. The effectiveness of eLSPI as a diagnostic tool was tested by measuring changes in tissue perfusion resultant from tourniquet application, and intra-articular epinephrine. eLSPI produced real-time perfusion video images of tissue blood flow in the knee joint. Tourniquet applications produced consistent decreases in mean perfusion index measurements (29.3% +/- 5.1% in meniscus; 39.5% +/- 8.2% in synovium with an intra patient variability of 6%-9%). A dose-dependent vasoconstrictive response to the administration of intra-articular epinephrine was measured, with maximum dose producing a mean decrease in perfusion of 31.0%-9.3% in meniscus and 41.2%-10.9% in synovium. eLSPI consistently detects decreases in articular tissue blood flow resultant from tourniquet inflation or from the administration of increasing concentrations of epinephrine. These are the first in vivo results indicating physiologic changes in articular tissue as a function of two commonly applied practices in endoscopic joint surgery.

A comparison of two laser-based methods for determination of burn scar perfusion: laser Doppler versus laser speckle imaging.

UNLABELLED: Laser Doppler perfusion imaging (LDI) is an established technique for early assessment of burn depth to help determine a course of treatment. Laser speckle perfusion imaging (LSPI) is an alternative laser based, non-invasive perfusion monitoring technique that offers rapid and high resolution images of tissue. We have evaluated the ability of the LSPI instrument in determining and monitoring burn scar perfusion over time and compared it with the LDI instrument as a standard. METHODS: Ten patients with hypertrophic burn scars (time since injury: 1-8 months) were recruited. Burn scars were scanned with both instruments (LSPI and LDI) monthly over a period of 11 months. Clinical grading of the burn scars was assessed on every scan date using the Vancouver burn scar scale. RESULTS: Comparison of the perfusion values determined by each instrument shows a strong positive correlation, r2=0.86 (n=63). Each instrument's output also correlated significantly with the clinical grading of the scar, indicating the expected decrease in perfusion as the clinical condition of the scars improved with time. SIGNIFICANCE: The new LSPI instrument compared favorably with the established LDI instrument, yielding similar results. The considerably faster scan time and higher resolution of the LSPI method provides a distinct clinical advantage, both in terms of patient comfort and for reliably matching perfusion characteristics to their associated anatomical features. The fast temporal response of the LSPI instrument could be used to monitor near real-time responses to mechanical or pharmacological interventions to study dynamic vascular changes to burn damaged tissues.

Endoscopic laser imaging of tissue perfusion: new instrumentation and technique.

BACKGROUND AND OBJECTIVES: New instrumentation, based on a previously established laser speckle perfusion imaging (LSI) technique is evaluated for its ability to capture and generate blood flow images during endoscopic surgery. STUDY DESIGN/MATERIALS AND METHODS: Investigations are detailed in an in-vitro blood flow model simulating physiological properties of vascularized tissue, and in-vivo in rabbit joint capsule tissue. RESULTS: In-vitro measurements showed a linear response of the instrument to blood flow in the range of 0-800 microl/minute, where data points were significantly correlated with an r(2) value of 0.96. In-vivo measurements showed a 58.7% decrease to the medial collateral ligament during occlusion of the femoral artery. CONCLUSIONS: Blood flow images demonstrate that the endoscopic LSI technique is capable of measuring relative tissue blood flow changes at high resolutions and rapid response times and incorporates well with endoscopic surgeries.

Comparison of laser speckle and laser Doppler perfusion imaging: measurement in human skin and rabbit articular tissue.

Laser Doppler perfusion imaging (LDI) is currently used in a variety of clinical applications, however, LDI instruments produce images of low resolution and have long scan times. A new optical perfusion imager using a laser speckle measurement technique and its use for in vivo blood flow measurements are described. Measurements of human skin and surgically exposed rabbit tissue made using this instrument were compared with a commercial laser Doppler perfusion imaging instrument. Results from blood flow measurements showed that the laser speckle imager measured an 11-67% decrease in blood flow under arterial occlusion. Under similar conditions, the laser Doppler imager measured blood flow decreases of 21-63%. In comparison with LDI, it was observed that the higher temporal resolution of the laser speckle imager was more sensitive to measuring the hyperaemic response immediately following occlusion. This in vivo study demonstrated some of the several advantages laser speckle imaging has over conventional LDI, making the new instrument more versatile in a clinical environment.

Evaluation of laser-Doppler perfusion imaging for measurement of blood flow in cortical bone.

Most techniques currently available to measure blood flow in bone are time consuming and require destruction of the tissue, but laser-Doppler technology offers a less invasive method. This study assessed the utility of laser-Doppler perfusion imaging (LDI) to measure perfusion in cortical bone. Twelve mature New Zealand White rabbits were assigned to one of three groups: normal control, constriction (norepinephrine), or dilatation (nitroprusside). The left and right medial tibiae were consecutively scanned at red (634-nm) and near-infrared (810-nm) wavelengths to examine the repeatability of LDI output. The pharmacological intervention groups were injected with the respective drug, and LDI measurements at 810 nm were obtained concurrently with colored microsphere-determined flow in all of the groups. LDI effectively quantified blood flow in cortical bone and detected physiologically induced changes in perfusion. A significant positive correlation was found between microsphere-determined flow and LDI output (r = 0.6, P < 0.05). Repeatability of consecutive LDI measurements was within 5%. The effectiveness of LDI to measure perfusion in bone suggests this method has potential for investigating the role of blood flow in bone metabolism and remodeling.

6-Acylamino-2-[(alkylsulfonyl)oxy]-1H-isoindole-1,3-dione mechanism-based inhibitors of human leukocyte elastase.

A study of various 2-[(alkylsulfonyl)oxy]-6-substituted-1H-isoindole-1,3-diones' inhibition of chymotrypsin compared to inhibition of HLE reveals that acylamino substitution in the 6-position increases selectivity and potency of these inhibitors for HLE. The best HLE inhibitor in this series was 6-(methylglutaryl)amino-2-[(ethylsulfonyl)oxy]-1H-isoindole-1,3-di one with a kobs/[I] = 220,000 M(-1) s(-1).

Dynamic measurement of bone blood perfusion with modified laser Doppler imaging.

Although the mechanisms are not clearly defined, blood flow may play an important role in moderating skeletal adaptation. Most techniques currently available to measure blood flow in bone are time-consuming and require destruction of the tissue, but laser Doppler technology offers a less invasive method. The present study assessed whether laser Doppler perfusion imaging could detect changes in perfusion in cortical bone. By use of modified laser Doppler perfusion imaging with an adjustable, incorporated, near infrared-laser gain photodetection system, perfusion of blood in the mid-diaphyseal tibial cortex of New Zealand White rabbits (n = 5) was measured before, during, and after occlusion of the femoral artery. During occlusion, perfusion decreased 69% compared with control levels; removal of the arterial clip caused flux values to return to near normal. Laser Doppler perfusion imaging provides a two-dimensional image related to blood flow, and the results of this pilot study suggest that it may be an effective technique for imaging in vivo dynamic changes in perfusion in cortical bone.

The expanded role of acute care nurses: the issue of liability.

For a variety of reasons the activities performed by health care professionals are ever increasing, however the legislative process regulating these specialised workers is not keeping up with the practice realities. While competency statements and credentialling mechanisms are developing, they are not uniformly in place for specialists. Therefore activities completed by these practitioners may be legally controversial. This study documents a variety of tasks completed independently by nurses employed in three acute care hospitals that legally require medical orders and then examines the legal implications of this role extension. Credentialling is one mechanism by which nurses and other health care professionals can ensure they possess the levels of knowledge and skill required to perform the advanced activities required of them and consequently avoid negative legal repercussions.

Differential effects of p53 mutants on the growth of human bronchial epithelial cells.

We investigated the effects of five different p53 mutants on the growth of primary cultures of normal human bronchial epithelial (NHBE) cells. The five defective viral pZIP-Neo constructs contained the following mutations at mutational hot-spots found in human cancers: codons 143ala, 175his, 248trp, 249ser, and 273his. NHBE cells were infected with the p53 mutants, wild-type p53, or the pZIP-Neo vector control. The 143ala, 248trp, and 273his mutants, as well as wild-type p53, decreased the colony-forming efficiency and inhibited the growth of NHBE cells. The 175his mutant did not significantly change the growth rates. In NHBE cells from three donors, the 249ser mutant conferred a substantial growth advantage to the NHBE cells in a colony-forming-efficiency assay. In NHBE cells isolated from one donor, the 249ser mutant also produced a significant life span extension. These cells grew rapidly through 80 population doublings and entered an apparent "crisis" in passage 14. Karyotypic analyses of one culture at multiple passages revealed aneuploid populations with alterations of chromosomes 5, 11, and 13; quantitative DNA analysis detected aneuploidy in late passages from that culture and two other primary cultures. These data demonstrated that the codon 249ser mutation could provide a growth advantage to bronchial epithelial cells and suggest that this mutant protein can induce genomic instability.

Factors influencing the appropriateness of restorative dental treatment: an epidemiologic perspective.

OBJECTIVES: An epidemiology analysis was performed to identify patient and dentist factors influencing over- and undertreatment of restorative services in a sample of insured adults. METHODS: At baseline, 681 Washington State employees and their spouses, aged 20 to 34 years and residing in the Olympia or Pullman areas, were interviewed by telephone. Oral assessments were conducted to measure personal characteristics, oral disease, and restoration quality. Adults were followed for two years to measure use of restorative services from dental insurance claims. Each adult's baseline and claims data were linked with provider and practice variables collected from the dentist who provided treatment. RESULTS: For overtreatment, 39 percent of adults received one or more replacement restorations in nondecayed teeth with satisfactory fillings at baseline, while 18 percent of adults had one or more restorations placed in teeth with no decay and fillings. An adult's probability of overtreatment was higher if the adult had more fillings at baseline, or if an adult's dentist was younger, had a busy practice, advertised, charged higher fees, had less continuing education, or had a solo practice. For undertreatment, about 16 percent of adults either received no replacement restorations in teeth with unsatisfactory fillings at baseline, or had decayed teeth at baseline that were not filled or crowned. An adult's probability of undertreatment was higher if an adult had less decayed or more missing surfaces at baseline, or if an adult's dentist believed in sharing information with patients, had a busy practice, or reported not placing fillings when radiographic evidence of new caries was present. CONCLUSIONS: A minority of adults aged 20 to 34 experienced potential over- or undertreatment of restorative services, which are influenced by both patient and dentist factors.

In vivo comparison of scanning technique and wavelength in laser Doppler perfusion imaging: measurement in knee ligaments of adult rabbits.

At present, there are only two laser Doppler perfusion imaging systems (LDIs) manufactured for medical applications: a 'stepwise' and a 'continuous' scanning LDI. The stepwise scanning LDI has previously been investigated and compared with coloured microsphere determined standardised flow. The continuous scanning LDI is investigated and compared with the stepwise scanning LDI for its ability to measure in vivo, hypoaemic, ligament tissue blood flow changes. The continuous scanning system was supplied with two lasers, red and near infrared (NIR), allowing for additional assessment of the effect of wavelength on imaging ligament perfusion. Perfusion images were obtained from surgically exposed rabbit medial collateral ligaments (MCL). Continuous and stepwise LDI scans were compared using correlation and linear regression analysis of image. averages and standard deviations. Using the same method of analysis, LDI measurements using red and NIR lasers indicated a high degree of correlation, at least over the ranges of perfusion assessed, indicating that red and NIR lasers measure similar regions of flow in the rabbit MCL. These experiments confirm that both LDI techniques provide a valid in vivo measure of dynamic changes in connective tissue perfusion and could have significant impact on the understanding and treatment of joint injury and arthritis.

Molecular analysis of the cyclin-dependent kinase inhibitor genes p15INK4b/MTS2, p16INK4/MTS1, p18 and p19 in human cancer cell lines.

Cyclin-dependent kinase-4 inhibitor genes (INK4) regulate the cell cycle and are candidate tumor-suppressor genes. To determine if alterations in the coding regions of the p18 and p19 genes, which are novel members of the INK4 family and if they correlate with the development of human cancer, 100 human cancer cell lines were analyzed. Two other INK4 gene family members, p15INK4b/MTS2 and p16INK4/MTS1 genes were also analyzed. Homozygous deletions of the p15INK4b/MTS2 gene were detected in 29 cancer cell lines. Thirty-five homozygous deletions and 7 intragenic mutations of the pl6INK4/MTS1 gene were also detected in these cell lines. Neither homozygous deletions nor intragenic mutations of the p18 and p19 genes were found except in an ovarian cancer cell line, SKOV3, harboring a single base pair deletion in exon 1 of p19. In p16INK4/MTS1 expression analysis, 5 cell lines with both authentic and alternative spliced p16INK4/MTS1 mRNA had no detectable p16INK4/MTS1 protein. These results suggest the hypotheses that either post-translational modification or enhanced degradation may be responsible for the lack of detection of the p16INK4/MTS1 protein. Using Western blot analysis, subsets of 26 human cancer cell lines were examined for p18 expression and 39 cell lines for p19 expression. All of these cell lines expressed the p18 or p19 protein, with the exception of SKOV3, which did not express p19. Therefore, the INK4 gene family may be divided into 2 groups. One group includes p15INK4b/MTS2 and p16INK4/MTS1, in which genetic and epigenetic alterations might contribute to the development of human cancers. The other group includes p18 and p19, in which somatic mutations are uncommon in many types of human cancer, and their role in human carcinogenesis and cancer progression is uncertain.

Tissue-specific growth suppression and chemosensitivity promotion in human hepatocellular carcinoma cells by retroviral-mediated transfer of the wild-type p53 gene.

Selective expression of cytotoxic gene products in tumor cells is one of the goals of gene therapy for treating cancer. We are developing such a strategy for the treatment of human hepatocellular carcinoma (HCC) by linking the wild-type p53 (WT-p53) gene with HCC-associated transcriptional control elements (TCE) to achieve selective growth inhibition of retrovirally transduced HCC cells. Replication-defective, amphotrophic retroviruses were constructed containing a WT-p53 complementary DNA (cDNA) that is transcriptionally regulated by the HCC-associated alpha-fetoprotein (AFP) gene TCE. Expression of exogenous WT-p53 from this retroviral vector was limited to AFP-producing cells. Introduction of WT-p53 into AFP-positive HCC cells by retroviral infection markedly inhibited their clonal growth in monolayer and soft agar cultures, and increased the sensitivity of these cells to the chemotherapeutic drug, cisplatin. Therefore, restoration of WT-p53 expression in HCC cells, in combination with chemotherapeutic drugs, can be considered as a strategy for the therapy of human liver cancer.

Education in emergency nursing: a new beginning. Part 2.

Continuing from the previous issue, this paper discusses the process of developing the content of new courses under the umbrella of the Graduate Diploma/Masters in Emergency Care Nursing offered at Griffith University Gold Coast, Australia. Part 2 of this paper includes information on course structure, implementation, content, assessing methods and competences and the logistics of the course.