Human placental villi contain stromal macrovesicles associated with networks of stellate cells.

Placental function is essential for fetal development and establishing the foundations for lifelong health. The placental villous stroma is a connective tissue layer that supports the fetal capillaries and villous trophoblast. All the nutrients that cross the placenta must also cross the stroma, and yet little is known about this region. This study uses high-resolution three-dimensional imaging to explore the structural complexity of this region within the placental villi. Serial block-face scanning electron microscopy and confocal microscopy were used to image the placental villous stroma in three-dimensions. Transmission electron microscopy (TEM) was used to generate high resolution two-dimensional images. Stereological approaches were used to quantify volumes of stromal constituents. Three-dimensional imaging identified stromal extracellular vesicles, which constituted 3.9% of the villous stromal volume. These stromal extracellular vesicles were ovoid in shape, had a median length of 2750 nm (range 350-7730 nm) and TEM imaging confirmed that they were bounded by a lipid bilayer. Fifty-nine per cent of extracellular vesicles were in contact with a fibroblast-like stellate cell and these vesicles were significantly larger than those where no contact was observed. These stellate cells formed local networks with adherent junctions observed at contact points. This study demonstrates that the villous stroma contains extracellular macrovesicles which are considerably larger than any previously described in tissue or plasma. The size and abundance of these macrovesicles in the villous stroma highlight the diversity of extracellular vesicle biology and their roles within connective tissues.

Development, commissioning, and evaluation of a new intensity modulated minibeam proton therapy system.

PURPOSE: To invent, design, construct, and commission an intensity modulated minibeam proton therapy system (IMMPT) without the need for physical collimation and to compare its resulting conformity to a conventional IMPT system. METHODS: A proton therapy system (Hitachi, Ltd, Hitachi City, Japan; Model: Probeat-V) was specially modified to produce scanned minibeams without collimation. We performed integral depth dose acquisitions and calibrations using a large diameter parallel-plate ionization chamber in a scanning water phantom (PTW, Freiburg, Germany; Models: Bragg Peak ionization chamber, MP3-P). Spot size and shape was measured using radiochromic film (Ashland Advanced Materials, Bridgewater NJ; Type: EBT3), and a synthetic diamond diode type scanned point by point in air (PTW Models: MicroDiamond, MP3-P). The measured data were used as inputs to generate a Monte Carlo-based model for a commercial radiotherapy planning system (TPS) (Varian Medical Systems, Inc., Palo Alto, CA; Model: Eclipse v13.7.15). The regular ProBeat-V system (sigma ~2.5 mm) TPS model was available for comparison. A simulated base of skull case with small and medium targets proximal to brainstem was planned for both systems and compared. RESULTS: The spot sigma is determined to be 1.4 mm at 221 MeV at the isocenter and below 1 mm at proximal distances. Integral depth doses were indistinguishable from the standard spot commissioning data. The TPS fit the spot profiles closely, giving a residual error maximum of 2.5% in the spot penumbra tails (below 5% of maximum) from the commissioned energies 69.4 to 221.3 MeV. The resulting IMMPT plans were more conformal than the IMPT plans due to a sharper dose gradient (90-10%) 1.5 mm smaller for the small target, and 1.3 mm for the large target, at a representative central axial water equivalent depth of 7 cm. CONCLUSIONS: We developed, implemented, and tested a new IMMPT system. The initial results look promising in cases where treatments can benefit from additional dose sparing to neighboring sensitive structures.

A pilot hole does not reduce the strains or risk of fracture to the lateral cortex during and following a medial opening wedge high tibial osteotomy in cadaveric specimens.

AIM: It has been suggested that the use of a pilot-hole may reduce the risk of fracture to the lateral cortex. Therefore the purpose of this study was to determine the effect of a pilot hole on the strains and occurrence of fractures at the lateral cortex during the opening of a high tibial osteotomy (HTO) and post-surgery loading. MATERIALS AND METHODS: A total of 14 cadaveric tibias were randomized to either a pilot hole (n = 7) or a no-hole (n = 7) condition. Lateral cortex strains were measured while the osteotomy was opened 9 mm and secured in place with a locking plate. The tibias were then subjected to an initial 800 N load that increased by 200 N every 5000 cycles, until failure or a maximum load of 2500 N. RESULTS: There was no significant difference in the strains on the lateral cortex during HTO opening between the pilot hole and no-hole conditions. Similarly, the lateral cortex and fixation plate strains were not significantly different during cyclic loading between the two conditions. Using a pilot hole did not significantly decrease the strains experienced at the lateral cortex, nor did it reduce the risk of fracture. CONCLUSIONS: The nonsignificant differences found here most likely occurred because the pilot hole merely translated the stress concentration laterally to a parallel point on the surface of the hole.Cite this article: K. Bujnowski, A. Getgood, K. Leitch, J. Farr, C. Dunning, T. A. Burkhart. A pilot hole does not reduce the strains or risk of fracture to the lateral cortex during and following a medial opening wedge high tibial osteotomy in cadaveric specimens. Bone Joint Res 2018;7:166-172. DOI: 10.1302/2046-3758.72.BJR-2017-0337.R1.

Avascular necrosis in children with cerebral palsy after reconstructive hip surgery.

PURPOSE: Progressive hip displacement is one of the most common orthopaedic pathologies in children with cerebral palsy (CP). Reconstructive hip surgery has become the standard treatment of care. Reported avascular necrosis (AVN) rates for hip reconstructive surgery in these patients vary widely in the literature. The purpose of this study is to identify the frequency and associated risk factors of AVN for reconstructive hip procedures. METHODS: A retrospective analysis was performed of 70 cases of reconstructive hip surgery in 47 children with CP, between 2009 and 2013. All 70 cases involved varus derotation osteotomy (VDRO), with 60% having combined VDRO and pelvic osteotomies (PO), and 21% requiring open reductions. Mean age at time of surgery was 8.82 years and 90% of patients were Gross Motor Function Classification System (GMFCS) 4 and 5. Radiographic dysplasia parameters were analysed at selected intervals, to a minimum of one year post-operatively. Severity of AVN was classified by Kruczynski's method. Bivar- iate statistical analysis was conducted using Chi-square test and Student's t-test. RESULTS: There were 19 (27%) noted cases of AVN, all radio- graphically identifiable within the first post-operative year. The majority of AVN cases (63%) were mild to moderate in severity. Pre-operative migration percentage (MP) (p = 0.0009) and post-operative change in MP (p = 0.002) were the most significant predictors of AVN. Other risk factors were: GMFCS level (p = 0.031), post-operative change in NSA (p = 0.02) and concomitant adductor tenotomy (0.028). CONCLUSION: AVN was observed in 27% of patients. Severity of displacement correlates directly with AVN risk and we suggest that hip reconstruction, specifically VDRO, be performed early in the 'hip at risk' group to avoid this complication.

Muscle density predicts changes in bone density and strength: a prospective study in girls.

OBJECTIVE: We sought to determine whether muscle density, an index of skeletal muscle fat content, was predictive of 2-year changes in weight-bearing bone parameters in young girls. METHODS: Two-year prospective data from 248 girls, aged 8-13 years at baseline. Peripheral quantitative computed tomography was used to measure changes in bone strength indices (bone strength index [BSI, mg(2)/mm(4)] and strength-strain index [SSIp, mm(3)]) and volumetric bone mineral density [vBMD, mg/cm(3)] at distal metaphyseal and diaphyseal regions of the femur and tibia, as well as calf and thigh muscle density (mg/cm(3)), and muscle cross-sectional area (MCSA, mm(2)), indices of skeletal muscle fat content and muscle force production, respectively. RESULTS: After controlling for potential confounders, greater gains in femur BSI (44%, P<0.002), total femur vBMD (114%, P<0.04) and femur trabecular vBMD (306%, P<0.002) occurred in girls in the lowest versus the highest groups of baseline thigh muscle density. Greater gains in tibial BSI (25%, P<0.03) and trabecular vBMD (190%, P<0.002) were also observed in the lowest versus the highest baseline calf muscle density groups. CONCLUSION: Baseline muscle density is a significant predictor of changes in bone density and bone strength in young girls during a period of rapid skeletal development.

Isolation and characterization of human osteoblasts from needle biopsies without in vitro culture.

SUMMARY: We isolate and characterize osteoblasts from humans without in vitro culture. These techniques should be broadly applicable to studying the pathogenesis of osteoporosis and other bone disorders. INTRODUCTION: There is currently no data regarding the expression of specific genes or pathways in human osteoblasts that have not been subjected to extensive in vitro culture. Thus, we developed methods to rapidly isolate progressively enriched osteoblast populations from humans and characterized these cells. METHODS: Needle bone biopsies of the posterior iliac crest were subjected to sequential collagenase digests. The cells from the second digest were stained with an alkaline phosphatase (AP) antibody, and the AP+ cells were isolated using magnetic cell sorting. RESULTS: Relative to AP- cells, the AP+ cells contained virtually all of the mineralizing cells and were enriched for key osteoblast marker genes. The AP+ cells were further purified by depletion of cells expressing CD45, CD34, or CD31 (AP+/CD45/34/31- cells), which represented a highly enriched human osteoblast population devoid of hematopoietic/endothelial cells. These cells expressed osteoblast marker genes but very low to undetectable levels of SOST. We next used high-throughput RNA sequencing to compare the transcriptome of the AP+/CD45/34/31- cells to human fibroblasts and identified genes and pathways expressed only in human osteoblasts in vivo, but not in fibroblasts, including 448 genes unique to human osteoblasts. CONCLUSIONS: We provide a detailed characterization of highly enriched human osteoblast populations without in vitro culture. These techniques should be broadly applicable to studying the pathogenesis of osteoporosis and other bone disorders.

Fundamental radiological and geometric performance of two types of proton beam modulated discrete scanning systems.

PURPOSE: The purpose of this investigation was to compare and contrast the measured fundamental properties of two new types of modulated proton scanning systems. This provides a basis for clinical expectations based on the scanned beam quality and a benchmark for computational models. Because the relatively small beam and fast scanning gave challenges to the characterization, a secondary purpose was to develop and apply new approaches where necessary to do so. METHODS: The following performances of the proton scanning systems were investigated: beamlet alignment, static in-air beamlet size and shape, scanned in-air penumbra, scanned fluence map accuracy, geometric alignment of scanning system to isocenter, maximum field size, lateral and longitudinal field uniformity of a 1 l cubic uniform field, output stability over time, gantry angle invariance, monitoring system linearity, and reproducibility. A range of detectors was used: film, ionization chambers, lateral multielement and longitudinal multilayer ionization chambers, and a scintillation screen combined with a digital video camera. Characterization of the scanned fluence maps was performed with a software analysis tool. RESULTS: The resulting measurements and analysis indicated that the two types of delivery systems performed within specification for those aspects investigated. The significant differences were observed between the two types of scanning systems where one type exhibits a smaller spot size and associated penumbra than the other. The differential is minimum at maximum energy and increases inversely with decreasing energy. Additionally, the large spot system showed an increase in dose precision to a static target with layer rescanning whereas the small spot system did not. CONCLUSIONS: The measured results from the two types of modulated scanning types of system were consistent with their designs under the conditions tested. The most significant difference between the types of system was their proton spot size and associated resolution, factors of magnetic optics, and vacuum length. The need and benefit of mutielement detectors and high-resolution sensors was also shown. The use of a fluence map analytical software tool was particularly effective in characterizing the dynamic proton energy-layer scanning.

Associations of physical activity duration, frequency, and load with volumetric BMD, geometry, and bone strength in young girls.

SUMMARY: More efficacious physical activity (PA) prescriptions for optimal bone development are needed. This study showed that PA duration, frequency, and load were all independently associated with bone parameters in young girls. Increased PA duration, frequency, and load are all important osteogenic stimuli that should be incorporated into future PA interventions. INTRODUCTION: This study evaluated the associations of physical activity (PA) duration, frequency, load, and their interaction (total PA score = duration × frequency × load) with volumetric bone mineral density, geometry, and indices of bone strength in young girls. METHODS: Four hundred sixty-five girls (aged 8-13 years) completed a past year physical activity questionnaire (PYPAQ) which inquires about the frequency (days per week) and duration (average minutes per session) of leisure-time PA and sports. Load (peak strain score) values were assigned to each activity based on ground reaction forces. Peripheral quantitative computed tomography was used to assess bone parameters at metaphyseal and diaphyseal sites of the femur and tibia of the non-dominant leg. RESULTS: Correlations across all skeletal sites between PA duration, frequency, load and periosteal circumference (PC), bone strength index (BSI), and strength-strain index (SSI) were significant (p ≤ 0.05), although low (0.10-0.17). A 2.7-3.7% greater PC across all skeletal sites was associated with a high compared to a low PYPAQ score. Also, a high PYPAQ score was associated with greater BSI (6.5-8.7%) at metaphyseal sites and SSI (7.5-8.1%) at diaphyseal sites of the femur and tibia. The effect of a low PYPAQ score on bone geometric parameters and strength was greater than a high PYPAQ score. CONCLUSIONS: PA duration, frequency, and load were all associated with bone geometry and strength, although their independent influences were modest and site specific. Low levels of PA may compromise bone development whereas high levels have only a small benefit over more average levels.

Clinical characterization of a proton beam continuous uniform scanning system with dose layer stacking.

A proton beam delivery system on a gantry with continuous uniform scanning and dose layer stacking at the Midwest Proton Radiotherapy Institute has been commissioned and accepted for clinical use. This paper was motivated by a lack of guidance on the testing and characterization for clinical uniform scanning systems. As such, it describes how these tasks were performed with a uniform scanning beam delivery system. This paper reports the methods used and important dosimetric characteristics of radiation fields produced by the system. The commissioning data include the transverse and longitudinal dose distributions, penumbra, and absolute dose values. Using a 208 MeV cyclotron's proton beam, the system provides field sizes up to 20 and 30 cm in diameter for proton ranges in water up to 27 and 20 cm, respectively. The dose layer stacking method allows for the flexible construction of spread-out Bragg peaks with uniform modulation of up to 15 cm in water, at typical dose rates of 1-3 Gy/min. For measuring relative dose distributions, multielement ion chamber arrays, small-volume ion chambers, and radiographic films were employed. Measurements during the clinical commissioning of the system have shown that the lateral and longitudinal dose uniformity of 2.5% or better can be achieved for all clinically important field sizes and ranges. The measured transverse penumbra widths offer a slight improvement in comparison to those achieved with a double scattering beam spreading technique at the facility. Absolute dose measurements were done using calibrated ion chambers, thermoluminescent and alanine detectors. Dose intercomparisons conducted using various types of detectors traceable to a national standards laboratory indicate that the measured dosimetry data agree with each other within 5%.

Laboratory tumor lysis syndrome complicating LBH589 therapy in a patient with acute myeloid leukaemia.

LBH589 is a novel cinnamic hydroxamic acid analog (HAA) pan-histone deacetylase inhibitor (HDACi) currently in early phase clinical development.

Stability of antioxidants in an apple polyphenol-milk model system.

The stability of antioxidants in an apple polyphenol-milk model system was examined. The model system consisted of skim milk fortified with pH-neutralised apple polyphenols (AP, 0-200mg per 100ml milk), with or without ascorbic acid (100mg per 100ml milk). Physical and chemical changes were evaluated after thermal treatment (120°C, 5min) and oxidative storage (20°C and 38°C, up to 12 weeks). Antioxidant capacity was determined using both oxygen radical absorbance capacity (ORAC) assay and ferric reducing antioxidant power (FRAP) assay. Significant antioxidant capacity was detected in the presence of milk. Antioxidant capacity was retained during thermal treatment but decreased slowly during storage. The concentration of ascorbic acid decreased rapidly, and was close to zero after 2-week storage at 38°C or 10-week storage at 20°C. The brownness of the polyphenol-milk system increased over storage duration of 0-12 weeks; this effect was retarded by the addition of ascorbic acid. This high polyphenol-milk has demonstrated good physical stability.

Radiologic validation of a fast neutron multileaf collimator.

Teletherapy with high linear energy transfer radiations (LET), perhaps more than with low LET types, requires careful beam collimation to limit effects to normal structures. Intensity modulated techniques may also hold promise in this regard. Accordingly, a remote computer-controlled, high-resolution multileaf collimator (MLC) is placed into service at the Gershenson Radiation Oncology Center's fast neutron therapy center of the Karmanos Cancer Institute, Detroit, Michigan. Prior to clinical application the basic radiological properties of the fast neutron MLC are studied. Complicating the evaluation is the mixed neutron and gamma radiation field environment encountered with fast neutron beams. As a reference the MLC performance is compared to an existing multirod collimator (MRC) used at the facility for more than ten years. The MLC aggregate transmission is found to be about 4%, slightly outperforming the MRC. The measured gamma component for a closed collimator is 1.5 times higher for the MLC, compared with the MRC. The different materials used for attenuation, steel and tungsten, respectively account for the difference. The geometry for the MRC is double focused whereas that for the MLC is single focused. The resulting penumbrae agree between the focused axis of the MLC and both axes of the MRC. Penumbra differences between the focused and unfocused axes were not observable at small field sizes and a maximum of about 1 cm for a 25 x 25 cm2 field at 2.5 cm depth in water. For a 10 x 10 cm2 field the focused penumbra is 9 mm, and the unfocused is 12 mm. The many benefits of the fully automatic MLC over the semimanual MRC are considered to justify this compromise.

Experiences with an application of industrial robotics for accurate patient positioning in proton radiotherapy.

BACKGROUND: Protons beams deliver targeted radiation doses with greater precision than is possible with electrons or megavoltage X-ray photons, but to retain this advantage, patient positioning systems at proton clinics must meet tighter accuracy requirements. For this and other reasons, robots were incorporated into the treatment room systems at MPRI. METHODS: The Midwest Proton Radiotherapy Institute (MPRI) is the first radiotherapy facility in the United States to use commercial robots with six degrees of freedom for patient positioning, rather than a traditional bed with four degrees of freedom. RESULTS: This paper outlines the ways in which robots are used at MPRI and attempts to distil insights from the experience of treating over 200 radiotherapy patients with a robotic system from February 2004 to late 2006. CONCLUSIONS: The system has performed well, and with great reliability, but there is room for future improvement, especially in ease of use and in reducing the time to get patients into position.

Patient-specific margins for proton therapy of lung.

Lung cancer treatment presents a greater treatment planning and treatment delivery challenge in proton beam therapy compared to conventional photon therapy due to the proton beam's energy deposition sensitivity to the breathing-induced dynamic tissue density variations along the beam path. Four-dimensional computed tomography (4D-CT) has been defined as the explicit inclusion of temporal changes of tumor and normal organ mobility into an image series. It allows more accurate delineation of lung cancer target volumes by suppression of any breathing motion artifacts present in the CT images. It also allows analysis of the tumor's 3D spatial movement within a breathing phase cycle. The motivation for this study was to investigate dosimetric errors caused by lung tumor motion in order to find an optimal method of design for patient compensators and apertures for a passive scattering beam delivery system and treatment of the patient under free breathing conditions. In this study, the maximum intensity projection (MIP) method was compared to patient-specific internal margin designs based on a single breathing phase at the end-of inhale (EOI) or middle-of-exhale (MOE). It was found that MIP method provides superior tumor dose distribution compared to patient-specific internal margin designs derived from 4D-CT.

Compact multileaf collimator for conformal and intensity modulated fast neutron therapy: electromechanical design and validation.

The electromechanical properties of a 120-leaf, high-resolution, computer-controlled, fast neutron multileaf collimator (MLC) are presented. The MLC replaces an aging, manually operated multirod collimator. The MLC leaves project 5 mm in the isocentric plane perpendicular to the beam axis. A taper is included on the leaves matching beam divergence along one axis. The 5-mm leaf projection width is chosen to give high-resolution conformality across the entire field. The maximum field size provided is 30 x 30 cm2. To reduce the interleaf transmission a 0.254-mm blocking step is included. End-leaf steps totaling 0.762 mm are also provided allowing opposing leaves to close off within the primary radiation beam. The neutron MLC also includes individual 45 degrees and 60 degrees automated universal tungsten wedges. The automated high-resolution neutron collimation provides an increase in patient throughput capacity, enables a new modality, intensity modulated neutron therapy, and limits occupational radiation exposure by providing remote operation from a shielded console area.

Tuberculosis and HIV co-infections in Jamaican children

BACKGROUND: There has been a worldwide increased prevalence of tuberculosis (TB) in recent years, with a similar trend observed in Jamaica and more recently in children admitted to the University Hospital of the West Indies, Jamaica. Data regarding paediatric TB, especially as it relates to all aspects of HIV co-infection, are needed from developing countries in diverse geographic settings to enhance prevention and treatment policies (National Institutes of Health, Office of AIDS Research, FY 2005 Budgetary Planning Meeting, March 11, 2003, Washington, DC). OBJECTIVE: To determine associated factors and outcomes of tuberculosis in HIV-infected and noninfected children in Jamaica. METHOD: We reviewed records of children aged 0 - 12 years attending the University Hospital of the West Indies during January 1999 to December 2002. Associated factors and outcomes in HIV-infected and HIV-negative cases with TB were compared using exact statistical methods to account for the small number of children and an adjustment for multiple testing. TB diagnosis was determined using modified World Health Organization (WHO) criteria. RESULTS: There was a significant increase of active TB cases from 1999 to 2002 with 24 children diagnosed over this period All 24 children (100) had received the Bacillus-Calmette-Guerin (BCG) vaccine. Eleven (46) of these were HIV-infected, all via mother-to-child transmission. HIV-infected children were statistically more likely to be older than non-infected children (mean 4.2 vs 2.6 years), and also to have failure to thrive, digital clubbing, hepatomegaly, splenomegaly, generalized adenopathy and negative Mantoux tests. Appropriate in-hospital anti-TB therapy was given. Hospital stay was longer (median 7.4 vs. 2.8 months) and death was more likely (7/11 vs 2/13) in HIV-infected vs non-infected children. Triple antiretroviral therapy was given in three of the 11 HIV-infected cases and this markedly improved outcome. Household family members with active TB were identified in twelve cases. CONCLUSIONS: HIV and TB co-infection is an increasing problem in Jamaican children. Severity of illness and death is greater in HIV-infected children, despite appropriate anti-TB therapy. Antiretroviral drugs must be made available to this population. Efforts must be enhanced to reduce mother-to-child-transmission of HIV/AIDS and to strengthen the public health management of TB (contact tracing and completion of TB therapy by directly observed therapy)

Thermoluminescence dosimetry in mixed neutron/gamma radiation beam.

Miniature CaF2:Tm (TLD-300) were adopted for dosimetry in a mixed neutron/gamma beam produced by the Detroit superconducting cyclotron. A method utilizing the linear relationship between the ratios of the areas under the low and high temperature peaks of the glow curve and the ratios of gamma dose to total dose (Dgamma/DT) in d(48.5)+Be fast-neutron beam was applied for both the gamma component and the total dose measurements. The detectors were grouped based on their response to uniform 60Co dose. Within each group the ratio of the peak areas of the glow curve demonstrated uniformity and stability better than 1.5% (one standard deviation). The parameters of the linear regression between the fractional gamma component in the neutron beam and the ratio of peak areas were derived from measurements at locations with predetermined Dgamma/DT. The method was applied to measure the transmission through the existing multi-rod and newly designed multi-leaf collimators.

Reduction of Cr(VI) by "palladized" biomass of Desulfovibrio desulfuricans ATCC 29577.

A novel catalytic activity of palladium [Pd(0)]-coated cells of Desulfovibrio desulfuricans ATCC 29577 ["bio-Pd(0)"] is demonstrated. Reduction of 700 microM Cr(VI) occurred within 24 h using formate (25 mM) or hydrogen (1 atm) as the electron donor, under conditions whereby cells lacking bound Pd(0), or palladium metal manufactured via chemical reduction of soluble Pd(II), did not reduce Cr(VI). The biomass-bound Pd(0) also functioned in the continuous removal of 400 microM Cr(VI) from a 1 mM solution under H(2) (flow residence time approximately 5 h), where chemically prepared Pd(0) was ineffective. This demonstrates a new type of active bioinorganic catalysis, whereby the presence of biomass bound to Pd(0) confers a novel catalytic capability not seen with Pd base metal or biomass alone.

Tuberculosis and HIV co-infections in Jamaican children.

BACKGROUND: There has been a worldwide increased prevalence of tuberculosis (TB) in recent years, with a similar trend observed in Jamaica and more recently in children admitted to the University Hospital of the West Indies, Jamaica. Data regarding paediatric TB, especially as it relates to all aspects of HIV co-infection, are needed from developing countries in diverse geographic settings to enhance prevention and treatment policies (National Institutes of Health, Office of AIDS Research, FY 2005 Budgetary Planning Meeting, March 11, 2003, Washington, DC). OBJECTIVE: To determine associated factors and outcomes of tuberculosis in HIV-infected and noninfected children in Jamaica. METHOD: We reviewed records of children aged 0 - 12 years attending the University Hospital of the West Indies during January 1999 to December 2002. Associated factors and outcomes in HIV-infected and HIV-negative cases with TB were compared using exact statistical methods to account for the small number of children and an adjustment for multiple testing. TB diagnosis was determined using modified World Health Organization (WHO) criteria. RESULTS: There was a significant increase of active TB cases from 1999 to 2002 with 24 children diagnosed over this period All 24 children (100%) had received the Bacillus-Calmette-Guerin (BCG) vaccine. Eleven (46%) of these were HIV-infected, all via mother-to-child transmission. HIV-infected children were statistically more likely to be older than non-infected children (mean 4.2 vs 2.6 years), and also to have failure to thrive, digital clubbing, hepatomegaly, splenomegaly, generalized adenopathy and negative Mantoux tests. Appropriate in-hospital anti-TB therapy was given. Hospital stay was longer (median 7.4 vs. 2.8 months) and death was more likely (7/11 vs 2/13) in HIV-infected vs non-infected children. Triple antiretroviral therapy was given in three of the 11 HIV-infected cases and this markedly improved outcome. Household family members with active TB were identified in twelve cases. CONCLUSIONS: HIV and TB co-infection is an increasing problem in Jamaican children. Severity of illness and death is greater in HIV-infected children, despite appropriate anti-TB therapy. Antiretroviral drugs must be made available to this population. Efforts must be enhanced to reduce mother-to-child-transmission of HIV/AIDS and to strengthen the public health management of TB (contact tracing and completion of TB therapy by directly observed therapy) to eliminate the spread