To observe the impact of N and P concentrations on utilization of

To observe the impact of N and P concentrations on utilization of iron by bioreporter Palr0397-luxAB, a series of and concentrations in Fraquil medium with three

Fe3+ concentrations were set to determine the response of luciferase activities to the concentrations of N and P. In Fraquil medium with 10 or 100 nM Fe3+, luciferase activity of bioreporter Palr0397-luxAB was enhanced with the increase in concentration and decreased slightly (remaining at a high level) with ranging from 100 to 900 μM (Fig. 3a); similarly, its luciferase activity increased significantly when increased from 0.1 to 1 μM and varied a little with further increase in concentration (Fig. 3b). In Fraquil medium with 1000 nM Fe3+, its luciferase activity increased slightly with the increased N and P concentrations. When the concentrations of N and check details P were high enough (e.g. 100 μM and 10 μM in this study), further increases in N and P concentrations had little influence on the luciferase activity, showing that iron utilization might not be affected by the uptake of N and P in cells. The variation of N and P concentrations had no effect on luciferase activity of bioreporter in 1000 nM Fe3+ concentration condition, which also indicated that iron utilization was not directly related with the uptake of N and P in cells. Fur acts as a transcriptional repressor of iron-regulated promoters by virtue

of its iron-dependent DNA-binding activity to regulate expression of several genes involved in iron homeostasis (Escolar P450 inhibitor et al., 1999). At high concentrations, Co2+ and Mn2+ presumably

mimic Fe2+ (Bagg & Neilands, 1987; Hantke, 1987), and Zn2+ can also activate Fur in vitro (Bagg & Neilands, 1987). So these metals could possibly interfere with iron detection. In addition, other metals such as Cu2+ can compete with iron to chelate dissolved organic siderophores secreted by cells, thus decreasing iron availability (Nicolaisen et al., 2008). The concentrations of metals in lakes greatly varied and are easily affected by surrounding environments. Take the concentration ranges of Co2+, Zn2+, and Cu2+ in Wuhan City (China) for instance; they were 3.7–4.9, 13.1–181.2, and 18.4–83.8 nM in Donghu Lake located in a scenic area, were 7.8–17.6, 1.2–285.1, and 43.1–916.7 nM in Moshui Lake situated in an industrial area, and were 4.9–6.8, 0–0.9, and 58.4–67.7 nM in Houguan Lake in the suburbs (Yu et al., 2007). In an attempt to determine the influence of Co2+, Mn2+, Zn2+, and Cu2+ concentrations on iron bioavailability, luciferase activity of bioreporter Palr0397-luxAB at six concentrations of Co2+, Mn2+, Zn2+, and Cu2+ was, respectively, measured in Fraquil medium with three Fe3+ concentrations. The increase in Mn2+ concentration had no effect on luciferase activity of the bioreporter (Fig. 3d).

Almost 1 million Canadians travel annually to malaria endemic are

Almost 1 million Canadians travel annually to malaria endemic areas, with several hundred cases reported each year.[1] Travelers who visit friends and relatives (VFRs) are well known to be at increased risk for malaria.[2, 3] Anecdotally, cases of malaria at Winnipeg Children’s Hospital (WCH) appeared to be increasing over time. The aim of this study was to review the aspects of malaria at WCH in both travelers and immigrants, and to identify possible gaps in management. Charts for all cases of malaria buy Olaparib in children (≤18 years), identified by ICD-9 code and hematology lab record review

and confirmed by positive thick or thin smears, as reviewed by a hematopathologist, presenting to WCH from January PARP phosphorylation 1, 1989,

to December 31, 2008, were retrospectively reviewed. Data were collected by way of a collection tool. Our hospital is the only tertiary pediatric center for the province of Manitoba, northwestern Ontario, eastern Saskatchewan, and southern Nunavut. There are an estimated 50,000 outpatient visits to the emergency department (ED) per year. Data were analyzed using Microsoft Excel (Microsoft, Seattle, WA, USA). The review was approved by the University of Manitoba Bannatyne Research Ethics Board. Statistical comparisons were done using Fisher’s exact test. From 1989 to 2008, 38 cases of pediatric malaria were identified in patients presenting to WCH. The mean age of cases was 8.4 ± 4.6 years, and 50% were male. Most cases occurred in older Astemizole children, with 24 cases (63%) > 6 years of age. On average, two cases of malaria were identified per year. Twelve cases occurred in pediatric travelers from malaria non-endemic areas (11 from Canada,

1 from UK), 11 of which were among VFRs (children born in Canada or overseas, returning to family’s nation of origin to visit friends and relatives). Six VFRs traveled to India, and five to sub-Saharan Africa. One child traveled to the Solomon Islands with family on business. The mean time from date of return to Canada to diagnosis was 123.3 days. The remaining 26 cases occurred among new immigrants and refugees, with a mean time from arrival in Canada to diagnosis of 92.3 days. Only 4 immigrants emigrated from India or Pakistan, while 22 emigrated from sub-Saharan Africa (Nigeria and Mozambique most commonly). All but two (93%) of the immigrant/refugee cases presented from 2000 onwards, whereas only four (33%) of the travel-related cases occurred in the same time period (Figure 1). From the traveler’s group, information about pre-travel counseling was available for 10 patients of whom 6 consulted a clinician prior to travel, none via a travel clinic. Only one child was prescribed appropriate malarial prophylaxis for the area of travel, and the parents of that child forgot to administer it overseas (two cases not specified, one given nothing, two given chloroquine inappropriately).

There is a relative paucity of data on the morbidity/mortality as

There is a relative paucity of data on the morbidity/mortality associated with bacterial pneumonia in the era of GSI-IX potent cART in those with higher CD4 cell counts, although several cohort studies have indicated declining rates of morbidity and mortality associated with cART use and, in some studies, pneumococcal polysaccharide vaccine (PPV-23) [9–11]. In a Danish study exploring

the risks for hospitalization with pneumonia (including viral pneumonia but excluding PcP), HIV-1-infected patients with a nadir CD4 count >300 cells/μL had a rate of bacterial pneumonia of 1.25 per 100 PY [5]. In the Strategies for Management of Anti-Retroviral Therapy (SMART) study, which enrolled participants with baseline CD4 count ≥350 cells/μL [12], patients on continuous

cART had a rate of bacterial pneumonia of 1.3 per 100 PY. The clinical benefits of intermittent recombinant interleukin-2 (rIL-2) in HIV-1-infected adults on cART have been explored in two major Phase III international studies. In Evaluation of Subcutaneous Interleukin-2 in a Randomized International Trial (ESPRIT), HIV-1-infected adults on or starting cART, with CD4 count ≥300 cells/μL, were randomized to intermittent rIL-2 with cART (IL-2 arm) or cART alone (control arm or non-IL-2 arm) [13]. The primary endpoints, ADI and death, were reported in both study arms for the duration of follow-up. The main results of ESPRIT have been reported [14]. In summary, the receipt of rIL-2 conferred no clinical benefit with respect to ADI and all-cause mortality despite a significant

difference in CD4 count compared with the control arm, with CD4 count averaged over follow-up being 159 cells/μL [95% confidence interval (CI) 145–174 cells/μL; P<0.001] higher in the IL-2 arm than in the control arm. Recombinant IL-2 used in the oncology and/or HIV setting [15] has been associated with an increased risk of some bacterial infections, including cellulitis, osteomyelitis, Clostridium difficile infection [16], bacteraemia and bacterial pneumonia; the mechanism of the association is unclear. The ESPRIT cohort offered an opportunity to explore both the rate of bacterial pneumonia over several years (≈7 years) in a large cohort of cART-treated HIV-1-infected adults with moderate levels of immunodeficiency and the relationship between rIL-2 exposure and bacterial pneumonia. The methods [13] and main results [14] of ESPRIT have been reported previously. Key inclusion criteria included CD4 count ≥300 cells/μL and on/commencing cART. Centers for Disease Control category C patients could be enrolled provided that there was no active ADI for ≥12 months. Patients randomized to the IL-2 arm received three dosing cycles of rIL-2 [7.5 MIU subcutaneously twice a day for five consecutive days every 8 weeks] as induction in year 1.

Good estimates for the number of coinfected persons actually acce

Good estimates for the number of coinfected persons actually accessing care are not available. The only available data relate to the Province of Ontario, where approximately 65% of persons diagnosed with HIV have accessed care at least once (defined as having at least one HIV viral load measurement after diagnosis), whereas only 51% can be said to be in regular medical follow-up [17]. Thus, the 955 cohort participants probably represent close to 20% of all coinfected patients receiving

treatment in Canada. We have provided a comprehensive picture of the extent of vulnerabilities that present challenges to effective care and prevention of serious morbidity and mortality in this population. There are extremely high rates of social

instability, poverty, mental illness and alcohol and drug use. Aboriginals are disproportionately represented in our cohort. Whereas they comprised 3.8% of the Canadian population in 2006 and 8% of prevalent HIV infections [18], 15% of our cohort overall and 33% in British Columbia self-identified as aboriginal and a very high proportion of these were women (62%). The impact of these combined vulnerabilities on the health of the coinfected population appears Aurora Kinase inhibitor to be appreciable. Despite 82% of participants in the cohort receiving ART, only 71% were virologically suppressed. Another 6% had interrupted treatment at baseline. While these results are not dissimilar from those of other studies in IDU populations, these viral suppression rates are lower than those reported generally in HIV-infected persons [19-21]. Together, our results highlight that a significant proportion of participants Adenosine triphosphate have difficulty with treatment adherence and consequently are at risk for developing viral resistance and experiencing HIV-related disease progression. Indeed, the rate of AIDS was very high, at twice the reported

rate in a US HIV-infected population for the period 2003–2007 [22]. Incomplete HIV suppression may also have implications for HIV transmission, especially given the high percentage of participants that report sharing injection equipment and risky sexual behaviours. Finally, treatment interruptions have also been associated with increased risk for non-AIDS-related adverse outcomes, including liver disease progression and death, particularly among coinfected persons [23, 24]. HCV infection is a chronic infection which if left untreated follows a slow clinical course progressing to ESLD and hepatocellular carcinoma [25]. HIV increases chronicity and accelerates the natural history of HCV [7, 26]. This impact is mirrored in our cohort: the median age of the population and time since HCV infection suggest that we are poised for a peak of chronic liver disease and its consequences. Indeed, at baseline many participants already had significant fibrosis and ESLD.

The total population examined within the study period was from Ma

The total population examined within the study period was from March 2006 to August 2009.

In the total population examined, the prevalence of Selleck R788 PE was 2.2% [11]. In addition to the 76 HIV-positive cases included in the study, there were three HIV-positive women who developed PE (3.9%) and who were excluded from the study because this number was too small to allow valid comparisons of the prevalence of PE to be made between HIV-positive and HIV-negative women. None of the selected controls developed PE and all pregnancies resulted in the live birth of phenotypically normal neonates. In normal pregnancy the measured UtA-PI is affected by fetal crown–rump length, maternal age, body mass index, racial group and parity. In comparing normal with pathological pregnancies, the values of UtA-PI are expressed as multiples of the median (MoM) of the normal after appropriate adjustment for the above variables [11]. Normality of the data distribution was examined with the Kolmogorov–Smirnov test and probability plots. Data were expressed as mean ± standard deviation or as median and interquartile range (IQR) for normally and non-normally distributed data, respectively. Comparisons between groups were performed using the t-test or Mann–Whitney U-test for numerical data and the χ2 test for categorical data. Univariate regression analyses were performed where appropriate.

Power analysis indicated

that a sample of 76 HIV-positive and 2280 HIV-negative women would have more than 80% power (α 0.05) for Ruxolitinib clinical trial the detection of a mean difference of 0.26 in the mean UtA-PI (MoM) between the groups. As there are no previous data in pregnant women with HIV infection, the effect size was estimated from data presented in previous publications for pregnant women with known increased resistance in the uterine arteries, such as those who eventually develop PE [11]. The statistical analyses were performed using the Statistical Package for Social Sciences (Version 12.0; Carbohydrate SPSS, Chicago, IL, USA). The demographic and pregnancy characteristics and outcomes for the 76 HIV-positive and 2280 HIV-negative women are given in Table 1. In the HIV-positive group, 33 women (43.4%) were on antiretroviral treatment, including 14 (42.4%) on nucleoside reverse transcriptase inhibitors (NRTIs) and a protease inhibitor, 18 (54.5%) on NRTIs and a nonnucleoside reverse transcriptase inhibitor (NNRTI) and one (3.1%) on monotherapy. The median duration of treatment prior to the first trimester ultrasound scan was 22 months (IQR 7.5–39.5 months) and the majority of the women (n=29) were on antiretroviral treatment at the time of conception. Compared with the HIV-negative women, the HIV-positive women were more likely to be heavier, to be of African racial origin, to be nonsmokers and to deliver earlier and have smaller neonates.

To minimize the influences of procedural learning, on the first d

To minimize the influences of procedural learning, on the first day the naive subjects were given a few tens of practice trials with suprathreshold differences in stimulus orientation. Orientation discrimination thresholds were measured with a standard one-up, two-down staircase

procedure converging at 70.7% correct responses. The orientation step size of the staircase Y 27632 was 0.05 log units. Each staircase (i.e. a block of trials) consisted of eight reversals. The geometric mean of the last six reversals was calculated as the threshold. A typical staircase comprised 35–50 trials. The subjects compared a difference in orientation between two successively presented stimuli. Thirteen naive subjects were randomly assigned to practice under either the congruent condition (left panels in Fig.1A; Group I subjects, n = 6) or the incongruent condition (right panels in Fig. 1A; Group II subjects, n = 7). After the training,

their thresholds were measured under both the congruent and incongruent conditions, and at the trained 55° orientation as well as at an untrained orientation of 140° (Fig. 1B). Note that the two stimuli in learn more a trial occupied two different retinal locations, but these two retinal locations were the same in the congruent and incongruent conditions. Therefore, the congruent and incongruent spatial

relations of the two stimuli were in terms of a spatiotopic, rather than a retinotopic, reference frame (referred to as the spatiotopic stimulus relation throughout the text). As in many perceptual learning tasks, training decreased the subjects’ thresholds for orientation discrimination by approximately a factor of two in Group I subjects trained under the congruent condition (pre-training threshold 7.62° ± 0.48° vs. post-training Astemizole threshold 4.07° ± 0.3°, t = 6.41, P = 0.001, paired t-test) and in Group II subjects trained under the incongruent condition (pre-training threshold 7.44° ± 1.00° vs. post-training threshold 3.71° ± 0.32°, t = 4.35, P = 0.002). However, when the spatiotopic stimulus relation was switched from trained to untrained without changing the stimulus location on the retina, there was a significant elevation of the mean thresholds at the trained 55° orientation in both Group I subjects (t = 5.06, P = 0.004; left panel in Fig. 1B, compare the two bars corresponding to the 55° condition) and Group II subjects (t = 4.33, P = 0.005; right panel in Fig. 1B, compare the two bars corresponding to the 55° condition), indicating spatiotopic location specificity of the learning. This observation suggests that spatiotopic processing mechanisms can be tuned in favor of the trained spatiotopic stimulus relation.

4) Gingipain and hemagglutination activities were also restored

4). Gingipain and hemagglutination activities were also restored in FLL350c (Figs 3 and 4), thus confirming a role for PG0162 in virulence regulation in P. gingivalis W83. It is noteworthy that although FLL354 had the lowest gingipain

activity, its hemolysin profile was similar to the wild-type strain. Taken together, this is consistent with previous observations suggesting that hemolysin and gingipain activities can be distinct from each other (Deshpande & Khan, 1999). Collectively, our study showed that ECF sigma factors PG0162 and PG1660 play an important role in the regulation Selleck MK-3475 of gingipain, hemolytic, and hemagglutination activities, and could likely modulate the virulence potential of P. gingivalis. Because the exterior surface structures and factors of the infectious bacteria are the first to come in contact

with the host and must respond and adapt to the host environment, our observations are consistent with the role of ECF sigma factors in the regulation of virulence-associated genes (reviewed in Brooks & Buchanan, 2008). The activity of ECF sigma factors is most often negatively regulated by direct interaction with cognate antisigma factors, which prevent their association with the core RNA polymerase or facilitate holoenzyme dissociation (reviewed in Staron et al., 2009). While PG1660 appear to have a putative cognate antisigma factor PG1659 (,, a similar putative component is missing for PG0162. It is noteworthy

that the regulation of PG0162 on virulence observed in this study is occurring at the post-transcriptional level. It is likely that PG0162 may be involved in a unique and complex regulatory mechanism, and this requires further evaluation. This work was supported by Loma Linda University and Public Health Grant DE13664 and DE019730 from NIDCR (to H.M.F.). ”
“A technique based on an inverted Petri dish system was developed for the growth and isolation of soil oxalotrophic bacteria able to disperse on fungal mycelia. The method is related to the ‘fungal highways’ dispersion theory in which mycelial fungal networks allow active movement of bacteria in soil. Quantification of this phenomenon showed that bacterial dispersal occurs preferentially STK38 in upper soil horizons. Eight bacteria and one fungal strain were isolated by this method. The oxalotrophic activity of the isolated bacteria was confirmed through calcium oxalate dissolution in solid selective medium. After separation of the bacteria–fungus couple, partial sequencing of the 16S and the ITS1 and ITS2 sequences of the ribosomal RNA genes were used for the identification of bacteria and the associated fungus. The isolated oxalotrophic bacteria included strains related to Stenotrophomonas, Achromobacter, Lysobacter, Pseudomonas, Agrobacterium, Cohnella, and Variovorax. The recovered fungus corresponded to Trichoderma sp.

We found that the preferred spatial and temporal frequencies, spa

We found that the preferred spatial and temporal frequencies, spatial resolution and high temporal frequency cutoff of area MT neurons were reduced in aged monkeys, and were accompanied by the broadened tuning width of spatial frequency, elevated spontaneous activity, and decreased

signal-to-noise ratio. These results showed that, for neurons in area MT, aging significantly changed both the spatial and temporal frequency response tuning properties. Such evidence provides new insight into the changes occurring at the electrophysiological level that may be related to the aging-related visual deficits, especially in processing spatial and temporal information. selleck compound
“During neuronal maturation, the neuron-specific K–Cl co-transporter KCC2 lowers the intracellular chloride and thereby renders GABAergic transmission hyperpolarizing. Independently of its role as a co-transporter, KCC2 plays a crucial role in the maturation of dendritic spines, most probably via an interaction with the cytoskeleton-associated protein 4.1N. In this study, we show that neural-specific overexpression of KCC2 impairs the development of the neural tube- and neural crest-related structures in mouse embryos. At early

stages (E9.5–11.5), the transgenic embryos had a thinner learn more neural tube and abnormal body curvature. They displayed a reduced neuronal differentiation and altered neural crest cell pattern. At later stages (E11.5–15.5), the transgenic embryos had smaller brain structures and a distinctive cleft

palate. Similar results were obtained using overexpression of a transport-inactive N-terminal-deleted variant of KCC2, implying that the effects were not dependent on KCC2′s role as a K–Cl co-transporter. Interestingly, the neural tube of transgenic embryos had an aberrant cytoplasmic distribution of 4.1N and actin. This was corroborated in a neural stem cell line with ectopic expression of KCC2. Embryo phenotype and cell morphology were unaffected by a mutated variant of KCC2 which is unable to bind 4.1N. These results point to a role of KCC2 in neuronal differentiation BCKDHA and migration during early development mediated by its direct structural interactions with the neuronal cytoskeleton. KCC2 is a neuron-specific isoform of the K–Cl co-transporters. Its developmental upregulation is temporally associated with maturation of postsynaptic GABAergic inhibition in central neurons (Rivera et al., 1999; reviewed in Blaesse et al., 2009). Functional expression of KCC2 during neuronal development leads to a decrease in the intraneuronal Cl− concentration and, consequently, to a hyperpolarizing shift in the reversal potential of GABAA receptor-mediated currents (EGABA) from depolarizing values that are characteristic for immature neurons.

Interestingly, IAA addition upregulates genes encoding a type VI

Interestingly, IAA addition upregulates genes encoding a type VI secretion

system (T6SS), a kind of secretion system that has been specifically implicated in bacterium–eukaryotic host interactions. Moreover, many transcription factors showed altered expression in the different treatments, indicating that the regulatory machinery of the bacterium is altered in response to IAA (Van Puyvelde et al., 2011). Increasing evidence indicates that NO is a key signaling molecule that is involved in a wide range of functions in plants (Creus et al., 2005; Molina-Favero et al., 2008). It has been demonstrated that NO plays an important role in auxin-regulated signaling cascades, influencing root growth and development (Pagnussat et al., 2003). NO is produced by A. brasilense Sp245 under aerobic Ibrutinib conditions, mainly owing to the activity of periplasmic nitrate reductase (Nap) (Steendhoudt et al., 2001). A nap A. brasilense mutant produces only 5% of the NO produced by the wild type and is not able to promote lateral Selleckchem Small molecule library and adventitious root formation and plant development like the wild type (Molina-Favero et al., 2008). The relationship

between NO and IAA production in A. brasilense is still to be elucidated. However, a recent study revealed that a nap mutant of A. brasilense possesses a reduced ability to induce root hair formation and nodulation by rhizobia in vetch roots. Moreover, vetch roots inoculated with this mutant secreted less nod gene inducers than roots inoculated with wild-type A. brasilense, and the indole content of the growth

solution of napA-inoculated plants was reduced at a lower rate than those of wild-type-inoculated plants (Star et al., 2011). A wide variety of taxonomically different groups of microorganisms within the Bacteria and Archaea domains produce intracellular homopolymers or copolymers containing different alkyl groups at the β position, described Nintedanib (BIBF 1120) as polybetahydroxyalkanoates (PHAs). These polymers are used as energy and carbon storage compounds (Madison & Huisman, 1999). In A. brasilense, PHAs are major determinants for overcoming periods of carbon and energy starvation (Fig. 2). Increased survival upon starvation in phosphate buffer was observed in A. brasilense Sp7 relative to a phaC (PHA synthase) mutant defective in PHA production (Kadouri et al., 2002, 2003, 2005; Castro-Sowinski et al., 2010) (Fig. 2). The abilities of A. brasilense phaC and phaZ (PHA depolymerase) mutants to tolerate and survive to various stresses, including UV-irradiation, heat, osmotic shock, desiccation, and oxidative stress, were significantly impaired as compared with wild-type cells (Kadouri et al., 2003, 2005). In addition, PHA accumulation in A. brasilense was shown to support chemotaxis, motility, and cell multiplication. Therefore, it is well established that production of PHAs in A.

Decompression Illness is a useful aid for the diver and diving me

Decompression Illness is a useful aid for the diver and diving medic, which provides a ready reference of essential knowledge of DCI. The main chapters include: 1. Nitrogen update and elimination and bubble formation; 2. Decompression illness; 3. Patent foramen ovale; 4. Oxygen first aid; and 5. The realities

of diving accidents in remote places. Chapters are consistently represented with a number of chapters including case studies, which nicely illustrate clinical issues. The booklet is hard to fault. The only possible suggestion is to expand the information on basic first aid for divers; however, there is mention of the “DRSABCD” and life-saving procedures.[2] The absence check details of an index may also be a barrier for someone wanting to quickly find information, but the limited glossary contains useful definitions of some terms commonly used in association with DCI. Decompression Illness is written by John Lippmann, who has 40 years’ experience in diving and 30 years’ experience in researching, teaching, and consulting on safe diving, decompression, and accident management. It states in “About the Author” that John is “Executive Director and Director of Training of the Divers

Alert Network (DAN) Asia-Pacific, which he helped to found in 1994” (p. 5). Decompression Illness gives concise coverage on an important diving-associated illness. It ITF2357 cell line is an essential reference for diving organizations, clinics specializing in diving medicine, and those health professionals managing DCI. ”
“We present a case of Plasmodium vivax infection in a soldier, 4 months after returning from Afghanistan. Primary care physicians should be reminded of the possible delay in presentation of P. vivax when evaluating fever and the importance of terminal prophylaxis with primaquine to prevent relapse following return from malarious regions. A 32 year-old man presented to a regional hospital complaining of 5 days of high nocturnal fever, drenching sweat, chills, severe body ache, intermittent left upper quadrant pain, and headaches. He had been previously deployed with the Army for 11 months Ergoloid in the area surrounding Jalalabad, in

northeast Afghanistan near the Pakistan border, where he reported exposure to mosquitos, fleas, ticks, and lice. He took doxycycline for malaria prophylaxis, with brief supply interruptions while in the field. After he returned to the United States, he did not continue doxycycline or take primaquine, and was healthy for 4 months until the onset of the current illness. On examination, the temperature was 39°C and there was left upper quadrant tenderness. The rest of the examination was normal. The white blood cell count was 1,800 cells/mm3(segmented 21%, bands 28%, lymphocytes 31% and abnormal lymphocytes 11%), hemoglobin was 16.3 g/dL, and platelets were 54,000/mm3. Malaria smears were negative, and abdominal imaging revealed mild splenomegaly.