Anti-Vel alloimmunization and severe hemolytic disease of the fetus and newborn.

Only rare cases of anti-Vel-associated mild-to-moderate hemolytic disease of the fetus and newborn have been previously reported. No case of fetal anemia requiring prenatal therapy has been noted to date. We report such a case recently encountered at our Fetal Center. Strategies are discussed for managing pregnancy complicated with alloimmunization to an antibody to a high-prevalence antigen, including sources of red blood cells for intrauterine transfusions.Only rare cases of anti-Vel­associated mild-to-moderate hemolytic disease of the fetus and newborn have been previously reported. No case of fetal anemia requiring prenatal therapy has been noted to date. We report such a case recently encountered at our Fetal Center. Strategies are discussed for managing pregnancy complicated with alloimmunization to an antibody to a high-prevalence antigen, including sources of red blood cells for intrauterine transfusions.

Trypanosoma cruzi screening in Texas blood donors, 2008-2012.

Chagas disease is an important emerging disease in Texas that results in cardiomyopathy in about 30% of those infected with the parasite Trypanosoma cruzi. Between the years 2008 and 2012, about 1/6500 blood donors were T. cruzi antibody-confirmed positive. We found older persons and minority populations, particularly Hispanic, at highest risk for screening positive for T. cruzi antibodies during routine blood donation. Zip code analysis determined that T. cruzi is associated with poverty. Chagas disease has a significant disease burden and is a cause of substantial economic losses in Texas.

Development of multiplex PCR-ligase detection reaction assay for detection of West Nile virus.

We have developed a novel multiplex reverse transcription-PCR ligase detection reaction (RT-PCR/LDR) assay for the detection of West Nile virus (WNV) in both clinical and mosquito pool samples. The method relies on the amplification of three different genomic regions, one in the coding sequence of nonstructural protein NS2a and two in nonstructural protein NS5, to minimize the risk of detection failure due to genetic variation. The sensitivity of the PCR is complemented by the high specificity of the LDR step, and the detection of the LDR products can be achieved with capillary electrophoresis (CE) or a universal DNA microarray. We evaluated the limit of detection by both one-step and two-step multiplex RT-PCR/LDR/CE approaches, which reached, respectively, 0.005 and 0.017 PFU. The assay demonstrated 99% sensitivity when mosquito pool samples were tested and 100% sensitivity with clinical samples when the one-step approach was used. The broad strain coverage was confirmed by testing 34 WNV isolates belonging to lineages 1 and 2, and the high specificity of the assay was determined by testing other flaviviruses, as well as negative mosquito pool and clinical samples. In summary, the multiplex RT-PCR/LDR assay could represent a valuable complement to WNV serological diagnosis, especially in early symptomatic patients. In addition, the multiplexing capacity of the technique, which can be coupled to universal DNA microarray detection, makes it an amenable tool to develop a more comprehensive assay for viral pathogens.

Isolation of Stachybotrys from the lung of a child with pulmonary hemosiderosis.

Recently, Stachybotrys atra, a toxigenic fungus, has been implicated as a potential cause of pulmonary hemorrhage/hemosiderosis in infants living in water-damaged homes. Although epidemiologic evidence supports this association, neither the organism nor its toxic products has ever been recovered from humans. We report the first case in which Stachybotrys was isolated from the bronchoalveolar lavage fluid of a child with pulmonary hemorrhage. Stachybotrys was also recovered from his water-damaged home. The patient recovered completely after his immediate removal from the environment and subsequent cleaning of his home. This case provides further evidence that this fungus is capable of causing pulmonary hemorrhage in children.

Disinfection of eyelid speculums for retinopathy of prematurity examination.

OBJECTIVE: To evaluate the effectiveness of 70% isopropyl alcohol swabs in disinfecting eyelid speculums after examination for retinopathy of prematurity. METHODS: Two phases. Phase 1: 46 autoclave-sterilized eyelid speculums randomized into either a cleaned or control group following examination for retinopathy of prematurity. Speculums in the cleaned group were disinfected with a 70% isopropyl alcohol swab while control speculums were not cleaned. Bacterial and fungal cultures were then obtained. Phase 2: 20 autoclave-sterilized eyelid speculums inoculated with a clinically relevant dilution of adenovirus serotype 5 or herpes simplex virus type 2. Inoculated speculums were randomized into either a cleaned or control group. RESULTS: Phase 1: 17 (70.8%) of 24 cultures from the cleaned group yielded bacteria compared with 21 (95.5%) of 22 controls. Fungi were isolated from only 1 control and from no cleaned speculums. Phase 2: all speculums inoculated with adenovirus supported growth of the organism irrespective of cleaning with 70% isopropyl alcohol swabs. None of 5 cleaned speculums inoculated with herpes simplex virus type 2 supported viral growth, compared with 3 (60%) of 5 cultures positive for growth in the control group. CONCLUSION: Cleaning eyelid speculums with 70% isopropyl alcohol swabs provided inadequate disinfection against bacteria following examination for retinopathy of prematurity and against adenovirus in a laboratory simulation.

Oral manifestations of pediatric vertical HIV infection.

To assess the prevalence and prognostic significance of the history of oral manifestations in children with human immunodeficiency virus infection (HIV), a cohort study of 73 children with vertical HIV infection was conducted. The study subjects were examined every 6 months for oral manifestations. The period prevalence of oral manifestations ranged from a low of 1% for submandibular enlargement and 3% for hairy leukoplakia to a high of 36% for xerostomia and 51% for cervical lymphadenopathy. The occurrence of oral manifestations did not change significantly over time from 1995 to 1998. Finally, the odds of occurrence of cervical lymphadenopathy, xerostomia, and oral candidiasis were greater among children in whom these manifestations had been diagnosed in the preceding 6-18 months than in children without prior diagnosis. Oral manifestations are significant clinical outcomes in pediatric vertical HIV infection, particularly for children diagnosed previously with an oral manifestation.

Dental caries in HIV-infected children: a longitudinal study.

PURPOSE: The purpose of this descriptive longitudinal clinical study was to determine primary and permanent dentition caries status in HIV-infected children, and to compare caries status with the CD4 percentage (CD4%) and immune suppression category. MATERIALS AND METHODS: 73 children up to 9 years of age with vertical HIV transmission were evaluated for caries in the primary dentition at baseline and at 6 month intervals over a 30 month period; while 19 HIV-infected children between 5 and 11 years of age had their permanent dentition evaluated for caries at baseline and at 6 month intervals over a 24 month period. Caries status was also compared with CDC CD4 percentage (> 25%, 15-24%, < 15%), and CDC immune suppression categories (immune suppression: none, moderate, severe). With primary dentition caries, comparisons were made among all children (2-9 yr-olds, N = 73), < 2 yr-olds (N = 28), 2 to 4 yr-olds (N = 20), and 5 to 9 yr-olds (N = 25), and compared with NHANES III data. Caries-free status was also determined. RESULTS: During the 30-month period, there was an almost two-fold increase in primary tooth surface caries for the 2 to 9 year-olds. Caries-free status in the primary dentition declined from 60% at baseline to 37% at the 30-month period. With 5 to 11 years-olds, DMFS and DMFT remained relatively stable, while the proportion of caries-free individuals declined from 72% at baseline to 50% at 18 months. Caries in the primary dentition was increased substantially for those in the low CDC CD4 percentage categories and CDC moderate to severe immune suppression categories. CONCLUSION: Primary dentition caries status in HIV-infected children is considerably greater than that for the US pediatric population, and increases with decreasing CD4 percentage and moderate to severe immune suppression. HIV-infected children with caries-free primary dentitions are less frequent than in the US pediatric population, and caries-free status decreases with age, lower CD4 percentage and moderate to severe immune suppression.

Detection of fungal organisms in saliva from HIV-infected children: a preliminary cytologic analysis.

PURPOSE: Fungal infections in HIV-infected individuals are associated with advancement of disease. In pediatric HIV infection, symptomatic children have a significantly higher incidence of clinical candidiasis and persistent drug-resistant candidiasis than do asymptomatic HIV-infected children. The purpose of this preliminary cytologic study was to determine the prevalence of fungal organisms in whole unstimulated saliva from children with vertically acquired HIV infection. METHODS: The subjects included 27 HIV-infected and 11 HIV-exposed, but uninfected, children. Whole unstimulated saliva was obtained for cytologic evaluation (hematoxylin and eosin, silver stains) with selected samples evaluated by electron microscopy. RESULTS: Yeast and hyphae were identified cytologically in 19% of HIV-infected (22% symptomatic HIV-infected, 11% asymptomatic HIV-infected) and 9% of HIV-exposed, but uninfected, children. Fungal organisms were found more frequently in HIV-infected with moderate (18%) and severe (27%) suppression. Fungi were more frequent with antiretroviral therapy (22%) vs no antiretroviral therapy (0%) and no antifungal therapy (20%) vs. antifungal therapy (7%). Yeast and hyphal fungal forms are more prevalent in symptomatic HIV-infection with moderate and severe suppression, and those receiving antiretroviral agents, but no antifungal medications. CONCLUSION: Fungal organisms in the saliva may reflect oral carriage or mucosal colonization, which may influence the development of clinically significant candidiasis in these immunocompromised children.

Breastmilk cultures and infection in extremely premature infants.

OBJECTIVE: As expressed mother's milk (MM) is known to be colonized by microbial species, it is occasionally considered as a source of infection in premature infants, prompting some clinicians to obtain milk bacterial culture results before infant feeding. To determine whether serial microbial cultures of MM predict infection in premature infants. STUDY DESIGN: Milk microbial flora was determined by plate counts from aliquots of MM obtained from 161 mothers of infants born <30 weeks gestation (n = 209). Pathogens isolated from the same infant were tabulated. RESULT: Milk samples (n = 813) yielded 1963 isolates. There were no relationships between microbial counts and maternal age, ethnicity, education, skin-to-skin contact and infant infection. In 64 infants, milk and pathological isolates had presumptively the same Gram-positive organism, yet the odds of infection before or after exposure to milk containing that Gram-positive organism were not significant (1.18; 95% confidence interval=0.51, 2.76). In eight infants, milk and pathological isolates had presumptively the same Gram-negative organism, which appeared sporadically in milk, either before or after isolation in the infant. CONCLUSION: Results of initial milk cultures do not predict subsequent culture results. Random milk cultures, even if obtained at any time during hospitalization, are not predictive of infection in premature infants. The sporadic nature of the appearance of certain isolates, however, suggests common exposure of both mother and infant. Routine milk cultures do not provide sufficient data to be useful in clinical management.

Dematiaceous fungi are an increasing cause of human disease.

The dematiaceous fungi appear to be an increasing cause of human disease. At The Methodist Hospital, in Houston, Texas, five cases of serious disease caused by these fungi occurred between 1987 and 1992. Cerebral abscesses with Xylohypha bantiana followed treatment for lymphoma. An infection of the lower extremity with Exophiala jeanselmei var. castellanii followed cardiac surgery. Peritoneal growth of Alternaria tenuissima was a complication of peritoneal dialysis. Cerebral abscesses with Dactylaria gallopava occurred in a liver transplantation patient. A traumatic ankle wound contaminated with dirt led to an infection with Phialophora repens. All patients except the last were immunocompromised at the time of the infection; diabetics and patients on steroids may be at particular risk.

Trichoderma longibrachiatum infection in a pediatric patient with aplastic anemia.

Trichoderma longibrachiatum infection of the skin in an 11-year-old child with severe aplastic anemia and prolonged neutropenia is reported. The patient received systemic antifungal therapy and underwent bone marrow transplantation. To our knowledge, this is the first description of T. longibrachiatum infection in a pediatric patient. It also is the first case successfully treated with medical therapy. A review of the literature suggests that Trichoderma spp. are recognized as human pathogens with increasing frequency, particularly for immunocompromised patients, and should be considered in the differential diagnosis of fungal infections in the pediatric population.

Infantile cytomegalovirus-associated autoimmune hemolytic anemia.

Autoimmune hemolytic anemia (AIHA) is a hematologic disorder that is rarely seen in infants and young children. Most cases are associated with viral or bacterial infection, but the immunologic events leading to hemolysis are poorly understood. We describe two infants with severe cytomegalovirus (CMV)-associated warm antibody AIHA. One case was immunohematologically analyzed and showed suggestive evidence that endogenous anti-CMV IgG antibodies were the pathogenic antibodies leading to hemolysis, implicating a possible causal relationship between AIHA and CMV infection. Both patients were ultimately treated with intravenous CMV immune globulin, with subsequent improvement. These cases suggest that investigation for the presence of CMV in infantile AIHA is warranted and that CMV immune globulin should be considered as a therapeutic option.

Saliva collection technique for cytologic, microbiologic and viral evaluation in pediatric HIV infection.

Acquisition of saliva for biologic, immunologic and chemical analyses has been extremely difficult in infants and young children due to lack of cooperation and motor skills necessary for expectorating adequately. The purpose of this study was to investigate a technique for obtaining satisfactory quantities of whole, unstimulated saliva in the typical dental operatory setting for cytologic, microbiologic and viral evaluation, while requiring minimal cooperation and motor skills from pediatric patients. A low vacuum-assisted aspiration device was utilized to obtain samples from infants and children who were at risk for vertically acquired HIV-infection (age-range 6 mos to 8 yrs). Adequate saliva samples were collected in 175 of 196 (89 percent) attempts in 88 of 89 (99 percent) children (2.3 samples/child). Saliva was not obtained in twenty-one attempts primarily due to xerostomia (62.5 percent). Saliva sample volume obtained was variable, ranging from 1.2 to 3.6 mls with a collection time of approximately three to five minutes. Cell block preparations were made from the saliva, which allowed for cytologic evaluation of sloughed superficial squamous cells, evaluation of oral flora, and detection of yeast and hyphal fungal forms. Adequate volumes of supernate were also available for microbiologic and viral cultures, immunologic studies and PCR study for various viral agents shed in the saliva. Use of a vacuum-assisted collection device for whole unstimulated saliva in infants and young children in the dental operatory setting provides adequate saliva for multiple analyses, which may provide information regarding HIV disease status and early diagnosis of opportunistic infections.

Isolation of Lautropia mirabilis from oral cavities of human immunodeficiency virus-infected children.

Lautropia mirabilis, a pleomorphic, motile, gram-negative coccus, has been isolated from the oral cavities of 32 of 60 (53.3%) children infected with human immunodeficiency virus (HIV) and 3 of 25 (12.0%) HIV-uninfected controls; the association of L. mirabilis isolation with HIV infection is significant (P < 0.001). All children in the study, both HIV-infected children and controls, were born to HIV-infected mothers. The presence of this bacterium was not associated with clinical disease in these children. The HIV-infected children with L. mirabilis did not differ from the HIV-infected children without L. mirabilis in immunological status, clinical status, or systemic medications. The role of HIV infection itself or concomitant factors in the establishment of L. mirabilis in the oral cavity remains to be elucidated.