Hemorrhagic Herpes Simplex Virus Type 1 Nephritis: An Unusual Cause of Acute Allograft Dysfunction.

Interstitial nephritis due to viruses is well-described after solid organ transplantation. Viruses implicated include cytomegalovirus; BK polyomavirus; Epstein-Barr virus; and, less commonly, adenovirus. We describe a rare case of hemorrhagic allograft nephritis due to herpes simplex virus type 1 at 10 days after living donor kidney transplantation. The patient had a favorable outcome with intravenous acyclovir and reduction of immunosuppression.

A comparative analysis of experimental selection on the stickleback pelvis.

Mechanisms of natural selection can be identified using experimental approaches. However, such experiments often yield nonsignificant effects and imprecise estimates of selection due to low power and small sample sizes. Combining results from multiple experimental studies might produce an aggregate estimate of selection that is more revealing than individual studies. For example, bony pelvic armour varies conspicuously among stickleback populations, and predation by vertebrate and insect predators has been hypothesized to be the main driver of this variation. Yet experimental selection studies testing these hypotheses frequently fail to find a significant effect. We experimentally manipulated length of threespine stickleback (Gasterosteus aculeatus) pelvic spines in a mesocosm experiment to test whether prickly sculpin (Cottus asper), an intraguild predator of stickleback, favours longer spines. The probability of survival was greater for stickleback with unclipped pelvic spines, but this effect was noisy and not significant. We used meta-analysis to combine the results of our mesocosm experiment with previously published experimental studies of selection on pelvic armour. We found evidence that fish predation indeed favours increased pelvic armour, with a moderate effect size. The same approach found little evidence that insect predation favours reduced pelvic armour. The causes of reduced pelvic armour in many stickleback populations remain uncertain.

Genome-wide Gene-by-Sex Interaction Studies Identify Novel Nonsyndromic Orofacial Clefts Risk Locus.

Risk loci identified through genome-wide association studies have explained about 25% of the phenotypic variations in nonsyndromic orofacial clefts (nsOFCs) on the liability scale. Despite the notable sex differences in the incidences of the different cleft types, investigation of loci for sex-specific effects has been understudied. To explore the sex-specific effects in genetic etiology of nsOFCs, we conducted a genome-wide gene × sex (GxSex) interaction study in a sub-Saharan African orofacial cleft cohort. The sample included 1,019 nonsyndromic orofacial cleft cases (814 cleft lip with or without cleft palate and 205 cleft palate only) and 2,159 controls recruited from 3 sites (Ethiopia, Ghana, and Nigeria). An additive logistic model was used to examine the joint effects of the genotype and GxSex interaction. Furthermore, we examined loci with suggestive significance (P < 1E-5) in the additive model for the effect of the GxSex interaction only. We identified a novel risk locus on chromosome 8p22 with genome-wide significant joint and GxSex interaction effects (rs2720555, p2df = 1.16E-08, pGxSex = 1.49E-09, odds ratio [OR] = 0.44, 95% CI = 0.34 to 0.57). For males, the risk of cleft lip with or without cleft palate at this locus decreases with additional copies of the minor allele (p < 0.0001, OR = 0.60, 95% CI = 0.48 to 0.74), but the effect is reversed for females (p = 0.0004, OR = 1.36, 95% CI = 1.15 to 1.60). We replicated the female-specific effect of this locus in an independent cohort (p = 0.037, OR = 1.30, 95% CI = 1.02 to 1.65), but no significant effect was found for the males (p = 0.29, OR = 0.86, 95% CI = 0.65 to 1.14). This locus is in topologically associating domain with craniofacially expressed and enriched genes during embryonic development. Rare coding mutations of some of these genes were identified in nsOFC cohorts through whole exome sequencing analysis. Our study is additional proof that genome-wide GxSex interaction analysis provides an opportunity for novel findings of loci and genes that contribute to the risk of nsOFCs.

High Schools Struggle to Adopt Evidence Based Practices for the Management of Exertional Heat Stroke.

CONTEXT: Exertional heat stroke (EHS) deaths can be prevented by adhering to best practices. OBJECTIVE: We investigated the adoption of policies and procedures for the recognition and treatment of EHS and the factors influencing the adoption of a comprehensive policy. DESIGN: Cross Sectional. SETTING: Online questionnaire. PATIENTS OR OTHER PARTICIPANTS: Athletic trainers (ATs) practicing in the high school (HS) setting. MAIN OUTCOME MEASURE(S): Using the NATA Position Statement: Exertional Heat Illness, an online questionnaire was developed and distributed to ATs to ascertain their schools' current written policies for the use of rectal temperature and cold-water immersion (CWI). The Precaution Adoption Process Model (PAPM), allowed for responses to be presented across the various health behavior stages ("Unaware if have the policy", "Unaware for the need for the policy", "Unengaged", "Undecided", "Decided Not to Act", "Decided to Act", "Acting", and "Maintaining"). Additional questions included perceptions of facilitators and barriers. Data are presented as proportions. RESULTS: A total of 531 ATs completed this questionnaire. Overall, 16.9% (n=62) report adoption of all components for proper recognition and treatment of EHS. The policy component with the highest adoption was "cool first transport second" with 74.1% (n=110) of ATs reporting "Acting" or "Maintaining." The most variability in the PAPM responses was for a rectal temperature policy, with 28.7% (n=103) of ATs reporting "Decided not to Act" and 20.1% (n=72) reporting "Maintaining." The most commonly reported facilitator and barrier for rectal temperature included state mandate from state HS athletics association (n=274,51.5%) and resistance or apprehension from parents or legal guardians (n=311,58.5%), respectively. CONCLUSIONS: ATs in the HS setting appear to be struggling to adopt a comprehensive EHS strategy, with rectal temperature continuing to appear as the biggest undertaking. Tailored strategies based on health behavior, facilitators and barriers may aid in changing this paradigm.

Monoclonal antibodies against human T cell leukemia-lymphoma virus (HTLV) p24 internal core protein. Use as diagnostic probes and cellular localization of HTLV.

Four monoclonal antibodies, human T cell leukemia-lymphoma virus (HTLV) 6, 7, 8, and 9, which react with the 24,000 dalton internal core protein of HTLVI, have been developed. These monoclonal antibodies reacted with only HTLV-infected cells and not with a broad spectrum of normal, neoplastic, mitogen-stimulated, or virus-infected cells and tissues. HTLV 6, 7, 8, and 9 identified at least two different antigenic determinants on HTLV p24 that were also recognized by antibodies present in HTLV+ patient sera. Monoclonal antibodies HTLV 6, 7, 8, and 9 reacted in indirect immunofluorescence assays with HTLV p24 localized at the cell surface of 5-d cultures of HTLV-infected T cells and, as well, reacted with T cells infected with HTLVII, a new type of HTLV isolated from a patient (MO) with a T cell variant of hairy cell leukemia. Thus, HTLV 6, 7, 8, and 9 should prove to be useful diagnostic reagents in the identification of HTLV- and HTLVII-infected T cells.

Developmental arrest during larval life and life-span extension in a marine mollusc.

The length of larval life in the nudibranch Phestilia sibogae is determined by a chance encounter with a specific metamorphic stimulus associated with the post-larval benthic habitat. A developmental hiatus begins at the onset of larval metamorphic competence and ends at metamorphosis; aging is suspended during this hiatus. Because the duration of post-larval life is unaffected by the duration of larval life, total life-span varies with the length of the larval period. Developmental control of the timing of expression of life-history stages is an important factor regulating aging and senescence in animals with complex life cycles.

TDP-43 accumulation in inclusion body myopathy muscle suggests a common pathogenic mechanism with frontotemporal dementia.

TAR DNA binding protein-43 (TDP-43) is found in ubiquitinated inclusions (UBIs) in some frontotemporal dementias (FTD-U). One form of FTD-U, due to mutations in the valosin containing protein (VCP) gene, occurs with an inclusion body myopathy (IBMPFD). Since IBMPFD brain has TDP-43 in UBIs, we looked for TDP-43 inclusions in IBMPFD muscle. In normal muscle, TDP-43 is present in nuclei. In IBMPFD muscle, TDP-43 is additionally present as large inclusions within UBIs in muscle cytoplasm. TDP-43 inclusions were also found in 78% of sporadic inclusion body myositis (sIBM) muscles. In IBMPFD and sIBM muscle, TDP-43 migrated with an additional band on immunoblot similar to that reported in FTD-U brains. This study adds sIBM and hereditary inclusion body myopathies to the growing list of TDP-43 positive inclusion diseases.

Executive functioning in low birth weight children entering kindergarten.

OBJECTIVES: Poor executive functioning is associated with life-long difficulty. Identification of children at risk for executive dysfunction is important for early intervention to improve neurodevelopmental outcomes. STUDY DESIGN: This study is designed to examine relationships between birthweight and executive functioning in US children during kindergarten. Our hypothesis was that children with higher birthweights would have better executive function scores. We evaluated data from 17506 US children from the Early Childhood Longitudinal Study-Kindergarten 2011 cohort. Birthweight and gestational age were obtained by parental survey. Executive functions were directly assessed using the number reverse test and card sort test to measure working memory and cognitive flexibility, respectively. Teacher evaluations were used for additional executive functions. Data were analyzed using SAS to run all linear and logistical regressions. RESULTS: For every kilogram of birthweight, scores of working memory increased by 1.47 (P<0.001) and cognitive flexibility increased by 0.28 (P<0.001) independent of gender, gestational age, parental education, and family income. Low birthweight infants were 1.5 times more likely to score in the bottom 20% of children on direct assessment OR=1.49 (CI 1.21-1.85) and OR=1.55 (CI 1.26-1.91). CONCLUSIONS: Infants born low birthweight are at increased risk of poor executive functioning. As birthweight increases executive function scores improve, even among infants born normal weight. Further evaluation of this population including interventions and progression through school is needed.

Associations between birthweight and overweight and obesity in school-age children.

BACKGROUND: Relationships between birthweight and future obesity risk remain unclear. OBJECTIVE: To assess associations between birthweight and later obesity in a nationally representative cohort of early school-aged children. METHODS: We used linear and logistic regression to evaluate 10 186 term- or preterm children in the Early Childhood Longitudinal Study-Kindergarten Cohort 2011 for relationships between birthweight and later obesity and change in BMI z-score from kindergarten-to-second grade. All analyses were adjusted for sex, race/ethnicity, parental education and household income. RESULTS: Compared to children born normal birthweight (NBW), high birthweight (HBW) term children and large-for-gestational-age (LGA) preterm children had significantly greater BMI z-scores from kindergarten-to-second grade (p < 0.001). Term children born HBW had higher odds of obesity by kindergarten (adjusted odds ratios [aOR] 1.91, p < 0.0001). Among preterm children, odds of obesity was higher among LGA children starting in first grade (aOR 2.34, p < 0.05) and among small-for-gestational age children in second grade (aOR 2.26, p < 0.05). Compared to NBW children, HBW children had greater change in BMI z-score between kindergarten-first grade (p < 0.01). CONCLUSIONS: High birthweight term and LGA preterm children had increased adjusted odds of obesity in school-age compared to their NBW counterparts. Physicians may provide counselling early in life for families of large infants to help prevent future obesity.

Scanning electron microscopy studies in muscular dystrophy.

Scanning electron microscopy of unmanipulated erythrocytes from patients with myotonic dystrophy or Duchenne dystrophy and patients who were Duchenne carriers showed a large increase in the number of stomatocytes over the number in normal controls. No specific morhologic changes that would differentiate any of the dystrophic patients from one another were seen. Adverse conditions such as washing before fixation or extreme pH produced a greater change in erythrocytes from these patients than in those from normal controls.

A direct comparison of procedures for the detection of mycoplasma in tissue culture.

Mycoplasma contamination of cell cultures has been shown to perturb a number of immunologic parameters. Because such contamination is almost always introduced in the laboratory, the immunologist requires a procedure to screen his cell lines frequently for mycoplasma. Two procedures recently described for the detection of mycoplasma in cell cultures, the uridine-uracil incorporation procedure and a direct fluorescent assay, were compared with the standard procedures of agar culture and transmission electron microscopy. The results with uridine-uracil incorporation were totally non-concordant with those of any of the other 3 procedures and, moreover, were inconsistent through serial assays on the same cell culture. In contrast, the direct fluorescent assay, using the fluorochrome Hoechst 33258, yielded consistent results in full agreement with the agar culture data. Since the fluorescent assay is rapid and has discriminatory capability at least equivalent to that of agar culture, it would appear to be the method of choice for routine screening of cell cultures for mycoplasma.

Identification of viral infection by confocal microscopy.

Confocal microscopy is a valuable adjunct to electron microscopy in the fields of diagnostic and investigative virology. Confocal imaging can be used to examine large amounts of tissue stained by a variety of methods for evidence of viral infection. Areas thus identified can then be processed for ultrastructural study, allowing a highly focused search for viral pathogens. With the possible exception of the vibrating microtome, all of the equipment and reagents necessary for the preparation of specimens for confocal scanning are available in any well-stocked histology laboratory. Although originally developed to facilitate viral diagnosis by EM, the methods described herein can be applied to the ultrastructural study of any focal pathologic process.

Electron microscopic diagnosis of human flavivirus encephalitis: use of confocal microscopy as an aid.

The distinction between intracranial viral infections and inflammatory conditions requiring immunosuppression is important. Although specific laboratory reagents are readily available for some viruses, diagnosis of arbovirus infection is more difficult. Transmission electron microscopy (TEM) theoretically allows identification of viral particles independent of reagent availability, but it has limited sensitivity. We report two cases of human flavivirus encephalitis diagnosed by TEM. Laser scanning confocal microscopy (LSCM) was used in one case to survey unembedded tissue slices for focal abnormalities, from which fragments smaller than 1 mm2 were excised for epoxy embedding. This facilitated TEM identification of intracytoplasmic, budding, 35-40 nm spherical virus particles, confirmed by serology as St. Louis encephalitis. In contrast to mosquitoes and newborn mice, in which high viral loads are associated with minimal tissue responses, these biopsies showed florid angiodestructive inflammation and microgliosis, with rare virions in necrotic perivascular cells and astrocytes. To our knowledge, this represents the first ultrastructural study of St. Louis encephalitis in humans, indicating the potential value of LSCM-aided TEM.

Diagnosis and management of BK polyomavirus interstitial nephritis in renal transplant recipients.

BACKGROUND: Interstitial nephritis caused by BK polyomavirus is a recognized complication of renal transplantation. A study of renal transplant recipients at Duke University Medical Center was undertaken to evaluate diagnostic modalities and assess clinical outcomes in transplant polyomavirus infections. METHODS: Polyomavirus nephritis was identified in 6 of 240 patients who received renal transplants between January 1996 and June 1998 and an additional patient who underwent transplantation in 1995. The clinical records of these seven patients were reviewed, as were all renal biopsy and nephrectomy specimens. Electron microscopy (EM) was performed on negatively stained urine samples from 6 patients with polyomavirus infection and 23 patients with other diagnoses. RESULTS: Patients with polyomavirus infection shared several clinical features, including ureteral obstruction (5/7 patients), lymphocele (3/7), bacterial urinary tract infection (3/7), hematuria (3/7), cytomegalovirus infection (3/7), and immunosuppression with mycophenolate mofetil (6/7). All patients experienced elevations in serum creatinine, which stabilized or decreased in four patients with altered or decreased immunosuppression. The diagnosis of polyomavirus infection was established by renal biopsy and EM of urine in five patients, by biopsy alone in one, and by EM alone in one. Sequential examinations of urine by EM were used to monitor the course of infection in six patients. CONCLUSIONS: Interstitial nephritis due to BK polyomavirus occurred in 2.5% of patients receiving renal transplants at our center since 1996. Polyomavirus infection can cause transplant dysfunction and graft loss, but progression of the infection can frequently be abrogated with alterations in immunosuppressive therapy. Both renal biopsy and EM of urine samples are useful in the diagnosis and monitoring of polyomavirus infections.

Characterization of a novel family of ciliary body glycoproteins.

PURPOSE: To isolate and characterize ciliary body epithelial antigens reactive with a monoclonal antibody, 2B4.14.1. METHODS: A mouse monoclonal antibody generated against human corneal endothelium, 2B4.14.1 reacts with nonpigmented epithelium of human and guinea pig ciliary bodies. The ciliary body proteins reactive with 2B4.14.1 were identified by Western blotting and were partially purified by affinity chromatography with 2B4.14.1 coupled to a solid support matrix. Carbohydrate components of the antigenic molecules were analyzed by lectin chromatography and by digestion with the enzymes N-glycosidase F and endoglycosidase F. The cellular and subcellular distribution of the antigens was examined by immunoperoxidase staining and by immunoelectron microscopy of ultracryotome sections of ciliary body. RESULTS: 2B4.14.1 reacted with families of guinea pig and human ciliary body glycoproteins with estimated molecular weights ranging from 250 to 325 kD. In Western blots of samples reduced before electrophoresis, the high molecular weight bands were replaced by weakly reactive bands at 115 to 130 and 210 kD, indicating that the 2B4.14.1 ligands have disulfide bonds. 2B4.14.1 ligands from both guinea pig and human ciliary body were bound by immobilized lectins, including concanavalin A, Datura stramonium lectin, and Lens culinaris hemagglutinin, which recognize components of N-linked oligosaccharides. Guinea pig ciliary body antigens digested with N-glycosidase F and endoglycosidase F failed to react with 2B4.14.1 in Western blots, confirming the presence of N-linked oligosaccharide chains and indicating that they form an integral part of the 2B4.14.1-reactive antigenic site. Molecular weight shifts of glycosidase-digested antigens were consistent with the presence of two to four N-linked oligosaccharide units. In immunoperoxidase-stained sections of guinea pig and human ciliary body, 2B4.14.1 reacted primarily with nonpigmented epithelial cells. Staining of guinea pig epithelial cells was fairly uniform; staining of human epithelial cells was concentrated on the basal surface. By immunoelectron microscopy, a majority of the 2B4.14.1 antigenic reactivity was localized immediately external to the nonpigmented epithelial cell plasma membrane. CONCLUSIONS: 2B4.14.1 reacts with a novel family of high molecular weight glycoproteins associated with the nonpigmented epithelial cell surface in guinea pig and human ciliary body.

Expression of a copper-containing amine oxidase by human ciliary body.

PURPOSE: To examine the molecular structure and ultrastructural distribution of a novel amine oxidase in human ciliary body. METHODS: Human ciliary bodies were solubilized with a nonionic detergent. The solubilized material was subjected to affinity chromatography with 2B4.14.1, a monoclonal antibody which recognizes a family of ciliary body glycoproteins. Proteins eluted from the affinity column were further separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Peptides produced from a 2B4.14. 1-reactive protein with an approximate molecular weight of 100 kDa were analyzed by Edman degradation. The protein thus identified was further examined by Western blotting and immunoelectron microscopy with anti-peptide antisera. RESULTS: Peptide sequences from the 100 kDa ciliary body protein were identical to the predicted protein sequence of an amine oxidase identified recently in a human placental cDNA library. The identity of the ciliary body protein was confirmed by Western blotting with rabbit antiserum generated against the predicted carboxy-terminal peptide of human placenta amine oxidase. Western blotting under nonreducing conditions and following glycosidase digestion indicated that the native enzyme is a disulfide-linked homodimer with multiple N-linked oligosaccharide side chains. By immunoelectron microscopy, the ciliary body amine oxidase was localized to the plasma membranes of inner epithelial cells. CONCLUSIONS: Human placenta amine oxidase is present on the plasma membranes of ciliary body inner epithelial cells. This finding provides a potential explanation for amine oxidase enzyme activity detected in previous studies of anterior segment tissues. Though the functional role of human placenta amine oxidase in the eye is unclear, it may contribute to the production of H2O2 in aqueous humor.

Special techniques in diagnostic electron microscopy.

The power of electron microscopy as a diagnostic tool can be amplified considerably by the application of ancillary preparative and analytic methods. Subcellular chemistry and structure can be examined by various forms of microprobe analysis and by special staining methods, including cytochemical, immunocytochemical, and negative staining. Qualitative ultrastructural examination can be augmented by morphometric analysis. Correlative microscopic survey methods can be used as a means of targeting ultrastructural investigations. This article provides an overview of the use of these special techniques in the diagnosis and classification of tumors and other selected pathologic processes.

Community respiratory viral infection in adult lung transplant recipients.

STUDY OBJECTIVE: To define the epidemiology, clinical manifestations, and long-term complications of respiratory viral infections in adult lung transplant recipients. DESIGN: Retrospective review of the records of 122 adult lung transplant recipients over a 5-year period at one institution. RESULTS: Ten episodes of infection with respiratory syncytial virus, parainfluenza, influenza, or adenovirus were identified. All patients presented with symptoms of respiratory tract infection. Two patients died acutely and four patients subsequently had development of obliterative bronchiolitis (OB). CONCLUSIONS: These data suggest community respiratory viral infections cause significant morbidity and mortality in lung transplant recipients. Further prospective studies are warranted to clarify the relationship between respiratory viral infection and OB and to define the optimal therapy for these viral infections.

Six-year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada.

To identify the prevalence, seasonality and demographic characteristics of patients with viral gastroenteritis, we reviewed 6 years of retrospective data on viral agents of gastroenteritis screened by electron microscopy at 10 centers in the United States and Canada. From 52,691 individual electron microscopic observations, a virus was detected in 16% of specimens, and the yearly positive detection rate among centers ranged from 8 to 34%. Rotavirus was the agent most commonly observed (26 to 83%), followed by adenoviruses (8 to 27%, respiratory and enteric combined), and small round viruses (SRVs) (0 to 40%) which were second most common at two of the centers. Rotavirus and astrovirus detections occurred more often in the winter but seasonal trends in detection were not apparent for the other viruses. Of all astroviruses detected 64% were found in infants (less than 1 year); unlike the other agents studied SRVs were detected in a large percentage of infants (48%) and older children and adults (20%). Among hospitalized patients a majority of all astroviruses, caliciviruses and SRVs were detected 7 days or more after admission in contrast to both rotaviruses and adenoviruses which were more likely to be detected earlier. The data suggest that SRVs are common agents of gastroenteritis and may be important causes of nosocomial infections. Because of the relative insensitivity of direct electron microscopy as a screening method for SRVs, astroviruses and caliciviruses, these data probably underestimate the true prevalence of disease caused by these agents.