Molecular Proof Research Articles

Mucopolysaccharidosis type I: founder effect of the p.P533R mutation in North Africa

BackgroundMucopolysaccharidosis type I is a lysosomal storage disease resulting from a deficiency in alpha-L-iduronidase (IDUA), which causes the accumulation of partially degraded dermatan sulfate and heparan sulfate. This retrospective study, spanning eight years, analyzed data from 45 MPSI patients. The report aimed to explore the potential origin of the p.P533R mutation in the Maghrebin population by constructing a single-nucleotide polymorphism haplotype around the IDUA gene, in order to propose a molecular proof of a founder effect of the MPSI/p.P533R allele.Patients and methodsAll of the studied patients were from Libya (2), Mauritania (1) Morocco (21) and Tunisia (21) with first cousins being the most frequent union. The diagnosis of MPSI patients often involves the combination of urinary screening, leukocyte IDUA activity determination, and DNA molecular analysis. In our study, to identify the common p.P533R mutation, we performed both DNA sequencing and tetra-primer ARMS PCR assay. Additionally, Haploview was used to determine the specific haplotype that cosegregates with the p.P533R mutation. Controls were genotyped to ensure that all the SNPs were in Hardy–Weinberg equilibrium.ResultsIn the present report we confirmed the very strong impact of consanguinity on the incidence of MPSI disease. Furthermore, studied families of mixed ancestry shared common and specific haplotype, which was observed in studied populations, suggesting the presence of a founder effect in the North Africa.ConclusionThe p.P533R missense mutation was identified in each patient originated from Libya, Mauritania, Morocco and Tunisia. Furthermore, these MPSI patients exhibited the same IDUA haplotype. The occurrence of a shared AAGGGTG haplotype, among North African populations may be attributed to substantial historical gene exchange between these groups, likely stemming from migration, inter-ethnic marriage, or other forms of interaction throughout history.

Paternal peri-conceptional physical activity and the risk of congenital heart disease in offspring: A case-control study.

Genetic and environmental factors have been shown to contribute to the development of congenital heart disease (CHD). To date, the focus of scientific articles has primarily centered on genetics and maternal environmental factors, with comparatively less attention given to paternal risk factors. This study aims to investigate the potential association between paternal pre-conceptional physical activity levels (PA), along with paternal peri-conceptional smoking and alcohol consumption, and the risk of CHD in offspring. An observational case-control study was conducted in Lebanon, with 279 participants, aiming to investigate potential risk factors for CHD. We included children with confirmed CHD, born between 2012 and 2022. Controls born in the same timeframe were selected randomly from the general population using online questionnaire forms. Mean age of children included was 6 years old (0-10). The pre-conceptional PA was assessed using the Global Physical Activity Questionnaire validated in Arabic. In addition, paternal smoking, alcohol consumption, and maternal risk factors were collected. The study included 128 CHD cases (45.9%) and their parents, as well as 151 healthy infants (54.1%) and their parents. There were no statistically significant variations in the alcohol consumption noted between the fathers in the case and control groups (p=0.18). The paternal involvement in recreational-related PA during the peri-conception period was associated with a reduced risk of the CHD development in offspring by 46.9% (OR=0.531, 95% CI: 0.301-0.936, p=0.029). Additionally, increasing paternal total sitting time by 1 h above the average, which was approximately 260 min (4h), increased the risk of CHD in offspring by 0.4% (p=0.001). Moreover, paternal smoking exhibited an apparent association with a 56% increased risk of offspring developing CHD, notwithstanding that the confidence intervals included the null (OR=1.56, 95% CI: 0.86-2.8, p=0.136). This observational study is the first to report a potential association between paternal PA, and CHD in offspring. This study aligns with previous reports, advocating for the paternal engagement in PA and the adoption of healthy lifestyle habits, especially during the critical stages of conception. Such practices are strongly recommended to enhance fertility and promote optimal health for offspring. However, due to the subjectivity in reporting PA and lack of molecular proof, additional prospective and molecular studies are required to validate these findings.

Comments and Illustrations of Ultrasound Findings in Extrapulmonary Tuberculosis Manifestations.

This review describes the appearance of extrapulmonary tuberculosis manifestations in comprehensive and multiparametric ultrasound imaging. The aim is to increase awareness of typical ultrasound findings regarding extrapulmonary tuberculosis, correlate those with pathological features, and facilitate differential diagnosis. Point of care ultrasound protocols can be used as a screening method in high-risk populations, although the negative findings do not exclude tuberculosis. Conversely, the diagnosis of extrapulmonary tuberculosis can never be made using ultrasound alone, as many ultrasound findings in extrapulmonary tuberculosis are non-specific. However, ultrasound-based sampling techniques can significantly facilitate the collection of samples for microbiological or molecular proof of tuberculosis, as well as facilitating the establishment of alternative diagnoses.

To be or not to be a superpredator: a multidisciplinary assessment of the Iberian lynx in a reintroduction social scenario

Since 2015, following reintroductions, an Iberian lynx population in Vale do Guadiana (Portugal) has been breeding and expanding, bringing changes to both ecosystems and residents’ perceptions. We describe how ecological monitoring, genetic analysis and social surveys contribute to assess this new scenario of coexistence with ecological and social repercussions. Departing from a specific case of a monitored lynx family, we present, for the first time, molecular proof of interspecific competition between lynxes and other carnivores. We assessed and compared knowledge of local key actors about the superpredator effect of the lynx previous to and following reintroduction. Data on damage experienced by livestock breeders with foxes and perceptions about it are integrated here, demonstrating how important this lynx role can be for local actors. We present proof of the killing of two foxes and a genet by lynxes through the amplification of a specific lynx DNA region and other molecular analyses carried out on saliva samples. Local actors, who previously were skeptical about the lynx’s ecological effects, do recognize its effect over other wild carnivores presently. This is a major benefit, since social perceptions have been conditioning the acceptance of the lynx, its future expansion, and the whole process of reintroduction. We also present the first documented case of a natural migrant from Doñana (SW Spain) effectively integrated and reproducing in the Vale do Guadiana population. This case study demonstrates the importance of multidisciplinary knowledge in conservation programmes and how genetics and social surveys provide complementary information for monitoring in reintroduction and, generally, conservation programs of an umbrella species.

Molecular detection of bacteria from fish lakes in Wasit province

Fish products are very susceptible to microbial contamination, rendering them perishable thus challenging the customers health. The goal of this study was to determine the molecular identification and bacterial isolates diversity obtained from infected fish skin, gills, and fins obtained from lakes Alzarkan village in Wasit Province of Iraq during the period (October 2021- May 2022). A total of 50 fish were collected and 100 samples of organ lesions were taken. For bacterial identification, the tissuees of the fish were as cut with a sterile knife and blended in the buffered peptone water. Molecular identification and the obtained PCR product were are subjected to using the primers for Flavobacterium columnare, Edwardsiella ictaluri and Aeromonas salmonicida. Molecular analyseis revealed bacterial strains high proportion of Flavobacterium columnare identification was 20/50 (40%). The higher identification rates of Flavobacterium columnare in skin 10/20 (5%), in gills 7/20 (35%) and in fins 3/20 (15%). Accurate identification of the 20 isolated Flavobacterium columnare were confirmed by PCR. For identifying certain diseases, molecular approaches provided sensitive, rapid, and precise data without the need for time-consuming traditional techniques. The Edwardsiella ictaluri and Aeromonas salmonicida were not identified. While Flavobacterium columnare termed to be first molecular proof as a farmed fish pathogen in Iraq. Keywords: Molecular detection, Bacteria, Fish lakes, Wasit province, Iraq.

Corticosteroids in COVID-19: pros and cons.

The wide and rapid spread of the COVID-19 pandemic has placed an unanticipated burden on the global healthcare sector. This necessitated a swift response from the international community to reach a solution. Efforts were made in parallel to develop preventative and therapeutic modalities. Since then, drug repurposing has blossomed as a potentially rapid resolution and has included various agents with anti-viral and anti-inflammatory properties. Corticosteroids, being potent anti-inflammatory agents, have been placed under extensive investigation. Various trials have recorded the beneficial outcome of corticosteroids in decreasing the mortality and morbidity of COVID-19. With the high pace of escalating events, the quality and study design of clinical trials are varied. Therefore, this study aims to explore the role of corticosteroids in COVID-19 disease. It inspects the molecular, pharmacologic, and clinical proof behind this theory.

Mitigation of aircraft noise-induced vascular dysfunction and oxidative stress by exercise, fasting, and pharmacological α1AMPK activation: molecular proof of a protective key role of endothelial α1AMPK against environmental noise exposure.

Environmental stressors such as traffic noise represent a global threat, accounting for 1.6 million healthy life years lost annually in Western Europe. Therefore, the noise-associated health side effects must be effectively prevented or mitigated. Non-pharmacological interventions such as physical activity or a balanced healthy diet are effective due to the activation of the adenosine monophosphate-activated protein kinase (α1AMPK). Here, we investigated for the first time in a murine model of aircraft noise-induced vascular dysfunction the potential protective role of α1AMPK activated via exercise, intermittent fasting, and pharmacological treatment. Wild-type (B6.Cg-Tg(Cdh5-cre)7Mlia/J) mice were exposed to aircraft noise [maximum sound pressure level of 85 dB(A), average sound pressure level of 72 dB(A)] for the last 4 days. The α1AMPK was stimulated by different protocols, including 5-aminoimidazole-4-carboxamide riboside application, voluntary exercise, and intermittent fasting. Four days of aircraft noise exposure produced significant endothelial dysfunction in wild-type mice aorta, mesenteric arteries, and retinal arterioles. This was associated with increased vascular oxidative stress and asymmetric dimethylarginine formation. The α1AMPK activation with all three approaches prevented endothelial dysfunction and vascular oxidative stress development, which was supported by RNA sequencing data. Endothelium-specific α1AMPK knockout markedly aggravated noise-induced vascular damage and caused a loss of mitigation effects by exercise or intermittent fasting. Our results demonstrate that endothelial-specific α1AMPK activation by pharmacological stimulation, exercise, and intermittent fasting effectively mitigates noise-induced cardiovascular damage. Future population-based studies need to clinically prove the concept of exercise/fasting-mediated mitigation of transportation noise-associated disease.

Molecular evidence of hookworms in public environment of Bangladesh

Accurate diagnosis by precise identification of causative agents is essential for the effectiveness of any control interventions. Despite high zoonotic potential, available literature on hookworms in Bangladesh is still scarce and nonspecific. The objective of this study was to determine the occurrence of hookworms in public locations across northeastern Bangladesh (Sylhet metropolitan area) using integrated parasitological and molecular assays. A total of 130 samples (80 soil and 50 environmental canine feces) were collected and examined using modified flotation technique and formalin-ether sedimentation methods. Modified plate culture was used to isolate larvae. The identification was made based on morphometric features and confirmed by amplifying the ITS region of the nuclear rDNA. Overall, 66.2% (86/130) of examined samples were positive for hookworms infection. Characteristic eggs (61–68 × 29–37 μm) and/or larvae of hookworms were observed in 73.8% (59/80) soils and 54.0% (27/50) environmental fecal samples. Rhabditiform larvae (0.48–0.54 × 0.04–0.07 mm) were observed in cultured samples. Genetic analysis of rDNA sequences revealed the presence of Ancylostoma caninum and Ancylostoma ceylanicum. In this study, hookworms' contamination of the public environment was substantial. To the best of our knowledge, this is the first molecular proof of A. caninum and A. ceylanicum observed in urban public environment in Bangladesh.

Calreticulin mutations affect its chaperone function and perturb the glycoproteome.

Calreticulin (CALR) is an endoplasmic reticulum (ER)-retained chaperone that assists glycoproteins in obtaining their structure. CALR mutations occur in patients with myeloproliferative neoplasms (MPNs), and the ER retention of CALR mutants (CALR MUT) is reduced due to a lacking KDEL sequence. Here, we investigate the impact of CALR mutations on protein structure and protein levels in MPNs by subjecting primary patient samples and CALR-mutated cell lines to limited proteolysis-coupled mass spectrometry (LiP-MS). Especially glycoproteins are differentially expressed and undergo profound structural alterations in granulocytes and cell lines with homozygous, but not with heterozygous, CALR mutations. Furthermore, homozygous CALR mutations and loss of CALR equally perturb glycoprotein integrity, suggesting that loss-of-function attributes of mutated CALR chaperones (CALR MUT) lead to glycoprotein maturation defects. Finally, by investigating the misfolding of the CALR glycoprotein client myeloperoxidase (MPO), we provide molecular proof of protein misfolding in the presence of homozygous CALR mutations.

A 21-Year-Old Woman with Suspecting Gitelman Syndrome

<p><strong>Introduction: </strong>Gitelman syndrome (GS), is a hereditary condition marked by a specific kidney function impairment. GS symptoms do not normally manifest until beyond the age of six years. We present a rare case of a 21-year-old woman with a background in generalized weakness muscle cramps with tetany, fever, and dyspnea. The goal of this study is to give a case report of Gitelman syndrome suspicion.</p><p><strong>Result:</strong> Gitelman syndrome is identified through clinical indicators and lab testing, such as low potassium and magnesium levels in the blood, metabolic alkalosis, and low calcium levels in the urine. We do a variety of physical and laboratory tests to determine the disease and rule out other possibilities. Molecular proof is difficult to get due to a lack of resources and the high expense of genetic testing, which has become one of our case report's flaws. This patient was diagnosed as suspecting Gitelman syndrome and treated for electrolyte imbalances. The patients were given replacement therapy, which comprised intravenous potassium, sodium, and chloride therapy, as well as calcium therapy. In addition, we also provided some symptomatic therapy. She was discharged five days later in fine condition.</p><p><strong>Conclusion :</strong> This is one of the few reports of a 21-year-old lady being suspected of having Gitelman syndrome (GS). We conduct a battery of physical and laboratory tests to diagnose the disease and rule out other possibilities. We provide the appropriate therapies in accordance with the guidelines, and the outcomes are favorable.<em></em></p>

Molecular detection of Coxiella-like endosymbionts and absence of Coxiella burnetii in Amblyomma mixtum from Veracruz, Mexico.

Ticks are obligate ectoparasites associated with a wide range of vertebrate hosts, including domestic animals. Moreover, ticks are capable of transmitting many pathogens such as Coxiella. To date, Coxiella burnetii, the etiological agent of coxiellosis or Q fever, is the only valid species of the genera. Nevertheless, a wide range of agents denominated Coxiella-like have been detected in recent studies, mainly associated with ticks. The pathogenicity of these Coxiella-like agents is controversial as some of them can infect both birds and humans. In Mexico, knowledge about Q fever is scarce and limited to historical serological records, and there is an overall lack of molecular proof of any agent of the genus Coxiella circulating in the country. Therefore, the aim of this study was to detect the presence of Coxiella in ticks associated with cattle in all 10 regions of Veracruz, Mexico. To accomplish this objective, first, we identified ticks collected from cattle and horses in Veracruz. Then, for Coxiella detection, DNA extraction from ticks and PCR amplification of the 16S-rDNA of Coxiella was performed. Finally, we performed a phylogenetic reconstruction to determine the Coxiella lineages detected. From the 10 regions sampled we collected 888 ticks grouped in 180 pools, and only five Amblyomma mixtum from the locality of Castán, and one from Los Angeles from Tuxpan were found positive, which represents a frequency of 20% for each locality. This study represents the first attempt at molecular detection of Coxiella in ticks associated with cattle in the state of Veracruz, the major livestock producer in the country. The findings of the present study are relevant as they establish a precedent regarding the circulation of Coxiella-like agents, as well as the absence in three municipalities of the state of Veracruz of C. burnetii, an abortive agent of livestock importance.

Synthesis, physicochemical properties, theoretical and electrochemical studies of tetraglycidyl methylenedianiline

In the current study, a tetra-functional aromatic epoxy pre-polymer namely, tetraglycidyl methylenedianiline (TGMDA) was synthesized and characterized using 1H NMR (Nuclear Magnetic Resonance) and FT-IR (Fourier Transform Infrared Spectroscopy) spectral techniques. The rheological properties were tested using an advanced rheometer in isopropyl alcohol. The effect of temperature and solution concentration on the viscosity of the aromatic pre-polymer epoxy solution was evaluated. The study of shear rheology was performed at different temperatures of solutions of the epoxy pre-polymer in isopropyl alcohol on solutions of mass concentration (%; 0.5, 1.0, 1.5, 2.5, and 5%) and temperatures (298–328 K). The energies of the frontier molecular orbitals, as calculated by B3LYP method with 6-311++G(d, p) and 6-311++G (3d, 3p) basis sets permit the prediction of the chemical reactivity and stability of the TGMDA molecule. The thermochemical estimations of the studied compounds were also calculated and discussed. The heat of formation and Gibbs's free energy of the reactants and products were determined and analyzed. The study, likewise, showed that the reactions are endothermic and spontaneous in the reverse direction at 298.15 K. Non-linear optics (NLO) study concluded the existence of the opposite relationship between the βtot and the HOMO-LUMO energy gap values (Egap). The anti-corrosive property of TGMDA for carbon steel (CS) corrosion in 1 M HCl solution was evaluated using several electrochemical and theoretical methods. The TGMDA showed highest efficiency of 96.5% at 10−3M concentration. Monte Carlo (MC) and Molecular Dynamics (MD) simulations showed that the response significantly affected the metal surface-inhibitor interaction, giving molecular proof to the geometry and the adsorption energy.

Concurrent infection of a novel genotype of hepatopancreatic parvovirus and Enterocytozoon hepatopenaei in Penaeus vannamei in Taiwan.

Hepatopancreatic parvovirus (HPV) and Enterocytozoon hepatopenaei (EHP) are emerging and reemerging pathogens in shrimps. In the present study, a novel genotype of HPV concurrently infected with EHP in Penaeus vannamei in Taiwan leading to severe atrophy and damage of hepatopancreas were confirmed by histopathology, in situ hybridization, and PCR. The novel genotype of HPV exhibited 66%-69.5% sequence identities with all known HPVs and carried unique amino acid deletions and insertions in the VP gene. According to phylogenetic analysis, the Taiwan HPV isolates were classified as the genotype IV. The present study not only provided the histopathological and molecular proof of HPV and EHP co-infection in Taiwan, but also revealed the importance of investigating the geographical expansion of novel HPV genotypes.

Tuberculosis: Stop It with Effective Treatment

Aim: Tuberculosis is one of the oldest human diseases, dating back over 17,000 years based totally on molecular proof. Despite modern diagnostic and treatment approaches, people continue to suffer from tuberculosis, and it tops the list of ten deadly infectious diseases in the world, second only to the Human Immunodeficiency Virus. TB is a worldwide pandemic, as per World Health Organization (WHO).It tops the causes of death among HIV-positive individuals. In this review we assess the challenges faced due to tuberculosis and its management and the strategies adopted to tackle it.Because of the dearth of number one health-care infrastructure in rural regions of several states which includes Health care provided privately not regulated nicely, which ends up in First- and second-line anti-TB drugs are widely used irrationally; infection with human immune deficiency virus; loss of political will; and, The most crucial, the corrupt management are all fundamental demanding situations in India's combat against tuberculosis. Another rising danger to TB eradication is multidrug-resistant tuberculosis (MDR-TB), which is the outcome of a failing or deteriorating TB control programme. The World Health Organization's "STOP TB" policy aims to eradicate tuberculosis as a public health hazard by 2050.

Metagenomic analysis reveals mixed Mycobacterium tuberculosis infection in a 18th century Hungarian midwife

The Vác Mummy Collection comprises 265 well documented mummified individuals from the late 16th to the early 18th century that were discovered in 1994 inside a crypt in Vác, Hungary. This collection offers a unique opportunity to study the relationship between humans and pathogens in the pre-antibiotic era, as previous studies have shown a high proportion of tuberculosis (TB) infections among the individuals. In this study, we recovered ancient DNA with shotgun sequencing from a rib bone sample of a 18th century midwife. This individual is part of the collection and shows clear skeletal changes that are associated with tuberculosis and syphilis. To provide molecular proof, we applied a metagenomic approach to screen for ancient pathogen DNA. While we were unsuccessful to recover any ancient Treponema pallidum DNA, we retrieved high coverage ancient TB DNA and identified a mixed infection with two distinct TB strains by detailed single-nucleotide polymorphism and phylogenetic analysis. Thereby, we have obtained comprehensive results demonstrating the long-time prevalence of mixed infections with the sublineages L4.1.2.1/Haarlem and L4.10/PGG3 within the local community in preindustrial Hungary and put them in context of sociohistorical factors.

Molecular Proof of a Clinical Concept: Expression of Estrogen Alpha-, Beta-Receptors and G Protein-Coupled Estrogen Receptor 1 (GPER) in Histologically Assessed Common Nevi, Dysplastic Nevi and Melanomas.

Background and Objectives: Epidemiologic data show significant differences in melanoma incidence and outcomes between sexes. The role of hormonal receptors in the pathogenesis of melanocytic lesions remains unclear, thus we performed this study aiming to assess estrogen receptors expression in different melanocytic lesions. Materials and Methods: We performed a cross-sectional study that included 73 consecutively excised melanocytic lesions. Estrogen receptor alpha (ERα), beta (ERβ), and G-protein coupled estrogen receptor (GPER) expression was analyzed in melanocytes and keratinocytes of common nevi, dysplastic nevi, melanoma, healthy skin margin, and in sebaceous and sweat gland cells. Results: ERβ expression was higher in dysplastic nevi margin melanocytes compared to common nevi (p = 0.046) and in dysplastic nevi keratinocytes compared to melanoma keratinocytes (p = 0.021). ERβ expression was significantly higher in margin melanocytes compared to melanoma melanocytes (p = 0.009). No difference in ERβ expression was shown between melanocytes of three types of lesions. GPER expression was higher in nuclei and cytoplasm of dysplastic nevi (p = 0.02 and p = 0.036 respectively) and at the margin compared to melanoma. GPER expression was lower in sebaceous glands of tissue surrounding common nevi (p = 0.025) compared to dysplastic nevi. GPER expression was higher in skin margin tissue melanocytes (p = 0.016 nuclear, p = 0.029 cytoplasmic) compared to melanoma melanocytes. There were no differences in ERα expression between the melanocytic lesions. Conclusion: Further large-scale studies are warranted to investigate the potential role of ERβ and GPER in the pathogenesis of melanocytic lesions.

Quantification of in-plane flexoelectricity in lipid bilayers

Lipid bilayers behave as 2D dielectric materials that undergo polarization and deformation in the presence of an electric field. This effect has been previously modeled by continuum theories which assume a polarization field oriented normal to the membrane surface. However, the molecular architecture of the lipids reveals that the headgroup dipoles are primarily oriented tangential to the membrane surface. Here, we perform atomistic and coarse-grained molecular dynamics simulations to quantify the in-plane polarization undergone by a flat bilayer and a spherical vesicle in the presence of an applied electric field. We use these predictions to compute an effective in-plane flexoelectric coefficient for four different lipid types. Our findings provide the first molecular proof of the in-plane polarization undergone by lipid bilayers and furnish the material parameter required to quantify membrane-electric field interactions.

Chronic active non-lethal human-type tuberculosis in a high royal Bavarian officer of Napoleonic times-a mummy study.

In paleopathology, morphological and molecular evidence for infection by mycobacteria of the M. tuberculosis complex (MTC) is frequently associated with early death. In the present report, we describe a multidisciplinary study of a well-preserved mummy from Napoleonic times with a long-standing tuberculous infection by M. tuberculosis senso stricto who died at the age of 88 years of focal and non-MTB related bronchopneumonia. The well-preserved natural mummy of the Royal Bavarian General, Count Heinrich LII Reuss-Köstritz (1763–1851 CE), was extensively investigated by macro- and histomorphology, whole body CT scans and organ radiography, various molecular tissue analyses, including stable isotope analysis and molecular genetic tests. We identified signs for a long-standing, but terminally inactive pulmonary tuberculosis, tuberculous destruction of the second lumbar vertebral body, and a large tuberculous abscess in the right (retroperitoneal) psoas region (a cold abscess). This cold abscess harboured an active tuberculous infection as evidenced by histological and molecular tests. Radiological and histological analysis further revealed extensive arteriosclerosis with (non-obliterating) coronary and significant carotid arteriosclerosis, healthy bone tissue without evidence of age-related osteopenia, evidence for diffuse idiopathic skeletal hyperostosis and mild osteoarthrosis of few joints. This suggests excellent living conditions correlating well with his diet indicated by stable isotope results and literary evidence. Despite the clear evidence of a tuberculous cold abscess with bacterioscopic and molecular proof for a persisting MTC infection of a human-type M. tuberculosis strain, we can exclude the chronic MTC infection as cause of death. The detection of MTC in historic individuals should therefore be interpreted with great caution and include further data, such as their nutritional status.

Pediatric Enteric Diagnostic Stewardship: The Right Test in the Right Context.

In this issue of Pediatrics , Cotter et al1 offer valuable analysis of the utility of molecular (nucleic acid amplification) enteric pathogen diagnostic technology in hospitalized children. These tests return highly worthwhile information for some infections, but many of their results are prone to misinterpretation. However, diagnostic guidelines based on classic microbiologic techniques do not perform well either.2 Molecular enteric microbiology is, therefore, a model opportunity for diagnostic stewardship to maximize worth and minimize wasteful expense. Cotter et al1 find that only 3.7% of tested hospitalized children benefitted from molecular diagnosis in terms of decreased length of stay and/or initiation of appropriate treatment. These data reinforce previous findings: enteric diagnostic use increases briskly after molecular testing becomes available and the tests find many unexpected signals reflecting difficult-to-interpret and/or nonactionable microbes. The authors candidly acknowledge the study’s limitations: nongeneralizability (their findings do not apply to outpatients) and reliance on historic comparison data. The benefit they cite applies to hospitalized children, and such populations are probably enriched for severe bacterial infections for which rapid and accurate molecular diagnosis has the greatest value; one would expect different yields in emergency facilities and ambulatory settings. … Address correspondence to Gillian A. M. Tarr, PhD, School of Public Health, University of Minnesota, MMC 807, 420 Delaware St SE, Minneapolis, MN 55455. E-mail: gtarr{at}umn.edu

The Prenatal Origin of Childhood Leukemia: Potential Applications for Epidemiology and Newborn Screening.

Childhood leukemias are heterogeneous diseases with widely differing incident rates worldwide. As circulating tumors, childhood acute leukemias are uniquely accessible, and their natural history has been described in greater detail than for solid tumors. For several decades, it has been apparent that most cases of childhood acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) initiate in utero. Circumstantial evidence in support of this contention includes the young age of onset and high rate of concordance among identical twins. “Backtracking” of leukemic somatic mutations, particularly gene translocations, to cord blood and dried blood spots collected during the perinatal period has provided molecular proof of prenatal leukemogenesis. Detection of a patient's leukemia translocation in easily accessible birth samples, such as dried blood spots, is straightforward with the knowledge of their idiosyncratic breakpoints. However, to translate these findings into population-based screening and leukemia prevention requires novel methods able to detect translocations at all possible breakpoints when present in a low frequency of cells. Several studies have attempted to screen for leukemic translocations, mainly the common ETV6-RUNX1 translocation, in cord blood samples from healthy children. Most studies have reported finding translocations in healthy children, but estimates of prevalence have varied widely and greatly exceed the incidence of leukemia, leading to concerns that technical artifact or contamination produced an artificially inflated estimate of translocation prevalence at birth. New generation techniques that capture the presence of these translocations at birth have the potential to vastly increase our understanding of the epidemiology of acute leukemias. For instance, if leukemic translocations are present at birth in a far higher proportion of children than eventually develop acute leukemia, what are the exposures and somatic molecular events that lead to disease? And could children with translocations present at birth be targeted for prevention of disease? These questions must be answered before large-scale newborn screening for leukemia can occur as a public health initiative. Here, we review the literature regarding backtracking of acute leukemias and the prevalence of leukemic translocations at birth. We further suggest an agenda for epidemiologic research using new tools for population screening of leukemic translocations.