Infection with Leptospira kirschneri Serovar Mozdok: First Report from the Southern Hemisphere

Leptospirosis is a global zoonosis caused by pathogenic Leptospira spp. In this study, we characterized two Leptospira kirschneri serogroup Pomona serovar Mozdok isolates, one obtained from a dog and the other from a patient with severe leptospirosis, 4 years later. Histopathological analysis showed that both isolates caused severe tissue damage when used to infect hamsters. While L. kirschneri serogroup Pomona serovar Mozdok is endemic in animals in Europe, there is only one report of human leptospirosis in the literature. Although strains belonging to L. kirschneri serogroup Pomona have been identified in cases of human leptospirosis in Europe, serovar Mozdok has not yet been implicated. The 4-year interval between isolations and the fact that this is the first report of serovar Mozdok as the causative agent of human leptospirosis in the southern hemisphere, demonstrates its epidemiological importance to public health. Moreover, the presence of serovar Mozdok in Brazil has the potential to affect vaccine and diagnostic test development.Leptospirosis is a reemerging zoonotic disease, and the global burden is showing an upward trend. The original estimates in 1999¹ predicted some 500,000 annual cases compared with the latest prediction of 873,000 cases and 49,000 mortalities per year, a 74.6% increase over 15 years.² Accurate laboratory diagnosis continues to be a limiting factor, meaning that the true global burden of leptospirosis is likely to be much higher.³ In Latin America, the prevalence of severe leptospirosis is high (10,000 cases a year) due to the tropical climate and lack of appropriate sanitation.³ Although the city of Pelotas has a subtropical climate, > 50 cases of human leptospirosis per 100,000 inhabitants are reported each year, one of the highest rates in southern Brazil.⁴ The infection rate in Pelotas is higher than the Brazilian average for the same period (3.5/100,000) and other regions with similar climatic conditions (> 10/100,000).⁵At present, there are 10 pathogenic Leptospira spp. classified into > 260 serovars⁶ and Leptospira interrogans, Leptospira borgpetersenii, and Leptospira kirschneri are most commonly associated with human leptospirosis.⁷ In Brazil, L. interrogans serogroup Icterohaemorrhagiae serovars Icterohaemorrhagiae and Copenhageni are the main cause of urban leptospirosis and have been widely studied,³ whereas rural leptospirosis and the associated serovars have been largely neglected. To the best of our knowledge, L. kirschneri serogroup Pomona serovar Mozdok has only been implicated in a case of human leptospirosis in Cuba.⁸ Serovar Mozdok is endemic to Croatia where it is prevalent in wild rodents. Human leptospirosis caused by serogroup Pomona is common in that region and while serovar Mozdok has not been implicated in any human cases,⁹ it is a causative agent of canine leptospirosis in Europe.¹⁰We report the isolation and characterization of two isolates of serovar Mozdok recovered from cases of canine and human leptospirosis in Pelotas, southern Brazil. The canine strain was isolated in 2009 during a municipal dog neutering campaign. Urine samples were aseptically collected from the bladder during ovarian hysterectomy, via aspiration using an insulin 30-G needle and syringe (BD Biosciences, Franklin Lakes, NJ). The urine was immediately inoculated into unsupplemented Ellinghausen-McCullough-Johnson-Harris (EMJH; Difco, Sparks, MD) medium (100 μL urine/5 mL EMJH), incubated for 1 hour and then subcultured into EMJH containing 10% of a commercial supplement (Difco). The dog from which the strain was isolated was asymptomatic and was released after the surgical procedure. The second isolate was obtained from the blood culture of a 56-year-old female patient from a rural area of the city. The patient presented with headache, myalgia, fever, vomiting, fatigue, sleepiness, and arthralgia and reported contact with dogs, rats, pigs, cattle, and flood water. The isolate was cultured in EMJH medium as described for the canine isolate. Both isolates were identified as L. kirschneri by means of secY gene sequencing.¹¹ Multilocus sequence typing (MLST)⁷ further characterized the isolates as L. kirschneri serogroup Pomona serovar Mozdok (ST 117). All sequencing procedures were performed using the paired-end technology on an Illumina Solexa platform (Illumina, San Diego, CA; GenBank accession numbers for sequences are shown in

Potent neutralization of severe acute respiratory syndrome (SARS) coronavirus by a human mAb to S1 protein that blocks receptor association

Effective prophylaxis and antiviral therapies are urgently needed in the event of reemergence of the highly contagious and often fatal severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) infection. We have identified eight recombinant human single-chain variable region fragments (scFvs) against the S1 domain of spike (S) protein of the SARS-CoV from two nonimmune human antibody libraries. One scFv 80R efficiently neutralized SARS-CoV and inhibited syncytia formation between cells expressing the S protein and those expressing the SARS-CoV receptor angiotensin-converting enzyme 2 (ACE2). Mapping of the 80R epitope showed it is located within the N-terminal 261-672 amino acids of S protein and is not glycosylation-dependent. 80R scFv competed with soluble ACE2 for association with the S1 domain and bound S1 with high affinity (equilibrium dissociation constant, Kd=32.3 nM). A human IgG1 form of 80R bound S1 with a 20-fold higher affinity of 1.59 nM comparable to that of ACE2 (Kd=1.70 nM), and neutralized virus 20-fold more efficiently than the 80R scFv. These data suggest that the 80R human monoclonal antibody may be a useful viral entry inhibitor for the emergency prophylaxis and treatment of SARS, and that the ACE2-binding site of S1 could be an attractive target for subunit vaccine and drug development.     

No Association Between NRG1 and ErbB4 Genes and Psychopathological Symptoms of Schizophrenia

Neuregulin 1 (NRG1) and v-erb-a erythroblastic leukemia viral oncogene homolog 4 (ErbB4) have been extensively studied in schizophrenia susceptibility because of their pivotal role in key neurodevelopmental processes. One of the reasons for the inconsistencies in results could be the fact that the phenotype investigated has mostly the diagnosis of schizophrenia per se, which is widely heterogeneous, both clinically and biologically. In the present study we tested, in a large cohort of 461 schizophrenia patients recruited in Scotland, whether several SNPs in NRG1 and/or ErbB4 are associated with schizophrenia symptom dimensions as evaluated by the Positive and Negative Syndrome Scale (PANSS). We then followed up nominally significant results in a second cohort of 439 schizophrenia subjects recruited in Germany. Using linear regression, we observed two different groups of polymorphisms in NRG1 gene: one showing a nominal association with higher scores of the PANSS positive dimension and the other one with higher scores of the PANSS negative dimension. Regarding ErbB4, a small cluster located in the 5′ end of the gene was detected, showing nominal association mainly with negative, general and total dimensions of the PANSS. These findings suggest that some regions of NRG1 and ErbB4 are functionally involved in biological processes that underlie some of the phenotypic manifestations of schizophrenia. Because of the lack of significant association after correction for multiple testing, our analyses should be considered as exploratory and hypothesis generating for future studies.     

Temporal mapping of the closure of the anterior fontanelle and contiguous sutures using computed tomography,in silico models of modern infants

The aim of this study is to quantify and statistically model the age-related decline in the fibrous connective tissue interface of the anterior fontanelle in modern Australian infants, using three-dimensional, semi-automated computed-assisted design protocols. Non-linear regression with variance models, using power functions, combined with quantile regression of the 5th and 95th population percentiles, were utilised to assess absolute anterior fontanelle surface area (AFSA) as a function of age, using multi-slice cranial computed tomography scans obtained from 256 infants aged < 30 months (males: n = 126, females: n = 109) from Brisbane children’s hospitals. Normalised AFSA (NFSA), standardised for variation in cephalic size, followed a progressive decline from birth, the greatest velocity change occurring between the 3–6 and 6–9 month cohorts. Growth of the neurocranium is the most significant within the first 8 months postpartum, with a mean increase of 19.03 mm in maximum cranial length and 10.04 mm in breadth. Directionality of fontanelle closure, quantified using spline curves refutes fundamental assumptions that the anterior fontanelle is consistent with a quadrilateral, and contiguous sutures exhibit constant velocity of closure. The present study provides normative values for fontanelle size and diameters as well as new predictive non-linear models for age substantiation, screening of developmental abnormalities and indicators of suspected child maltreatment in modern infants aged birth to 30 months.     

IFNL cytokines do not modulate human or murine NK cell functions

The interferon-lambda (IFNL) cytokines have been shown to be important in HCV infection with SNPs in the IFNL3 gene associated with both natural and treatment induced viral clearance. We have recently shown that rs1299860 (an IFNL3 associated SNP) and an NK cell gene, KIR2DS3, synergised to increase the odds of chronic infection in a homogenous cohort of Irish women infected with HCV. To characterise a biological basis for the genetic synergy, we investigated for any evidence that IFNL cytokines regulate NK cell functions. Using a range of functional responses, we did not find any evidence of NK cell activation by IFNL3, IFNL1 or IFNL2 cytokines. Similar results were found using human and murine NK cells. In addition, and in contrast to our preliminary study, we did not find any evidence that IFNL cytokines inhibited NK cell cytokine production; thus, the biological basis for the genetic synergy remains to be discovered.     

Point mutation of the autophosphorylation site or in the nuclear location signal causes protein kinase A RII beta regulatory subunit to lose its ability to revert transformed fibroblasts.

The RII beta regulatory subunit of cAMP-dependent protein kinase (PKA) contains an autophosphorylation site and a nuclear location signal, KKRK. We approached the structure-function analysis of RII beta by using site-directed mutagenesis. Ser114 (the autophosphorylation site) of human RII beta was replaced with Ala (RII beta-P) or Arg264 of KKRK was replaced with Met (RII beta-K). ras-transformed NIH 3T3 (DT) cells were transfected with expression vectors for RII beta, RII beta-P, and RII beta-K, and the effects on PKA isozyme distribution and transformation properties were analyzed. DT cells contained PKA-I and PKA-II isozymes in a 1:2 ratio. Over-expression of wild-type or mutant RII beta resulted in an increase in PKA-II and the elimination of PKA-I. Only wild-type RII beta cells demonstrated inhibition of both anchorage-dependent and -independent growth and phenotypic change. The growth inhibitory effect of RII beta overexpression was not due to suppression of ras expression but was correlated with nuclear accumulation of RII beta. DT cells demonstrated growth inhibition and phenotypic change upon treatment with 8-Cl-cAMP. RII beta-P or RII beta-K cells failed to respond to 8-Cl-cAMP. These data suggest that autophosphorylation and nuclear location signal sequences are integral parts of the growth regulatory mechanism of RII beta.