Effects of blastomere biopsy on post-natal growth and behavior in mice.

STUDY QUESTION: Does blastomere biopsy (BB) of preimplantation embryos induce long-term effects on their growth and post-natal behavior? SUMMARY ANSWER: BB induces long-term effects on body weight and behavior in male mice. WHAT IS KNOWN ALREADY: BB is an essential technique for performing preimplantation genetic diagnosis (PGD), a screening test that can detect genetic abnormalities of embryos before their transfer in utero. There is limited understanding of the post-natal consequences and safety of BB. STUDY DESIGN, SIZE, DURATION: Offspring who had a BB performed as embryos, as well as control offspring, were examined for body and neurological development and subjected to a screening battery of behavioral tests, designed to model symptoms of psychiatric disorders. At least 12 mice were used for each test over the course of 16 weeks. PARTICIPANTS/MATERIALS, SETTING, METHODS: Embryos were subjected to a single BB at the 8-cell stage and then cultured in vitro until the blastocyst stage (BB group). Two control groups were created, one consisting of embryos cultured in vitro without any manipulation (in vitro control (IVC) group) and one of embryos developed entirely in vivo (in vivo group). Embryos from in vitro groups (BB and IVC) were transferred to pseudo-pregnant female mice at the blastocyst stage. Body growth parameters and developmental landmarks of the resulting offspring were observed during their entire lifespan. Furthermore, validated behavioral tests were used to assess early communicative functions, startle reflex, and anxiety- and depression-like behaviors. MAIN RESULTS AND THE ROLE OF CHANCE: We found that male mice derived from BB exhibited peculiar behavioral alterations and changes in body weight. BB-derived male mice showed increased body weight with respect to both controls as early as the second week of life. Adult males displayed decreased times of immobility in the tail suspension test (P < 0.05) and deficits in habituation to, and pre-pulse inhibition of, the startle reflex (P < 0.05). BB did not affect communicative skills and anxiety-like responses. LIMITATIONS, REASONS FOR CAUTION: Extrapolation of these results to humans requires caution as the culture protocols used in human clinics could be better established than in mice research. Furthermore species-specific neurodevelopmental features could be a source of differences between mice and humans in the effects of BB. WIDER IMPLICATIONS OF THE FINDINGS: Our data demonstrate that BB affects long-term programming of post-natal development and behavior in mice, suggesting that PGD procedures could be a risk factor for late-onset, neurodevelopmental and metabolic disease predisposition. Thus, in light of our observations, long-term follow-up in humans or other primates generated after BB is needed. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the European Research Council (FP7/2007-2013)/Programme IDEAS GA no. 210103 to G.E.P. European Research Council - Programme FP7-KBBE-2012.1.3-04, GA no. 312097 Acronym: FECUND to G.E.P.; MIUR/CNR, Programme FIRB. GA n. B81J12002520001 Acronym: GenHome to P.L. The authors are participating in the COST action FA 1201 'Epiconcept' Epigenetic and Periconception Environment. No competing interests are declared.

Grand-paternal age and the development of autism-like symptoms in mice progeny.

Advanced paternal age (APA) contributes to the risk of autism spectrum disorders (ASDs) in children. In this study, we used a mouse model to investigate the effects of APA on behavioral features related to autistic syndromes (that is, social deficits, communication impairments and stereotypic/repetitive behaviors). We also examined whether such effects are transmitted across generations. To do this, males aged 15 months (APA) and 4 months (control) were bred with 4-month-old females, and the resulting offspring (F1) and their progeny (F2; conceived by 4-month-old parents) were tested for the presence and severity of ASD-like behaviors. Our results indicate that APA resulted in offspring that displayed distinctive symptoms of ASD. We found that both F1 conceived from old fathers and F2 derived from old grandfathers displayed increased ultrasound vocalization (USV) activity, decreased sociability, increased grooming activity and increased anxiety-like responses. Moreover, such abnormalities were partially transmitted to the second generation of mice, having APA grandfathers. In conclusion, our study suggests that the risk of ASD could develop over generations, consistent with heritable mutations and/or epigenetic alterations associated with APA.

Towards storage of cells and gametes in dry form.

We review published data on cell/gamete lyophilization. Most studies have utilized the same established protocols for cryopreservation (storage in liquid nitrogen) as for cell lyophilization (dehydration of frozen samples by water sublimation). Surveying natural lyoprotectants, we suggest trehalose and late embryogenesis abundant (LEA) proteins as ideal candidates for the reversible desiccation of mammalian cells/gametes. We find that despite the numerous water subtraction techniques, scientists have relied almost exclusively on lyophilization. There is thus room for improvement in both medium formulation and water subtraction strategies for dry cell/gamete storage. We believe the development of dry processing protocols for use in biobanks of cells/gametes, at reduced cost and with minimal carbon footprint, is within our grasp.

Longitudinal study in HIV/HCV-coinfected HAART-naive patients and role of HCV genotype.

To evaluate the impact of highly active antiretroviral therapy (HAART) on the course of hepatitis C (HCV) infection, we studied the biological and virological characteristics of 23 HCV/HIV-coinfected HAART-naive patients. The HCV genotype, HCV and HIV viral loads, serum alanine aminotransferase, CD4+ and CD8+ cell/mm3 were determined at baseline, 1 month, 6 months and 12 months after initiation of HAART. Results were analyzed both in terms of total population and of HCV genotype. The study of the total population suggests that this therapy did not determine a significant alteration of HCV viremia and levels of ALT, while a significant decrease in HIV viremia (-1.7log10 at one year from the start of HAART) and increase in CD4+ counts was observed (P < 0.005). The biological and virological parameters of HCV/HIV coinfection differed according to the HCV genotype. In particular, only genotype 4 showed a significant inverse correlation between HCV and HIV viral loads.

Drug-resistant tuberculosis in HIV-infected persons: Italy 1999-2000.

BACKGROUND: Patients with HIV infection may be a valuable target for assessing the impact of drug-resistant tuberculosis (TB). PATIENTS AND METHODS: An observational, prospective study was conducted in 96 infectious disease hospital units in Italy during 1999-2000. A total of 140 HIV-infected patients with diagnosis of TB and with an isolate tested for drug susceptibility entered the analysis. Drug resistance (DR) was defined as resistance to either isoniazid (INH) or rifampin (RIF), while multidrug resistance (MDR) was defined as resistance to INH and RIF. RESULTS: A total of 117 (83.6%) episodes of TB were classified as new cases and 23 (16.4%) as previously treated cases. Prevalence of resistance to INH or RIF was 12.8% and 4.3% among new cases, and 17.4% and 26.1% among previously treated cases, respectively. Prevalence rates of DR and MDR were 14.5% and 2.6% among new cases and 30.4% and 12.5% among previously treated cases, respectively. No statistically significant risk factors associated with DR or MDR TB emerged in this analysis. CONCLUSION: High prevalence rates of DR and MDR are present among HIV-infected TB patients in Italy, in particular among previously treated cases.

Chlamydia pneumoniae antibodies and angiographically demonstrated coronary artery disease in a sample population from Italy.

Recent reports suggest an association between Chlamydia pneumoniae and chronic coronary heart disease. This case-control study investigates the relationship between the presence of immunoglobin G (IgG) and immunoglobin A (IgA) when measured by means of microimmunofluorescence (MIF) and angiographically diagnosed coronary disease. Cases (n = 150) were angiography patients with at least one coronary artery lesion occupying at least 50% of the luminal diameter. Controls (n = 49) were angiography patients with no detectable signs of coronary artery disease and patients (n = 56) without signs or symptoms of coronary disease and with normal ECG results. No significant differences were revealed between the seroprevalence of IgG and IaA and geometric mean titers (GMT) as measured in cases and controls. When cases were compared with controls whose angiographic results were normal, after adjusting for established risk factors (cholesterol, smoking, hypertension, diabetes, age, gender and family history), the estimated risk of coronary artery disease was 0.79 (95% confidence interval (C.I.), 0.31-1.99) for the presence of IgG and was 0.94 (95 C.I., 0.37-2.39) for IgA. When cases were compared with controls with normal ECG results, the adjusted odds ratio (O.R.) for coronary artery disease was 1.17 (95%, C.I., 0.52-2.62) for the presence of IgG and 0.82 195% C.I., 0.36-1.86) for the presence of IgA. These results do not support an association between C. pneumoniae infection and coronary disease.