Recovery of a medieval Brucella melitensis genome using shotgun metagenomics

Shotgun metagenomics provides a powerful assumption-free approach to the recovery of pathogen genomes from contemporary and historical material. We sequenced the metagenome of a calcified nodule from the skeleton of a 14th-century middle-aged male excavated from the medieval Sardinian settlement of Geridu. We obtained 6.5-fold coverage of a Brucella melitensis genome. Sequence reads from this genome showed signatures typical of ancient or aged DNA. Despite the relatively low coverage, we were able to use information from single-nucleotide polymorphisms to place the medieval pathogen genome within a clade of B. melitensis strains that included the well-studied Ether strain and two other recent Italian isolates. We confirmed this placement using information from deletions and IS711 insertions. We conclude that metagenomics stands ready to document past and present infections, shedding light on the emergence, evolution, and spread of microbial pathogens

Apoptosis in spermatozoa of infertile men, clinical correlations

The methods for evaluation of male infertility include not only routine investigations, standardized by the WHO, but also complementary techniques, developed over the last years, in order to improve the predictive value of seminal analysis for natural conception and assisted reproduction. With reference to these new methods, studies suggest that sperm with certain levels of DNA fragmentation serve as a strong predictor of reduced male fertility. We studied subjects who underwent seminal fluid evaluation, because of an infertility condition, at the Department of Biomedical Sciences of the University of Sassari.The samples collected by masturbation were evaluated according to the World Health Organisation (1999).The samples was washed twice in PBS and cytocentrifuged for 10 min at 1800 rpm on polylysine-coated slides that were fixed in methanol at room temperature. The apoptosis was evaluated using the TUNEL (In Situ Cell Death Detection Kit, Fluorescein, Roche, Cat.No. 1 684 795). At fluorescent miscroscopy are counted at least 300 cells. Quantitative evaluation of apoptosis by the TUNEL method confirmed that apoptosis did not seem to be correlated with sperm concentration or morphology; however, we found a higher apoptotic rate in semen from patients affected by andrologic diseases, such as varicocele, than from those with alteration of semen characteristics. Apoptosis analysis might be used in infertile patients in order to understand the etiology of unexplained infertility and to improve therapeutic effectiveness

Educational methods in neuroanatomy: a review

Among the basic sciences providing relevant medical awareness, human anatomy, which includes gross and neuroanatomy, represents a crucial science in medical schools (Verhoeven et al., 2002). Although numerous changes have been performed in medical curricula worldwide, the anatomic background is deemed a cornerstone for approaching clinical medicine (McLachlan and Patten, 2006). In the last decades, several modifications in anatomy and, in particular, in neuroanatomy education have been done and numerous strategies have been identified to improve the skills of the students (Rizzolo et al, 2010). We carried out a non-systematic review focusing our interest on the transformation of traditional teaching methodology of neuroanatomy. Articles on neuroanatomy education were selected if the study described an approach for teaching/learning neuroanatomy and if the neuroanatomy course was aimed to all medical students but dental or healthcare students. PubMed database was used to select manuscripts with a predefined combined list of search terms: “neuroanatomy”, “anatomy”, “teaching”, “learning”, “methods and strategies”. Our search included only English language manuscripts published from December 1990 to January 2012. Abstracts and unpublished studies were excluded. References of all relevant retrieved articles, of relevant recent review articles, and abstracts published in proceedings were also evaluated manually in order to find additional articles. For data extraction an electronic form was prepared. Only eighteen of the fifty (36%) selected articles met the inclusion criteria. Two described the dissection as the primary educational method, whereas the majority used electronic tools providing three-dimensional information. Some authors highlighted that students could be more enthusiastic when they have the chance to study with cadavers deeming this approach near to clinical medicine (Zurada et al, 2011); however the comprehension of topography and spatial relationships of various structures is equally achieved with the use of electronic means. The post-mortem examination represents an opportunity to evaluate the variability of human anatomy; however, logistical, ethical, and safety issues hamper the implementation of this method in medical schools (Winkelmann, 2007). The method used for anatomy and neuroanatomy teaching is essential to augment the interest of the medical students towards medicine but several obstacles can hinder the implementation and scale-up of the best educational method

Anatomical variations in ancient sardinian populations

Archaeological excavations carried out in different burial sites of north-western Sardinia allowed to observe anatomical variations in ancient skeletal remains. A case of brachymetatarsia, consisting of bilateral abnormal shortness of the fourth metatarsal bone, was detected in an adult female uncovered from the Medieval village of Geridu (Sassari), dated back to the late 13th or the first half of the 14th century; such a rare deformity has a clinical incidence of 0.02% to 0.05% (1). Several anatomical variations were diagnosed in individuals brought to light from the plague cemetery of 16th century Alghero (Sassari). The skeleton of a 9-10-year-old child showed a skull malformation due to premature bilateral closure of the coronal suture, diagnosed as non-syndromic brachycephaly (2). Posterior schisis of the first cervical vertebra, consisting of failure of the midline fusion of the two hemiarches with a small gap, was seen in a male aged 20-30 years; this type of anomaly has a current occurrence of approximately 4%. Occipitalization of the atlas associated with posterior spondiloschisis was observed in a male aged 35-45 years. There is complete fusion of the superior articular facets of the first cervical vertebra with the occipital condyles; this congenital anomaly has a current incidence of 0.14 to 0.75% of the pop- ulation. The small number of published osteoarchaeological cases of anatomical variations makes any report important

Human The Distal Perforators Of Posterior Tibial Artery. A Study For The Correct Planning Of Medial Lower Leg Flaps

 The adipofascial flap, introduced by Lin in 1994 (1), has many advantages compared to fasciocutaneous or free flaps for lower limb reconstruction. Its dissection is relatively easy and fast with low donor site morbidity and it doesn’t alter the shape of the leg. The aim of this dissection study is to evaluate the anatomic localization of the most distal perforator of the posterior tibial vessels to provide an anatomical rationale for the safe harvesting of distally based medial adipofascial flaps of the leg. Thirty lower limbs from 15 cadavers were used for this study. In all the limbs, the femoral artery was cannulated. Different techniques of injection were employed. 26 specimens were injected with latex to fill the entire vascular network and ease dissection. 3 specimens were injected with a mixture of barium sulphate and gelatin and underwent a CT scan. CT scan dataset were used to make 3D reconstruction. They underwent anatomical dissection after the scan. 1 Specimen was injected with red epoxy resin mixed with barium sulphate (2). This technique is aimed to perform comparative anatomical vascular studies between CT three-dimensional reconstructions and traditional dissection techniques. A distal perforator was found in all specimens; the mean caliber was 0.77 mm. In all cases, the perforator artery passed in the septum between flexor hallucis longus m. and flexor digitorum longus m. and was accompanied by two veins. In our series, the distance between the lowest perforator and the medial malleolus ranged from 3.5 to 8.2 cm. The median was 6.75 cm, the 5th percentile 4 cm and the 95th percentile 8.1 cm. The mean distance of the perforator from the medial tibial border was 1.23 cm. The mean ratio between the distance of perforator from the medial malleolus and the total leg length was 21%. Compared to all previous researches (3), our study has found more distal perforators from posterior tibial perforator artery. This fact may have important clinical consequences because the anteromedial adipofascial flap would cover more distal soft tissue defects. Moreover, our data suggest some safety parameters to make the rising of a medial adipofascial leg flap safer in surgical practice

Is the corpus callosum degeneration a predictor of long-term disability in Multiple Sclerosis patients?

Axonal injury and loss of white matter has been well documented in corpus callosum (CC) of patients with multiple sclerosis (MS), an inflammatory demyelinating disorder of the of Central Nervous System (CNS). The CC is the largest compact white matter fiber bundle of the brain connecting the two cerebral hemispheres. It seems that CC continue to mature structurally from infancy to adulthood (Muetzel RL, 2008). The topographical organization of its fibers allows the association of its abnormalities with those of specific cortical regions. It is one of the few white matter tracts that can be adequately evaluated by conventional Magnetic Resonance Imaging (MRI), having sharply demarcated two-dimensional limits on a mid-saggital T1W imaging (Fiqueira FF, 2007). So, as a consequence of these anatomic characteristics, it is reasonable to assume that CC morphometrics might be used as a possible marker for the analysis of its integrity. Conventional and non-conventional MRI techniques have been used to characterize pathological damages of the CC; for instance, diffusion tensor imaging (DTI) can show white matter changes undetected by conventional MRI (Bester M, 2008). We carried out a review focusing our interest on the diagnostic efficacy of MRI to predict MS disability. PubMed search engine was used to select manuscripts with predefined search terms: “corpus callosum”, “multiple sclerosis”, “disability” and “magnetic resonance imaging”. Only manuscripts in English language, published from January 2003 to January 2013, were selected. Abstracts and unpublished studies were excluded. References of all relevant retrieved articles, of review articles, were also evaluated manually in order to find additional articles. For data extraction an electronic form was prepared. The findings of the review suggest that CC atrophy is a more sensitive marker of disease activity than the global atrophy measures, predicting clinically definite multiple sclerosis (CDMS)conversion as early as 6 months of clinically isolated syndrome (CIS) (Kalincik T, 2012). Furthermore, the occurrence or growth of lesions over the first year in the splenium of corpus callosum, together with the cerebellum and the thalamus, was associated with cognitive worsening at year 5. CC atrophy seems to be a simple and an accurate predictor of disability, mostly for secondary progressive MS (Fiqueira FF, 2007), and it seems helpful for routine clinical activities (Vaneckova M, 2012)

Neuroanatomy, the Achille's heel of medical students. A systematic analysis of educational strategies for the teaching of neuroanatomy

Neuroanatomy has been deemed crucial for clinical neurosciences. It has been one of the most challenging parts of the anatomical curriculum and is one of the causes of “neurophobia,” whose main implication is a negative influence on the choice of neurology in the near future. In the last decades, several educational strategies have been identified to improve the skills of students and to promote a deep learning. The aim of this study was to systematically review the literature to identify the most effective method/s to teach human neuroanatomy. The search was restricted to publications written in English language and to articles describing teaching tools in undergraduate medical courses from January 2006 through December 2017. The primary outcome was the observation of improvement of anatomical knowledge in undergraduate medical students. Secondary outcomes were the amelioration of long‐term retention knowledge and the grade of satisfaction of students. Among 18 selected studies, 44.4% have used three‐dimensional (3D) teaching tools, 16.6% near peer teaching tool, 5.55% flipped classroom tool, 5.55% applied neuroanatomy elective course, 5.55% equivalence‐based instruction‐rote learning, 5.55% mobile augmented reality, 5.55% inquiry‐based clinical case, 5.55% cadaver dissection, and 5.55% Twitter. The high in‐between study heterogeneity was the main issue to identify the most helpful teaching tool to improve neuroanatomical knowledge among medical students. Data from this study suggest that a combination of multiple pedagogical resources seems to be the more advantageous for teaching neuroanatomy

Hyaluronan Esters Drive Smad Gene Expression and Signaling Enhancing Cardiogenesis in Mouse Embryonic and Human Mesenchymal Stem Cells

BACKGROUND: Development of molecules chemically modifying the expression of crucial orchestrator(s) of stem cell commitment may have significant biomedical impact. We have recently developed hyaluronan mixed esters of butyric and retinoic acids (HBR), turning cardiovascular stem cell fate into a high-yield process. The HBR mechanism(s) remain still largely undefined. METHODOLOGY/PRINCIPAL FINDINGS: We show that in both mouse embryonic stem (ES) cells and human mesenchymal stem cells from fetal membranes of term placenta (FMhMSCs), HBR differentially affected the patterning of Smad proteins, one of the major conductors of stem cell cardiogenesis. Real-time RT-PCR and Western blot analyses revealed that in both cell types HBR enhanced gene and protein expression of Smad1,3, and 4, while down-regulating Smad7. HBR acted at the transcriptional level, as shown by nuclear run-off experiments in isolated nuclei. Immunofluorescence analysis indicated that HBR increased the fluorescent staining for Smad1,3, and 4, confirming that the transcriptional action of HBR encompassed the upregulation of the encoded Smad proteins. Chromatin immune precipitation and transcriptional analyses showed that HBR increased the transcription of the cardiogenic gene Nkx-2.5 through Smad4 binding to its own consensus Smad site. Treatment of mouse ES cells and FMhMSCs with HBR led to the concomitant overexpression of both Smad4 and α-sarcomeric actinin. Smad4 silencing by the aid of lentiviral-mediated Smad4 shRNA confirmed a dominant role of Smad4 in HBR-induced cardiogenesis. CONCLUSIONS/SIGNIFICANCE: The use of HBR may pave the way to novel combinatorial strategies of molecular and stem cell therapy based on fine tuning of targeted Smad transciption and signaling leading to a high-throughput of cardiogenesis without the needs of gene transfer technologies