Complete genome sequence of a German isolate of spartina mottle virus supports its classification as a member of the proposed genus "Sparmovirus" within the family Potyviridae.

Spartina mottle virus (SpMV), an unassigned member of the family Potyviridae, has been known since 1980, when it was first described in England and Wales in symptomatic plants of the genus Spartina. In infected cells, flexuous particles and pinwheel inclusion bodies were found that resemble those of potyvirids. To date, the NCBI database contains only two partial sequences of a German (Nessmersiel) and an Italian (Assisi) isolate, suggesting that SpMV could be the first member of a new genus, called "Sparmovirus", in the family Potyviridae. In this study, the first complete genome sequence of the German SpMV isolate (SpMV Ger) was determined. The genome of SpMV is a single-stranded, monopartite, polyadenylated RNA consisting of 9376 nucleotides. Sequence analysis revealed a genome organization similar to that of classical potyviruses, including many conserved features. In phylogenetic analysis, SpMV could not be assigned to any of the known genera, but it showed the closest relationship to rymoviruses and common reed chlorotic stripe virus (CRCSV, unassigned). Sequence comparisons confirmed that a new genus should be established containing SpMV, CRCSV, and three Bermuda grass mosaic virus isolates, which are considered divergent strains of SpMV.

Molecular diversity of poleroviruses infecting cucurbit crops in four countries reveals the presence of members of six distinct species.

When 66 cucurbit samples with yellowing symptoms from fields in Mali, the Philippines, Thailand and Uzbekistan were screened by RT-PCR using universal polerovirus primers, 21 were identified as harboring polerovirus RNA. When these 21 samples were screened with specific primers for the known cucurbit-infecting poleroviruses, suakwa aphid-borne yellows virus and a recombinant strain of cucurbit aphid-borne yellows virus were detected for the first time in the Philippines and Thailand. However, seven polerovirus-positive samples did not react with any of the known species-specific primers. Sequencing of 1.4-kb universal polerovirus RT-PCR products revealed the presence of two poleroviruses that had not been described previously. These viruses, from Mali and Thailand, were provisionally named pepo aphid-borne yellows virus and luffa aphid-borne yellows virus, respectively.

First Report of Zucchini yellow mosaic virus Associated with Leaf Crinkle and Yellow Mosaic Diseases of Cucurbit Plants in Mali.

The aphid-transmitted Zucchini yellow mosaic virus (ZYMV; genus Potyvirus, family Potyviridae) has been reported to cause severe epidemics and yield losses in cucurbit crops worldwide (1). In Africa, ZYMV has been detected in Algeria, Egypt, Madagascar, Mauritius, Mayotte, Morocco, Nigeria, Reunion, South Africa, Sudan, Swaziland, and Tunisia (1). In April 2009, leaf yellowing, mosaic, crinkling, and curling were common on cucurbit plants in fields in Mali. Symptomatic leaf samples were collected from five cucumber (Cucumis sativus) plants in Kati, two watermelon (Citrullus lanatus) plants in Samanko, and one weedy melon (Cucumis sp.) plant in Baguineda. All samples tested positive for ZYMV and were negative for Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Papaya ringspot virus type W (PRSV-W), Watermelon mosaic virus (WMV), and Watermelon silver mottle virus (WSMoV) by double-antibody sandwich (DAS)-ELISA. They also tested negative for Melon yellow spot virus (MYSV) by indirect ELISA. Antibodies against ZYMV and WMV were obtained from DSMZ, Braunschweig, Germany, and those against CGMMV, MYSV, PRSV-W, and WSMoV were provided by Shyi-Dong Yeh, National Chung Hsing University, Taichung, Taiwan. Six ZYMV ELISA-positive samples (three cucumber, two watermelon, and the weedy melon sample) were also tested by reverse transcription (RT)-PCR using the potyvirus universal primer pair Sprimer1/Oligo(dT) (2). The expected 1.6-kb viral cDNA was amplified from all six samples and each was sequenced. All sequences obtained from cucumber (GenBank Accession Nos. HM005307, HM005308, and HM005309), watermelon (GenBank Accession Nos. HM005311 and HM005312), and weedy melon (GenBank Accession No. HM005310) isolates were 1,684 nucleotides (nt) long excluding the 3' poly-A tails. They comprised the 3'-terminal of the NIb region (1 to 633 nt), the coat protein region (634 to 1473 nt), and the 3'-untranslated region (1,474 to 1,684 nt). Because the sequences shared high nucleotide identity (98.3 to 99.7%), these isolates were considered to be the same virus species. When the sequences were compared by BLASTn searching in GenBank and analyzed by DNAMAN Sequence Analysis Software (Lynnon Corporation, St-Louis, Pointe-Claire, Quebec, Canada), they were found to have the greatest nucleotide identity (97.4 to 98.0%) with the Connecticut strain of ZYMV (ZYMV-Connecticut; GenBank Accession No. D00692), within a clade of isolates from China, Italy, Japan, and the United States. When assessed separately, their coat protein (97.7 to 98.3% nucleotide and 98.9 to 99.6% amino acid identity) and 3'-untranslated regions (96.7 to 97.2% identity) also had greatest homology with ZYMV-Connecticut. To our knowledge, this is the first report of ZYMV infecting cucurbit plants in Mali. ZYMV should be taken into consideration when breeding cucurbit crops for this region, and managing viral diseases. References: (1) C. Desbiez et al. Plant Pathol. 46:809, 1997. (2) W. S. Tsai et al. Plant Dis. 94:378, 2010.

Application of Phi29 DNA polymerase in identification and full-length clone inoculation of tomato yellow leaf curl Thailand virus and tobacco leaf curl Thailand virus.

Tomato plants grown in greenhouses in Thailand developed typical symptoms of a tomato yellow leaf curl Thailand virus (TYLCTHV) infection. After confirmation by ELISA, a Phi29 DNA polymerase approach was chosen for further molecular analysis of TYLCTHV. Total DNA purified from infected tomato leaves was subjected to rolling-circle amplification (RCA) of DNA-A and DNA-B of TYLCVTHV. In addition, a new monopartite geminivirus with a putative recombinant background was identified by RCA and tentatively named tobacco leaf curl Thailand virus (TbLCTHV). To confirm the composition of both geminiviruses, full-length clones were established and used for inoculation of Nicotiana benthamiana by particle bombardment or agroinfection. When TYLCTHV DNA-A and DNA-B were applied together by particle bombardment or agroinfection, severe stunting, yellowing, and leaf curling were observed. Whereas TYLCTHV DNA-A and TbLCTHV revealed no infection after'particle bombardment, similar symptoms in N. benthamiana, like leaf upward curling and yellowing were observed following agroinfection.DNA components of TYLCTHV DNA-A and DNA-B were excised from their respective plasmids, ligated, and amplified by Phi29 DNA polymerase. The ability of viral concatamere inoculation was evaluated in particle co-bombardment experiments on N. benthamiana. Thus, particle bombardment of RCA-derived multimeric products proved to be at least as effective as inoculation with a partial repeat construct and tenfold as effective as inoculation with excised unit-lengths of DNA-A and DNA-B of TYLCVTHV when using each DNA component in an amount of 5 ng.

The complete nucleotide sequence of a capsicum chlorosis virus isolate from Lycopersicum esculentum in Thailand.

The complete nucleotide sequence of a tospovirus isolated from Lycopersicum esculentum in Thailand was determined. The L RNA comprises of 8912 nt and codes for the RNA-dependent RNA-polymerase (RdRp) (2877 aa). Two ORFs are located on the M RNA (4823 nt) encoding the non-structural (NSm) protein (308 aa) and the viral glycoprotein precursors (Gn/Gc) (1121 aa) separated by an intergenic region of 433 nt. ORFs coding for the non-structural (NSs) and nucleocapsid (N) protein, 439 aa and 275 aa, respectively, were identified on the S RNA (3477 nt) separated by an intergenic region of 1202 nt. The N protein of the Thailand isolate was most closely related to that of capsicum chlorosis virus (CaCV), sharing an amino acid sequence identity of 92.7%. Additionally, multiple sequence analyses revealed significant similarities to tospoviruses of the species Watermelon silver mottle virus and to several putative tospovirus entries in GenBank. Based on these alignments it is proposed to refer to all these different viruses as isolates of CaCV.

Ceratothripoides claratris, a New Vector of a Capsicum chlorosis virus Isolate Infecting Tomato in Thailand.

ABSTRACT Ceratothripoides claratris, the predominant thrips species on tomato in Thailand, was tested for vector competence and efficiency to transmit Capsicum chlorosis virus (CaCV) (isolate AIT) to tomato. The efficiency of adult-stage transmission was influenced by the larval stage at which virus was acquired. Adult C. claratris showed 69% transmission efficiency after acquiring the virus as freshly emerged (<1 h) first-instar larvae. However, when just molted (<1 h) second-instar larvae acquired the virus, the percentage of adult transmitters significantly decreased (48%). Transmission efficiency of up to 47% was detected with second-instar larvae of C. claratris which had acquired the virus as freshly emerged first-instar larvae. Transmission efficiency did not significantly differ between adult males and females, irrespective of the larval stage at which the virus was acquired. Highest transmission efficiency for CaCV was recorded in adult C. claratris derived from second-instar larvae collected from infected tomato plants in a greenhouse. Lowest transmission efficiency was observed in adults directly collected from infected tomato plants in the greenhouse. The spread of CaCV on tomato plants in greenhouses showed a close association with thrips infestations.

Predictive value of proton magnetic resonance spectroscopy in pediatric closed head injury.

We studied 26 infants (1-18 months old) and 27 children (18 months or older) with acute nonaccidental (n = 21) or other forms (n = 32) of traumatic brain injury using clinical rating scales, a 15-point MRI scoring system, and occipital gray matter short-echo proton MRS. We compared the differences between the acutely determined variables (metabolite ratios and the presence of lactate) and 6- to 12-month outcomes. The metabolite ratios were abnormal (lower NAA/Cre or NAA/Cho; higher Cho/Cre) in patients with a poor outcome. Lactate was evident in 91% of infants and 80% of children with poor outcomes; none of the patients with a good outcome had lactate. At best, the clinical variables alone predicted the outcome in 77% of infants and 86% of children, and lactate alone predicted the outcome in 96% of infants and 96% of children. No further improvement in outcome prediction was observed when the lactate variable was combined with MRI ratios or clinical variables. The findings of spectral sampling in areas of brain not directly injured reflected the effects of global metabolic changes. Proton MRS provides objective data early after traumatic brain injury that can improve the ability to predict long-term neurologic outcome.

Scanning electron microscopy of arteriovenous malformations.

Although arteriovenous malformations (AVMs) have been known to have direct communications between arteries and veins without interposing capillaries, the exact location of arterial and venous junctions have not been defined. Utilizing microscopic and endoscopic observations, Yamada and associates identified shunting arterioles (50 mu-250 mu) directly connected to the AVM core vessels. While dissecting the AVMs in functional areas of the brain, shunting arterioles were sectioned to interrupt the arterial blood supply. This technique allowed cleavage formation between the core vessels and surrounding brain, thus avoiding brain tissue removal and preserving microcirculation to functionally critical brain. We demonstrate histologically for the first time by scanning electron microscopy shunting arterioles and communicating venules (20 mu-200 mu).

Mild head injury in children: identification, clinical evaluation, neuroimaging, and disposition.

Pediatric head injury presents in various degrees of severity. Early intervention in the patient with a severe head injury is the key to preventing secondary central nervous system damage. Patients with a head injury are easily identified, often by clinical examination alone. However, patients with a mild head injury present a challenge to practitioners, particularly in identification, knowing what is important in the clinical evaluation, deciding whether to use neuroimaging, and knowing where to send the child for observation. Use of the Glasgow Coma Score, primary survey, and identification of historic and clinical features that are suggestive of severe head injury may guide pediatric nurse practitioners in providing appropriate medical care and disposition.

[Increase of prilocaine-induced methemoglobinemia following anesthesia induction]. / Verstärkung der prilocain-induzierten Methämoglobin-ämie durch Narkoseeinleitung?

Although the local anaesthetic prilocaine is less cardio- and neurotoxic than lidocaine, it bears the disadvantage of the formation of methaemoglobin by the metabolite o-toluidine. Prilocaine is often successfully used, especially for the blockade of the brachial plexus, but one problem of this technique is the failure rate of 3-10%, with the consequence that general anaesthesia after administration of prilocaine is frequently necessary. Methaemoglobin formation after prilocaine administration has been thoroughly investigated. Nothing is known, however, about the interactions of prilocaine and the induction of general anaesthesia relative to methaemoglobinaemia. CASE REPORT. Two patients (47 and 52 years old) each received 500 mg prilocaine for the axillary blockade of the brachial plexus. After 100 and 120 min respectively, it was necessary to induce general anaesthesia, for which 350 mg thiopental, 1 mg alfentanil and 45 mg atracurium were used. At 15 min after induction, methaemoglobin levels had increased by 70% and 25%, respectively, from baseline before general anaesthesia. CONCLUSION. It is not possible to explain these findings conclusively with the present method. To check whether displacement of o-toluidine from the plasma protein binding might have been responsible, we provoked methaemoglobinaemia in vitro by adding o-toluidine to heparinised blood. Thiopental was then added to half the specimens. Subsequently, methaemoglobin levels were lower in the samples with thiopental. Three explanations seem plausible: (1) Thiopental blocks the hydroxylase of the endoplasmic reticulum, with the result that o-toluidine cannot be further metabolised, leading to higher o-toluidine and methaemoglobin levels. (2) Isoflurane improves the blood supply of the liver. This results in increased metabolism of prilocaine to o-toluidine. (3) The results were accidental. To clarify which of these explanations is correct, further investigation is necessary.

Can somatosensory evoked potential monitoring predict energy dynamics during controlled hypotension?

Somatosensory evoked potentials and redox (reduction/oxidation ratio) of cytochrome a,a3 were studied simultaneously before, during, and after controlled hypotension used for arteriovenous malformation resection. These studies were also conducted before and after ED-IC (external-internal carotid) bypass procedures for treatment of patients with transient ischaemic attacks. The former served as an acute model and the latter as a chronic model of low blood flow (ischaemia). The use of non-invasive reflection spectrophotometry (for redox studies) in conjunction with somatosensory evoked potentials (SSEP) has demonstrated that metabolic and electrophysiological changes parallel each other during "controlled" hypotension. The authors conclude that an analysis of SSEP changes are valuable in the study of metabolic and functional states of the brain during controlled hypotension.

Direct approach to arteriovenous malformations in functional areas of the cerebral hemisphere.

The natural history of patients with arteriovenous malformations (AVM's) suggests that serious morbidity associated with AVM's in functional areas is likely to be much greater than in silent areas. Various modes of treatment of AVM's in functional areas, including direct surgical intervention, embolization, and irradiation, have been considered to carry high risks. The authors advocate direct surgical intervention to these AVM's via a microsurgical technique based on knowledge of the hemodynamic anatomy of AVM's. The technique is designed to circumscribe the AVM without removing any surrounding cortical tissue or white matter and to preserve microcirculation in the functional area. Controlled hypotension (mean arterial blood pressure 40 to 60 mm Hg) is appropriate to enhance the safety of surgical procedures without causing metabolic and electrophysiological dysfunction. Another means to prevent neurological complications is multi-staged resection of larger AVM's, which permits obliteration compartment by compartment. This technique has the advantage of maintaining circulatory sufficiency in the functional area. There was no mortality among 56 patients who underwent the surgical procedure described. Of those, 55 patients resumed their preoperative occupation and one patient became self-sufficient.

Choroid plexus tumors in infants.

Choroid plexus tumors continue to be a challenge in their diagnosis, treatment, and associated conditions that occur in perioperative management. Improved radiographic techniques have recently aided surgical planning, and recent developments in operating room technology are having an impact on the favorable outcome of these patients. Cerebrospinal fluid dynamics often add to the complexity of the perioperative management.

Pediatric choroid plexus neoplasms: MR, CT, and pathologic correlation.

Choroid plexus papillomas are rare, constituting approximately 0.5% of all intracranial neoplasms. Four benign choroid plexus papillomas and one choroid plexus carcinoma were retrospectively reviewed in patients aged 4-20 months who had been examined with magnetic resonance (MR) imaging with a field strength of 0.5 T or 1.0 T and with computed tomography (CT) before and after the administration of contrast material. In general, the tumors were of intermediate signal intensity on T1-weighted images and of either intermediate or increased signal intensity (T2 lengthening) with T2 weighting. All demonstrated variable areas of internal signal void interpreted as signifying regional blood flow, calcification, or old hemorrhage. CT findings included relatively uniform contrast enhancement. Microscopic pathologic changes of the benign lesions mimicked the appearance of normal choroid plexus and confirmed the highly vascular nature of these tumors. MR imaging, with its high-resolution multiplanar techniques, offers direct visualization of these lesions in relation to normal anatomy and better discrimination and confirmation of their intraventricular location, facilitating surgery and postoperative follow-up.

Safety limits of controlled hypotension in humans.

Controlled hypotension is a safe and convenient means of allowing a surgeon to perform intracranial aneurysm, arteriovenous malformation and vascular tumor surgery. The mean arterial pressure between 40 and 60 mmHg induces diminished pulsatile arterial pressure, thus preventing rupture of these abnormal vasculatures. It is still possible to maintain cerebral metabolism in the functional level within this 40-60 mmHg blood pressure range. This statement is based on our experience of the physical and neurological outcome of patients after surgery, and on analyses of somatosensory evoked potential and redox of cytochrome a, a3 in the mitochondria of the cerebral cortical cells.

Lumbosacral intramedullary myolipoma. Case report.

Intradural myolipoma is an unusual tumor. A case is described in which extramedullary and intramedullary striated muscle fibers caused gross contraction of a lipomatous tumor as well as of the spinal cord during electrical stimulation.

Acute spontaneous cerebellar hemorrhage.

Acute spontaneous cerebellar hemorrhage presenting with ataxia, dysarthria, vomiting, dizziness, and coma is commonly the result of hypertension. Early diagnosis is possible, and appropriate treatment, if timely executed, may be lifesaving.

Stereotactic localization of deep-seated arteriovenous malformations in the functional area.

The authors approached 5 arteriovenous malformations (AVMs) located deep in functional areas of the cortex with major draining veins found in the mesial hemisphere by using the stereotactic method. This procedure is a useful adjunct to total resection of these AVMs. No mortality or significant morbidity was noted after the surgical procedure. The authors believe that direct attack on AVMs in areas of critical function is the procedure of choice in order to preserve neurological function.