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Biomedical subjects

D Knierim

Publications and source records attributed to D Knierim.

10 recordsLinked to original sources

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.

Bacillus Phages↗

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.

Amino Acid Motifs↗

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).

Arterioles↗

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.

Animals↗

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.

Adolescent↗

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.

Acute Disease↗

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.

Cerebral Angiography↗

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.

Adolescent↗