Mild traumatic brain injury is the most prevalent neurological insult and frequently results in neurobehavioural sequelae. However, little is known about the pathophysiology underlying the injury and how these injuries change as a function of time. Although diffusion tensor imaging holds promise for in vivo characterization of white matter pathology, both the direction and magnitude of anisotropic water diffusion abnormalities in axonal tracts are actively debated. The current study therefore represents both an independent replication effort (n = 28) of our previous findings (n = 22) of increased fractional anisotropy during semi-acute injury, as well as a prospective study (n = 26) on the putative recovery of diffusion abnormalities. Moreover, new analytical strategies were applied to capture spatially heterogeneous white matter injuries, which minimize implicit assumptions of uniform injury across diverse clinical presentations. Results indicate that whereas a general pattern of high anisotropic diffusion/low radial diffusivity was present in various white matter tracts in both the replication and original cohorts, this pattern was only consistently observed in the genu of the corpus callosum across both samples. Evidence for a greater number of localized clusters with increased anisotropic diffusion was identified across both cohorts at trend levels, confirming heterogeneity in white matter injury. Pooled analyses (50 patients; 50 controls) suggested that measures of diffusion within the genu were predictive of patient classification, albeit at very modest levels (71% accuracy). Finally, we observed evidence of recovery in lesion load in returning patients across a 4-month interval, which was correlated with a reduction in self-reported post-concussive symptomatology. In summary, the corpus callosum may serve as a common point of injury in mild traumatic brain injury secondary to anatomical (high frequency of long unmyelinated fibres) and biomechanics factors. A spatially heterogeneous pattern of increased anisotropic diffusion exists in various other white matter tracts, and these white matter anomalies appear to diminish with recovery. This macroscopic pattern of diffusion abnormalities may be associated with cytotoxic oedema following mechanical forces, resulting in changes in ionic homeostasis, and alterations in the ratio of intracellular and extracellular water. Animal models more specific to the types of mild traumatic brain injury typically incurred by humans are needed to confirm the histological correlates of these macroscopic markers of white matter pathology.

BACKGROUND: Multimedia Messaging Service (MMS) is used by neurosurgical residents to transmit scan images to the attending neurosurgeon in conjunction with telephone consultation. This service has been well received by the attending neurosurgeons, who felt that after viewing scan images on their phones, they felt increased confidence in clinical decision making and that it reduced the need for recall to the hospital. OBJECTIVE: The use of MMS can be extended to junior doctors making referrals from regional hospitals with no neurosurgical cover. This study aims to validate the competency of non-neurosurgically trained junior doctors in selecting optimal images to transmit via MMS to the attending neurosurgeon on call. METHODS: Ten junior doctors with no formal neurosurgical training and five neurosurgical residents were interviewed. They were shown the full complement of images together with relevant clinical history and assessment. They were then asked to make the radiological diagnosis and then select two images for MMS transmission to the attending neurosurgeon that they thought would best aid the neurosurgeon in clinical decision making. The attending neurosurgeon was asked to comment, on each image, whether his management plan would differ if he was shown the entire series of the images. RESULTS: All the images chosen are deemed appropriate, and the decision made based on the MMS images would be similar if the entire series of images were available to the neurosurgeon. However, 7 of 10 junior doctors were unable to read magnetic resonance images of lumbar spine. There was no significant difference in the images chosen by the neurosurgical residents and the junior doctors. CONCLUSION: It is feasible and safe for junior doctors to utilize MMS to transmit computed tomographic images to a neurosurgeon while making an urgent referral. The images selected are representative of the disease pathology and facilitate clinical decision making.

A method for exploring protein-protein interactions using hydrogen/deuterium exchange coupled to mass spectrometry is described. The method monitors the exchange of backbone (amide) hydrogens in solutions of deuterated water that primarily occur on portions of the protein exposed to solvent. In the presence of a protein binding partner, regions that experience reduced exchange are either part of the protein-protein interaction interface or undergo conformational changes to reduce accessibility to solvent. This method has the advantage of being used under physiological conditions with unmodified proteins. In this chapter, we describe an approach suitable for probing interactions among relatively large proteins using conventional mass spectrometry systems. The interaction between human transferrin and the Neisseria meningitidis receptor protein, transferrin binding protein B, provides a challenging system as an example.

HASH(0x2b1624a93ed0)

Pathogenic bacteria in the Neisseriaceae possess a surface receptor mediating iron acquisition from human transferrin (hTf) that consists of a transmembrane iron transporter (TbpA) and a surface-exposed lipoprotein (TbpB). In this study, we used hydrogen/deuterium exchange coupled to mass spectrometry (H/DX-MS) to elucidate the effects on hTf by interaction with TbpB or derivatives of TbpB. An overall conserved interaction was observed between hTf and full-length or N-lobe TbpB from Neisseria meningitidis strains B16B6 or M982 that represent two distinct subtypes of TbpB. Changes were observed exclusively in the C-lobe of hTf and were caused by the interaction with the N-lobe of TbpB. Regions localized to the 'lip' of the C1 and C2 domains that flank the interdomain cleft represent sites of direct contact with TbpB whereas the peptides within the interdomain cleft that encompass iron binding ligands are inaccessible in the closed (holo) conformation. Although substantial domain separation upon binding TbpB cannot be excluded by the H/DX-MS data, the preferred model of interaction involves binding hTf C-lobe in the closed conformation. Alternate explanations are provided for the substantial protection from deuteration of the peptides encompassing iron binding ligands within the interdomain cleft but cannot be differentiated by the H/DX-MS data.

Of the 834 Salmonella spp. isolates collected from 2002 to 2004 in the New Territories East Cluster hospitals in Hong Kong, 21% were resistant to 3-14 antibiotics and 30% were resistant to nalidixic acid with reduced susceptibility to ciprofloxacin. A class 1 integron was present in 105 (13%) isolates, of which more than 50% were Salmonella Typhimurium. Class 2 or 3 integrons were not detected in any of the isolates. DNA sequencing showed that there were 16 different integrons, of which the most common, detected in 40% of isolates, was dfrA12-orfF-aadA2. Almost half of the isolates had lost the qacEdelta1-sulI 3' conserved segment (3'CS). Four isolates had an orf513 gene downstream of the 3'CS. There was probably one predominant clone among each heterogeneous population of integron-harboring S. Typhimurium, S. Virchow, and S. Derby isolates in our community. This is the first thorough study of the prevalence and characteristics of integrons in Salmonella isolates in Hong Kong and the first to report a class 1 integron in S. Birkenhead, S. Landwasser, S. Litchfield, and S. Reading.

1. PCR and RT-PCR methods for 5-hour and 3-hour detection, respectively, of salmonellae and Vibrio cholerae in stool, food and environmental water samples have been developed. 2. Such methods can be used in routine laboratories for rapid detection of salmonellae and V cholerae and are essential for infection control purposes.

A total of 182 Salmonella enterica serotype Typhi isolated from three hospitals in Hong Kong from 1986 to 1992 were tested for their susceptibility to 21 antimicrobial agents. Four percent or less were resistant to chloramphenicol, ampicillin, some of the cephalosporins, nalidixic acid, tetracycline and trimethoprim and 6% to 1024 mg/l sulfamethoxazole. All were susceptible to the aminoglycosides and the 4-quinolones. Nineteen isolates were resistant to at least 1, and up to 9, antibiotics. Of 8 chloramphenicolor multiply-resistant isolates studied, only 3 could transfer their resistances while resistance of one could only be mobilized. Four of 5 ampicillin-resistant strains produced a beta-lactamase of pI 5.5. Antibiotic resistances were mediated by plasmids of 106, 116 or 221 kb of incompatibility groups H, I1 and K. Three resistant isolates did not harbour any plasmid. A total of 43 (24%) S. Typhi harboured plasmids ranging in size from 4.3 to 221 kb. Plasmids of 106 kb and 8.5 kb were found in 17 and 10 isolates, respectively. Restriction enzyme digestion of these two plasmids showed that each could be differentiated into 3 types. Of 89 isolates that were phage typed, 38% were untypable, while 17% and 12% were of phage types E1 and A, respectively, and the rest belonged to 17 other types.

A selective and sensitive method of microemulsion EKC (MEEKC) with electrochemical detection (ED) was developed for separation and determination of 14 flavonoids. In order to obtain the better stability for the studied flavonoids, oil (ethyl acetate) with low interfacial surface tension was employed as organic solvent. A running buffer composed of 0.9%(w/v, 30 mM) SDS, 0.9%(w/v, 21 mM) sodium cholate (SC), 0.9%(w/v, 121 mM) butan-1-ol, 0.6%(w/v, 68 mM) ethyl acetate, and 98.2% v/v 10 mM Na(2)B(4)O(7)-20 mM H(3)BO(3) buffer (pH 7.5) was applied for the separation of flavonoids. Under the optimum conditions, the relationship between peak currents and analyte concentrations was linear over about 1.3 and 1.7 orders of magnitude with detection limits (defined as S/N = 3) ranging from 0.02 to 0.5 mug/mL for all analytes. This method was applied for the determination of flavonoids in real samples with simple extraction procedures, and the assay results were satisfactory.