Twelve tips on how to survive PBL as a medical student.

BACKGROUND: Starting medical school can be both exciting and daunting. This is particularly the case when the style of learning is different from that which has been experienced previously. For many students, their first experience of learning through a problem-based learning (PBL) approach is when they commence their medical student programme. AIM: This article provides 12 tips on how to survive PBL as a medical student. METHODS: The tips have been based on the authors' experience of PBL and the current literature evidence base. A chronological order was used for the tips to guide the reader, whether student or PBL facilitator, through tips for the various stages of the PBL process. RESULTS: These 12 tips provide students and PBL facilitators with 12 practical tips to help them to realise the learning process and rationale for PBL. The tips commence with surviving the initial PBL sessions and continue through the process, finishing with the use of PBL in the clinical setting where the written scenarios are replaced by patient case histories. CONCLUSION: Using a PBL approach facilitates the learning of clinical and science knowledge in context through clinical scenarios, whilst working and learning together as a group. It is envisaged that these tips will be beneficial for PBL facilitators working with students that are new to PBL, and for the PBL students themselves.

Effects of intraarticular IL1-Ra for acute anterior cruciate ligament knee injury: a randomized controlled pilot trial (NCT00332254).

OBJECTIVE: To evaluate the clinical effectiveness of intraarticular IL-1 receptor antagonist (IL-1Ra) for anterior cruciate ligament (ACL) tear. METHODS: Eleven patients with acute ACL tear confirmed by magnetic resonance imaging (MRI) were randomized to receive a single intraarticular injection of IL-1Ra (anakinra 150 mg, n = 6) or equal volume of saline placebo (1 ml, n = 5). The double-blinded treatment was administered a mean 2 weeks after injury. Synovial fluid (SF) (n = 9 patients) and sera (all patients) were available at baseline (prior to injection) and immediately prior to surgery (mean 35 days later) and analyzed for SF IL-1α, IL-1ß, IL-1Ra and serum hyaluronan (HA), an indicator of synovial inflammation. The primary outcome, standardized Knee Injury and Osteoarthritis Outcome Score (KOOS) questionnaire, was obtained at 0 (baseline), 4, and 14 days after injection. RESULTS: Compared with placebo, the IL-1Ra group had substantially greater improvement in key outcomes over 14 days (KOOS pain P = 0.001; activities of daily living P = 0.0015; KOOS sports function P = 0.0026; KOOS quality of life (QOL) P = 0.0048; and total KOOS P < 0.0001). There were no adverse reactions in either group. SF IL-1α (P = 0.05) and serum HA (P = 0.03), but not IL-1ß, or IL-1Ra, decreased significantly in the IL-1Ra but not the placebo treated patients. Compared with placebo, IL-1α was borderline significantly different in the IL-1Ra treated group (P = 0.06). CONCLUSIONS: Administered within the first month following severe knee injury, IL-1Ra reduced knee pain and improved function over a 2-week interval. This promising proof of concept study provides a new paradigm for studies of acute joint injury and suggests that a larger follow-up study is warranted.

Forensic investigation of cross platform massively multiplayer online games: Minecraft as a case study.

Minecraft, a Massively Multiplayer Online Game (MMOG), has reportedly millions of players from different age groups worldwide. With Minecraft being so popular, particularly with younger audiences, it is no surprise that the interactive nature of Minecraft has facilitated the commission of criminal activities such as denial of service attacks against gamers, cyberbullying, swatting, sexual communication, and online child grooming. In this research, there is a simulated scenario of a typical Minecraft setting, using a Linux Ubuntu 16.04.3 machine (acting as the MMOG server) and Windows client devices running Minecraft. Server and client devices are then examined to reveal the type and extent of evidential artefacts that can be extracted.

Metabolism of Hydroxy Fatty Acids in Developing Seeds in the Genera Lesquerella (Brassicaceae) and Linum (Linaceae).

Members of the genus Lesquerella produce seed oil that contains a high proportion of hydroxy fatty acids (HFAs). There are three groups of Lesquerella species that are distinguished by their most abundant seed oil fatty acid: lesquerolic acid (20:1OH; e.g. Lesquerella fendleri), densipolic acid (18:2OH; e.g. Lesquerella kathryn), and auricolic acid (20:2OH; e.g. Lesquerella auriculata). To investigate the biochemistry of HFA production in Lesquerella species, the conversion of putative radiolabeled intermediates of HFA biosynthesis, including 18:1, 20:1,18:1OH, 18:2OH, and 20:1OH, was examined in developing embryos of L. fendleri, L.kathryn, and L. auriculata. The results are consistent with (a) 18:1OH formation by hydroxylation of 18:1, (b) elongation and desaturation of 18:1OH to produce 20:1OH and 18:2OH, respectively, and (c) desaturation of 20:1OH to produce 20:2OH. The desaturation of 20:1OH was also found to occur in developing embryos of high, but not low, linolenic acid flax. This suggests that the desaturation is catalyzed by the extraplastidial linoleate desaturase. Confirming this suggestion, it was notable that 18:1OH and 18:2OH were found in low and high linolenic flax (Linum usitatissimum) seeds, respectively, at levels of 0.2 to 1%.

Induction of Lipid and Oleosin Biosynthesis by (+)-Abscisic Acid and Its Metabolites in Microspore-Derived Embryos of Brassica napus L.cv Reston (Biological Responses in the Presence of 8[prime]-Hydroxyabscisic Acid).

Microspore-derived (MD) embryos of Brassica napus L. cv Reston were used to test the effects of (+)-abscisic acid ([(+)-ABA]) and its metabolites, 8[prime]-hydroxyabscisic acid (8[prime]-OH ABA) and (-)-phaseic acid (PA), on the accumulation of very long-chain monounsaturated fatty acids (VLCMFAs) and induction of genes encoding a 19-kD oleosin protein and a [delta]15 desaturase during embryogenesis. Developing early to mid-cotyledonary MD embryos at 16 to 19 d in culture were treated with 10 [mu]M hormone/metabolite for 4 d. At various times during incubation, embryos and medium were analyzed to determine levels of hormone/metabolite, VLCMFAs, and oleosin or [delta]15 desaturase transcripts. The VLCMFAs, 20:1 and 22:1, primarily in triacylglycerols, increased by 200% after 72 h in the presence of (+)-ABA and 8[prime]-OH ABA relative to the control. In contrast, treatment with PA for 72 h had little effect (20% increase) on the level of VLCMFAs. The first 24 to 72 h of (+)-ABA treatment were critical in the induction of VLCMFA biosynthesis, with 8[prime]-OH ABA lagging slightly behind (+)-ABA in promoting this response. The accumulation of VLCMFAs was positively correlated with an increase in elongase activity. (+)-ABA and its 8[prime]-OH ABA metabolite induced the accumulation of a 19-kD oleosin transcript within 2 to 4 h in culture. In addition, both (+)-ABA and 8[prime]-OH ABA induced the same level of [delta]15 desaturase transcript by 8 h. PA had no effect on the induction of either oleosin or [delta]15 desaturase transcripts. To our knowledge, this is the first report of the biological activity of 8[prime]-OH ABA and of stimulatory effects of (+)-ABA and 8[prime]-OH ABA on lipid and oleosin biosynthesis.

Microsomal Lyso-Phosphatidic Acid Acyltransferase from a Brassica oleracea Cultivar Incorporates Erucic Acid into the sn-2 Position of Seed Triacylglycerols.

Developing seeds from Brassica oleracea (L.) var botrytis cv Sesam were examined for the ability to biosynthesize and incorporate erucic acid into triacylglycerols (TAGs). Seed embryos at mid-development contained a high concentration of erucic acid in diacylglycerols and TAGs, and substantial levels were also detected in free fatty acids, acyl-coenzyme A (CoA), phosphatidic acid, and phosphatidylcholine. Homogenates and microsomal fractions from seeds at mid-development produced [14C]eicosenoyl- and [14C]erucoyl-CoAs from [14C]oleoyl-CoA in the presence of malonyl-CoA and reducing equivalents in vitro. These fatty acids were incorporated into TAGs via the Kennedy pathway. However, unlike most Brassicaceae, the B. oleracea was able to insert significant erucic acid into the sn-2 position of TAGs. It was shown that the lyso-phosphatidic acid acyltransferase (LPAT) incorporated erucic acid into the sn-2 position of lyso-phosphatidic acid. The erucoyl-CoA:LPAT activity during seed development and the sn-2 erucic acid content of the TAG fraction in mature seed were compared to those in B. napus, Tropaeolum majus, and Limnanthes douglasii. There was a correlation between the in vitro erucoyl-CoA:LPAT activity and the sn-2 erucic acid content in seed TAGs. To our knowledge, this is the first member of the Brassicaceae reported to have an LPAT able to use erucoyl-CoA. This observation has important implications for efforts being made to increase the erucic acid content in B. napus, to supply strategic industrial feedstocks.

Cloning and molecular characterization of an Arabidopsis thaliana RING zinc finger gene expressed preferentially during seed development.

The RING (Really Interesting New Gene) finger is a zinc-binding domain that is found in proteins from a variety of species. This paper reports the cloning and characterization of, as yet, only the second RING finger protein gene from plants, A-RZE, in Arabidopsis thaliana. In addition to the RING-finger motif, A-RZF also contains a putative nuclear localization signal. A-RZF is encoded by a single copy gene with an intron of 595 bp interrupting the 5' leader sequence and the coding region. Northern blot analysis indicated that A-RZF is expressed preferentially during seed development. The RING-finger motif, putative nuclear localization signal, and unique expression pattern, predict an important role during seed development for A-RZF.

Facial thermal input to the trigeminal spinal nucleus of rabbits and rats.

In rabbits and rats under urethane anesthesia, a systematic survey was made of the caudal trigeminal nucleus, using glass coated tungsten microelectrodes. This revealed many neurons in the marginal layer sensitive either to warming or cooling the facial skin. The majority of these neurons were specifically temperature sensitive. In rats, a somatotopic arrangement of cold receptive fields was evident within the marginal layer of the trigeminal nucleus, with the ophthalmic division of the trigeminal nerve represented most laterally, the mandibular most medially and the maxillary division represented over most of the recording region. This arrangement is similar to the mediolateral distribution seen in cats. In rabbits, the distribution was less rigorous but the ophthalmic division tended to be represented caudally and the mandibular division rostrally. The maxillary division was represented over most of the recording region. All receptive fields were ipsilateral and showed spatial convergence of input. Both rabbits and rats possessed a concentration of thermal receptive-fields around the nose, whisker pad and mouth. At steady skin temperatures, marginal units gave bell-shaped intensity functions which were very similar to those reported for the cold and warm receptors. With rapid changes of temperature, neurons responded with a dynamic outburst which corresponded to the equivalent receptor response.

Measurement of lysozyme by an enzyme-linked immunosorbent assay.

In this study an enzyme-linked immunosorbent assay has been developed for the determination of lysozyme in saliva, serum and urine. The assay relies on the detection of specific protein rather than lytic activity, a property which has been shown to be most suitable for the quantitation of lysozyme in mucin containing substances. Our results indicate that no pretreatment is necessary for the immunochemical method. The assay is sensitive to concentrations as low as 1 microgram lysozyme/l. The intra-assay and inter-assay coefficients of variation were 5.9% and 15.8% respectively. The lysozyme level in whole saliva was 55.53 +/- 30.35 mg/l, in serum the level was 0.64 +/- 0.15 mg/l and in urine it was 0.17 +/- 0.22 mg/l. Comparisons between immunochemical determination and lytic assays showed a good correlation (serum, r = 0.79, P less than 0.01; saliva, r = 0.85, P less than 0.005; treated saliva, r = 0.96, P less than 0.001).

Lipid composition of metacestodes of Taenia taeniaeformis and lipid changes during growth.

A lipid analysis was performed on developing metacestodes of Taenia taeniaeformis removed from the livers of rats at times varying from 3 to 35 weeks post infection. Lipid accounted for 7-21% of the dry weight of the parasites. The highest proportions were found at the earlier stages. The distribution was as follows; neutral lipid 27-45%; glycolipid 5-11%; and phospholipid 50-61%. The major neutral lipid was cholesterol, and minor neutral lipids were sterol esters, triglycerides, diglycerides and monoglycerides. Hydrocarbons were present throughout development, but in the highest amounts at the earlier stages. Five different glycolipids were found, all of which were identified as glycosphingolipids. An increase in the proportion of more complex glycolipids was noted as parasites grew older. Ten different phospholipids were identified, with the major components being phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine. Other phospholipids were: lysophosphatides, phosphatidylinositol, phosphatidic acid, diphosphatidylglycerol, sphingomyelin, and an unknown phospholipid component. Changes in the relative amounts of the two major phospholipids were found when the early and late stages were compared. Two lipids found throughout development were identified as glycosylated dolichol phosphates, and they comprised between 1 and 3% of the total phospholipid fraction. Nineteen fatty acids were detected, and the fatty acid distribution for each lipid class at each stage was determined. Seven major fatty acids were common to each. These were: hexadecanoic, octadecanoic, oleic, linoleic, arachidonic, docosanoic, and docosahexaenoic.

Papillary urothelial hyperplasia. A precursor to papillary neoplasms.

Precursor lesions of papillary urothelial neoplasms have not been well characterized. We reviewed the surgical pathology files of the Johns Hopkins Hospital and three regional hospitals from 1992 to present. Sixteen cases of papillary hyperplasia, defined as undulating urothelium arranged into thin mucosal papillary folds, were identified (in 11 men and five women: age range, 40-89 years). Relative to the diagnosis of papillary hyperplasia, nine patients had a history of papillary urothelial neoplasms; in one of these cases, the patient also had subsequent papillary urothelial neoplasms, and two of these patients had concurrent papillary urothelial neoplasms with papillary hyperplasia. In one of these nine cases, papillary hyperplasia arose in the scar of a prior papillary urothelial neoplasm. In two cases, the patients had concurrent, yet no prior history, of papillary urothelial neoplasms. Of these 11 cases, three had multiple resections showing papillary hyperplasia over time. In case 12, the patient had a history of moderate urothelial atypia. The remaining four patients had no history of papillary urothelial neoplasms or urothelial atypia. We describe papillary hyperplasia as a well-defined entity that is usually asymptomatic and generally found on routine follow-up cystoscopy for papillary urothelial neoplasms. Papillary hyperplasia appears to be a precursor lesion of low-grade papillary urothelial neoplasms.

Radiolabeling, stability, and body distribution in rats, of low molecular weight polylactide homopolymer and polylactide-polyethyleneglycol copolymer.

In order to study its fate in vivo, a low molecular-weight polylactide homopolymer was derivatized with a p-methoxyphenyl moiety, so as to make it susceptible to radiolabeling with 125I. A low molecular weight polylactide-polyethyleneglycol copolymer capped with ap-methoxyphenyl residue was also synthesized. The derivatized polymers were successfully [125I]iodinated in organic medium. The radiolabeled products were freed from [125I]iodide by dialysis and shown to be stable for 24 h on incubation at 37 degrees C in buffered saline or in blood. On longer incubation at 37 degrees C in buffered saline the radiolabeled polylactide released [125I]iodide and [125I]iodinated 3-(p-methoxyphenyl)propionic acid. The radiolabeled copolymer was more stable on incubation at 37 degrees C in buffered saline, but some [125I]iodide was released. The tissue distribution of radioactivity was determined 5 min, 1, 5 and 24 h after injecting male rats with 125I-labeled homopolymer or copolymer. Intravenous, intraperitoneal and subcutaneous injection routes were employed. Further rats were injected with [125I]iodide, to aid interpretation of the data. After administration of labeled homopolymer, a high concentration of radioactivity was found in the liver tissue. The levels slowly decreased over 24 h, and the polymer was successively found in the small and large intestine and the faeces. This is probably indicative of excretion via the bile. Concurrently radioactivity was excreted in the urine. After administration of labeled copolymer, a high concentration of radioactivity was found in the liver and the residual soft tissue, the latter fraction containing two-thirds of the radioactivity one hour after injection. The precise tissue location that this result indicates was not identified. After 1 h radioactivity was excreted in the faeces, again probably via the bile, and in the urine. Tissue distributions after intraperitoneal or subcutaneous injections were concordant with the above results and interpretations, with the additional factor of slow clearance from the injection site.

Steric stabilization of microspheres with grafted polyethylene oxide reduces phagocytosis by rat Kupffer cells in vitro.

Sterically stabilized polyethylene oxide-polystyrene copolymer microspheres, (PS-PEO) and charge stabilized polystyrene (PS) microspheres of similar size (1 micron) were prepared in order to compare their uptake by cultured rat Kupffer cells isolated by centrifugal elutriation. The uptake of the sterically stabilized particles was found to be much less than that for the charge stabilized control. The uptake of microspheres stabilized with covalently grafted PEO was lower or equivalent to that of control microspheres stabilized by the adsorption of the non-ionic PEO-polypropylene oxide (PPO-PEO) surfactant Poloxamer 238 or Methoxy-PEO. Phagocytic uptake by Kupffer cells at low and body temperature (8 degrees C and 37 degrees C) demonstrated that PS-PEO particles showed both low adherence and low metabolic uptake. The adsorption of PEO, as Poloxamer 238, to particles with covalently attached or grafted PEO resulted in a synergistic reduction in uptake that was greater than the individual effects of grafting and adsorption alone (P less than or equal to 0.001). It is suggested that this combination produces a more effective steric barrier on the particle surface with the Poloxamer adsorbing to the surface between the grafted PEO chains. The relevance to drug targeting/carrier systems is discussed.

Influence of surface coverage with poly(ethylene oxide) on attachment of sterically stabilized microspheres to rat Kupffer cells in vitro.

The attachment to rat Kupffer cells of polymeric microspheres, sterically stabilized with different amounts of pendant poly(ethylene oxide) (PEO), was assessed in vitro. Four types of copolymer polystyrene (PS) microspheres were synthesized by variation of four possible monomer ratios that included styrene, methoxy-PEO-methacrylate (750 and 2000 mol. wt PEO) and allylurea. This produced poly(styrene-(methoxy-PEO)methacrylate) microspheres with hydrophilic side-groups of either urea (PS-U-PEO) and/or mixed molecular weight (750/2000 mol. wt) PEO (PS-U-M-PEO, PS-M-PEO), or single molecular weight (2000) PEO (PS-PEO) at their surfaces. The hypothesis was tested that increasing the total content of PEO comprising the steric barrier reduces attachment to cell surfaces. Attachment of PEO microspheres bearing the urea spacer and/or mixed molecular weight PEO was found to be intermediate between charge stabilized control PS and PEO (2000 mol. wt) bearing particles. Post-adsorption of different Poloxamer (PEO-poly(propylene oxide)-PEO) surfactants to the microspheres further decreased attachment. Significant negative linear correlations between surface PEO content, measured by electron spectroscopy for chemical analysis (ESCA), and attachment to Kupffer cells were demonstrated. Decreases in attachment also resulted with all graft PEO particles bearing adsorbed sodium dodecyl sulphate (SDS), whilst the attachment of SDS-treated PS control particles increased. It is proposed that trains of adsorbed graft PEO are displaced by the SDS to increase the effective fraction of graft PEO within the steric layer. Overall, increasing the amount of hydrophilic PEO in the steric layer, from graft and adsorbed sources, reduces the attachment of these particles to Kupffer cells in vitro.

Duplex ultrasound in the assessment of vascular disease in clinical hypertension.

Duplex ultrasound combines B-mode imaging and pulsed Doppler flow analysis and is rapidly becoming the most useful and widely applied method for the evaluation of atherosclerotic arterial disease. Recent technical improvements including color flow Doppler allow its application to practically all vascular beds in the body. Duplex is noninvasive and in many areas its accuracy in the detection of arterial stenosis and occlusion rivals angiography. Duplex ultrasound has become the preferred method of evaluation of patients with cerebrovascular disease. It provides information about the degree of stenosis, location of disease, and plaque characterization, and in many cases replaces standard cerebral arteriography. Duplex ultrasound has been used to detect renal artery stenosis in patients suspected of having renovascular hypertension with an accuracy of 93% in the diagnosis of hemodynamically significant stenosis. It also has important applications in the assessment of mesenteric artery stenosis, peripheral vascular disease, and venous disease. Because it is accurate, reasonably inexpensive, and noninvasive, duplex ultrasound can be used to follow the progression of untreated atherosclerotic lesions throughout the body. Used in this fashion, it provides important information about the natural history of atherosclerotic lesions, their clinical prognosis, and the effect of interventions, either medical or surgical, on lesions.

Biotypes of Haemophilus parainfluenzae from the respiratory secretions in chronic bronchitis.

A total of 2401 isolates of Haemophilus parainfluenzae was isolated from respiratory secretions of 36 healthy adults and 128 patients with chronic bronchitis over a period of 1 year. The isolates were allocated to eight biotypes, by their production of indole, urease and ornithine decarboxylase. Biotypes I and II constituted most of the isolates of H. parainfluenzae from the oropharynx of controls (75%) and chronic bronchitics (c. 90%). Among the patients, there was no difference in the isolation rate between oropharyngeal swabs and sputum specimens. Biotypes III, IV, VI, VII and VIII were isolated less frequently, as was a new taxon defined here as biotype V which does not produce indole, urease or ornithine decarboxylase. Biotype III was isolated significantly less frequently from cases of chronic bronchitis than from controls, whereas biotype II was isolated somewhat more frequently from the patients, especially during acute episodes.

The use of fluorescent tracers in the peripheral sensory nervous system.

We have developed an injection technique which simplifies the use of fluorescent tracers in investigating the projections of peripheral nerves both in rats and pigeons, and avoids the difficulties experienced with the cut nerve technique. The tracers are injected into the peripheral nerve and transported to cell bodies in the dorsal root ganglia (DRG). By a careful choice of tracer it is possible to investigate the projections of two or more nerves in the same preparation. We have used two combinations of tracer which can be viewed simultaneously in the cell at the same excitation wavelength (405 nm): (1) propidium iodide (PI) and bisbenzimide (Bb) which produce an orange cytoplasmic fluorescence and blue nuclear fluorescence respectively; and (2) Fast blue (FB) and Nuclear yellow (NY) which label the cytoplasm blue and the nucleus yellow. This distribution of tracers is similar to that reported in central neurones but differs from that reported in the efferent peripheral nervous system. Using these tracers we have observed double-labelled neurones in the DRG of both rats and pigeons, which is an indication for the dichotomizing of peripheral fibres. The existence of two tracers in double-labelled cells has been confirmed microspectrophotometrically.

Somatization and the vocabulary of everyday bodily experiences and concerns: a community study of adolescents.

OBJECTIVE: To describe the frequency of everyday bodily experiences and health concerns in a general population of adolescents 12 to 16 years of age in Ontario and to explore whether the concept of "somatization," identified from those youths with many of these symptoms, is meaningful and related to other variables. METHOD: A representative sample of the population was obtained by stratified random sampling. Children with a chronic medical condition were excluded. Parents and their adolescent children filled out a series of questionnaires to measure health concerns, complaints, and more dramatic losses of function. Information was also collected on certain background factors, psychiatric problems, and impairments in adaptive functioning. RESULTS: Parents and youths endorsed the items with the same rank order of frequency, but there was virtually no agreement between parents and youths on the presence or absence of individual somatic symptoms. Users of medical services did not tend to have many more health concerns than others, and there was a weak relationship between the number of health concerns reported by a youth and both impairment in adaptive functioning and psychiatric problems. CONCLUSION: These data suggest that the concept of somatization has limited general value over and above a relationship with other psychiatric problems.

Long lasting inhibition of horseradish peroxidase (HRP) transport in sensory nerves induced by capsaicin pretreatment of the receptive field.

Capsaicin, when injected into the scrotal skin of adult rats was seen to produce a long-lasting desensitization. To investigate the possible effect of this treatment on the sensory neurones of the pudendal nerve, rats were injected with horseradish peroxidase (HRP) at different time intervals following pretreatment with capsaicin. The HRP was applied in two different ways. In one group of rats HRP was injected into both sides of the scrotal skin between 2 and 12 days after capsaicin pretreatment of one side of the scrotal skin. More HRP labelled neurones were seen in the S1 and L6 dorsal root ganglia (DRG) on the non-treated side than on the pretreated side. Similar results were obtained when the HRP was injected directly into the pudendal nerve. A marked decrease in the number of HRP labelled neurones on the pretreated side was observed even when HRP was injected into the nerve up to 1 year after the capsaicin pretreatment. It appears therefore that at least part of the effect of capsaicin on peripheral sensory neurones is due to an effect of capsaicin on the axoplasmic transport processes, and that this effect is long lasting.