A slip above a slip: retrolisthesis of the motion segment above a spondylolytic spondylolisthesis.

OBJECTIVE: The purpose of this study is to assess the degenerative changes in the motion segments above a L5S1 spondylolytic spondylolisthesis and to view these in light of the retrolisthesis in the segment immediately above the slip. A spondylolytic spondylolisthesis causes an abnormal motion and predisposes to degenerative changes at the L5S1 disc. Degenerative changes in the adjacent segments would influence the symptomatology and natural history of the disease and the treatment options. The extent of degenerative changes in the levels immediately above a L5S1 spondylolytic spondylolisthesis is not well documented in the literature. We have noted retrolisthesis at this level, but this has not been previously reported or assessed. MATERIALS AND METHODS: Thirty-eight patients with a symptomatic L5S1 spondylolytic spondylolisthesis with a mean age of 52.8 years (95% CI 47.2-58.4); 55.3% (n = 21) females and 44.7% (n = 17) males. We assessed the lumbar lordosis, slip angle, sacral slope, grade of the slip, facet angles at L34 and L45 on both sides, facet degenerative score (cartilage and sclerosis values), disc degenerative score (Pfirrmann) at L34, L45 and L5S1 and the presence of retrolisthesis at L45. RESULTS: We noted that 29% (11) had a retrolisthesis at L45. The degenerative scores reduced significantly from L5S1 through L45 and L34. Slip angle and L45 disc degenerative score were the only factors that occurred consistently in patients with a retrolisthesis. CONCLUSIONS: There is a cascade of degenerative changes that involve both the disc and the facet joints at the levels above a spondylolytic spondylolisthesis. The degenerative changes at the L45 disc and a higher slip angle are consistent findings in patients with a retrolisthesis at the level above the slip.

Benign gastric outlet obstruction--spectrum and management.

Intrinsic or extrinsic obstruction of the pyloric channel or duodenum either by benign or malignant diseases leads to gastric outlet obstruction. With improvement in science and technology, the spectrum of gastric outlet obstruction has changed from peptic ulcer disease to corrosives and malignant diseases. Newer investigations like computerized tomography and endoscopy have supplemented the previous clinical tests like saline load test and barium series. Improvised treatment modalities like endoscopic balloon dilatation and endoscopic incision have circumvented the use of surgery which was the gold standard for management of gastric outlet obstruction. Newer modalities like biodegradable stents have an upcoming role in the management.

Oxidative stability and shelf-life evaluation of selected culinary oils.

Four out of eight 'healthier' oils-namely, almond oil, avocado oil, hazelnut oil and macadamia nut oil-studied were rich sources of monounsaturated fatty acids like olive oil. Grape seed oil, rice barn oil (marketed recently), toasted sesame oil and walnut oil contained high levels of essential fatty acids. The order of oxidative stability determined by Rancimat measuring of the induction period at four temperatures (90 degrees C, 100 degrees C, 110 degrees C, and 120 degrees C) was found to be macadamia oil > rice bran oil approximately toasted sesame oil > avocado oil > almond oil > hazelnut oil > grape seed oil > walnut oil. High-level monounsaturated fatty acid oils gave a linear relationship between 100 times the reciprocal of the induction period against the total unsaturated fatty acid content obtained as %C18:2 + 0.08 x C18:1 + 2.08 x %C18:3, while the polyunsaturated fatty acid oils gave an exponential relationship. In the case of rice bran and hazelnut oils, shelf-life prediction from the extrapolation of the Arrhenius plots and the Q(10) factors was compared well with that of storage time given by the oil producers. In the cases of the other oils (with an exception of macadamia nut oil), the predicted shelf-lives were significantly lower than that of the storage times; especially, walnut oil (very prone to oxidation) gave 15-20 times lower shelf-life than the best-before storage life.

New UK graduates' knowledge of training and service provision within restorative dentistry - a survey.

Objective To assess new UK graduates' knowledge of training and service provision within restorative dentistry.Design A national descriptive cross-sectional survey.Subjects and methods An online survey assessing clinicians' knowledge of restorative dentistry, who had graduated within the last four years in the UK, was distributed across the UK via postgraduate dental deaneries. One-hundred responses were accepted as a sample of a potential population of 4,000.Main outcome measure How well respondents understood the service provision and training aspects of the specialty of restorative dentistry.Results The responses were received from graduates from a variety of dental schools across the UK. Of those respondents, 41 reported receiving career guidance within restorative dentistry. 45 new graduates were confident in their understanding of the specialty, while 53 were confident in the differences between restorative dentistry and monospecialty training. The respondents appeared unaware regarding treatment priorities within restorative dentistry departments. Most respondents felt that receiving teaching on restorative dentistry as a specialty and career pathway would be beneficial.Conclusion The results suggest that new graduates may benefit from clarification regarding the specialty of restorative dentistry, however, caution must be taken due to the limitations of the study.

The importance of the patient voice in vaccination and vaccine safety-are we listening?

Much has been written about the patient-physician relationship over the years. This relationship is essential in maintaining trust in the complex arena of modern diagnostic techniques, treatment and prevention, including vaccines and vaccine safety. However, a great deal of this material was written from the viewpoint of clinicians and academics. The patient voice may be positive or negative, fragmented or complex. Information sources are weighed and treated differently, according to the value system and risk perceptions of the individual. In post-trust societies, when people have less confidence in health authorities, communication needs to be more than a paternalistic top-down process. Notions of empowerment and individual patient choice are becoming crucial in medical care. The 'voice of the patient', which includes healthy individuals receiving vaccines, needs to be heard, considered and addressed. With respect to childhood immunizations, this will be the voice of the parent or caregiver. The key to addressing any concerns could be to listen more and to develop a communication style that is trust-based and science-informed. Regulatory agencies are encouraging clinical and patient-reported outcomes research under the umbrella of personalized medicine, and this is an important step forward. This paper attempts to reflect the paradigm shift towards increasing attention to the patient voice in vaccination and vaccine safety.

An active-site carboxyl group in liquefying alpha-amylase: specific chemical modification.

Bacillus amyloliquefaciens alpha-amylase activity is pH-dependent and the plot log (Vmax/Km) versus pH implicated a carboxyl group of aspartic acid/glutamic acid at the active site. Chemical modification of alpha-amylase with EDC confirmed this view. Further, analysis of inactivation kinetics showed that modification of a single carboxyl group led to complete loss of the enzymic activity.

Short-chain peptide analysis by high-performance liquid chromatography coupled to electrospray ionization mass spectrometer after derivatization with 9-fluorenylmethyl chloroformate.

Resolution and characterization of short-chain peptides (M(r) = 200-1000) and free amino acids were demonstrated by the use of precolumn derivatization with 9-fluorenylmethyl chloroformate (Fmoc) followed by reverse-phase high-performance liquid chromatography (RP-HPLC) interfaced with an electrospray ionization mass spectrometer (ESI-MS). At pH 10, in addition to derivatization at the N terminus, epsilon-NH(2) and OH groups of lysine and tyrosine residues, respectively, were also derivatized. Fmoc derivatives showed at least 2 orders of magnitude higher ionization potential in the presence of trifluoroacetic acid. The detection levels for both the free amino acid and peptide derivatives were in a few hundred picomoles compared to 10-50 nmol for the underivatized samples. The mass spectra of the peptides before or after derivatization showed the presence of only singly charged ions. However, collision-induced dissociation of the derivatized peptides showed predominance of b-type ions that are relatively less complicated in assigning the peptide sequence.

Isolation and characterization of 2S cocoa seed albumin storage polypeptide and the corresponding cDNA.

The amine pool of cocoa is known to be an essential component for the development of the typical cocoa flavor. To better understand and to produce an intense in vitro cocoa flavor, identification of the polypeptides that are the source of the amine flavor precursor pool is essential. Chromatographic analysis of the polypeptide profile of unfermented cocoa resulted in identification of a novel storage polypeptide of M(r) 8515. The N-terminal sequence of the first 34 residues of the purified polypeptide shows similarity to 2S storage albumins of cotton and Brazil nut and sweet protein, Mabinlin. To identify the corresponding cDNA of the putative cocoa 2S albumin, 18 randomly chosen clones from the cDNA library of immature Theobroma cacao seed mRNA were sequenced, and a full-length cDNA clone encoding a protein harboring the N-terminal sequence of the novel polypeptide was selected. The open reading frame of the clone encodes a polypeptide of M(r) 17125. Comparison of the translated amino acid sequence of the precursor protein or the mature polypeptide against the Swiss-Prot and TrEMBL databases shows high sequence similarity (>52%) and identity (>38%) to many plant 2S albumins. Tryptic peptide mass fingerprinting of the purified polypeptide by high-performance liquid chromatography-electrospray ionization mass spectrometry shows 10 masses that match the expected tryptic peptides of the deduced sequence. Together with the published work on plant 2S albumin processing, the results presented here suggest that post-translational processing yields a 73-residue polypeptide (residue positions 78-150) corresponding to the 9 kDa subunit of the mature cocoa 2S albumin protein.

Primary structure of the abundant seed albumin of Theobroma cacao by mass spectrometry.

The most abundant albumin present in seeds of Theobroma cacao was purified to apparent homogeneity as judged by high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS), sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and NH(2)-terminal sequence analysis. Tryptic peptide mass fingerprinting of the purified protein by HPLC/ESI-MS showed the presence of 16 masses that matched the expected tryptic peptides corresponding to 95% of the translated amino acid sequence from the cDNA of the 21 kDa cocoa albumin. Collision-induced dissociation MS/MS analysis of the C-terminal peptide isolated from the CNBr cleavage products provided unequivocal evidence that the mature cocoa albumin protein is nine amino acid residues shorter than expected from the reported cDNA of its corresponding gene. The experimentally determined M(r) value of 20234 was in excellent agreement with the truncated version of the amino acid sequence. The purified cocoa albumin inhibited the catalytic activities of bovine trypsin and chymotrypsin. The inhibition was stoichiometric with 1 mol of trypsin or chymotrypsin being inhibited by 1 mol of inhibitor with apparent dissociation constants (K(i)) of 9.5 x 10(-8) and 2. 3 x 10(-6) M, respectively, for inhibitor binding at pH 8.5 and 37 degrees C. No inhibition of the catalytic activities of subtilisin, papain, pepsin, and cocoa endoproteases was detected under their optimal reaction conditions.

Characterization of peptides formed during fermentation of cocoa bean.

Analysis by SDS-PAGE and GPC-MS of fermented cocoa extracts shows changes in the amount and composition of the major proteins, accompanied by formation of complex distributions of peptides. MS/MS studies and application of SEQUEST sequencing software have allowed identification of two related peptides, a hexapeptide and a nonapeptide, formed from vicilin, one of the cocoa storage proteins. Time course studies of the two peptides show different abundance profiles and indicate, in part, production of the hexapeptide from the nonapeptide.

Abdominal tuberculosis. Diagnosis by fine needle aspiration cytology.

One hundred five cases of tuberculosis of the abdomen (excluding genitourinary involvement) diagnosed initially by fine needle aspiration cytology between 1988 and 1991 were analyzed retrospectively. The diagnosis of tuberculosis was suspected clinically in 60% of the cases prior to aspiration. The anatomic locations of the lesions were ileocecal in 69 cases, lymph node in 18, colon in 10, jejunoileal in 2, duodenojejunal in 1, peritoneal in 1 and liver in 4. Cytomorphologic analysis of the aspirates from these 105 cases showed only necrotic material in 19 cases, necrotic material with epithelioid granuloma in 36 cases and epithelioid granuloma without necrosis in 50 cases. Ziehl-Neelsen staining for acid-fast bacilli was positive in 47 of 105 cases (45%). Cultures for mycobacteria were positive in 5 of 24 cases. Histologic confirmation was available in five cases only. The cytodiagnosis correlated well with the radiologic findings and response to antituberculosis chemotherapy.

Parathyroid cyst: a rare cause of an anterior neck mass.

Parathyroid cysts are rare. Most of them present in the anterior neck as cystic neck swellings. A case of cervical parathyroid cyst is presented, along with a brief review of the literature regarding the aetiology, clinical features, diagnosis and management of this condition.

Tuberculosis of the parotid gland.

Tuberculosis of the parotid gland is a rare entity. Only about a hundred cases have been reported till date, mostly from parotidectomy specimens. The present case was diagnosed by fine needle aspiration and treated successfully by short-course antitubercular chemotherapy. An early diagnosis can avoid parotidectomy, which can be a hazardous procedure in a medically treatable condition.

Bio-artificial liver from cultured human foetal hepatocytes: feasibility and prospects.

BACKGROUND: Conventional therapy for acute liver failure has not been able to improve survival beyond 40%. Apart from liver transplantation, the most promising development in this field is the utilization of cultured hepatocytes to make 'bio-artificial liver support systems' as a 'bridge to transplantation' or ideally as a 'bridge to total recovery'. This study examines the feasibility of culturing foetal hepatocytes without the use of growth factors and formulating a bio-artificial liver support device in our set-up. METHODS: Foetal hepatocytes were harvested from the liver obtained from mid trimester abortions at Armed Forces Medical College and Command Hospital (SC), Pune. The liver was perfused with Phosphate Buffered Saline (PBS) and collagenase type IV and was cut with a pair of sterile scissors into tiny pieces. Cells so separated, were washed with PBS plus foetal calf serum and stirred to disperse the cell aggregates. Filtered cell suspensions were inoculated in polystyrene flasks containing hepatocyte culture medium (MEM E: 75%, M199: 25%, BSA: 0.1%, Bovine Insulin 5 micrograms/ml, FCS: 10%, Penicillin: 10 i.u., Streptomycin 50 micrograms/ml, Hydrocortisone 5 micrograms/ml and incubated at 37 degrees C. The functional capabilities of the cultured hepatocytes were analyzed by studying production of albumin and a foetoprotein. Structural integrity of hepatocytes was assessed by light and electron microscopy. RESULTS: The hepatocyte yield varied from 2 to 60 x 10(6) cells/L with an average of 38 x 10(6) cells/L in the eight consecutive experiments. Initial hepatocyte viability varied from 25% to 90% with an average of 61%. The yield and the viability of hepatocytes were adversely affected by the condition of foetus at birth and use of intra-amniotic injections for inducing abortions. Hepatocyte monolayers and colonies formed in 75% experiments. The cultures could be maintained in incubation without the use of epidermal or hepatocyte growth factors for 2-25 days with a mean survival of 8.9 days. The cells in culture were found to be structurally normal and functionally active and could be cryo-preserved. These hepatocytes were inoculated into a hollow fiber module to formulate bio-artificial liver support device. The cultures ultimately developed either cellular disintegration or bacterial infections despite use of antibiotics in the culture medium. CONCLUSIONS: We conclude that it is feasible to maintain foetal hepatocyte cultures without the use of expensive growth factors for over 8 days. Bio-artificial liver formulated with cultured foetal hepatocytes is now a step closer to clinical trials.

Efficacy of endoscopic wire guided biliary brushing in the evaluation of biliary strictures.

Endoscopic brush cytology is a valuable technique for the evaluation of biliary strictures. The sensitivity of this technique varies from 30% to 83%, however, it can have specificity of 100%. We retrospectively evaluated the usefulness of wire-guided biliary brush cytology in biliary strictures in our hospital over a 3 years period from 1997 to 2000. Brushings from 58 biliary strictures were obtained during endoscopic retrograde cholangiography. They were compared with histological proof obtained by surgical biopsy or percutaneous fine-needle aspiration cytology and/or clinical findings. These were reported as benign or malignant. Eleven patients were excluded due to incomplete data. Eighteen patients had benign brushings. Fourteen of the 29 patients in whom histological confirmation of malignancy brushings was obtained were also reported as malignant. The sensitivity of endoscopic brushings was 48.2%, specificity 100% and diagnostic accuracy 55.2%. No major complications were seen in our study group.

CT in nonvisualised vertebral levels caudal to the complete block on lumbar myelogram.

CT was performed in 51 patients at and caudal to the level of lumbar myelographic block due to disc herniation with or without lumbar canal stenosis. The causes of complete or almost complete myelographic block included disc herniation (27 patients)disc herniation with spinal stenosis (12 Patients), spinal canal stenosis (11 patients) and posterior osteophytes (1 patient). CT helped in detecting the non-visualised vertebral levels, demonstrated a surgically treatable lesion below the level of myelographic block and provided information for preoperative planning.

Fetal gastrointestinal malformations.

In an audit to evaluate fetal gastrointestinal (GIT) malformations, case sheets of all mothers who gave birth to newborns with GIT malformations were analysed regarding the maternal history, prenatal ultrasound and the postnatal structural malformations and perinatal management. In 1999, eleven babies were born with gastrointestinal malformations, one was a still birth. Nine had associated malformations of other systems, two had trisomy 21. Eight babies were operated in the immediate newborn period, 5 survived. Ultrasound was not very accurate in the diagnosis of GIT anomaly in this series. In view of associated chromosomal and structural anomaly a fetal ECHO and genetic amniocentesis is warranted. Most of these babies require immediate surgery after birth, so they need to be delivered in a centre which is equipped with good neonatal and pediatric surgery care.

Cyclodialysis cleft diagnosed by conventional ultrasonography.

This case report describes the diagnosis and management of a traumatic cyclodialysis cleft, in the absence of newer investigating modalities like ultrasonic biomicroscopy (UBM). This entity is frequently difficult to recognize clinically owing to the extreme hypotony and media haze that these patients usually present with, which makes gonioscopy particularly taxing. Adding to the diagnostic dilemma is the difficulty in delineating the lesion ultrasonically due to the extreme anterior location of the cleft. In these situations, high frequency ultrasound has been found to be extremely helpful, but the availability of such sophistication is difficult. Presented here is demonstration of the cleft with a microconvex ultrasound probe of 9 MHz frequency. The patient was successfully managed conservatively. This modality could prove to be a valuable adjunct to the diagnosis of such cases even in the situations of nonavailability of high frequency ultrasonography.

Lysine- and threonine-sensitive aspartokinase isoenzymes from spinach leaves share common antigenic determinants.

The lysine- and threonine-sensitive isoenzymes of aspartate kinase were purified to homogeneity from spinach leaves and polyclonal antibodies were raised in rabbits. The antibodies were characterized by various immunological tests like Ouchterlonys-double-diffusion, titrations of the inhibition of enzyme activity and ELISA. The antibodies against the lysine-sensitive isoenzyme could recognise as little as 50 ng of the pure antigen protein and that against the threonine-sensitive form could recognise 200 ng of the protein in the ELISA tests. The immunological tests have also shown that the lysine and threonine sensitive isoenzymes of aspartate kinase share some common antigenic determinants and differ in others.

Fine needle aspiration biopsy of pancreas.

Percutaneous fine needle aspiration biopsy (FNAB) were performed in 22 cases of pancreatic lesions. FNAB was done with the help of ultrasonographic guidance in 14 cases and intraoperatively in 2 cases. In rest of the cases it was done without any radiological guidance. Diagnosis was confirmed either by histopathology or exploratory laparotomy by follow-up data in rest of the cases. There was no false positive or negative diagnosis. Nuclear crowding, nuclear enlargement and prominent nucleoli were the important cytological criteria for pancreatic adenocarcinoma. Cytology smears of the islet cell tumour show round to oval monomorphic cells with central to eccentric nuclei and fine reddish granulations in cytoplasm. There was no complication following aspiration. Thus FNAB of pancreas is a relatively safe, reliable and quick technique which can be performed under ultrasound guidance.