Hiding in Plain Sight: A Case of Ornithine Transcarbamylase Deficiency Unmasked Post-Liver Transplantation.

Ornithine transcarbamylase deficiency represents the most common inherited defect of the urea cycle. This enzyme, predominantly found in the liver, plays a crucial role in recycling free ammonia, with deficiencies often leading to fatal complications. Here, we present the case of a 63-year-old man with alcoholic cirrhosis who underwent orthotopic liver transplantation, gradual worsening of his mental status, and progressive elevation of ammonia levels. Liver allograft function was deemed normal, raising concern for a donor-derived metabolic disorder of the urea cycle. Evaluation of the donor patient's blood revealed that the donor was heterozygous for the OTC gene. Posttransplantation changes in mental status should prompt a clinician to consider the most likely causes; however, once these have been ruled out, it is important to consider the less common causes of metabolic derangements. The rarity of these disorders makes expertise of diagnosis, standardization of evaluation, and treatment strategies challenging.

Process standardization for the manufacture of Thabdi Peda.

Traditional dairy products in India are manufactured using age old methods. Such methods varies from place to place. For industrial production of such products a standardized process is needed. Present study was designed to arrive at a method of manufacture for Thabdi Peda, a very popular sweet in Saurashtra region of Gujarat. Range of process parameters like fat percent of milk (4 to 8 %), rate of sugar addition (6 to 10 %) and duration of final heat desiccation (20 to 60 min) were studied and optimum values determined using Response Surface Methodology (RSM) with central composite rotatable design (CCRD). The samples obtained from trials were analyzed for sensory, physicochemical, compositional and textural attributes. The optimized process developed with 10 kg batch of milk having 6 % fat, 8.33 % rate of sugar addition and 34 min duration of heating produced most acceptable product. Standardized Thabdi Peda was found to contain on an average 16.80 % fat, 17.48 % moisture, 11.25 % protein, 20.95 % lactose, 29.99 % sucrose, 3.53 % ash and it gave 28.75 % yield. The pH, water activity and HMF (µ Mole/100 g) content were 6.42, 0.807 and 121.91 respectively. Standard plate count, Yeast and Mold counts were observed to be 3.68 log cfu/g, and 2.51 log cfu/g respectively. No coliforms were observed in Thabdi Peda.

Flavonol dioxygenase chemistry mediated by a synthetic nickel superoxide.

Nature exploits transition metal centers to enhance and tune the oxidizing power of natural oxidants such as O2 and H2O2. The design and interrogation of synthetic metallocomplexes with similar reactivity to metalloproteins provides one strategy for gaining insight into the mechanistic underpinnings of oxygen-activating enzymes such as oxidases, oxygenases, and dioxygenases like Ni-quercetinase (Ni-QueD). Ni-QueD catalyzes the oxidative ring opening of the polyphenol quercetin, a natural product with antioxidant properties. Herein, we report the synthesis and characterization of Ni(13-DOB), a Ni(II) species complexed by an N4-macrocycle that has been characterized by single crystal X-ray crystallography. Ni(13-DOB) forms a Ni-superoxide intermediate (Ni(13-DOB)O2•-) upon treatment with H2O2 and Et3N, as verified by resonance Raman spectroscopy. We demonstrate through UV/vis and LCMS that Ni(13-DOB)O2•- is capable of the 1-electron oxidation of flavonols, including both 3-hydroxyflavone (3-HF, the simplest flavonol) and quercetin itself. Incorporation of two O-atoms into the flavonol radical via superoxide from Ni(13-DOB)O2•- precedes oxidative cleavage of the flavonol scaffold in each case, consistent with quercetinase ring cleavage by Ni-QueD in Streptomyces sp. FLA. Conversion of 3-HF into 2-hydroxybenzoylbenzoic acid was accomplished with catalytic turnover of Ni(13-DOB) at ambient temperature, as confirmed by HPLC timecourses and GCMS analysis of isotopic labeling studies. The Ni(13-DOB)-mediated oxidative cleavage of quercetin to the corresponding biomimetic phenolic ester was also verified through 18O-isotopic labeling studies. Through the HPLC characterization of both on- and off-pathway products of flavonol dioxygenation by Ni(13-DOB)O2•-, the stringent reaction pathway control provided by enzyme active sites is highlighted.

Vibrio cholerae non-O1 infection in cirrhotics: case report and literature review.

Vibrio species are ubiquitous in the marine environment and can cause severe infections in cirrhotic patients. Patients with liver disease should be warned about the potential dangers of consuming raw or undercooked seafood, and avoiding exposure of wounds to seawater. We report a case of severe sepsis from Vibrio cholerae non-O1 in a patient with cirrhosis awaiting orthotopic liver transplant. This case is aimed to advise clinicians about the importance of V. cholerae subtypes, and non-cholera Vibrio species infections in cirrhotic patients, highlighting the need to educate these patients to stay away from undercooked seafood.

Paclitaxel sensitivity of breast cancer cells with constitutively active NF-kappaB is enhanced by IkappaBalpha super-repressor and parthenolide.

The transcription factor nuclear factor-kappaB (NF-kappaB) regulates genes important for tumor invasion, metastasis and chemoresistance. Normally, NF-kappaB remains sequestered in an inactive state by cytoplasmic inhibitor-of-kappaB (IkappaB) proteins. NF-kappaB translocates to nucleus and activates gene expression upon exposure of cells to growth factors and cytokines. We and others have shown previously that NF-kappaB is constitutively active in a subset of breast cancers. In this study, we show that constitutive activation of NF-kappaB leads to overexpression of the anti-apoptotic genes c-inhibitor of apoptosis 2 (c-IAP2) and manganese superoxide dismutase (Mn-SOD) in breast cancer cells. Furthermore, expression of the anti-apoptotic tumor necrosis factor receptor associated factor 1 (TRAF1) and defender-against cell death (DAD-1) is regulated by NF-kappaB in certain breast cancer cells. We also demonstrate that NF-kappaB-inducible genes protect cancer cells against paclitaxel as MDA-MB-231 breast cancer cells modified to overexpress IkappaBalpha required lower concentrations of paclitaxel to arrest at the G2/M phase of the cell cycle and undergo apoptosis when compared to parental cells. The effect of NF-kappaB on paclitaxel-sensitivity appears to be specific to cancer cells because normal fibroblasts derived from embryos lacking p65 subunit of NF-kappaB and wild type littermate embryos were insensitive to paclitaxel-induced G2/M cell cycle arrest. Parthenolide, an active ingredient of herbal remedies such as feverfew (tanacetum parthenium), mimicked the effects of IkappaBalpha by inhibiting NF-kappaB DNA binding activity and Mn-SOD expression, and increasing paclitaxel-induced apoptosis of breast cancer cells. These results suggest that active ingredients of herbs with anti-inflammatory properties may be useful in increasing the sensitivity of cancers with constitutively active NF-kappaB to chemotherapeutic drugs. Oncogene (2000) 19, 4159 - 4169

The effects of shear stress on endothelial cell retention and function on expanded polytetrafluoroethylene.

We evaluated the adherence of indium 111-radiolabeled endothelial cells to fibronectin-treated expanded polytetrafluoroethylene surfaces exposed to high (437 s-1) vs low (218 s-1) shear and the influence of shear on prostacyclin production. Canine jugular vein factor VIII-positive endothelial cells in passages 3 through 6 were incubated with 111Indium-oxine, and labeled cells were seeded onto fibronectin-treated expanded polytetrafluoroethylene patches. Patches with confluent cells were exposed to shear in a Weissenberg rheogoniometer for intervals ranging up to 60 minutes. Percent endothelial cell retention was determined by gamma counting of patches and media and by histologic evaluation. Prostacyclin production (tritiated radioimmunoassay of 6-keto-prostaglandin F1 alpha) was assayed on perfusing media. Results showed no differences in 6-keto-prostaglandin F1 alpha production between shear rates or time periods. Endothelial cell retention did not differ between the shear rates. Rotational shear caused persistent cell loss over time in either high- or low-shear conditions. This persistent cell loss in response to steady rotational shear differs from that in response to identical rates of pulsatile linear shear in our laboratory where cell loss approached zero after 15 minutes.

An in vitro slice preparation of the cat lateral geniculate nucleus.

A slice preparation of the cat thalamus containing the lateral geniculate nucleus and the terminal portion of the optic tract is described. Ultrastructurally the slices remain relatively normal for only a short time after cutting. Indeed most cellular elements deteriorate quickly with time but patches of relatively intact tissue were still present even 10 h after cutting and maintenance in a storage bath. However, for 4-5 h after cutting long-lasting intracellular recordings of high quality and stability were obtained, and intrasomatic injection of horseradish peroxidase used for the morphological identification of recorded neurones as X or Y cells.

Surgical management of localized amyloidosis.

OBJECTIVE: To demonstrate the role of two-dimensional reconstruction images on computed tomography (CT) in the treatment planning for laryngeal amyloidosis. To discuss the treatment for isolated laryngeal amyloidosis and compare the role of endoscopic versus an open surgical approach to management. STUDY DESIGN: Retrospective review. METHODS: The medical records from 1984 to the present with the diagnosis of localized respiratory tract amyloidosis at Geisinger Medical Center were reviewed. RESULTS: Five previously unpublished cases of localized laryngeal amyloidosis were identified with the supraglottic region the major site of involvement. Hoarseness and airway compromise were the presenting symptoms. CT two-dimensional reconstruction imaging was used to evaluate two cases with extensive laryngeal involvement that required an external surgical approach to relieve symptoms. CONCLUSIONS: Localized laryngeal amyloidosis is a rare disease that requires surgical management when symptomatic. CT two-dimensional reconstruction can be helpful in detailing the extent of disease and planning surgery. A lateral external supraglottic approach has been found to be successful in treating patients with large supraglottic masses.

Use of a rapid intraoperative parathyroid hormone assay in the surgical management of parathyroid disease.

OBJECTIVE: To evaluate the utility of a rapid intraoperative parathyroid hormone (PTH) immunoradiometric assay in the surgical management of parathyroid disease, particularly with reference to limiting extent of cervical exploration. DESIGN: Nonrandomized prospective study. SETTING: Academic tertiary care center. PATIENTS: Forty-three consecutive patients undergoing parathyroid exploration for adenoma or hyperplasia had rapid PTH assays performed from blood drawn at induction and 7 minutes after resection of all hyperfunctioning parathyroid tissue. OUTCOME MEASURES: Excision of all hyperfunctioning parathyroid tissue as assessed by bilateral neck exploration, postoperative normalization of serum calcium and PTH levels, and resolution of clinical symptoms. RESULTS: The intraoperative rapid PTH assay accurately reflected whether all hyperfunctioning parathyroid tissue was excised in every patient. In 41 patients, all hyperfunctioning parathyroid tissue was resected at the time of surgery and confirmed by a corresponding decrease in the intraoperative postexcision rapid PTH determination as well as by subsequent normalization of postoperative serum calcium and PTH levels and resolution of clinical symptoms. In 2 patients, the postexcision rapid PTH assay determination was not consistent with removal of all hyperfunctioning parathyroid disease and both patients demonstrated persistent hyperparathyroidism postoperatively. CONCLUSIONS: The intraoperative rapid PTH assay may be of significant benefit in permitting directed unilateral parathyroid explorations for adenoma when combined with preoperative localization with a technetium-99m sestamibi scan. Additionally, the rapid PTH assay has proved to be of benefit in confirming excision of all hyperfunctioning parathyroid tissue in patients with multiple gland hyperplasia, particularly those who may harbor ectopic parathyroid tissue.

Determination of polyphenols and free radical scavenging activity of Tephrosia purpurea linn leaves (Leguminosae).

BACKGROUND: Leaves of Tephrosia purpurea Linn. (sarpankh), belonging to the family Leguminaceae, are used for the treatment of jaundice and are also claimed to be effective in many other diseases. This research work was undertaken to investigate the in vitro antioxidant activity of aqueous and ethanolic extracts of the leaves. METHOD: The therapeutic effects of tannins and flavonoids can be largely attributed to their antioxidant properties. So, the quantitative determinations were undertaken. All the methods are based on UV-spectrophotometric determination. RESULT: The total phenolic content of aqueous and ethanolic extracts showed the content values of 9.44 ± 0.22% w/w and 18.44 ± 0.13% w/w, respectively, and total flavonoid estimation of aqueous and ethanolic extracts showed the content values of 0.91 ± 0.08% w/w and 1.56 ± 0.12%w/w, respectively, for quercetin and 1.85 ± 0.08% w/w and 2.54 ± 0.12% w/w, respectively, for rutin. Further investigations were carried out for in vitro antioxidant activity and radical scavenging activity by calculating its percentage inhibition by means of IC(50)values, all the extracts' concentrations were adjusted to fall under the linearity range and here many reference standards like tannic acid, gallic acid, quercetin, ascorbic acid were taken for the method suitability. CONCLUSION: The results revealed that leaves of this plant have antioxidant potential. The results also show the ethanolic extract to be more potent than the aqueous decoction which is claimed traditionally. In conclusion, T. purpurea Linn. (Leguminosae) leaves possess the antioxidant substance which may be responsible for the treatment of jaundice and other oxidative stress-related diseases.

HPTLC Method for the Simultaneous Estimation of Valsartan and Hydrochlorothiazide in Tablet Dosage Form.

A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the simultaneous estimation of valsartan and hydrochlorothiazide in combined dosage forms. The stationary phase used was precoated silica gel 60F(254). The mobile phase used was a mixture of chloroform: methanol: toluene: glacial acetic acid (6:2:1:0.1 v/v/v/v). The detection of spots were carried out at 260 nm. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 300 to 800 ng/spot for valsartan and 100 to 600 ng/spot for hydrochlorothiazide. The limit of detection and the limit of quantification for the valsartan were found to be 100 and 300 ng/spot respectively and for hydrochlorothiazide 30 and 100 ng/spot respectively. The proposed method can be successfully used to determine the drug content of marketed formulation.

Stability-indicating Simultaneous HPTLC Method for Olanzapine and Fluoxetine in Combined Tablet Dosage Form.

A rapid, selective and stability-indicating high performance thin layer chromatographic method was developed and validated for the simultaneous estimation of olanzapine and fluoxetine in combined tablet dosage form. Olanzapine and fluoxetine were chromatographed on silica gel 60 F(254) TLC plate using methanol:toluene (4:2 v/v) as the mobile phase and spectrodensitometric scanning-integration was performed at a wavelength of 233 nm using a Camag TLC Scanner III. This system was found to give compact spots for both olanzapine (R(f) value of 0.63+/-0.01) and fluoxetine (R(f) value of 0.31+/-0.01). The polynomial regression data for the calibration plots showed good linear relationship with r(2)=0.9995 in the concentration range of 100-800 ng/spot for olanzapine and 1000-8000 ng/spot for fluoxetine with r(2)=0.9991. The method was validated in terms of linearity, accuracy, precision, recovery and specificity. The limit of detection and the limit of quantification for the olanzapine were found to be 30 and 100 ng/spot, respectively and for fluoxetine 300 and 1000 ng/spot, respectively. Olanzapine and fluoxetine were degraded under acidic, basic and oxidation degradation conditions which showed all the peaks of degraded product were well resolved from the active pharmaceutical ingredient. Both drugs were not further degraded after thermal and photochemical degradation. The method was found to be reproducible and selective for the simultaneous estimation of olanzapine and fluoxetine. As the method could effectively separate the drugs from their degradation products, it can be employed as a stability-indicating method.

Chronic gastric volvulus: report of a case and review of literature.

A patient with chronic gastric volvulus is reported and the related literature reviewed. The clinical presentation, diagnosis, and treatment are discussed. Features of acute and chronic volvulus are compared.

Ascites in agnogenic myeloid metaplasia: association with peritoneal implant of myeloid tissue and therapy.

The case of a patient with agnogenic myeloid metaplasia and myelofibrosis engrafted upon a longstanding treated case of polycythemia rubra vera and presenting with ascites due to peritoneal implants of myeloid tissue is presented. Comments on the differential diagnosis of ascites in general, and especially in myelofibrosis, are entered. The specific simple methodology for the confirmation of this diagnosis through microscopic examination of the sediment of the ascitic fluid for the detection of megakaryocytes and erythroblasts is presented. Radiotherapy in moderate amounts offers a very effective and long-lasting form of this entity.

Histaminergic H1 and H2 excitatory receptors in the guinea pig uterus and taenia coli.

Histamine, 2-(2-pyridyl)ethylamine (PEA), a specific H1 receptor agonist, and 4-methylhistamine (4-MH), a specific H2 receptor agonist, produced a dose-dependent contraction of estrogen-primed guinea pig uterus and taenia coli. Mepyramine (10(-7) M), a specific H1 receptor antagonist, competitively blocked the responses to histamine and PEA in both the preparations. Metiamide (10(-6) M), a specific H2 receptor antagonist, inhibited the responses to histamine and 4-MH in the guinea pig uterus and taenia coli. Our data suggest the presence of both H1 and H2 receptors in the guinea pig uterus and taenia coli. Both receptors appear to be excitatory in nature.

Comparative studies of the effects of 5-hydroxytryptamine and noradrenaline on the rat anococcygeus muscle.

The effects of 5-hydroxytryptamine (5HT) and noradrenaline (NA) have been studied on rat anococcygeus muscle. 1. 5HT and NA produced a dose-dependent contraction of rat anococcygeus muscle. Cyproheptadine (1.0 X 10(-6) M), a specific 5HT receptor blocker, failed to inhibit the responses to either 5HT or NA. 2. However, phentolamine, a specific alpha receptor antagonist competitively blocked the responses to 5HT and NA. 3. The responses to 5HT were inhibited in the reserpinized (5 mg/kg i.p. 24 h) and 6-hydroxydopamine (6-OHDA) pre-treated preparations. 6-OHDA produced a leftward shift of the dose-response curve of NA. Reserpine pre-treatment potentiated lower doses of NA and the threshold dose of NA was significantly decreased. 4. Nialamide (2.2 x 10(-6) M), the mono-amine oxidase inhibitor produced a significant leftward shift of the dose-response curve of both 5HT and NA. Pyrogallol (2.3 x 10(-5) M), the catechol-o-methyl transferase inhibitor also potentiated the responses to both 5HT and NA, but the potentiation was significant at lower doses of 5HT and NA. 5. Our data suggest that 5HT- and NA-induced contractions in rat anococcygeus muscle are mediated through common alpha adrenoceptors. 5HT actions are probably indirect, mediated through the release of NA.

Effect of terminal heat sterilization on the stability of phospholipid-stabilized submicron emulsions.

Parenteral emulsions similar in composition to the commercially available fat emulsions were prepared using a Microfluidizer. The relationship of pH and terminal heat sterilization by autoclaving on the stability of phospholipid-stabilized emulsions was evaluated. The stability of these formulations was determined by using droplet size and zeta potential measurements. It was found that the zeta potentials of emulsions stabilized by more purified phospholipid were lower in magnitude compared to those stabilized by less purified phospholipid. Furthermore, acidic emulsions (between pH 5.0 and 7.0) showed an increased droplet size upon autoclaving, whereas the emulsions adjusted to an alkaline pH (pH 8.0 to 10.0) did not show a significant change in their droplet sizes upon autoclaving. The zeta potentials of all the emulsions "increased" (in magnitude) upon autoclaving and achieved values between -40 and -50 mV. The results of our studies could be explained on the basis of hydrolysis of phosphatidylcholine at different pH values, leading to a breakdown of the film surrounding the oil droplet, which results in coalescence and/or cracking of the emulsions.