Feasibility and impact of a one-stop thyroid clinic in a low- and middle-income country.

BACKGROUND: The study was done to evaluate the feasibility, safety and outcomes of a one-stop thyroid clinic (OSTC) in a low- and middle-income country (LMIC) setting. METHODS: This was a prospective non-randomised case control study consisting of patients with thyroid nodules evaluated and managed at a tertiary referral centre in an LMIC between February 2019 and January 2020. Patients were divided into two groups based on the kind of preoperative evaluation protocol: OSTC group (n = 118) - OSTC protocol, and control group (CG, n = 108) - routine protocol. RESULTS: Baseline clinical characteristics of the two groups including median age (p = 0.13) and gender distribution (p = 0.76) were comparable. The majority of patients in both groups belonged to a low-income group (46.6% vs 47.3%; p = 0.91), followed by a middle-income group (35.6% vs 30.5%; p = 0.41). The median number of outpatient department visits (1 vs 3 days; p = < 0.001), waiting time for neck ultrasonography (1 vs 3 days; p = < 0.0001), fine needle aspiration cytology (1 vs 2 days; p = < 0.0001), and out of pocket expenditure (INR 3 965 vs 6 624; p = < 0.001) was significantly less in the OSTC group compared to the CG. Patients in the OSTC group reported better satisfaction levels (p = < 0.0001). Accuracy of diagnosis did not differ between the two groups (p = 0.14). CONCLUSION: OSTC practice is feasible, provides comparative clinical outcomes and seems cost effective in an LMIC. This protocol can be adopted as a routine practice in any health system.

Pancreatic and peripancreatic tuberculosis presenting as hypoechoic mass and malignancy diagnosed by ultrasound-guided fine-needle aspiration cytology.

BACKGROUND: Pancreatic and peripancreatic tuberculosis is an extremely uncommon disease, presenting as hypoechoic mass on ultrasonography and imaging mimicking malignancy. Consequently, it represents a diagnostic challenge. AIMS: To study 14 unusual cases of pancreatic and peripancreatic tuberculosis undergoing ultrasound-/endoscopic-guided fine-needle aspiration cytology (FNAC) in the 5-year period from 2006 to 2010. MATERIALS AND METHODS: Endoscopic-guided FNAC was done in two cases, while ultrasound-guided FNAC was performed in 12 cases using 22-G needles via a percutaneous transabdominal approach. The aspirated material was quickly smeared onto glass slides, air dried, and wet fixed in 95% ethyl alcohol for subsequent Papanicolaou staining. RESULTS: All pancreatic and peripancreatic tuberculosis cases showed solid-cystic pancreatic mass. Smears showed epithelioid cell granulomas, multinucleated giant cells, mixed inflammatory cells and histiocytes against a necrotic background. The common anatomic locations were the head, peripancreatic, tail and body of the pancreas. CONCLUSIONS: Ultrasound-/endoscopic-guided FNAC is a safe, reliable and cost-effective method for preoperative diagnosis of pancreatic and peripancreatic tuberculosis. Clinical symptoms and accurate diagnostic approach by ultrasound-/endoscopic-guided FNAC of pancreatic and peripancreatic tuberculosis is needed to avoid performing redundant laparotomy. Despite its rarity, pancreatic and peripancreatic tuberculosis should be considered for differential diagnosis of pancreatic and peripancreatic cystic mass in endemic developing countries.

Caecal lymphangioma: a rare cause of gastrointestinal blood loss.

Lymphangioma is the malformation of the lymphatic system. Lymphangioma is a benign tumour and most colonic lymphangiomas do not cause symptoms and do not require treatment. Resection is required in patients with bleeding or intussusceptions. We report a case of intestinal lymphangiomatosis in a 31-year-old man who presented with recurrent melena and anaemia, which were diagnosed endoscopically and treated with surgical resection.

Endodontic management of badly broken down teeth using the canal projection system: two case reports.

AIM: Teeth that have been weakened by caries and require root canal treatment to maintain their functional integrity may present with minimal coronal tooth structure and are a challenge for isolation and restoration. The aim of this clinical report is to demonstrate the management of badly broken down teeth using the Projector Endodontic Instrument Guidance System (PEIGS). SUMMARY: The PEIGS is an adjunct to root canal treatment designed to enhance the ease of treatment delivery. Use of this system facilitates projection of canal orifices from the floor of the pulp chamber to the cavosurface, providing direct visualization of and physical access to the projected canals. This report demonstrates the use of this novel device for the management of two badly broken down teeth. KEY LEARNING POINTS: Use of the endodontic projection system has the following advantages: * 'Projects' the canal orifice from the floor of the pulp chamber to the cavosurface, thereby enhancing visualization and access to the canals. * The bonded coronal build up reduces the risk of interappointment crack initiation and coronal-radicular fracture of weakened tooth structure. * Permits individualization of canals especially when they lie in close proximity to each other on the chamber floor. * Isolation may be facilitated by ease of clamp retention, rendering many structurally debilitated teeth endodontically treatable.

Reduced pulmonary endothelium-bound angiotensin converting enzyme activity in diabetic rabbits.

We examined the effects of diabetes on pulmonary capillary endothelial cell (EC) function in diabetic rabbits. One, three and six weeks after alloxan treatment, rabbits were anesthetized and pulmonary endothelium-bound angiotensin converting enzyme (ACE) activity was estimated from the single-pass transpulmonary hydrolysis of benzoyl-Phe-Ala-Pro (BPAP), an ACE specific substrate. ACE activity significantly decreased in 1- and 3-week diabetic rabbits and returned to control levels at 6 weeks. Capillary dilation, parenchymal hemorrhage and erythrocyte clumping were maximal in 3-week diabetic rabbits. We conclude that in the alloxan-diabetic rabbit, there are transient functional and more persistent morphological alterations.

The rheumatological prodrome: an unusual inaugural manifestation of acute myeloblastic leukemia.

Uncommon patterns of presentation of acute leukemia pose diagnostic problems. A rheumatological prodrome in acute myeloblastic leukemia is very rare. We describe one such patient who had a normal haemogram. Bone marrow examination done later revealed acute myeloblastic leukemia. The case is discussed with reference to literature.

Intimal thickness and layering, and smooth muscle cell phenotypes in aorta of youth.

Proneness to the lesions of atherosclerosis varies along the length and circumferential topography of the aorta. Smooth muscle cells, in particular those of the 'modulated' synthetic phenotype which are able to proliferate and synthesize matrix proteins, are considered to play an important role in lesion progression. We report on a study of the aortic intima at a lesion-prone site from abdominal aorta and a lesion-resistant site from thoracic aorta in young humans to determine (1) whether the histologic structure and the smooth muscle cell composition show quantitative differences between lesion-prone and lesion-resistant aortic sites; (2) whether there are gender differences, and (3) whether any differences increase in degree with increasing age in this young population. Material for this study was obtained as part of the NIH-funded multicenter study on Pathobiological Determinants of Atherosclerosis in Youth (PDAY) from autopsies of male and female subjects between the ages of 15 and 34, victims of unexpected sudden death, usually from trauma. The samples consisted of strips of abdominal and thoracic aorta, all derived from the same anatomical sites standardized in the PDAY studies. The thickness of total intima (TI) and its elastic hyperplastic (EH) layer was measured. Smooth muscle cells of all types (SMC) and separately those of the synthetic phenotype (SynSMC) were quantified in each site using immunohistochemical procedures in replicate sections of uniform thickness. The intima of the atherosclerotic lesion-prone dorsal half of the abdominal aorta (AD) shows significant differences from the lesion-resistant ventral half of thoracic aorta (TV) in that (1) its EH layer is significantly thicker; (2) its EH layer has a comparatively higher number of both total SMC and SynSMC, even when adjusted for intimal thickness, and (3) the age-related increase in thickness of both TI and EH layer of AD is much greater than that of TV.

Determination of cis and trans isomers of monocrotophos in technical products by reversed-phase column liquid chromatography.

A simple reversed-phase column liquid chromatographic method for the determination of cis and trans isomers of monocrotophos (MCP) using a C18 column, aqueous acetonitrile as eluent and UV detection at 218 nm was developed. The method was used for quality assurance and to study the relative stabilities of cis and trans isomers in technical products of MCP.

Separation, identification and determination of sanguinarine in argemone and other adulterated edible oils by reversed-phase high-performance liquid chromatography.

A simple, rapid and reliable reversed-phase high-performance liquid chromatographic method for the separation and determination of sanguinarine in argemone and other edible oils has been developed. The separation has been achieved on a C18 column with CH3OH-CH3CN-tetrahydrofuran-water as mobile phase using diode array detection at 280 nm. The minimum detection limit of sanguinarine in the adulterated edible oils is 5 microg/g.

Conditional transformation of rat embryo fibroblast cells by a cyclin D1-cdk4 fusion gene.

Cyclin D1 gene overexpression is a frequent event in a number of human cancers. These observations have led to the suggestion that cyclin D1 alterations might play a role in the etiology of cancer. This possibility is supported by the finding that transfection of mammalian cells with cyclin D1 can accelerate progression through the G1 phase of the cell cycle. Moreover, cyclin D1 can function as an oncogene by cooperating with activated Ha-ras to transform primary rat embryo fibroblasts (REFs). In addition, cyclin D1 transgenics develop hyperplasia and neoplasia of the thymus and mammary gland. We have constructed a novel fusion gene consisting of full-length human cyclin D1 and cdk4 genes. This fusion gene was expressed in insect cells and the fusion protein was shown to be enzymatically active. The fusion gene was expressed in mammalian cells under the control of tet-repressor. This fusion gene immortalized primary REFs, and cooperated with activated Ha-ras to transform primary REFs, in terms of anchorage-independent growth in vitro and formation of tumors in vivo. Utilizing a tet-regulated gene expression system, we have shown that proliferation of stably transfected primary REFs in vitro and in vivo is dependent on the continued expression of the cyclin D1-cdk4 fusion gene. These cell lines could be useful in the discovery of novel cancer therapeutics to modulate cyclin D1.cdk4 activity.

Gas chromatographic-mass spectrometric separation, identification and determination of C6-C12 cyclic imides in thermally treated epoxy and alkyd resins to locate hot spots inside large electricity generators.

A simple and rapid GC-MS method for separation, identification and quantitative determination of long-chain cyclic imides in the 300 degrees C thermally treated epoxy and alkyd resins has been developed. The method provides a positive means of identifying C6-C12 cyclic imide derivatives by GC-MS and enables the specific area of overheating to be identified, thereby averting catastrophic failures of power generators in service.

Casein kinase 2 binds to and phosphorylates BRCA1.

The BRCA1 gene encodes a complex protein that appears to be involved in some aspects of DNA repair, transcription, or cell cycle regulation. The phosphorylation of BRCA1 is enhanced following episodes of DNA damage or during cell cycle progression, indicating that phosphorylation may be an important regulatory mechanism. Through a yeast two hybrid assay, we found that the beta-subunit of casein kinase 2 (CK2) associated with a carboxy-terminal region of BRCA1. This association was much weaker with the same fragment bearing a missense mutation (M1775R) that has been identified in breast tumors. The interaction was also evident in Sf9 cells. Subsequent studies showed that BRCA1 was phosphorylated in vitro by CK2. An analysis by site directed mutagenesis of BRCA1 showed that in vitro phosphorylation by CK2 required a serine at aa1572. These data implicate CK2 as a potential mediator of BRCA1 activity.

Malignant rhabdoid tumor of the duodenum.

A polypoid malignant rhabdoid tumor of the duodenum is presented. The pattern of metastatic spread in this 58-year-old man included multiple duodenal and small intestinal transmural tumor implants and a large peribronchial lymph node causing superior vena cava syndrome. Microscopically, the tumor was composed of a diffuse population of rhabdoid cells characterized by homogeneous globular cytoplasmic inclusions that tended to indent or displace eccentric, vesicular nuclei with nucleoli. No glandular features were noted. Immunohistochemical and ultrastructural evaluation revealed that these inclusions contained vimentin, an intermediate filament of the mesenchymal cytoskeleton. Phenotypic features of a rhabdoid tumor have been reported in 10 poorly differentiated malignancies of the gastrointestinal tract. This is the first case report of a malignant rhabdoid tumor of the small intestine. Regardless of the site of the lesion, tumors showing these features are generally associated with a poor prognosis.

Purification, characterization, and kinetic mechanism of cyclin D1. CDK4, a major target for cell cycle regulation.

The cyclin D1.CDK4-pRb (retinoblastoma protein) pathway plays a central role in the cell cycle, and its deregulation is correlated with many types of cancers. As a major drug target, we purified dimeric cyclin D1.CDK4 complex to near-homogeneity by a four-step procedure from a recombinant baculovirus-infected insect culture. We optimized the kinase activity and stability and developed a reproducible assay. We examined several catalytic and kinetic properties of the complex and, via steady-state kinetics, derived a kinetic mechanism with a peptide (RbING) and subsequently investigated the mechanistic implications with a physiologically relevant protein (Rb21) as the phosphoacceptor. The complex bound ATP 130-fold tighter when Rb21 instead of RbING was used as the phosphoacceptor. By using staurosporine and ADP as inhibitors, the kinetic mechanism of the complex appeared to be a "single displacement or Bi-Bi" with Mg2+.ATP as the leading substrate and phosphorylated RbING as the last product released. In addition, we purified a cyclin D1-CDK4 fusion protein to homogeneity by a three-step protocol from another recombinant baculovirus culture and observed similar kinetic properties and mechanisms as those from the complex. We attempted to model staurosporine in the ATP-binding site of CDK4 according to our kinetic data. Our biochemical and modeling data provide validation of both the complex and fusion protein as highly active kinases and their usefulness in antiproliferative inhibitor discovery.

Aflatoxin B1 contamination in maize samples collected from different geographical regions of India--a multicentre study.

Under a multicentre study conducted by the Indian Council of Medical Research, 2,074 samples of maize were collected from rural and urban areas of 11 states representing different geographical regions of the country. These samples were analysed for aflatoxin B1 using the AOAC method. Analytical quality assurance between various participating laboratories was ensured through analysis of check-samples. About 26% of maize samples collected from 11 states exceeded the permissible Indian regulatory limit of 30 micrograms kg-1. No statistically significant difference in percentage of samples contaminated (> 30 micrograms kg-1) was observed between pooled rural (27.8%) and urban (23.7%) data. The maximum level of contamination of 666 micrograms kg-1 was observed in the state of Haryana. The median level of < 5 micrograms kg-1 was observed in the states of Gujarat, Haryana, West Bengal, Andhra Pradesh and Karnataka. In all other states studied, the median level was found to vary between 10 and 35 micrograms kg-1.

Construction and characterization of a one-plasmid system for the controlled expression of genes in mammalian cells by tetracycline.

The two-plasmid system of Gossen and Bujard [Gossen and Bujard (1992) Proc. Natl. Acad. Sci. USA 89, 5547-5551] to express mammalian genes in a tetracycline-repressed fashion was combined into a single-plasmid system. Two variants of this single-plasmid system that differ in the multiple cloning site (MCS) region are described. These vectors were used to stably transfect raf kinase domain into the normal rat kidney epithelial cell line (NRKE) to obtain a conditionally transformed cell line. These vectors were also used to stably transfect wild-type and mutant human p53 into the human osteosarcoma cell line, SAOS-2. Tetracycline repressed gene expression in both cell lines; about 12-fold in NRKE and about 80-fold in SAOS-2 cell line.

Production of a novel polyketide through the construction of a hybrid polyketide synthase.

The lactone rings of the polyketides platenolide and tylactone are synthesized by condensation of acetate-, proprionate-, and butyrate-derived precursors. A hybrid tylactone/platenolide synthase was constructed to determine if the choice of substrate is programmed by the polyketide synthase and to ascertain if a substrate different than that normally used in the first step of platenolide synthesis could be incorporated into the final polyketide. In this work, we report the successful incorporation of a propionate in place of the acetate normally used in the first step of platenolide synthesis. This result demonstrates that polyketide synthases choose a particular substrate at defined steps and provides strong evidence that substrate choice is programmed by the acyl transferase domain of a large, multifunctional polyketide synthase.

Targets for cancer therapy in the cell cycle pathway.

Progression through the cell cycle is a complex process that is regulated at many levels by several proteins. We are just beginning to understand how these proteins accomplish this regulation. The p16-cyclin D1.cdk4-pRb pathway is one of the most important pathways, which is altered in a majority of cancers of different types. This review focuses on some of the proteins in this pathway that offer new opportunities for drug discovery. Some guidelines to evaluate the relevance of these proteins as targets for cancer therapy and the importance of developing a combination therapy targeting multiple pathways are also discussed.