Prognosis of Kidney Transplant Recipients With Pretransplantation Malignancy: A Nationwide Population-Based Cohort Study in Taiwan.

OBJECTIVE: The objective of this study was to evaluate the prognosis of kidney transplant recipients with pretransplantation malignancy and the incidence of recurrent malignancy in kidney transplant recipients using claims data from Taiwan's universal health insurance program. METHOD: A total of 4350 transplant recipients were retrospectively analyzed. The rates of pretransplantation or recurrent malignancy, which was defined by their inclusion in the catastrophic illness patient registry using the International Classification of Diseases, 9th Revision, were evaluated. Cox proportional hazard regression and Kaplan-Meier curves were used for the analyses. RESULTS: In total, there were 4350 kidney transplant recipients, 52.1% of patients were male, the mean age at transplantation was 45.8 years old, and the percentages of diabetes mellitus, hypertension, hepatitis B viral infection, and hepatitis C viral infection were 14%, 63.2%, 4.2%, and 2.4%, respectively. There were 95 patients (2.2%) with pretransplantation malignancy. The top 3 pretransplantation malignancies, in decreasing order, were urinary tract, kidney, and breast cancers. After kidney transplantation, 10 recipients had recurrent cancer. The overall cancer recurrence rate was 10.5%. These 10 cancers included urothelial carcinoma (n = 5), renal cell carcinoma (n = 3), breast cancer (n = 1), and thyroid cancer (n = 1). Eleven recipients had a secondary cancer. Patients without pretransplantation and post-transplantation malignancy had the best survival. Patients with pretransplantation malignancy had a greater occurrence of cancers and increased mortality regardless of whether or not they had recurrence of cancer. CONCLUSION: Our results suggest the higher risk of cancer, recurrent or secondary, and mortality after kidney transplantation. Adequate waiting time before transplantation and preventive strategies are strongly suggested in kidney transplant recipients with cancer history.

Influence of pretransplantation dialysis time and lupus activity on outcome of kidney transplantation in systemic lupus erythematosus.

BACKGROUND: Kidney transplantation (KT) has better outcome compared with dialysis in lupus patients. The duration lupus patients need to wait before KT remains debatable, especially in patients with lupus activity. We analyzed a renal transplantation database to elucidate if pretransplantation dialysis (PTD) time and lupus activity affected outcome. METHODS: From 1984 to 2012, 31 Chinese lupus nephritis patients underwent KT at our hospital. The lupus activity was defined as nonrenal systemic lupus erythematosus disease activity index (SLE-DAI) score. Biopsy-proven acute rejection/recurrent lupus nephritis (RLN) were recorded. Chronic allograft dysfunction (CAD) was defined as doubling of serum creatinine level. Graft failure was defined as return to dialysis. We calculated relative hazard ratios (HR) with 95% confidence intervals (CI) from Cox proportional-hazards regression models. RESULTS: In total, 31 lupus patients with KT (7 men and 24 women), with a mean age of 35.3 years at transplantation, were enrolled in this study. The mean follow-up duration was 8.2 years. The mean PTD time was 3.3 years. Both PTD time and lupus activity before transplantation had no effect on CAD and graft failure. Longer PTD time was associated with more acute rejection (HR = 1.20; 95% CI, 1.02-1.41). Also, maximal lupus activity after transplantation was associated with more CAD (HR = 6.44; 95% CI, 1.36-30.57). CONCLUSION: For Chinese lupus patients with KT, longer PTD time was associated with worse outcome. Patients should undergo KT immediately if a kidney is available for donation, even with active lupus disease. It is necessary to monitor lupus activity after transplantation due to its effect on outcome.

A comparison of the thresholding strategies of micro-CT for periodontal bone loss: a pilot study.

OBJECTIVES: Micro-CT provides three-dimensional details and has been widely used for biomedical assessments. This study aimed to determine the most appropriate threshold method for quantitatively assessing the dynamics of periodontal destruction. METHODS: Inflammation was induced by submerging a silk ligature in the sulcus of the maxillary second molars of rats, and the animals were killed prior to ligature placement and after 7 and 21 days. The maxillae were examined for the bone resorptive activities by micro-CT, histology and tartrate-resistant acid phosphatase staining. The imaging threshold was determined by CT phantom, global and local algorithms. A bone fraction measurement from each threshold-determining technique was compared with histomorphometry. The reliability and reproducibility were examined by the intraclass correlation coefficient (ICC) and the coefficient of variation. RESULTS: Significant reduction of inflammatory infiltration (p < 0.01) and active osteoclastic resorption (p < 0.05) from Day 7 to Day 21 were noted. High inter- and intraexaminer agreement were demonstrated in both histomorphometric and micro-CT assessments (ICC > 0.98). The algorithm-based technique demonstrated stronger correlation to histomorphometry than phantom-based thresholds, and the highest agreement was presented by the local algorithm (ICC > 0.96). This, however, was considerably computationally expensive. CONCLUSIONS: The local threshold-determining algorithm is suggested for examining inflammation-induced bone loss. Further investigation will be aimed at enhancing computational efficiency.

PDGF-simvastatin delivery stimulates osteogenesis in heat-induced osteonecrosis.

Heat generated during implant osteotomy might lead to osteonecrosis and delayed bone repair, thus resulting in impaired early osseointegration and fixation of bone-anchoring devices. In this study, we proposed to overcome heat-induced injury to bone by fabricating core-shell polymeric biodegradable microspheres encapsulating a mitogenic factor, platelet-derived growth factor (PDGF), and a differentiation factor, simvastatin, in a simultaneous or sequential release profile. Microspheres encapsulating bovine serum albumin (BSA), PDGF, simvastatin, PDGF-in-core with simvastatin-in-shell, and simvastatin-in-core with PDGF-in-shell were delivered to fill standardized osteotomy sites on edentulous ridges of rat maxillae under irrigated or non-irrigated conditions. In the absence of irrigation, significant reduction of cell viability and increase in inflammation and sequestrum formation without evidence of osteogenesis were observed. Both PDGF and simvastatin deliveries facilitated cell viability and reduced osteonecrosis. Localized osteogenesis was seen under simvastatin treatment, while generalized but primitive osteogenesis was noted in PDGF-treated osteotomy sites. In addition, sequential PDGF-simvastatin delivery further augmented osteogenesis and promoted bone maturation. The results suggested that sequential PDGF-simvastatin delivery was an effective modality to prevent heat-induced complications and facilitate bone apposition after implant osteotomy, potentially favoring the early fixation of bone-anchoring devices and oral implant osseointegration.

Análise das prevalências de doenças detectadas pelo programa nacional de triagem neonatal no município de Araraquara no ano de 2009 / Prevalence analysis of diseases detected by the National Neonatal Screening Program in Araraquara city (Brazil) in the year 2009

O Programa Nacional de Triagem Neonatal (PNTN) implantado no Brasil tem como objetivo, considerando cada fase de execução local, detectar doenças como fenilcetonúria, hipotereoidismo congênito, hemoglobinopatias e fibrose cística. O objetivo deste trabalho foi analisar, através de um estudo transversal e observacional, a prevalência das doenças detectadas pelo PNTN no município de Araraquara emitidas pela APAE de São Paulo no período compreendido entre abril e dezembro de 2009. Os resultados mostram que o município apresentou, no ano de 2009, prevalência de fenilcetonúria e hipotireoidismo congênito de 0,06% acima da média nacional, que é de 0,01% e 0,03%, respectivamente. Com relação às hemoglobinopatias, encontrou-se prevalência do traço para anemia falciforme de 2,15% abaixo da média nacional, que é de 2,6%. A prevalência do traço C no município foi de 0,57%, semelhante a valores nacionais disponíveis na literatura. FA BART´S confirmado apresentou valor de 0,13% abaixo da média de 0,38% da região do município de Araraquara. A realização do teste do pezinho e o aconselhamento aos cuidadores são fatores importantes para redução de morbidades relacionadas à evolução dessas doenças.

The National Neonatal Screening Program (NNSP) set up in all Brazil, aims, through planned phases of local implementation, to detect diseases such as phenylketonuria, congenital hypothyroidism, hemoglobinopathies and cystic fibrosis. The aim of this study was to assess, through a cross-sectional observational study, the prevalence of the diseases detected by the NNSP in the city of Araraquara, in records issued by the São Paulo APAE laboratory in the period between April and December 2009.The results show that Araraquara had a prevalence of phenylketonuria and congenital hypothyroidism 0.06% above the national averages of 0.01% and 0.03% respectively. With respect to hemoglobinopathies, the prevalence of sickle cell trait was 2.15% below the national average of 2.6%. The prevalence of Hb C in the city was 0.57%, similar to national values reported in the literature. Confirmed Hb Bart´s had a prevalence of 0.13% in Araraquara, below the average of 0.38% for the surrounding region. The neonatal screening by heel-prick test and counseling for caregivers are important factors in reducing morbidity related to the evolution of these diseases.

Topical anesthetic EMLA for postoperative wound pain in stereotactic gamma knife radiosurgery: a perspective, randomized, placebo-controlled study.

BACKGROUND: Patients who undergo stereotactic gamma knife radiosurgery (GKRS) need a rigid frame fixation for the stereotactic procedures. Many patients suffered from postoperative wound pain after frame removal. The present study investigated whether an additional application of a topical anesthetic prior to frame removal could reduce this discomfort. PATIENTS AND METHODS: 60 patients who underwent GKRS were enrolled in this study. Of these 60 patients, 30 were treated with a topical application of EMLA, a eutectic mixture of 2.5% lidocaine and 2.5% prilocaine; the remaining 30 were treated with a placebo. The nurses explained the definition of the visual analogue scale (VAS, scored from 0 to 10), and the patients evaluated their pain at 7 time points during the GKRS procedure by using the VAS. After each of these evaluations, the patients' vital signs (blood pressure, heart rate, and respiratory rate) were measured. RESULTS: There was no significant difference in the patients' age, gender, duration of frame fixation, and types of the lesions between the EMLA and placebo groups. The EMLA group reported significantly lower pain scores 20 and 60 min after frame removal than the placebo group (p=0.001 and p<0.001, respectively). Additionally, patients in the placebo group had significantly higher blood pressure readings compared with baseline data, during and after frame removal, thus indicating that postoperative wound pain caused them more discomfort after frame removal. CONCLUSION: EMLA when applied 60 min before frame removal has an anesthetic effect of reducing the postoperative wound pain in patients who undergo GKRS.

'Shame on you'--the psychosocial impact of genital warts.

We aimed to investigate whether patients with genital warts experience greater feelings of shame and lower self-esteem compared with controls. Sixty patients with genital warts were compared with 60 asymptomatic genitourinary (GU) medicine patients and 60 orthopaedic outpatients. The shame scores of those with warts (31.08) were significantly higher (P < 0.0001) than either control group (GU medicine controls 20.77; orthopaedic controls 19.00). The impact on health-related quality of life (HRQoL) by each of the individual emotional parameters of shame, low self-esteem, intrusive thoughts, avoidance behaviour and self-efficacy impact was examined in the wart sample group. Only internalized shame (P = 0.001) and intrusive thoughts (P < 0.0001) were significant in predicting HRQoL scores. There are emotional implications in having genital warts, which can have a profound effect on a patient's quality of life and these need addressing just as much as the physical warts.

Estudo de metodologia analítica para a determinação do cetoconazol em formulações farmacêuticas / Study of analytical methodology for ketoconazole determination in pharmaceutical preparation

Cetoconazol é um antifúngico sintético, derivado imidazólico de amplo espectro de ação, efetivo no tratamento de infecções superficiais e sistêmicas. Foram estudadas diferentes metodologias para análise do cetoconazol em especialidades farmacêuticas diversas usando espectrofotometria no ultravioleta, no infravermelho e análise térmica. Os resultados mostram que a espectrofotometria ultravioleta é um método rápido, prático e econômico e apontam que outros métodos como a espectrofotometria no infravermelho e análise térmica são uma alternativa à análise do cetoconazol em diferentes especialidades farmacêuticas.

Ketoconazole is a synthetic broad-spectrum oral and topical antifungal drug derived from imidazole, effective in the treatment of superficial mycoses and systemic infections. In this study we have tested several methods to analyze ketoconazole in various pharmaceutical products containing this drug, employing techniques such as UV and IR spectrophotometry and thermal analysis. The results showed that UV spectrophotometryis a fast, practical and economical method and indicated that other methods, such as IR spectrophotometry and thermal analysis, could be good alternative methods for ketoconazole analysis in certain pharmaceutical forms.

Dynamical structure factor and spin-density separation for a weakly interacting two-component Bose gas.

We show that spin-density separation in a Bose gas is not restricted to 1D but also occurs in higher dimensions. The ratio (alpha) of the intraspecies atom-atom interaction strength to the interspecies interaction strength strongly influences the dynamics of spin-density separation and the elementary excitations. The density wave is phononlike for all values of alpha. For alpha < 1, the spin wave is also phononlike. The spin waves have a quadratic dispersion in the alpha=1 coupling regime, while in the phase separated regime (alpha > 1) the spin waves are found to be damped. The dynamical structure factor reveals two distinct peaks corresponding to the density and spin waves for alpha < or =1. For alpha > 1 there is only one dynamical structure factor peak corresponding to the density wave.

Posttraumatic stress and co-morbidity following myocardial infarction among older patients: the role of coping.

This study aimed to investigate co-morbidity and coping strategies among older patients who suffer from different levels of posttraumatic stress disorder (PTSD) following myocardial infarction. Ninety-six older myocardial infarction (MI) patients were recruited from two general practices and completed the Posttraumatic Stress Diagnostic Scale (PDS), the General Health Questionnaire-28 (GHQ-28) and the COPE Scale. Ninety-two older patients with no previous MI experience constituted the control. Using the PDS, 30, 42 and 28% had full, partial and no-PTSD respectively. There were significant differences between the patient groups and the control on all GHQ-28 sub-scales. Significant differences were also identified between the patient groups in the following coping strategies: seeking emotional social support, suppression of competing activities, restraint coping, focusing on and venting of emotion, mental and behavioural disengagement. Controlling for bypass surgery, previous mental health difficulties, angioplasty, heart failure and angina, MANCOVA results did not change the overall results of the GHQ-28 but changed the results of coping in that seeking emotional social support and behavioural disengagement stopped being significant. Coping was a partial mediator between different levels of post-MI PTSD and co-morbidity. Depending on the severity of PTSD symptoms, co-morbidity and coping strategies can vary among older patients. Older patients with full-PTSD tend to use both maladaptive coping strategies as well as problem-focused coping.

Toxicidade de fármacos nitrofurânicos / Toxicity of nitrofuran drugs

Durante o planejamento estrutural de novos fármacos, é possível prever a influência de grupamentos específicos na atividade farmacológica. Entre estes, encontra-se o grupo nitro, que possui potencial atividade antimicrobiana, estando presente em diversos fármacos como o metronidazol, nitrofural, furazolidona, oxamniquina, cloranfenicol, entre outros. Também, a introdução do grupo nitro na molécula pode alterar as propriedades físico-químicas e eletrônicas da substância, estando presente em fármacos de outras classes terapêuticas como anti-úlcera, ansiolítico, antiinflamatório. Entretanto, restrições têm sido apontadas para o planejamento de novos fármacos contendo este grupo, devido à toxicidade relacionada. Este estudo trata-se da revisão sobre a toxicidadede compostos nitrofurânicos, bem como os possíveis mecanismos e a utilização do método de latenciação na diminuição desta toxicidade.

Determination of calcium and magnesium in hydroethanolic extracts of propolis by atomic absorption flame spectrophotometry

Propolis is a natural product collected by honeybees and has a large range of pharmacological activity, including antimicrobial, antitumoral, antioxidant andanti-inflammatory. Its use as a popular medicine is increasing all over the world, creating a need for quality control of the commercial products. In this study the levels of calcium and magnesium in commercial hydroalcoholic propolis extracts from varios states of Brazil were determined by atomic absorption flame spectrophotometry and different values were obtained for northern and southern states. This study can beextended to the analysis of metals that are harmfulto health. The results showed that the calibration curves were linear over a wide concentration range (0.5-4.0 miug.mL-1 for calcium and 0.05-0.4 miug.mL-1 formagnesium) with good correlation coefficients (0.999 and 0.988, respectively). Good analytical recovery (94%) was obtained. The proposed method showed adequate precision and relative standard deviation lower than 2%. The method is accurate and precise as well as having advantages such as simplicity and speed.

Rapid test for the evaluation of the activity of the prodrug hydroxymethylnitrofurazone in the processing of Trypanosoma cruzi messenger RNAs.

No fully effective treatment has been developed since the discovery of Chagas' disease by Carlos Chagas in 1909. Since drug-resistant Trypanosoma cruzi strains are occurring and the current therapy is effectiveness in the acute phase but with various adverse side effects, more studies are needed to characterize the susceptibility of T. cruzi to new drugs. Many natural and/or synthetic substances showing trypanocidal activity have been used, even though they are not likely to be turned into clinically approved drugs. Originally, drug screening was performed using natural products, with only limited knowledge of the molecular mechanism involved in the development of diseases. Trans-splicing, which is unusual RNA processing reaction and occurs in nematodes and trypanosomes, implies the processing of polycistronic transcription units into individual mRNAs; a short transcript spliced leader (SL RNA) is trans-spliced to the acceptor pre-mRNA, giving origin to the mature mRNA. In the present study, permeable cells of T. cruzi epimastigote forms (Y, BOL and NCS strains) were treated to evaluate the interference of two drugs (hydroxymethylnitrofurazone - NFOH-121 and nitrofurazone) in the trans-splicing reaction using silver-stained PAGE analysis. Both drugs induced a significant reduction in RNA processing at concentrations from 5 to 12.5 microM. These data agreed with the biological findings, since the number of parasites decreased, especially with NFOH-121. This proposed methodology allows a rapid and cost-effective screening strategy for detecting drug interference in the trans-splicing mechanism of T. cruzi.

Rapid test for the evaluation of the activity of the prodrug hydroxymethylnitrofurazone in the processing of Trypanosoma cruzi messenger RNAs

No fully effective treatment has been developed since the discovery of Chagas' disease by Carlos Chagas in 1909. Since drug-resistant Trypanosoma cruzi strains are occurring and the current therapy is effectiveness in the acute phase but with various adverse side effects, more studies are needed to characterize the susceptibility of T. cruzi to new drugs. Many natural and/or synthetic substances showing trypanocidal activity have been used, even though they are not likely to be turned into clinically approved drugs. Originally, drug screening was performed using natural products, with only limited knowledge of the molecular mechanism involved in the development of diseases. Trans-splicing, which is unusual RNA processing reaction and occurs in nematodes and trypanosomes, implies the processing of polycistronic transcription units into individual mRNAs; a short transcript spliced leader (SL RNA) is trans-spliced to the acceptor pre-mRNA, giving origin to the mature mRNA. In the present study, permeable cells of T. cruzi epimastigote forms (Y, BOL and NCS strains) were treated to evaluate the interference of two drugs (hydroxymethylnitrofurazone - NFOH-121 and nitrofurazone) in the trans-splicing reaction using silver-stained PAGE analysis. Both drugs induced a significant reduction in RNA processing at concentrations from 5 to 12.5 æM. These data agreed with the biological findings, since the number of parasites decreased, especially with NFOH-121. This proposed methodology allows a rapid and cost-effective screening strategy for detecting drug interference in the trans-splicing mechanism of T. cruzi.

Síntese e modelagem molecular do novo derivado indolinônico como candidato a antiinflamatório COX-2 seletivo / Synthesis and molecular modeling of a novel indolinone derivative as a candidate anti-inflammatory selectively inhibiting COX-2

Os fármacos antiinflamatórios são sabidamente os mais comercializados em todo o mundo, e apesar disto apresentam sérios efeitos colaterais, sobretudo no que se refere ao trato gastrintestinal. A descoberta de novos protótipos com atividade e segurança terapêutica melhoradas continua sendo uma busca constante. Com o adventoda química computacional torna-se mais fácil o estudo teórico do comportamento fisiológico de uma nova substância bem como a compreensão do possível mecanismo de ação destas novas moléculas. Assim, através de modelos matemáticos de moléculas e receptores estudou-se neste trabalho o composto I (1-(2,6- diclorofenil)indolin-2-ona) quanto à sua possibilidade deinibir seletivamente a isoforma COX-2 da enzima prostaglandina endoperóxido sintase (PGHS), e também as melhores posições para a introdução de grupamentos químicos e modificações moleculares.

Microondas doméstico na síntese de derivados ftalimídicos / Use of domestic microwave oven in the synthesis of phthalimide derivatives

Microondas são utilizadas na síntese orgânica desde 1986, e mostra-se vantajosa em diversos aspectos como possibilidade de maiores rendimentos, maior seletividade e menor decomposição térmica. A ftalimida e derivados, constituem-se em uma importante classe de compostos utilizados na química orgânica sintética, e do ponto de vista da Química Farmacêutica e Medicinal, é considerada um importante bióforo constituindo subunidade estrutural de caráter farmacofórico para uma série de compostos com diferentes atividades farmacológicas, entre elas, a anemia falciforme. O objetivo deste trabalho foi desenvolver metodologia alternativa na síntese de derivados ftalimídicos explorando a condensação de anidrido ftálico com aminas funcionalizadas sob radiação de microondas doméstico. Os resultados mostraram que os compostos ftalímidicos sintetizados podem ser obtidos em menores tempos reacionais (5-10 minutos) e maiores rendimentos(60-89%) quando comparados ao aquecimento convencional (refluxo), demonstrando o potencial da utilização do microondas na obtenção destas moléculas...

Interações medicamentosas de fitoterápicos e fármacos: Hypericum perforatum e Piper methysticum / Drug interactions between herbs and medicines: Hypericum perforatum and Piper methysticum

A utilização de produtos naturais na medicina popular é milenar e persiste até os dias atuais. Entretanto, a idéia de que estes produtos são isentos de toxicidade torna o uso de medicamentos fitoterápicos cada vez maior e indiscriminado. Este trabalho trata de uma revisão sobre as interações que podem ocorrer com a utilização concomitante de Hypericum perforatum L. (erva de são joão) e Piper methysticum F. (kava-kava) com fármacos, podendo levar a sérios efeitos tóxicos, incluindo a fatalidade.

Natural products in popular medicine have been used for hundreds of years and persists nowadays. However, the idea that these products are exempted of toxicity turns the use of herbs to be larger and indiscriminate. This work is a review of interactions that can happen with concomitant use of Hypericum perforatum L. (St. John's wort) and Piper methysticum F. (kava-kava) with medicines that can result in serious toxicological effects including fate.

Psychosocial benefits of solitary reminiscence writing: an exploratory study.

Claims have been made that reminiscence has benefits for older people's psychological well-being, and that writing memories may be a therapeutic process. This paper describes an exploratory study in which five nursing home residents engaged in a process of writing their memories by themselves, in a series of booklets containing memory prompts and photographs, over a period of four weeks. Each completed booklet was typed up by researchers and returned to participants the following week, with a bound copy provided to participants at the end of the study period. Analysis focuses on two sets of data: an in-depth case study of one participant, and a thematic analysis of field notes, researcher reflections, and the written material produced by the other study participants. The case study revealed three main themes: views on the past; sharing the past; and confidence in writing about the past. The field note analysis indicated the presence of four themes: proof and maintenance of skills; psychological or internal processes; social contact; and pleasure in reminiscence. The writing was seen as cathartic and provided a meaningful purpose, an opportunity to exercise writing skills and memory, and a focus for participants to share key stories with others. This exploratory study suggests that there is potential in using solitary writing within a reminiscence framework to improve psychological well-being in older people. However, caution should be exercised when encouraging older people to write their stories. Issues of confidentiality, audience, support, and appropriateness of the activity for the individual need consideration.

Proteomic investigation of metabolic shift in mammalian cell culture.

Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.

Purification and characterization of a variant of rhodocetin from Calloselasma rhodostoma (Malayan pit viper) venom.

Rhodocetin is a novel C-type lectin-related protein (CLP) purified from the venom of Calloselasma rhodostoma. Thus far, it is the only reported CLP whose alpha and beta subunits are not linked by an interdisulfide bond. We report here the isolation of a variant of rhodocetin from a different source of venom. This variant of rhodocetin exhibited a different elution profile in reverse-phase HPLC as compared to the rhodocetin reported in our original publication [Wang et al., (1999), Biochemistry 38, 7584-7593]. Specifically, the alpha subunit of the variant was eluted at a considerably lower percentage of acetonitrile, which suggested a less hydrophobic polypeptide chain as compared to the original rhodocetin. Using a combination of microcharacterization techniques such as peptide mapping, mass spectrometry, and amino acid sequencing, we identified an amino acid substitution, 163K, in the polypeptide chain that could account for the difference in elution behavior of the alpha subunit. In addition, we also found a conserved E88D substitution in the beta chain which was not apparent during reverse-phase HPLC. However, neither of these substitutions resulted in the alteration of the functional properties of the rhodocetin variant.