Role of lymphadenectomy for patients undergoing radical nephrectomy for renal cell carcinoma

The potential benefits of a lymph node dissection (LND) include more accurate staging, decreased local recurrence rates, and improved survival. However, only limited data support the potential benefits of routine, extensive LND for renal cell carcinoma (RCC). Because of lack of data, no clear practice standard has been established about whether to perform LND and, if so, to what extent. The value of LND for RCC is only relevant if the pattern of lymphatic spread is predictable, which it is not. However, although little evidence supports the value of LND for RCC, it is probable that an occasional patient will have very early metastasis confined to the area of the primary and secondary major lymphatic flow medially and perhaps will benefit therapeutically. In this case, a limited LND is supportable. No information currently available strongly supports the value of a more extensive and potentially more morbid LND for either staging or therapeutic value. Extensive investigation is necessary in order to establish LND as a standard component of RCC surgery.     

Inhibition of Mitochondrial Complex II by the Anticancer Agent Lonidamine

The antitumor agent lonidamine (LND; 1-(2,4-dichlorobenzyl)-1H-indazole-3-carboxylic acid) is known to interfere with energy-yielding processes in cancer cells. However, the effect of LND on central energy metabolism has never been fully characterized. In this study, we report that a significant amount of succinate is accumulated in LND-treated cells. LND inhibits the formation of fumarate and malate and suppresses succinate-induced respiration of isolated mitochondria. Utilizing biochemical assays, we determined that LND inhibits the succinate-ubiquinone reductase activity of respiratory complex II without fully blocking succinate dehydrogenase activity. LND also induces cellular reactive oxygen species through complex II, which reduced the viability of the DB-1 melanoma cell line. The ability of LND to promote cell death was potentiated by its suppression of the pentose phosphate pathway, which resulted in inhibition of NADPH and glutathione generation. Using stable isotope tracers in combination with isotopologue analysis, we showed that LND increased glutaminolysis but decreased reductive carboxylation of glutamine-derived α-ketoglutarate. Our findings on the previously uncharacterized effects of LND may provide potential combinational therapeutic approaches for targeting cancer metabolism.     

Lymph node dissection in patients with kidney cancer: when is it indicated?

It is necessary to consider the potential risks and benefits of performing a lymph node dissection (LND) at the time of radical nephrectomy. LND may lead to more accurate staging, a decrease in local recurrence, and an increase in survival for patients with metastatic disease limited to the resected lymph nodes. However, there are risks associated with the treatment that must be considered. The advantages of LND vary among patients, depending on the location and extent of disease progression. For patients with stage T3 or T4 disease, we recommend a limited LND. Patients with T3 or T4 disease with grossly positive nodes with or without hematogenous metastasis should undergo a more extensive LND.     

The Motor Disorder of Classic Lesch‐Nyhan Disease

Reports describing the neurological features of Lesch‐Nyhan disease (LND) vary widely, thereby implying the involvement of different neurological substrates. The movement abnormalities in 20 patients with LND were investigated. Dystonia was the most frequent and severe movement disorder. At rest, hypotonia was more frequent than hypertonia. These findings are compatible with basal ganglia dysfunction in LND.     

The motor disorder of classic Lesch-Nyhan disease

Reports describing the neurological features of Lesch-Nyhan disease (LND) vary widely, thereby implying the involvement of different neurological substrates. The movement abnormalities in 20 patients with LND were investigated. Dystonia was the most frequent and severe movement disorder. At rest, hypotonia was more frequent than hypertonia. These findings are compatible with basal ganglia dysfunction in LND.     

Nucleotide degradation products in cerebrospinal fluid (CSF) in inherited and acquired pathologies

CSF purines were grossly elevated compared with controls only in adenylosuccinate lyase (ADSL) deficiency and TB meningitis. The former representing low permeability, the latter severe damage to the normal blood/brain barrier. By contrast, the similarity to controls, with no difference between Lesch-Nyhan disease (LND) or LND variants, would exclude hypoxia as a factor in the severe neurological deficits in LND. Similar findings in purine nucleoside phosphorylase (PNP) deficiency (although nucleosides replace the normal bases) likewise exclude hypoxia in the aetiology of the albeit milder neurological deficits.     

Nucleotide Degradation Products in Cerebrospinal Fluid (CSF) in Inherited and Acquired Pathologies

CSF purines were grossly elevated compared with controls only in adenylosuccinate lyase (ADSL) deficiency and TB meningitis. The former representing low permeability, the latter severe damage to the normal blood/brain barrier. By contrast, the similarity to controls, with no difference between Lesch–Nyhan disease (LND) or LND variants, would exclude hypoxia as a factor in the severe neurological deficits in LND. Similar findings in purine nucleoside phosphorylase (PNP) deficiency (although nucleosides replace the normal bases) likewise exclude hypoxia in the aetiology of the albeit milder neurological deficits.     

Mitochondrial targeting drug lonidamine triggered apoptosis in doxorubicin-resistant HepG2 cells

Mitochondria play a crucial role in the induction and execution of apoptosis. Accordingly, recent suggestions have been made to use agents that directly act on mitochondria to trigger apoptosis so that drug-sensitive and-resistant tumour cells can be eliminated. To test this hypothesis, human hepatocarcinoma HepG2 and its derivative R-HepG2 with doxorubicin (Dox) resistance as a result of expression of P-glycoprotein were used to investigate the effect of lonidamine (LND), a new mitochondrial targeting drug, on the induction of apoptosis. Results from our study indicate that R-HepG2 cells were more sensitive to LND than parental cells in terms of cytotoxicity determined by alamar blue assay. Cell death induced by LND was associated with the hallmarks of apoptosis such as mitochondrial membrane depolarization, release of cytochrome c, phosphatidyl-serine externalization and DNA fragmentation. Moreover, combined treatment of cells with Dox and LND elicited more cell death. Taken together, our results suggest a potential use of LND as an anti-cancer drug to bypass drug resistance and to trigger tumour destruction through apoptosis in HepG2 and R-HepG2 cells.     

The impact of bird herbivory on macrophytes and the resilience of the clear-water state in shallow lakes: a model study

Land-use change associated with human development can alter aquatic habitat and imperil aquatic species. Fish are challenged when urban streams are altered, for example for stormwater conveyance, but little is known about how such activities influence the space use of individual fish. Electronic tagging and experimental displacement of fish can be used to explore site fidelity and homing behaviour of fish and can therefore be useful for testing hypotheses about space use and habitat selection. In this study, we used experimental displacement to determine how longnose dace (LND, Rhinichthys cataractae) utilize reaches within a watershed that have varying degrees of degradation. LND were tagged using passive integrated transponders (PIT tags), transported upstream, and released either into the natural stream reach, impaired stormwater drain reach, or at their confluence. Fixed PIT antennas were used to monitor movement of the PIT-tagged fish among the three reaches for a period of 3 weeks. LND exhibited dramatic and rapid selection against the stormwater drain. No LND moved into the drain and 97% of fish transported to the drain left within 24 h. LND were actively avoiding the stormwater drain, emphasizing the need for enhancement work to improve the biological connectivity of the system.     

Human HPRT1 gene and the Lesch–Nyhan disease: Substitution of alanine for glycine and inversely in the HGprt enzyme protein

Lesch–Nyhan disease (LND) is a rare X-linked inherited neurogenetic disorder of purine metabolism in which the enzyme, hypoxanthine-guanine phosphoribosyltransferase (HGprt) is defective. The authors report three novel independent mutations in the coding region of the HPRT1 gene from genomic DNA of (a) a carrier sister of two male patients with LND: c.569G>C, p.G190A in exon 8; and (b) two LND affected male patients unrelated to her who had two mutations: c.648delC, p.Y216X, and c.653C>G, p.A218G in exon 9. Molecular analysis reveals the heterogeneity of genetic mutation of the HPRT1 gene responsible for the HGprt deficiency. It allows fast, accurate detection of carriers and genetic counseling.     

Urinary Function following Laparoscopic Lymphadenectomy for Male Rectal Cancer

Objectives

Urinary function can be protected following open lateral node dissection (LND) with pelvic autonomic nerve preservation (PANP) for advanced rectal cancer. However data regarding urinary function after laparoscopic LND with PANP have not been reported. The goal of this study was to determine the effects of laparoscopic LND with PANP on urinary function in male patients with rectal cancer.

Methods

Urine flowmetry was performed using an Urodyn flowmeter. Patients were also asked to complete the standardized International Prostate Symptom Score (IPSS) questionnaire before surgery and 6 months after. In total, this study consisted of 60 males with advanced rectal cancer.

Results

No significant differences were seen in maximal urinary flow rate, voided volume or residual volume before and after surgery. The total IPSS score increased significantly after surgery and at least 41 patients (68.3%) reported there was no change in one of the seven IPSS questions.

Conclusions

Laparoscopic LND with PANP was relatively safe in preserving urinary function.     

Hypoxanthine-guanine phosphoribosyltransferase deficiency: biochemical and molecular findings in six Argentine patients

Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency is an inborn error of purine metabolism responsible for Lesch-Nyhan Disease (LND) and its partial phenotypes, HPRT-related hyperuricemia with neurologic dysfunction (HRND) and hyperuricemia alone. We report here the recognition of six Argentine patients, two with LND and four with HRND. All patients presented elevated excretion of uric acid, hypoxanthine, and xanthine and decreased HPRT enzyme activities <1 nmol/h/mg Hb. The molecular analysis demonstrated in the two LND patients a novel inherited transition mutation, c.203T >C (L68P), in one subject and a germline transition mutation, c.209G >A (G70E), in the other. In the HRND patients a novel transversion mutation, c.584 A >C (Y195S), was found in three related patients and an inherited transition mutation, c.143G >A (R48H), in the fourth subject.     

Immunohistochemical profile of some neurotransmitters and neurotrophins in the seminiferous tubules of rats treated by lonidamine

Lonidamine (LND) or [1-(2,4-dichlorobenzyl)-1H-indazole-3-carboxylic acid] is an anticancer and antispermatogenic drug that exerts a large number of effects on tumor cells and germ cells. Sexually mature male Sprague-Dawley rats were housed at 22 degrees C on a 12-h light/12-h dark cycle 1 week before the experiments, with free access to food and water. LND was suspended in 0.5% methylcellulose at a concentration of 10 mg/mL and administered orally at the dose of 10 mL/kg (b.w.) as a single dose. Control rats received an equal amount of vehicle. Testes were removed, fixed for 24 h in 2% glutaraldehyde and 2% paraformaldehyde in 0.1 M sodium phosphate (pH 7.2 at 22 degrees C), rinsed with the same buffer, and stored at room temperature. From each sample, a block of tissue was removed by sectioning through the organ. After dehydration in ethanol at increasing concentrations (70-100%), each block was embedded in paraffin and serial 5 mm thick sections were cut using a rotatory microtome. The immunoreactivity for NTs has been observed in spermatogonia of untreated rats, while the rats treated with LND showed an immunohistochemical localization in all the stages of germinal cells. The generally well-expressed immunoreactivity for the neurotrophins receptors in treated rats observed in our study is presumably attributable to alterations of the receptors' structure and/or expression leading to changes of the activity, affinity, localization or protein interactions that may depend on sensitization of ion channels (induced by LND). Neurotrophins (NTs) appear to be interesting proteins for the modulation of sperm maturation and motility with a prominent role for the nerve growth factor (NGF), that may exert an autocrine or paracrine role. We therefore investigated the location and distribution of immunoreactivity for some neurotransmitters (SP, VIP, CGRP, nNOS, Chat), neurotrophins (NGF, BDNF, NT-3) and their own receptors (TrKA, TrKB, TrKC, p75) in the seminiferous tubules of male rats treated by LND in the light of the literature on this topic.     

Hypoxanthine-Guanine Phosphoribosyltransferase Deficiency: Biochemical and Molecular Findings in Six Argentine Patients

Hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency is an inborn error of purine metabolism responsible for Lesch-Nyhan Disease (LND) and its partial phenotypes, HPRT-related hyperuricemia with neurologic dysfunction (HRND) and hyperuricemia alone. We report here the recognition of six Argentine patients, two with LND and four with HRND. All patients presented elevated excretion of uric acid, hypoxanthine, and xanthine and decreased HPRT enzyme activities <1 nmol/h/mg Hb. The molecular analysis demonstrated in the two LND patients a novel inherited transition mutation, c.203T >C (L68P), in one subject and a germline transition mutation, c.209G >A (G70E), in the other. In the HRND patients a novel transversion mutation, c.584 A >C (Y195S), was found in three related patients and an inherited transition mutation, c.143G >A (R48H), in the fourth subject.     

Molecular Characterization and Structure Analysis of Hprt in a Chinese Patient with Lesch-Nyhan Disease

Lesch-Nyhan disease (LND) is caused by deficiency of hypoxanthine guanine phosphoribosyltransferase (HPRT). The aim of the present study is to characterize the molecular deficiency of a clinical diagnosed Chinese patient with attenuated variant of LND. The coding region and the intron-exon boundaries of HPRT1 gene were sequenced by standard methods, and HPRT activity was assayed by HPLC method. Structure analysis was performed to estimate the consequence of the mutant of HPRT1 gene. A new mutation c.245T>G (p.Ile82Ser) was identified in this patient, and heterozygous mutation was found in the patient's mother. The activity of HPRT in the patient was completely undetectable. Structure study indicates that the mutation of p.Ile82Ser may lead to loss of hydrophobic side chain and disrupt its normal conformation of HPRT protein. It is helpful for diagnosis of LND that sequencing analysis of HPRT1 gene is performed in male infant and juvenile with hyperuricaemia and neurologic dysfunction in Chinese.     

Carrier and prenatal diagnosis of Lesch–Nyhan disease due to a defect in HPRT gene expression regulation

Lesch–Nyhan disease (LND) is caused by lack of hypoxanthine–guanine phosphoribosyltransferase (HPRT) activity. Mutations in HPRT1 gene show variability in type and location within the gene, and in certain patients the HPRT coding sequence is normal and the molecular defect cannot be found. These patients presented a decreased HPRT1 expression of unknown cause. This is the first report of a carrier and prenatal diagnosis of LND due to a defect in HPRT gene expression regulation.     

Partial characterization of the proteome of the mouse striatum

Many diseases of the mammalian CNS, including Parkinson's (PD) and Lesch Nyhan disease (LND), are associated with programmatic neurodegeneration or dysfunction of dopaminergic neurons in the mesencephalon, the nigrostriatal pathway, and its projections in the striatum [1-4]. Proteomic studies on brain tissue of both animal models and human PD patients have provided evidence for dysfunction and damage of many pathways, including oxidative stress-related damage, ubiquitin-proteasome dysfunction, mitochondrial energy metabolism deficiencies, and synaptic function [5-11]. To date no such proteomic studies have been reported in the related and rare basal ganglia disorder LND, a developmental rather than a neurodegenerative neurological disorder caused by deficiency of the enzyme hypoxanthine-guanine phosphoribosyltransferase (HPRT) that regulates a major step in the purine salvage pathway [12]. Many studies have demonstrated that the both human LND patients and a mouse knockout model of HPRT deficiency have significantly reduced levels and uptake of dopamine in the striatum [4, 13-16] that is likely to be the principal cause of the CNS disorder. The precise molecular and cellular mechanisms that underlie this neurotransmitter defect are unknown.     

The effect of a non-starch polysaccharide-hydrolysing enzyme (Rovabio® Excel) on feed intake and body condition of sows during lactation and on progeny growth performance

A total of 200 (Large White × Landrace) sows were used in a 39-day study to evaluate the effects of feeding a non-starch polysaccharide (NSP)-hydrolysing enzyme multicomplex (Rovabio® Excel) in conjunction with a high- or reduced nutrient-density diet during lactation on sow body condition, feed intake and progeny performance. Eight sows were selected each week for 25 weeks, blocked by parity and BW into groups of four, and within the block randomly assigned to one of the four treatments (n = 50/treatment). Treatments were: (1) LND: low energy (13.14 MJ of DE/kg), low CP (15%) diet; (2) LND + RE: LND with 50 mg/kg NSP-hydrolysing enzyme; (3) HND: high energy (14.5 MJ of DE/kg), high CP (16.5%) diet; and (4) HND + RE: HND with 50 mg/kg NSP-hydrolysing enzyme. Sows were fed treatment diets from day 109 of gestation until the day of subsequent service. Between weaning and re-service, Rovabio® Excel addition to LND diets resulted in an increase in energy intake; however, a reduction was observed when supplemented to the HND diet (P < 0.05). The inclusion of Rovabio® Excel increased feed and energy intake during week 3 (days 15 to 21) of lactation (P < 0.05). Sows fed diets supplemented with Rovabio® Excel had greater back-fat depth at weaning and service (P < 0.05); however, the magnitude of change in back-fat depth during lactation and from farrowing to service was not different between treatments. Feeding the HND diet increased energy intake before farrowing, throughout lactation and during the weaning to service interval (P < 0.01); however, overall, average daily feed intake tended to be reduced (P < 0.10). At service, sows fed the HND diet were heavier than sows fed the LND diet (P < 0.05); however, the magnitude of change in BW between treatments was not different. Feeding the HND diet to sows resulted in a tendency for heavier piglets at birth (P = 0.10) that tended to grow at a faster rate and be heavier at weaning than piglets from sows fed the LND diet (P = 0.06). These results indicate that NSP-degrading enzymes offer minimal benefit to sows and their progeny when fed before and during lactation; however, increasing energy intake of sows during lactation may beneficially affect progeny.     

PRTFDC1 is a genetic modifier of HPRT-deficiency in the mouse

Lesch-Nyhan disease (LND) is a severe X-linked neurological disorder caused by a deficiency of hypoxanthine phosphoribosyltransferase (HPRT). In contrast, HPRT-deficiency in the mouse does not result in the profound phenotypes such as self-injurious behavior observed in humans, and the genetic basis for this phenotypic disparity between HPRT-deficient humans and mice is unknown. To test the hypothesis that HPRT deficiency is modified by the presence/absence of phosphoribosyltransferase domain containing 1 (PRTFDC1), a paralog of HPRT that is a functional gene in humans but an inactivated pseudogene in mice, we created transgenic mice that express human PRTFDC1 in wild-type and HPRT-deficient backgrounds. Male mice expressing PRTFDC1 on either genetic background were viable and fertile. However, the presence of PRTFDC1 in the HPRT-deficient, but not wild-type mice, increased aggression as well as sensitivity to a specific amphetamine-induced stereotypy, both of which are reminiscent of the increased aggressive and self-injurious behavior exhibited by patients with LND. These results demonstrate that PRTFDC1 is a genetic modifier of HPRT-deficiency in the mouse and could therefore have important implications for unraveling the molecular etiology of LND.