Points to consider in seeking biosafety approval for research, testing, and environmental release of experimental genetically modified biocontrol products during research and development.

Novel genetically modified biological control products (referred to as "GM biocontrol products") are being considered to address a range of complex problems in public health, conservation, and agriculture, including preventing the transmission of vector-borne parasitic and viral diseases as well as the spread of invasive plant and animal species. These interventions involve release of genetically modified organisms (GMOs) into the environment, sometimes with intentional dissemination of the modification within the local population of the targeted species, which presents new challenges and opportunities for regulatory review and decision-making. Practices developed for GMOs, primarily applied to date for GM crops may need to be adapted to accommodate different types of organisms, such as insects, and different technologies, such as gene drive. Developers of new GM biocontrol products would benefit from an early understanding of safety data and information that are likely to be required within the regulatory dossier for regulatory evaluation and decision making. Here a generalizable tool drawing from existing GM crop dossier requirements, forms, and relevant experience is proposed to assist researchers and developers organize and plan their research and trialing. This tool requires considering specifics of each investigational product, their intended use, and country specific requirements at various phases of potential product development, from laboratory research through contained field testing and experimental release into the environment. This may also be helpful to risk assessors and regulators in supporting their systematic and rigorous evaluation of new biocontrol products.

Issues and gaps in international guidance and national regulatory systems affecting international live insect trade.

International trade in live insects involves the shipping of many different species, for various purposes, with a variety of handling requirements regulated by numerous authorities with varying objectives. The diversity of factors at play has both created and been subject to a complex regulatory landscape. A review of global production, shipping and use experiences from a range of perspectives has shown gaps and inconsistencies in international guidance and national implementation. Private carriers add another layer of uncertainty that is disproportionate to risks, resulting in variable practices and charges. Many benefits can come from international trade in insects, including pollinator services, control of pests and of disease vectors, and enhanced international scientific research and innovation. These benefits will be better achieved through a more evidence-based and efficient approach to regulating trade. This change in approach will in turn require an improved and widely accepted risk-management landscape for insect trade.

Le commerce international d'insectes vivants s'appuie sur les expéditions de nombreuses espèces différentes à diverses fins, assorties de multiples exigences relatives à la manutention régies par des autorités différentes poursuivant des objectifs distincts. La diversité des facteurs en jeu a rendu nécessaire la création d'un paysage normatif complexe, avec les contraintes qui lui sont associées. L'examen à l'échelle mondiale de la production, du transport et des expériences dans ce domaine depuis diverses perspectives a révélé un certain nombre de lacunes et d'incohérences au niveau des directives internationales et de leurs applications concrètes dans les pays. Les transporteurs privés ajoutent une dimension d'incertitude supplémentaire qui est disproportionnée par rapport aux risques, ce qui entraîne une forte variabilité des pratiques et des charges. Les échanges internationaux d'insectes génèrent nombre d'activités bénéfiques, parmi lesquelles les services de pollinisation, la lutte contre les ravageurs et les vecteurs de maladie, et l'accroissement de la recherche scientifique internationale et de l'innovation. Les objectifs attendus seront mieux atteints en adoptant une approche de la réglementation des échanges qui s'appuie davantage sur des éléments factuels et sur les gains d'efficacité. Ce changement méthodologique nécessitera à son tour la mise en place d'un cadre amélioré et plus largement accepté de la gestion des risques dans le domaine du commerce des insectes.

El comercio internacional de insectos vivos, que supone el transporte de muchas especies diferentes con fines diversos, se acompaña de requisitos de manipulación que dictan numerosas autoridades con todo tipo de objetivos. La diversidad de los factores que entran en juego ha sido a la vez causa y consecuencia de un complejo panorama reglamentario. Los autores, tras describir un estudio a escala mundial de la experiencia de producción, transporte y utilización de insectos vivos desde diferentes puntos de vista, exponen las carencias e incoherencias observadas en las directrices internacionales y su traslación a escala nacional. Los transportistas privados añaden otro factor de incertidumbre que tiene un peso desproporcionado en relación con el nivel de riesgo y se traduce en procedimientos y precios heterogéneos. El comercio internacional de insectos puede traer consigo muchos beneficios, en particular servicios de polinización, control de plagas y de vectores de enfermedad, y avances de la investigación e innovación científica a escala internacional. Pero obtener esos beneficios será más fácil si se aborda la regulación del comercio de manera más eficaz y científicamente fundamentada, operando un cambio de lógica que exigirá, a su vez, la aplicación al comercio de insectos de un régimen más sofisticado y ampliamente aceptado de gestión de los riesgos.

Renal support in acute kidney injury.

Acute kidney injury requiring renal replacement therapy (RRT) is associated with an unacceptably high mortality rate. Despite the identification of the modality, timing and intensity of dialysis, membrane biocompatibility, hollow fiber and catheter properties as potential modifying factors, there is little convincing evidence for the superiority of one over the other. However, the available data suggest that the early initiation of RRT may be beneficial. A focused review of clinical trials and meta-analysis of clinical trials of RRT is provided.

Intra-abdominal hypertension can be monitored with femoral vein catheters during CRRT and may cause access recirculation.

OBJECTIVES: Intra-abdominal hypertension (IAH) is an increasingly recognized disorder, and its diagnosis depends on accurate pressure monitoring. Bladder-based protocols are favored, but are not always clinically feasible. Abdominal venous (i.e. vena cava) pressure measurements are an alternative but are logistically challenging. We hypothesized that for patients suffering acute kidney injury, transducers built into renal replacement therapy (RRT) machines offer a simple opportunity to monitor pressures using catheters inserted via femoral veins. DESIGN: We performed in vitro testing of the accuracy of pressure transducers incorporated into continuous RRT devices, using highly calibrated instrumentation in the IAH-relevant range of 0 to +50 mmHg. We developed a protocol for using this modality in vivo, by stopping all pumps so as to allow equilibration of pressures: clinical application in a patient with femoral vein catheters and IAH was then described. RESULTS: In vitro analyses showed accuracy of the extracorporeal pressure transducers with an r² of 0.998, p < 0.001. In the patient case, the pressure transduced at the RRT device was identical to those obtained from bladder catheters. IAH also led to access recirculation and ineffective therapy. CONCLUSIONS: Pressure sensors incorporated into continuous RRT machines can be accurate in the IAH physiologic range, and thus may be used to easily measure intra-abdominal pressure via appropriate-length femoral vein-inserted access catheters. If not relieved, IAH can be an under-appreciated cause of access recirculation and ineffective clearance for any RRT modality (continuous or intermittent) using femoral catheters.

Sonic hedgehog delivered by an adeno-associated virus protects dopaminergic neurones against 6-OHDA toxicity in the rat.

Direct intracerebral administration of sonic hedgehog (SHH) reduces 6-OHDA and MPTP toxicity to nigral dopaminergic cells in rats and primates. To determine whether transfection of the DNA sequence for SHH using viral vectors also protects against 6-OHDA toxicity, a type 2 adeno- associated virus (AAV) incorporating 600 base pairs of N-terminal SHH DNA was generated to induce SHH expression in rat striatum.AAV-SHH was injected into the striatum, 3 weeks prior to the initiation of an unilateral partial 6-OHDA nigro-striatal lesion. Animals receiving 4x10(7) viral particles of AAV-SHH showed a reduction in (+)-amphetamine induced ipsilateral turning over 4 weeks, when compared to animals receiving vehicle or a LacZ encoding vector. Following vehicle or AAV-LacZ administration, 6-OHDA caused a marked loss of striatal dopamine content and nigral tyrosine hydroxylase (TH) immunopositive cells. Following treatment with 4x10(7) viral particles of AAV-SHH the loss of striatal dopamine content was reduced and there was marked preservation of nigral dopaminergic cells. However, administration of 4x10(8) particles of AAV-SHH did not cause a significant change in (+)-amphetamine-induced rotation, striatal dopamine levels or the number of nigral TH immunoreactive cells following 6-OHDA lesioning compared to vehicle or AAV-LacZ treated animals. The results show that SHH delivered via a viral vector can protect dopaminergic neurons against 6-OHDA toxicity and suggest that this could be developed into a novel treatment for PD. However, the effects maybe dose limited due to uncoupling of hedgehog receptor signalling at higher levels of SHH expression.

Tuberculosis of the spine (Pott's disease) presenting as 'compression fractures'.

STUDY DESIGN: Case reports and survey of literature. OBJECTIVE: Case reports of two women with tuberculosis (TB) of the spine (Pott's disease) presenting with severe back pain and diagnosed as compression fracture are described. Physicians should include Pott's disease in the differential diagnosis when patients present with severe back pain and evidence of vertebral collapse. SETTING: Ohio, USA METHODS: A review of the literature on the pathogenesis, pathophysiology, clinical presentation, diagnostic methods, treatment and prognosis of spinal TB was conducted. RESULTS: After initial delay, proper diagnosis of spinal TB was made in our patients. Microbiologic diagnosis confirmed M. tuberculosis, and appropriate medical treatment was initiated. CONCLUSIONS: Although uncommon, spinal TB still occurs in patients from developed countries, such as the US and Europe. Back pain is an important symptom. Vertebral collapse from TB may be misinterpreted as 'compression fractures' especially in elderly women. Magnetic resonance imaging scan (MRI) is an excellent procedure for the diagnosis of TB spine. However, microbiologic diagnosis is essential. Mycobacterium tuberculosis may be cultured from other sites. Otherwise, biopsy of the spine lesion should be done for pathologic diagnosis, culture and stain for M. tuberculosis. Clinicians should consider Pott's disease in the differential diagnosis of patients with back pain and destructive vertebral lesions. Proper diagnosis and anti-tuberculosis treatment with or without surgery will result in cure.

Behavioural and immunohistochemical changes following supranigral administration of sonic hedgehog in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated common marmosets.

Sonic hedgehog (SHH) has trophic actions on dopaminergic cell cultures and protects them from MPP(+) toxicity but its in vivo actions have not been explored. We now investigate the effects of unilateral supranigral administration of SHH on nigro-striatal function in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated common marmosets. SHH (0.1 or 1.0 microg) or vehicle was stereotaxically injected into the region of the right substantia nigra twice with an interval of 5 weeks between administrations. The first or second administration of low dose SHH (0.1 microg) did not significantly improve motor disability or locomotor activity compared to time-matched vehicle-treated animals. There was, however, an approximately 30% improvement in both motor disability and locomotor activity following the first administration of high dose SHH (1.0 microg). No further improvements occurred following the second high dose SHH treatment. Acute oral administration of L-3,4-dihydroxyphenylalanine (L-DOPA) produced a smaller increase in locomotor activity and greater reversal of motor disability in animals treated with SHH than occurred in vehicle-treated common marmosets. In the substantia nigra pars compacta, ipsilateral to SHH administration, the number of tyrosine hydroxylase-positive neurones was increased by 21% (P > 0.05) and 57% (P < 0.05) in low and high dose SHH groups respectively compared to the untreated contralateral hemisphere. There was no difference in the number of glial fibrillary acidic protein-positive cells. SHH may improve nigro-striatal function by restoring tyrosine hydroxylase positivity. This is reflected by an improvement in basal disability and a reduction in the lesion-induced response to L-DOPA.

6-Hydroxydopamine lesioning differentially affects alpha-synuclein mRNA expression in the nucleus accumbens, striatum and substantia nigra of adult rats.

The effect of a unilateral 6-hydroxydopamine (6-OHDA) lesion and/or repeated administration of levodopa (L-DOPA) to normal and 6-OHDA-lesioned rats on alpha-synuclein mRNA expression was investigated by in situ hybridization histochemistry. A 6-OHDA lesion of the nigro-striatal pathway alone, confirmed by the loss of nigral tyrosine hydroxylase mRNA expression, markedly decreased alpha-synuclein mRNA in the lesioned substantia nigra (SN). In contrast, the levels of alpha-synuclein mRNA in the denervated striatum and nucleus accumbens were not altered. Chronic administration of L-DOPA to normal or 6-OHDA-lesioned rats had no effect on alpha-synuclein mRNA expression in the SN, striatum or nucleus accumbens. These data confirm that alpha-synuclein is localized in the nigro-striatal tract but that its gene expression is not regulated by dopamine.

Transitional cell carcinoma manifesting as acute cor pulmonale: cause of microscopic tumor embolism.

Acute cor pulmonale is an uncommon manifestation of microscopic pulmonary tumor embolism. We describe the case of an 84-year-old man with a history of transitional cell carcinoma (TCC) of the urinary bladder who had acute cor pulmonale and died within a few hours after the onset of dyspnea. Autopsy showed that the right ventricle was dilated without hypertrophy. Microscopic examination of the lung showed that the small arteries, arterioles, and capillaries were filled with micrometastases of TCC. Microscopic pulmonary tumor embolism has rarely been reported with TCC, and to the best of our knowledge, such a fulminant course has not been previously described in the English language.

How important is history of unconsciousness in head injury patients?

A total of 100 patients of head injury were studied. They all underwent thorough clinical and neurological examination, skull radiography and CT scan of head. And with the help of all these parameters an evaluation was done to find out the importance of history of unconsciousness on risk of intracranial complications. A lot of controversies exist about detection of factors that make a patient of head injury a high risk candidate for developing intracranial complications. Young active population was the most commonly affected group in head injury with male preponderance at all ages. Roadside accident found to be the main cause of head trauma in adults while fall from height in paediatric age group. Although Glassgow Coma Index (GCI) was found to be a good predictor for intracranial complications as well as final outcome in patients with total score of 12 or less but it was not equally good in patients of minor head injury (GCI = 13-15). Similarly history of unconsciousness was not found to be correlating well with risk of intracranial complications but longer duration of unconsciousness was found to be a poor prognostic index.

Overexpression of the CstF-64 and CPSF-160 polyadenylation protein messenger RNAs in mouse male germ cells.

Messenger RNAs for several components of the transcriptional apparatus are greatly overexpressed in postmeiotic male germ cells in rodents (Schmidt and Schibler, Development 1995; 121:2373-2383). Because of the tight coupling of polyadenylation and transcription, we examined expression in germ cells of mRNAs for key polyadenylation factors. The mRNA for the 64 000 M(r) subunit of the cleavage stimulation factor (CstF-64) was expressed at least 250-fold greater in mouse testicular RNA than in liver RNA. RNA blot analysis showed that the mRNA for the 160 000 M(r) subunit of the cleavage and polyadenylation specificity factor was similarly overexpressed, as was the mRNA for the large subunit of RNA polymerase II. General transcription factors, such as the TATA-binding protein and transcription factor IIH, and splicing factors, such as components of the small nuclear ribonucleoproteins, were also expressed in meiotic and postmeiotic germ cells. The X-linked CstF-64 protein is expressed before and after but not during meiosis in the mouse (Wallace et al., Proc Natl Acad Sci U S A 1999; 96:6763-6768), which suggests that overexpression of mRNA transcription and processing factors plays an essential role in postmeiotic germ cell mRNA metabolism.

The gene for a variant form of the polyadenylation protein CstF-64 is on chromosome 19 and is expressed in pachytene spermatocytes in mice.

Many mRNAs in male germ cells lack the canonical AAUAAA but are normally polyadenylated (Wallace, A. M., Dass, B., Ravnik, S. E., Tonk, V., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., and MacDonald, C. C. (1999) Proc. Natl. Acad Sci. U. S. A. 96, 6763-6768). Previously, we demonstrated the presence of two distinct forms of the M(r) 64,000 protein of the cleavage stimulation factor (CstF-64) in mouse male germ cells and in brain, a somatic M(r) 64,000 form and a variant M(r) 70,000 form. The variant form was specific to meiotic and postmeiotic germ cells. We localized the gene for the somatic CstF-64 to the X chromosome, which would be inactivated during male meiosis. This suggested that the variant CstF-64 was an autosomal homolog activated during that time. We have named the variant form "tau CstF-64," and we describe here the cloning and characterization of the mouse tauCstF-64 cDNA, which maps to chromosome 19. The mouse tauCstF-64 protein fits the criteria of the variant CstF-64, including antibody reactivity, size, germ cell expression, and a common proteolytic digest pattern with tauCstF-64 from testis. Features of mtauCstF-64 that might allow it to promote the germ cell pattern of polyadenylation include a Pro --> Ser substitution in the RNA-binding domain and significant changes in the region that interacts with CstF-77.

Two distinct forms of the 64,000 Mr protein of the cleavage stimulation factor are expressed in mouse male germ cells.

Polyadenylation in male germ cells differs from that in somatic cells. Many germ cell mRNAs do not contain the canonical AAUAAA in their 3' ends but are efficiently polyadenylated. To determine whether the 64,000 Mr protein of the cleavage stimulation factor (CstF-64) is altered in male germ cells, we examined its expression in mouse testis. In addition to the 64,000 Mr form, we found a related approximately 70,000 Mr protein that is abundant in testis, at low levels in brain, and undetectable in all other tissues examined. Expression of the approximately 70,000 Mr CstF-64 was limited to meiotic spermatocytes and postmeiotic spermatids in testis. In contrast, the 64,000 Mr form was absent from spermatocytes, suggesting that the testis-specific CstF-64 might control expression of meiosis-specific genes. To determine why the 64,000 Mr CstF-64 is not expressed in spermatocytes, we mapped its chromosomal location to the X chromosome in both mouse and human. CstF-64 may, therefore, be absent in spermatocytes because the X chromosome is inactivated during male meiosis. By extension, the testis-specific CstF-64 may be expressed from an autosomal homolog of the X chromosomal gene.

Quantification of the expression of a temperature-sensitive pigment allele in sailfin mollies (Poecilia latipinna) by image analysis.

Image analysis was used to quantify the activity of a temperature-sensitive macromelanophore-determining allele in sailfin mollies as the percentage of the body surface area covered by macromelanophores. Fish heterozygous for the macromelanophore-determining allele produced very few macromelanophores when raised at either 25 or 28 degrees C, even after more than 200 days. In contrast, the mean percent coverage for genetically identical fish raised at 22 degrees C increased steadily throughout the course of the experiment. Production of macromelanophores was sex influenced, with greater expressivity seen in males. At 22 degrees C, the mean percent coverages had significantly diverged between males and females by the age of 201 days. From that point on, the percent macromelanophore coverage of the males was consistently significantly higher than that of the females. The tendency to produce greater melanization at cooler temperatures is not the result of a heat-sensitive tyrosinase enzyme, as is the case in mammals carrying the Himalayan allele. In mollies, the activity of tyrosinase increases between 22 and 29 degrees C. We hypothesize that production of macromelanophores is under the control of a proto-oncogene.

Chronic L-DOPA treatment increases striatal cannabinoid CB1 receptor mRNA expression in 6-hydroxydopamine-lesioned rats.

The effect of a unilateral 6-hydroxydopamine (6-OHDA) lesion of the left medial forebrain bundle and 3 weeks treatment with L-DOPA of normal and 6-OHDA lesioned rats on CB1r mRNA expression was investigated by in situ hybridization. A 6-OHDA lesion of nigrostriatal pathway alone, confirmed by the loss of nigral tyrosine hydroxylase mRNA, did not alter CB1r mRNA levels in the dopamine depleted striatum. Similarly, chronic L-DOPA treatment of normal rats had no effect on striatal CB1r mRNA expression. In contrast, chronic L-DOPA treatment of 6-OHDA-lesioned rats significantly increased CB1r mRNA expression in the denervated striatum. These results suggest that the CB1r activity may be altered by L-DOPA's action and this may be related to the treatment of Parkinson's disease.

Multiple diverticula of gall bladder.

Diverticula of the gall bladder are of no clinical significance unless associated with stones. Isolated location on Hartmann's pouch, body or neck have been explained on disordered embryogenesis but diffuse location all over the gall bladder is difficult to explain. We report a patient in whom multiple gall bladder diverticula were associated with a stone in the common bile duct.

Leiomyoblastoma of small intestine.

Gastrointestinal tumours are rare in children and leiomyoblastoma of small intestine is still rarer. We report a case of leiomyoblastoma of small intestine in a child who presented with acute intestinal obstruction.

Comparative evaluation of conservative management versus early surgical intervention in appendicular mass--a clinical study.

The management of appendicular mass seems to be taking turn with the availability of better antibiotics, care and anaesthesia. Quite a few workers turned to the other extreme of traditional conservative approach and reported acceptable results with immediate intervention during phlegmonous stage. But practically, surgeons still continue to adopt the same old regime. To alleviate this fear and circumvent the disadvantages of immediate intervention a midpath regime called early appendicectomy has been clinically evaluated. With this new approach in the present study preventing misdiagnosis (15%), no chance of recurrence as also of missing in the follow-up and shorter overall hospital stay with economy are some strong reasons to adopt this regime.