Renal tumor biopsy does not increase the risk of surgical complications of minimally invasive partial nephrectomy.

BACKGROUND: For patients with cT1 renal lesions, Partial Nephrectomy (PN) is the gold standard treatment. However, 20% of small renal masses are benign, situation in which the PN is an overtreatment. The percutaneous Renal Tumor Biopsy (RTB) may lower the risk of overtreatment as there is a 90% concordance rate on histotype between the RTB and the final pathology. It has been suggested that the RTB could increase the difficulty of the PN and increase the risk of surgical complications. OBJECTIVE: To compare surgical outcomes and complications of PN with or without previous RTB. DESIGN, SETTING, AND PARTICIPANTS: monocentric retrospective review of patients who underwent laparoscopic or robotic-assisted PN between January 2012 and December 2019. MEASUREMENTS: perioperative complications were recorded using Clavien-Dindo classification, peroperative data included operative time, clamping time and blood loss, and histological outcomes of RTB and PN. RESULTS AND LIMITATIONS: In total, 163 patients were included in our study. There were significantly less benign lesions in PN with prior RTB: 7% (4/56) vs. 20% (22/107) without prior RTB (P=0.03). There were no significant differences regarding Clavien-Dindo>2 perioperative complications with respectively 7% (4/56) vs. 10% (11/107) (P=0.57). Same goes for peroperative data such as duration of surgery (P=0.81), warm ischemia (P=0.07) and blood loss (P=0.13). CONCLUSIONS: RTB does not increase the risk of surgical complications of PN and may reduce the risk of small renal masses overtreatment.

[Intravesical adjuvant regimen of epitubicin for intermediate risk NMIBC: Feasability study from CC-AFU vessie]. / Instauration du traitement adjuvant endovésical par épirubicine des tumeurs de vessie n'infiltrant pas le muscle : premier retour national d'expérience du CC-AFU vessie.

INTRODUCTION: Mitomycin C is the gold standard intravesical adjuvant therapy for intermediate-risk non-muscle-invasive bladder cancer (NMIBC). Tensions in the supply of mitomycin have emerged in France since late 2019. The ANSM in agreement with the AFU proposed to use epirubicin, already available in other European countries in this indication. The objective of our study was to report the initial French experience with the use of epirubicin in adjuvant treatment of NMIBC. MATERIALS AND METHODS: We undertook a French multicenter retrospective descriptive study to collect, from the centers of the members of the CC-AFU bladder, the clinico-pathological data of the patients, the indications, the modalities of use (dose, indication, circuit in the pharmacy) and the tolerance data of epirubicin. The impact of the COVID-19 epidemic on treatment interruptions was also identified. Of the 20 centers contacted, 5 (25%) had implemented the epirubicin administration protocol developed by the CC-AFU bladder subcommittee. A total of 61 patients were treated with endovesical instillations of epirubicin between November 2019 and November 2020 for NMIBC at a single dose of 50mg. RESULTS: A total of 61 patients (mean age 67 years, 64-77 years) were treated with epirubicin, of which 45 (73.8%) were male. The patients had intermediate-risk NMIBC in 88.5%, the rest had high-risk disease. Induction therapy without or with maintenance was planned for 48 (78.7%) and 13 patients (21.3%), respectively. The preparation and administration of epirubicin was similar to that of mitomycin: central pharmacy preparation for same-day dispensing with immediate outpatient instillation. Unlike mitomycin, urinary alkalinization was not required. Of the 498 total instillations scheduled, 345 were performed (69.3%). The COVID-19 epidemic significantly impacted epirubicin delivery: one patient could not start treatment (1.6%), 8 patients (13.1%) had to discontinue it permanently; the rest of the patients underwent delayed instillations (18%). Other causes of discontinuation included infectious complications (9.8%). No major toxicities were reported. CONCLUSION: The implementation of an adjuvant epirubicin treatment protocol presented a good feasibility with low toxicity, without modifying the organization of the patients' care pathway. In the context of unpredictable mitomycin shortage, epirubicin represents a good therapeutic alternative in the endovesical adjuvant treatment of intermediate-risk NMIBC. LEVEL OF PROOF: 3.

Invasion and persistent intracellular colonization of erythrocytes. A unique parasitic strategy of the emerging pathogen Bartonella.

The expanding genus Bartonella includes zoonotic and human-specific pathogens that can cause a wide range of clinical manifestations. A productive infection allowing bacterial transmission by blood-sucking arthropods is marked by an intraerythrocytic bacteremia that occurs exclusively in specific human or animal reservoir hosts. Incidental human infection by animal-adapted bartonellae can cause disease without evidence for erythrocyte parasitism. A better understanding of the intraerythrocytic lifestyle of bartonellae may permit the design of strategies to control the reservoir and transmittable stages of these emerging pathogens. We have dissected the process of Bartonella erythrocyte parasitism in experimentally infected animals using a novel approach for tracking blood infections based on flow cytometric quantification of green fluorescent protein-expressing bacteria during their interaction with in vivo-biotinylated erythrocytes. Bacteremia onset occurs several days after inoculation by a synchronous wave of bacterial invasion into mature erythrocytes. Intracellular bacteria replicate until reaching a stagnant number, which is sustained for the remaining life span of the infected erythrocyte. The initial wave of erythrocyte infection is followed by reinfection waves occurring at intervals of several days. Our findings unravel a unique bacterial persistence strategy adapted to a nonhemolytic intracellular colonization of erythrocytes that preserves the pathogen for efficient transmission by blood-sucking arthropods.

Turning placenta into brain: placental mesenchymal stem cells differentiate into neurons and oligodendrocytes.

OBJECTIVE: We aimed to induce neural stem (NSC) and progenitor cells (NPC) from human placental tissues. STUDY DESIGN: Placental stem cells from first-trimester placental chorionic villi and term chorion were isolated. Neural differentiation was initiated with plating on collagen, retinoic acid, and/or human brain-derived neurotrophic factor and epidermal and fibroblast growth factor. Differentiation into neurons, oligodendrocytes, and astrocytes was monitored by immunohistochemistry. Two-dimensional polyacrylamide gel electrophoresis, high-performance liquid chromatography, and tandem mass spectrometry were used to identify proteins involved in the differentiation. RESULTS: Differentiated cells were mostly immediately postmitotic with some more but not fully mature postmitotic neurons. Neurons had dopaminergic or serotonergic character. Some cells differentiated into predominantly immature oligodendrocytes. Upon differentiation, neuron-specific proteins were up-regulated, whereas placental proteins were reduced. CONCLUSION: Stem cells derived from human placenta can be differentiated into neural progenitors.

Neurogenic characteristics of placental stem cells in preeclampsia.

OBJECTIVE: Preeclampsia is associated with perinatal brain injury. Autologous placenta stem cell transplantation represents a promising future treatment option for neuroregeneration. The aim of this study was to compare the neuroregenerative capacity of preeclampsia-placenta stem cells to previously characterized placentas from uncomplicated pregnancies. STUDY DESIGN: Placenta stem cells from amnion (epithelium, mesenchyme) and chorion were assessed for cell surface markers and the formation of neuronal-like cells, oligodendrocytes and their progenitors in culture. RESULTS: Markers of preeclampsia-placenta stem cells were different from uncomplicated pregnancies-placenta stem cells in amnion epithelium and chorion, but not in amnion mesenchyme. Similarly to uncomplicated pregnancies-placenta stem cells, preeclampsia-placenta stem cells derived from amnion and chorion differentiated preferably into nestin-positive stem/progenitor cells and Tuj-1-positive neurons. However, other important markers were varying after neurogenic differentiation of uncomplicated pregnancies- and preeclampsia-placenta stem cells. CONCLUSION: Surface marker expression patterns of preeclampsia-placenta stem cell's and uncomplicated pregnancies-placenta stem cell's differ. In vitro differentiation assays, however, provide evidence that both preeclampsia-placenta stem cells and uncomplicated pregnancies-placenta stem cells are comparably suitable for neuroregeneration purposes.

Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells.

Placental tissue draws great interest as a source of cells for regenerative medicine because of the phenotypic plasticity of many of the cell types isolated from this tissue. Furthermore, placenta, which is involved in maintaining fetal tolerance, contains cells that display immunomodulatory properties. These two features could prove useful for future cell therapy-based clinical applications. Placental tissue is readily available and easily procured without invasive procedures, and its use does not elicit ethical debate. Numerous reports describing stem cells from different parts of the placenta, using nearly as numerous isolation and characterization procedures, have been published. Considering the complexity of the placenta, an urgent need exists to define, as clearly as possible, the region of origin and methods of isolation of cells derived from this tissue. On March 23-24, 2007, the first international Workshop on Placenta Derived Stem Cells was held in Brescia, Italy. Most of the research published in this area focuses on mesenchymal stromal cells isolated from various parts of the placenta or epithelial cells isolated from amniotic membrane. The aim of this review is to summarize and provide the state of the art of research in this field, addressing aspects such as cell isolation protocols and characteristics of these cells, as well as providing preliminary indications of the possibilities for use of these cells in future clinical applications.

Manufacture of a cell-free amnion matrix scaffold that supports amnion cell outgrowth in vitro.

We manufactured a cell-free extracellular matrix scaffolds in order to obtain a support material for amnion cell outgrowth, eventually being used for repair of prematurely ruptured fetal membrane. Human preterm or term amnion tissue was separated into its collagenous extracellular matrix and cell components. The acellular scaffold was explored for its capacity to support regrowth of isolated human amnion epithelial or mesenchymal cells in vitro. The outgrowth of amnion cells on and in the scaffold was investigated by scanning and transmission electron microscopy, and confocal laser scanning microscopy. Cell-free amnion matrix scaffolds demonstrated a porous collagen fiber network similar as in native amnion. Inoculation of acellular amnion scaffolds with human amnion cells revealed that its property to support amnion cell outgrowth was retained. Amnion epithelial and mesenchymal cells were found to grow into dense layers on the surface of the scaffold within 3-4 days and 7-8 days, respectively, and to some extent, invaded the scaffold during the culture period. Manufactured acellular amnion matrix retains structural and functional properties required for cell outgrowth in vitro. It may become useful to repair prematurely ruptured fetal membranes.

Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration.

OBJECTIVE: Mesenchymal stem cells (MSCs) have a broad differentiation potential. We aimed to determine if MSCs are present in fetal membranes and placental tissue and to assess their potential to differentiate into neurogenic and mesodermal lineages. STUDY DESIGN: MSCs isolated from first and third trimester chorion and amnion and first trimester chorionic villi and characterized morphologically and by flourescence-activated cell sorting analysis. Their ability to mature under different culture conditions into various cells of mesodermal and neuroectodermal cell lines was assessed by immuno- and cytochemical staining. RESULTS: Independent of gestational age, cells isolated from fetal membranes and placenta showed typical MSC phenotype (positive for CD166, CD105, CD90, CD73, CD49e, CD44, CD29, CD13, MHC I; negative for CD14, CD34, CD45, MHC II) and were able to differentiate into mesodermal cells expressing cell markers/cytologic staining consistent with mature chondroblasts, osteoblasts, adipocytes, or myocytes and into neuronal cells presenting markers of various stages of maturation. The differentiation pattern was mainly dependent on cell type. CONCLUSION: Mesenchymal cells from chorion, amnion, and villous stroma can be differentiated into neurogenic, chondrogenic, osteogenic, adipogenic, and myogenic lineage. Placental tissue obtained during prenatal chorionic villous sampling or at delivery might be an ideal source for autologous stem cell graft for peripartum neuroregeneration and other clinical issues.

Differential influence of maternal and fetal pregnancy factors on the in-vitro induction of human regulatory T cells: a preliminary study.

PROBLEM: Given the important role of regulatory T cells (Treg) for successful pregnancy, the ability of soluble maternal and fetal pregnancy factors to induce human Treg was investigated. METHOD OF STUDY: Peripheral blood mononuclear cells (PBMCs) or isolated CD4+CD25‒ cells were cultured in the presence of pooled second or third trimester pregnancy sera, steroid hormones or supernatants from placental explants, and the numbers and function of induced CD4+CD25+FOXP3+ Treg were analysed. RESULTS: Third trimester pregnancy sera and supernatants of early placental explants, but not sex steroid hormones, induced an increase of Tregs from PBMCs. Early placental supernatant containing high levels of tumour necrosis factor-α, interferon-γ, interleukins -1, -6 and -17, soluble human leucocyte antigen-G, and transforming growth factor-ß1, increased the proportion of Treg most effectively and was able to induce interleukin-10-secreting-Treg from CD4+CD25‒cells. CONCLUSIONS: Compared with circulating maternal factors, placental- and fetal-derived factors appear to exert a more powerful effect on numerical changes of Treg, thereby supporting fetomaternal tolerance during human pregnancy.

Construction of versatile high-level expression vectors for Bartonella henselae and the use of green fluorescent protein as a new expression marker.

Expression vectors suitable for directing high levels of protein synthesis in Bartonella henselae have been constructed based on the mobilisable broad-host-range (IncQ) plasmidpMMB206. They confer kanamycin resistance and feature the taclac (tac-lacUV5 in tandem) promoters in front of a polylinker followed by the rrnB transcriptional stop point. While expression of genes fused to the taclac promoter is constitutive in one vector, the lacIq gene carried by the other vector allows a controlled, IPTG-inducible gene expression. These vectors were tested by subcloning a mutated gfp gene coding for the green fluorescent protein (GFP) from Aequorea victoria into the multiple cloning site and introducing the resulting plasmids into Escherichia coli and B. henselae. GFP expression was determined by measuring fluorescence via flow cytometry or directly by immunoblotting. Compared to E. coli, expression of GFP in B. henselae was more tightly controlled by lacIq and resulted in much higher levels of both IPTG-induced and constitutive gene expression. In vitro infection of endothelial cells indicated that GFP expression does not adversely affect the interaction of B. henselae with host cells. These data demonstrate that (i) the established expression vectors are useful for directing controlled or constitutive high-level protein synthesis in B. henselae and (ii) that GFP is a valuable expression marker which may has important applications in studying the bacterial genetics and cellular interactions of this emerging human pathogen.

Cardiac vein thrombosis and haemorrhagic myocardial necrosis; report of a case with review of the literature.

A 29-year-old woman, addicted to heroin since the age of 15 years, presented with a 4-day history of acute inspiratory chest pain, dyspnoea and vomiting associated with hypoventilation. She died 3 h after admission to the intensive care unit in spite of active resuscitative measures. The main autopsy findings were limited to the heart, which showed widespread cardiac vein thrombosis, and both ventricles and the atria were associated with multiple areas of haemorrhagic myocardial necrosis. We review the literature of this uncommon pathological entity and discuss its possible pathogenesis.

Isolation of a gene encoding cysteine synthase from Flavobacterium K3-15.

The cysteine synthase gene (cysK) from Flavobacterium K3-15 was cloned and sequenced. The gene exhibits 30-50% identity to known cysteine synthases on both the DNA and the amino acid levels. The pyridoxal phosphate binding site of the enzyme is part of a conserved motif comprising seven amino acids (SIKDRIA). The lys31 residue of the flavobacterial enzyme is conserved in all known cysteine synthases. The cysK gene from Flavobacterium K3-15 was heterologously expressed and the gene product identified by immunoblotting and determination of the enzyme activity.

[Immediate-acting oral morphine sulfate in patients with cancer pain]. / Sulfato de morfina oral de liberación inmediata en pacientes con dolor por cáncer.

In this article, we describe our experience with oral morphine for the treatment of patients with cancer pain who were referred to a Palliative Care Unit. Morphine was used in 3,399 days treatment with an average of 65 days per patient. In 50 patients (96%), there was a decreased intensity of pain from severe (Visual Analogue Scale average (VAS) 8.7) to mild (VAS average 1.9). Daily average dose was 95.6 mg. Only 26 patients (50%) required an increase in dosage. The side effects were minimal and controllable. Even though most authors recommend morphine q4h, in our population q6h was enough to achieve pain control in 83% of patients. It should be known by the medical population that morphine is essential therapy for pain in cancer patients. This study confirms that morphine is an excellent drug for the control of pain in cancer patients, with minimal dosages and controllable side effects.

Generation of an osteogenic graft from human placenta and placenta-derived mesenchymal stem cells.

The objective of the study was to determine the feasibility of generating a biodegradable, stem cell-loaded osteogenic composite graft from human placenta. Initially, a scaffold from human chorion membrane was produced. Human placenta mesenchymal stem cells (MSCs) derived from either first-trimester chorionic villi or term chorion membrane were differentiated osteogenically on this scaffold. Outgrowth, adherence, and osteogenic differentiation of cells were assessed by immunohistochemistry (IHC), scanning electron microscopy, protein expression, and real-time polymerase chain reaction (RT-PCR). Our results showed that a cell-free extracellular matrix scaffold can be generated from human chorion. Seeded MSCs densely adhered to that scaffold and were osteogenically differentiated. Calcium and alkaline phosphatase were detected in the cell-scaffold constructs as a proof of mineralization and findings were confirmed by IHC and RT-PCR results. This study shows for the first time that generation of an osteogenic composite graft using placental tissue is feasible. It might allow therapeutic application of autologous or allogeneic grafts in congenital skeletal defects by means of a composite graft.

Essential sulfhydryl groups in the catalytic center of the tonoplast H(+)-ATPase from coleoptiles ofZea mays L. as demonstrated by the biotin-streptavidin-peroxidase system.

Functional properties and the localization of essential SH-groups of the tonoplast H(+)-ATPase fromZea mays L. were studied. In contrast to the pyrophosphate-dependent H(+)-translocation activity of the tonoplast, the H(+)-ATPase activity was inhibited by SH-blocking agents, such as N-ethylmaleimide and iodoacetic acid. In the case ofp-hydroxymercuribenzoate, HgCl2 and oxidized glutathione, the inhibition could be reversed by adding reduced glutathione or dithiothreitol.Incubation of tonoplast vesicles with oxidized glutathione or N-ethylmaleimide in the presence of Mg·ADP-a competitive inhibitor of the ATP-dependent H(+) pump-avoided the inhibition of the H(+)-pumping activity. This effect is an indication for the occurrence of essential SH-groups at the catalytic site of the H(+)-ATPase.In order to characterize the active center these thiols were specifically labeled with maleimidobutyrylbiocytin. Subsequently, the membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to an immobilizing membrane. The maleimidobutyrylbiocytin-labeled active-center protein was detected by a biotin-streptavidin-peroxidase staining system and was shown to be a 70-kDa subunit of the tonoplast H(+)-ATPase. It is suggested that the oxidation state of the critical sulfhydryl groups within the active center of the enzyme and their reversible blocking by endogenous compounds might be of great importance for the regulation of the enzyme activity in vivo.

[Development of formerly preterm infants with a birth weight of below 1500 grams: concept and results of a follow-up program in up to school-aged children in central Switzerland]. / Entwicklung ehemaliger Frühgeborener mit einem Geburtsgewicht unter 1500 Gramm: Konzept und Resultate von Nachkontrollen bis zum Schulalter in der Zentralschweiz.

AIM: To document the psychomotor development and general health of former very low birthweight infants born between 1980 and 1986 from birth up to school age. We wished to evaluate the quality of neonatal intensive care in Central Switzerland over this time period and test the reliability of a patient-oriented follow-up programme. If successful, the latter could perhaps serve as a model for a national follow-up programme in Switzerland. METHODS: Information regarding three different developmental periods was collected. The medical records of the perinatal period were used to abstract details of labour and delivery and the neonatal period. The records of the infant follow-up programme were used to describe psychomotor development between 0 and 24 months of age. The current health status and school performance were evaluated using a questionnaire sent to parents and teachers. RESULTS: Of 139 infants born with a birthweight of < or = 1500 g, 102 were discharged home (mortality rate 26.6%). One third was not screened for hearing deficits or retinopathy of prematurity. Eighty-two were seen in the infant follow-up programme between 0 and 24 months of age. Seventy-seven percent of these infants were judged to be normal and discharged from the infant follow-up programme; 1/5 of these infants had had transient motor problems treated by physical therapy. Twenty-three percent of the infants seen in infant follow-up had persistent but mainly minor motor handicaps, and only two infants (2%) had multiple handicaps. At school age, data from 99 of the 102 surviving infants was collected. Ninety-six percent attended regular school, but almost half of them had significant school problems and required professional help. These problems correlated poorly with the results of examinations during early childhood (positive predictive value 67%). CONCLUSIONS: These long-term results of a population of preterm infants born in Central Switzerland in the 1980s are encouraging. To ensure completeness of early ophthalmological and audiological examinations of all former small preterm infants, neonatal follow-up programmes should adhere to uniform guidelines.

Characterization of Bartonella clarridgeiae flagellin (FlaA) and detection of antiflagellin antibodies in patients with lymphadenopathy.

Cat scratch disease (CSD) is a frequent clinical outcome of Bartonella henselae infection in humans. Recently, two case reports indicated Bartonella clarridgeiae as an additional causative agent of CSD. Both pathogens have been isolated from domestic cats, which are considered to be their natural reservoir. B. clarridgeiae and B. henselae can be distinguished phenotypically by the presence or absence of flagella, respectively. Separation of the protein content of purified flagella of B. clarridgeiae by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis indicated that the flagellar filament is mainly composed of a polypeptide with a mass of 41 kDa. N-terminal sequencing of 20 amino acids of this protein revealed a perfect match to the N-terminal sequence of flagellin (FlaA) as deduced from the sequence of the flaA gene cloned from B. clarridgeiae. The flagellin of B. clarridgeiae is closely related to flagellins of Bartonella bacilliformis and several Bartonella-related bacteria. Since flagellar proteins are often immunodominant antigens, we investigated whether antibodies specific for the FlaA protein of B. clarridgeiae are found in patients with CSD or lymphadenopathy. Immunoblotting with 724 sera of patients suffering from lymphadenopathy and 100 healthy controls indicated specific FlaA antibodies in 3.9% of the patients' sera but in none of the controls. B. clarridgeiae FlaA is thus antigenic and expressed in vivo, providing a valuable tool for serological testing. Our results further indicate that B. clarridgeiae might be a possible etiologic agent of CSD or lymphadenopathy. However, it remains to be clarified whether antibodies to the FlaA protein of B. clarridgeiae are a useful indicator of acute infection.

Range of global motion of the cervical spine: intraindividual reliability and the influence of measurement device.

Range of motion tests are often employed in the quantification of musculoskeletal impairment and in the assessment of the efficacy of therapeutic interventions. The aim of the present study was to compare the absolute values for, and the day-to-day reliability of, measures of cervical spinal mobility made with two computerised motion analysis devices. The ranges of cervical flexion, extension, lateral bending, axial rotation, and axial rotation in flexion and extension were determined for 19 volunteers using both the CA6000 Spine Motion Analyser and the Zebris CMS system; all measures were repeated on a second occasion 1-3 days later. The test-retest reliability was good for each instrument: there was no significant difference between the mean values derived on the two separate days (P>0.05), and the corresponding intraclass correlation coefficients were 0.75-0.93 for all pri-O mary movements and 0.57-0.93 for axial rotation in flexion or in extension. For each primary movement, a small but significant difference (1-10%; P<0.05) between the values derived from the two instruments was observed, the systematic nature of which was revealed by the excellent correlation coefficients between them. For the measures of axial rotation in flexion or in extension, however, there was not only a poor correlation between the data obtained from the two devices, but the mean values also differed significantly. Each device is highly reliable in itself and can be used with confidence in longitudinal studies. The establishment of 'normal' values for the primary motions should take account of the slight differences observed between devices. Normal values for rotation in flexion or extension cannot be established until the source of the device-dependent difference is identified.