Small colony variants constitute a slow-growing subpopulation of bacteria with distinctive phenotypic and pathogenic traits. Phenotypically, small colony variants have a slow growth rate, atypical colony morphology and unusual biochemical characteristics, making them a challenge for clinical microbiologists to identify. Clinically, small colony variants are better able to persist in mammalian cells and are less susceptible to antibiotics than their wild-type counterparts, and can cause latent or recurrent infections on emergence from the protective environment of the host cell. This review covered the phenotypic, genetic and clinical picture associated with small colony variants, with an emphasis on staphylococci, for which the greatest amount of information is available.

Small Colony Variants: a Pathogenic Form of Bacteria that Facilitates Persistent and Recurrent Infections

Porcine circovirus type 2 (PCV2) infections: clinical signs, pathology and laboratory diagnosis.

Publications Financial Statement pursuant to rule 154-ter paragraph 1 of Legislative Decree No. 58/1998 Annual Report Annual Report on Form 20-F for the Securities and Exchange Commission Fact Book (in Italian and English) Eni in 2011 (in English) Interim Consolidated Report as of June 30 pursuant to rule 154-ter paragraph 2 of Legislative Decree No. 58/1998 Corporate Governance Report pursuant to rule 123-bis of Legislative Decree No. 58/1998 (in Italian and English) Remuneration Report pursuant to rule 123-ter of Legislative Decree No. 58/1998 (in Italian and English)

Annual Report 2011 -個人別教育研究報告-

Respiratory disease in pigs is common in modern pork production worldwide and is often referred to as porcine respiratory disease complex (PRDC). PRDC is polymicrobial in nature, and results from infection with various combinations of primary and secondary respiratory pathogens. As a true multifactorial disease, environmental conditions, population size, management strategies and pig-specific factors such as age and genetics also play critical roles in the outcome of PRDC. While non-infectious factors are important in the initiation and outcome of cases of PRDC, the focus of this review is on infectious factors only. There are a variety of viral and bacterial pathogens commonly associated with PRDC including porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (MHYO) and Pasteurella multocida (PMULT). The pathogenesis of viral respiratory disease is typically associated with destruction of the mucocilliary apparatus and with interference and decrease of the function of pulmonary alveolar and intravascular macrophages. Bacterial pathogens often contribute to PRDC by activation of inflammation via enhanced cytokine responses. With recent advancements in pathogen detection methods, the importance of polymicrobial disease has become more evident, and identification of interactions of pathogens and their mechanisms of disease potentiation has become a topic of great interest. For example, combined infection of pigs with typically low pathogenic organisms like PCV2 and MHYO results in severe respiratory disease. Although the body of knowledge has advanced substantially in the last 15 years, much more needs to be learned about the pathogenesis and best practices for control of swine respiratory disease outbreaks caused by concurrent infection of two or more pathogens. This review discusses the latest findings on polymicrobial respiratory disease in pigs.

Polymicrobial respiratory disease in pigs

Clinical signs and pathological features are still the corner-stones to suspect and diagnose overt disease associated with PCV2 infection. The clinico-pathological scope of this viral infection has been expanded over time. From the initial description of postweaning multisystemic wasting syndrome, some enteric, respiratory and reproductive disorders have been subsequently linked with PCV2. Porcine dermatitis and nephropathy syndrome, an immunocomplex disease, has also been associated with infection by this virus. All together, these conditions have been grouped under the name of porcine circovirus diseases (PCVD) or porcine circovirus associated diseases (PCVAD). The precise mechanisms by which a PCV2 infected pig develops a PCV2 subclinical infection or a clinical PCVD/PCVAD are still to be fully elucidated, but inferences based upon clinical, gross and histologic findings from field cases of disease have been useful to suggest the pathogenesis of this viral infection. The objective of the present review is to update the current knowledge on the clinical and pathological scope of PCV2 infections, as well as on their diagnosis. Moreover, a proposal on a unified PCVD/PCVAD terminology and clearly defined diagnostic criteria for these conditions are also given. © 2011 Elsevier B.V. All rights reserved.

Porcine circovirus type 2 (PCV2) infections: Clinical signs, pathology and laboratory diagnosis


 
 
 Does revascularization of xenotransplanted human ovarian grafts occur equally efficient from both the cortical and medullary side of the cortex tissue?
 
 
 
 Murine vessels were established equally efficient from both sides of human ovarian grafts, demonstrating that the fibrous cortical side does not hinder the neovascularization process.
 
 
 
 There is a lack of consensus regarding the surgical approach for transplanting frozen-thawed ovarian tissue and various techniques are being used clinically worldwide. Some centers attach ovarian tissue with stitches or Interceed® on top of the decorticated remaining ovary to mimic normal ovarian architecture. Other centers transplant ovarian tissue in subcortical pockets made by longitudinal or transverse incisions in the remaining intact ovary to facilitate vascularization from both sides of the graft. While other centers transplant to sub-peritoneal pelvic sites only. It is unknown which techniques and transplantation sites provide the most efficient revascularization of the ovarian grafts.
 
 
 
 Three pieces of cryopreserved ovarian cortex (approximately 5x5x1 mm in size) were donated and thawed from 12 women and two of the pieces were transplanted to subcutaneous pockets on the dorsal side of ovariectomized immunodeficient female mice. The third piece was processed as an ungrafted control. Ovarian grafts were retrieved after 8 days to analyze the spatial distribution of graft-revascularization using immunohistochemical detection of murine CD31, and after 8 weeks to evaluate follicle density (follicles/mm3).
 
 
 
 Ovarian cortex was donated from 12 women aged 27-31 years having their tissue frozen for fertility preservation. The CD31 positive vessel area and density were quantified using a custom designed application (APP) from Visiopharm®. Three regions of interest (ROIs) were defined in each tissue section; the cortical side, the center, and the medullary side. Vessels were sub-divided into three categories according to size: micro-vessels (<300 µm2), small vessels (300-1000 µm2), and large vessels (1000-3000 µm2).
 
 
 
 After 8 days xenotransplantation, a statistical significant lower density of vessels was found in the center of the human ovarian grafts compared to the cortical and medullary sides (cortical side: 323 ± 14 vessels/mm2; center: 240 ± 12 vessels/mm2; medullary side: 301 ± 18 vessels/mm2; p < 0.001). The mean percentage of CD31 positive vessel area in the human ovarian grafts was similar in the three ROIs, but lowest in the center of the grafts (cortical side: 3.9% ± 0.2 (SEM); center: 3.5% ± 0.2; medullary side: 4.0% ± 0.3; p = 0.17). Micro-vessels comprised 89-91% of all vessels in the three ROIs, demonstrating that the vast majority of vessels were newly formed. Heatmaps were generated for each section based on the CD31 positive area to visualize the localization of vessels in the ovarian grafts and the maps showed highly variable vascularization patterns in the grafts from different patients. Overall, vascularization appeared to occur equally efficient in all peripherical areas of the ovarian grafts. The follicle density in ungrafted cortex tissue was 51.8 ± 17.3 follicles/mm3 and 14.7 ± 3.7 follicles/mm3 after 8 weeks xenografting, resulting in an average follicle survival rate of 28%.
 
 
 
 The current study was not designed to directly compare clinically used transplantation techniques, which should be studied in larger animal models, like sheep, in which the structure of ovarian tissue is similar to human.
 
 
 
 Revascularization was established equally efficient from both sides of xenotransplanted human ovarian cortex, suggesting that transplantation techniques ensuring revascularization from both sides of the ovarian graft, such as sub-cortical or peritoneal pockets, may facilitate a more efficient revascularization of the graft than techniques allowing revascularization from only the medullary side.
 
 
 
 not applicable


O-037 Revascularization of human ovarian grafts occurs equally efficient from both sides of the cortex tissue: implications for clinical practice

Differential hepatoprotective effects of semaglutide and lanifibranor in the GAN diet-induced obese and biopsy-confirmed mouse model of NASH with advanced fibrosis and HCC

Polycystic ovaries (PCO) contain antral follicles that arrest growing around 3–11 mm in diameter, perturbing the dominant follicle’s selection and the subsequent ovulatory process. Proteomic alterations of PCO follicular fluid (FF) (i.e., microenvironment in which the oocyte develops until ovulation) have been studied from large follicles in connection with oocyte pickup during ovarian stimulation. The present study aimed to detect proteomic alterations in FF from unstimulated human small antral follicles (hSAF) obtained from PCO. After performing deep-sequencing label-free proteomics on 10 PCO and 10 non-PCO FF samples from unstimulated hSAF (4.6–9.8 mm), 1436 proteins were identified, of which 115 were dysregulated in PCO FF samples. Pathways and processes related to the immune system, inflammation, and oxidative stress appeared to be upregulated in PCO, while extracellular matrix receptors interactions, the collagens-containing extracellular matrix, and the regulation of signaling were downregulated. The secreted proteins SFRP1, THBS4, and C1QC significantly decreased their expression in PCO FF, and this downregulation was suggested to affect future oocyte competence. In conclusion, our study revealed, for the first time, evidence of proteomic alterations occurring in the FF of PCO hSAF that may be related to the dysfunction of follicular growth and subsequent oocyte competence.

/pdf/proteomic-alterations-in-follicular-fluid-of-human-small-3w0hz37p.pdf

Proteomic Alterations in Follicular Fluid of Human Small Antral Follicles Collected from Polycystic Ovaries—A Pilot Study


 
 
 Can mechanical injury by needle puncturing of frozen thawed human ovarian cortex tissue prior to transplantation increase revascularization of xenografts and improve follicle survival?
 
 
 
 Mechanical injury caused by needle puncture prior to transplantation improved follicle survival 2-fold in human ovarian cortex tissue after 4 weeks xenografting.
 
 
 
 Cryopreservation and autotransplantation of ovarian tissue is an established fertility preservation method which has resulted in numerous live births worldwide. However, the efficiency of the method is hampered by a critical follicle loss after transplantation. Faster revascularization of the transplanted graft would shorten the ischemic period and could potentially improve follicle survival. Controlled tissue damage by laser or needles can induce local angiogenesis by initiating the wound healing mechanisms and has been applied clinically in cardiovascular surgery for decades. Whether controlled mechanical tissue damage could be used to improve grafting of human ovarian cortex tissue has not yet been investigated.
 
 
 
 A total of 32 frozen-thawed human ovarian cortex pieces were cut in halves; one half serving as the untreated control and the other half was punctured approximately 200 times with a 29G needle immediately after thawing. The cortical pieces were transplanted subcutaneously to 10 immunodeficient mice. Ovarian grafts were retrieved after 3, 6 and 10 days (n = 8) to analyse graft revascularization and gene expression, and after 4 weeks to assess follicle density (n = 8).
 
 
 
 Ovarian cortex was donated by 16 women (aged 24-36 years) undergoing ovarian tissue cryopreservation for fertility preservation at a University Hospital. Gene expression analysis of proinflammatory cytokines (Tnf-α), apoptotic factors (Bcl2 and Bax), and angiogenic factors (Vegfa, Angptl4, Ang1, and Ang2) were performed by qPCR. Follicle density after 4 weeks xenografting were evaluated histologically. Graft-revascularization was assessed using immunohistochemical detection of murine Cd31.
 
 
 
 Our results showed that the ovarian grafts which had been punctured with needles prior to transplantation contained a larger number of surviving follicles per mm3 compared to the untreated control (control group: 2.9 ± 0.6 (mean±SEM) follicles/mm3; needle group: 6.4 ± 2.4 follicles/mm3). Follicle densities between patients and groups varied from 0.8 to 26 follicles/mm3. Overall, a 2.3-fold increase in follicle density was found in the needle group compared to control, though this finding was not significantly different (p-value = 0.1182). The relative gene expression of Tnf-α and the Bax/Bcl2 ratio after 6 days grafting was similar in the needle and control group, indicating that the mechanical injury did not induce an additional inflammatory or apoptotic response in the grafts. The relative gene expression of Vegf, Angptl4, and Ang2 after 6 days grafting was highest in the needle group compared to the control group, though not significant (p>0.1). Results of the gene expression analysis from Day 3 and 10 are awaited, and so are the quantification of the Cd31 positive vessel area and vascular density from Day 3, 6, and 10, which will provide valuable insight into the initial graft revascularization.
 
 
 
 The ovarian follicles are highly heterogeneously distributed in the ovarian cortex and can vary 100-fold between cortex pieces from the same woman and between patients. A high variability in follicle density within and between treatment groups and patients was found in the current study. Thus, solid conclusions cannot be made.
 
 
 
 Our findings suggest a positive effect of mechanical injury on human ovarian grafts as needle puncturing prior to transplantation improved follicle survival after xenotransplantation. Clinically, this technique would be easy to implement since it is performed during the thawing process and does not involve any additional treatment of the patient.
 
 
 
 Not applicable


P-450 Mechanical injury improves follicle survival in xenotransplanted human ovarian cortex


 
 
 Does eight weeks of low dose hCG priming improve the outcome of ovarian stimulation for IVF/ICSI in women with low ovarian reserve.
 
 
 
 Significantly more oocytes were retrieved following eight weeks of low dose hCG priming, when compared to an identical IVF/ICSI treatment immediately before the priming period.
 
 
 
 Administration of androgens prior to ovarian stimulation for IVF might upregulate the FSH receptor level on granulosa cells making the follicles more responsive to exogenous FSH. LH and hCG stimulate the follicular androgen synthesis, acting through LH receptors on the theca cells, enabling their use as an intra-ovarian androgen priming method. Exogenous testosterone and DHEA priming have been shown to increase the number of retrieved oocytes and the live birth rate, but systemic androgen administration is associated with side effects. Long term ‘intra-ovarian’ priming with hCG without systemic effects in women with low ovarian reserve has not previously been investigated.
 
 
 
 A prospective, paired, non-blinded study including 20 women serving as their own controls conducted between January 2021 and July 2021 at The University Hospital Copenhagen Rigshospitalet, Denmark. Participants underwent two identical consecutive IVF/ICSI treatments, a Control cycle including elective freezing of all blastocysts and a Study cycle with fresh blastocyst transfer. The Control and Study cycle were separated by approximately eight weeks (two menstrual cycles) of androgen priming by daily injections of 260 IE hCG.
 
 
 
 Inclusion criteria were age 18-40 years, regular menstrual cycle 23-35 days and AMH <6.29 pmol/L. Control and Study IVF/ICSI cycles were performed in a fixed GnRH-antagonist protocol using a daily dose of 300 IU rFSH and GnRH antagonist 0.25 mg from stimulation day 5-6. Primary endpoint was the follicular output rate defined as the pre-ovulatory follicle count (>16 mm) on hCG trigger day divided by the antral follicle count (2-10 mm) on cycle day 2-3.
 
 
 
 20 women with a mean (SD) age of 36.8 (3.2) years and an AMH level of 3.7 (1.5) pmol/L were included and completed the study. The number of oocytes retrieved was significantly higher in the Study compared to Control Cycle: 4.7 (2.8) vs. 3.2 (1.7), (P = 0.007), and the number of blastocysts was 1.2 (1.6) vs. 0.6 (0.7) in the Study compared to Control cycle, (P = 0.11). The follicular output rate was 0.4 (0.4) in the Study cycle initiated immediately after eight weeks of hCG priming, which was similar to the follicular output rate of 0.38 (0.2) in the Control cycle performed just before initiating the priming period (P = 0.81). The number of follicles above 10 mm on trigger day was 7.3 (5.0) in the Study and 5.3 (2.9) in the Control cycle (P = 0.018). Although the duration of stimulation seemed longer in the Study compared to Control cycle, 10.1 (1.5) and 8.9 (2.7) days respectively (P = 0.05), the same trigger criterion was defined and used in the two cycles confirmed by a similar mean diameter of the two biggest follicles on trigger day: 18.5 (1.1) vs. 18.3 (1.1) mm in the Study and Control cycle, respectively (P = 0.91).
 
 
 
 The sample size calculation was based on the follicular output rate, and the study was not powered to look at oocyte retrieval rates. Bigger studies are needed to confirm this finding.
 
 
 
 Although no increase in the follicular output rate was demonstrated, significantly more oocytes were retrieved after eight weeks of hCG priming, which is an important finding in a population of women with low ovarian reserve. Benefits of long term low dose hCG priming must be confirmed in a larger study.
 
 
 
 NCT04643925


Susanne Elisabeth Pors

Papers

O-037 Revascularization of human ovarian grafts occurs equally efficient from both sides of the cortex tissue: implications for clinical practice

Differential hepatoprotective effects of semaglutide and lanifibranor in the GAN diet-induced obese and biopsy-confirmed mouse model of NASH with advanced fibrosis and HCC

Proteomic Alterations in Follicular Fluid of Human Small Antral Follicles Collected from Polycystic Ovaries—A Pilot Study

P-450 Mechanical injury improves follicle survival in xenotransplanted human ovarian cortex

P-626 Eight weeks of androgen priming by low dose hCG before IVF/ICSI in women with low ovarian reserve