Bacterial safety of cell-based therapeutic preparations, focusing on haematopoietic progenitor cells.

Bacterial safety of cellular preparations, especially haematopoietic progenitor cells (HPCs), as well as advanced therapy medicinal products (ATMPs) derived from stem cells of various origins, present a challenge for physicians, manufacturers and regulators. The article describes the background and practical issues in this area and illustrates why sterility of these products cannot currently be guaranteed. Advantages and limitations of approaches both for classical sterility testing and for microbiological control using automated culture systems are discussed. The review considers novel approaches for growth-based rapid microbiological control with high sensitivity and faster availability of results, as well as new methods for rapid bacterial detection in cellular preparations enabling meaningful information about product contamination within one to two hours. Generally, however, these direct rapid methods are less sensitive and have greater sampling error compared with the growth-based methods. Opportunities for pyrogen testing of cell therapeutics are also discussed. There is an urgent need for development of novel principles and methods applicable to bacterial safety of cellular therapeutics. We also need a major shift in approach from the traditional view of sterility evaluation (identify anything and everything) to a new thinking about how to find what is clinically relevant within the time frame available for the special clinical circumstances in which these products are used. The review concludes with recommendations for optimization of microbiological control of cellular preparations, focusing on HPCs.

Establishment of the first international repository for transfusion-relevant bacteria reference strains: ISBT working party transfusion-transmitted infectious diseases (WP-TTID), subgroup on bacteria.

BACKGROUND: Bacterial contamination of platelet concentrates (PCs) still remains a significant problem in transfusion with potential important clinical consequences, including death. The International Society of Blood Transfusion Working Party on Transfusion-Transmitted Infectious Diseases, Subgroup on Bacteria, organised an international study on Transfusion-Relevant Bacteria References to be used as a tool for development, validation and comparison of both bacterial screening and pathogen reduction methods. MATERIAL AND METHODS: Four Bacteria References (Staphylococcus epidermidis PEI-B-06, Streptococcus pyogenes PEI-B-20, Klebsiella pneumoniae PEI-B-08 and Escherichia coli PEI-B-19) were selected regarding their ability to proliferate to high counts in PCs and distributed anonymised to 14 laboratories in 10 countries for identification, enumeration and bacterial proliferation in PCs after low spiking (0·3 and 0·03 CFU/ml), to simulate contamination occurring during blood donation. RESULTS: Bacteria References were correctly identified in 98% of all 52 identifications. S. pyogenes and E. coli grew in PCs in 11 out of 12 laboratories, and K. pneumoniae and S. epidermidis replicated in all participating laboratories. The results of bacterial counts were very consistent between laboratories: the 95% confidence intervals were for S. epidermidis: 1·19-1·32 × 10(7) CFU/ml, S. pyogenes: 0·58-0·69 × 10(7) CFU/ml, K. pneumoniae: 18·71-20·26 × 10(7) CFU/ml and E. coli: 1·78-2·10 × 10(7) CFU/ml. CONCLUSION: The study was undertaken as a proof of principle with the aim to demonstrate (i) the quality, stability and suitability of the bacterial strains for low-titre spiking of blood components, (ii) the property of donor-independent proliferation in PCs, and (iii) their suitability for worldwide shipping of deep frozen, blinded pathogenic bacteria. These aims were successfully fulfilled. The WHO Expert Committee Biological Standardisation has approved the adoption of these four bacteria strains as the first Repository for Transfusion-Relevant Bacteria Reference Strains and, additionally, endorsed as a project the addition of six further bacteria strain preparations suitable for control of platelet contamination as the next step of enlargement of the repository.

Neonatal ovarian cyst: prenatal diagnosis and analysis of the cyst fluid.

Ovarian cysts of clinical significance are uncommon in the fetus and neonate. The authors diagnosed a large fetal cyst prenatally by ultrasound scan and removed it in the neonatal period. Analysis of the fluid obtained from the cyst demonstrated a capacity of the fetal ovary to synthesize estrogen and progesterone.

Sarcoid-like lesions associated with epithelial ovarian adenocarcinoma.

A woman with poorly differentiated endometrioid adenocarcinoma of the ovary had sarcoid-like lesions in the bone marrow. Sarcoid-like lesions may be misinterpreted as metastatic disease, resulting in inappropriate modification of therapy.

The clinical significance of blood transfusion at the time of radical hysterectomy.

A retrospective review was conducted to evaluate the clinical implications of perioperative blood transfusion in the surgical management of stage IB cervical cancer. The investigation focused on 126 patients treated with radical hysterectomy and retroperitoneal lymph node dissection who were found to have clear surgical margins, negative retroperitoneal lymph nodes, and no lymph-vascular space involvement in the hysterectomy specimen, and who had no perioperative radiation therapy, no history of immunosuppression with medication, and at least 18 months of follow-up. The distributions of age, weight, operative time, nodal yields, mean lesion diameters, median depths of invasion, and histologic subtypes were not statistically different between the transfused and untransfused groups. The average estimated blood loss among the transfused patients was 1104 mL, compared with 764 mL among the untransfused patients (P = .015). Among the 68 who received blood perioperatively, there were ten recurrences (14.7%), compared with two (3.4%) among the 58 patients who did not receive blood (P = .035). In this select population of patients, in which perioperative transfusion was isolated as a variable, transfusion adversely affected the outcome of surgical therapy.

Growth hormone treatment of breeding bulls used for artificial insemination improves fertilization rates.

To evaluate new therapeutical concepts for male subfertility, we tested the effects of exogenous recombinant bovine growth hormone (rbGH) on various endocrine and metabolic parameters both in blood and in seminal plasma of bulls. Sperm quality was assessed morphometrically and by monitoring the number of successful artificial inseminations (AIs) defined as non-return rates (NRR). Aliquots of 450 semen samples were used from each bull and each experimental period (4 wk before, 14 weeks during and 6 wk after treatment). Six out of ten sires (average age 8.4 years) were treated every two weeks with 640-mg depot formulated rbGH (Eli Lilly). Four bulls received vehicle only. Blood plasma bGH, IGF-I, insulin and glucose concentrations were increased with rbGH treatment. In seminal plasma there was no effect of rbGH treatment on fructose and citrate or on testosterone concentrations. With one exception, rbGH-treated bulls had greater IGFBP-3 concentrations in seminal plasma. Motility of spermatozoa after freezing and thawing was increased compared with pretreatment rates. Most interestingly, the number of successful AIs was increased by an average of 6.0% NRR when ejaculates from rbGH-treated bulls were used.

Postpartum femoral neuropathy.

Two cases of postpartum femoral neuropathy occurred. The literature on this subject is reviewed, and possible etiologic mechanisms are discussed. Complete functional recovery is typical of the excellent prognosis of puerperal femoral neuropathy. The importance of recognizing this complication is to predict a favorable prognosis and thus eliminate anxiety for both patient and physician.

[Characterization of secondary immune deficiency in patients with chronic glomerulonephritis]. / Kharakteristika vtorichnogo immunodefitsita u bol'nykh khronicheskim glomerulonefritom.

A study was made of the nature of secondary immuno-deficiency, peculiarities of disordered function of regulatory T-lymphocytes (helpers and suppressors), and their interrelationships in different clinical variants of chronic glomerulonephritis (CGN). Altogether 94 CGN patients were examined, of them 34 were with urinary syndrome and 60 with nephrotic syndrome. With relation to a clinical variant of disease the absolute amount of T-lymphocytes was significantly lowered, the amount and function of B-lymphocytes showed a tendency to an increase. Function of nonspecific Concanavalin A-induced T-suppressors using PHA as a mitogenic indicator (T-T-suppression) was insignificantly raised, and when lipopolysaccharide (T-B-suppression) was used it was considerably suppressed. T-lymphocyte helper activity in CGN patients was on an increase.

[Cross-reacting HLA antigens as a factor increasing the risk of childlessness in a family]. / Perekrestnoreagiruiushchie antigeny sistemy HLA kak fator, povyshaiushchii risk bezdetnosti v sem'e.

A comparative analysis of HLA phenotypes of fertile and infertile spouses revealed that among the infertile the sum of spouses showed compatibility in two or three HLA antigens. A detailed analysis also showed that 10 infertile couples exhibited identical and 2 antigens with a similar structure and function with so-called cross-reactions increasing the histocompatibility index to 75%. The significance of these facts for infertility risk factors is discussed.