Recruitment and screening policies and procedures used to establish a paid donor oocyte registry

We have reviewed the demographic characteristics of, and report abnormalities noted in, the de-novo growth and development of a paid oocyte donation programme. The personal profiles of all prospective oocyte donors were reviewed. Acceptance or rejection of candidates was based upon screening the results of medical, genetic and psychological testing. A total of 603 candidates initially responded to our advertisement. From this pool, 313 individuals were considered suitable and contacted by telephone. Following further conversation, 176 women were scheduled an entry interview. On completion of the formal screening process, 17.6% (n = 31) of those actually interviewed were denied entry. Thus, from the initial interested parties, only 23% of women wishing to participate in oocyte donation were considered suitable candidates. Given the high attrition rate, we concluded that the need for rigorous and thorough medical, psychological and genetic testing is mandatory for the establishment of a donor registry. Furthermore, professional counselling of prospective donors with respect to the results of tests and the implications of test results with respect to their future medical and reproductive health, are important parts of providing comprehensive care.      

The results of combined cataract extraction and trabeculectomy using separate incisions

A method of combined cataract extraction with posterior chamber intraocular lens and trabeculectomy using separate incisions was tested in 44 operations on 38 patients. The mean preoperative intraocular pressure (IOP) of 28.1 ± 11.7 (range 12 to 56) mmHg on maximum medication was lowered to 13.9 ± 3.4 (9 to 23) mmHg at one year, with half the eyes still requiring topical medication. The IOP was 40 mmHg or more preoperatively in eight eyes and 20 mmHg or more in only two patients at one year. There were no rises in IOP above 20 mmHg in the early postoperative period (days 1 and 2). Visual acuity was 6/9 or better in 27 and 6/12 in three eyes. There was an expulsive haemorrhage in one case, rupture of the posterior capsule in two eyes and a choroidal detachment in one eye, but no flat anterior chambers. The two-incision method allowed placement of an intraocular lens with good postoperative pressure control.     

Testicular ultrastructure in infertile men

The review shows typical ultrastructural alterations of germ, Sertoli, and Leydig cells in infertile men. Regardless of the cause of infertility, the disruption of the spermatogenic process usually occurs in the pachytene stage of meiotic prophase and in the stages of early spermatid maturation. The disturbances affect the cytoplasm more than the nucleus, and the synaptonemal complexes have shown significant stability even in the severely injured testes. An acrosome formation is found to be open to injury in more advanced germ cells during spermatid maturation. The manner of reaction of the Sertoli cells under different pathological conditions depends on the presence and degree of maturation of the neighboring germ cells. The appearance of immature Sertoli cells is accompanied by the loss of germ cells more advanced in their differentiation. In most pathologically altered testes, mature Sertoli cells reveal a universal manner of reaction of cell organelles. Leydig cell ultrastructure fluctuates considerably, and the alterations predominantly affect the sites of steroid synthesis, in spite of disease specificity. It becomes clear that a complex estimation of a real testicular state requires the application of new techniques as well as recognition of local control mechanisms. This will provide evidence toward elucidation of male infertility.     

Topoisomerase II beta expression level correlates with doxorubicin-induced apoptosis in peripheral blood cells

Anthracyclines are widely used in oncology. Both the response and side-effects of anthracyclines are individually variable, but determinants or predictive markers of this variability are not available. We investigated the variability in the expression of the anthracycline targets topoisomerases II (topo II) alpha and beta and its significance for the apoptotic response following exposure to the anthracycline doxorubicin. Only topo II beta protein expression was detected in peripheral blood cells. Usually considered a constitutively expressed protein, topo II beta varied 3-, 18-, and 16-fold on the mRNA, protein and activity levels, respectively, among the volunteers tested. In addition, the expression of topo II beta was modified by several mitogens, suggesting a role in the regulation of cell cycle. Strikingly, topo II beta activity correlated statistically significantly with the apoptotic response in peripheral blood leukocytes exposed to 1 μM doxorubicin. A longitudinal study in a subset of study subjects demonstrated that 30% of the topo II expression variability may be inherited. However, resequencing of the TOP2B gene in 48 unrelated individuals revealed only 8 gene variants, none of them with obvious effects on the expression or protein sequence of topo II beta. Taken together, the apoptotic response to doxorubicin in peripheral blood cells may be mediated by topo II beta. The expression level of topo II beta is intra- and inter-individually variable, and may in part determine the apoptotic response to doxorubicin and other anthracyclines.     

Pharmacogenetic screening for drug therapy: from single gene markers to decision making in the next generation sequencing era

Pharmacogenetics has substantially added to our understanding of the variability of drug response. A number of single gene markers have been established and are ready to use in clinical practice. Here we review the validity and utility of markers in a number of genes (CYP2D6, CYP2C19, CYP2C9, VKORC1, TPMT, UGT1A1, OATP1B1, KRAS and HLA locus) for therapy decisions. As drug response is a complex trait in the majority of cases, most of the identified functional variants will only explain a limited part of the variability of drug response. In this sense, a phenotype is the product of many low-penetrance variations. Technical progress has not only improved the cost-effectiveness of screening for single gene markers, but offers the possibility of generating vast amounts of genome-wide single nucleotide polymorphism (SNP) or sequence data for each patient. The latest challenge is to incorporate these amounts of data into pharmacogenetic decision support. We discuss here the challenges associated with choosing the correct therapy for patients who present to their physicians with personal genome data.     

Aetiological agents of ventilator-associated pneumonia and its resistance pattern – a threat for treatment

Background

Ventilator-associated pneumonia (VAP) is a common type of nosocomial pneumonia encountered in intensive care units. There are several aetiological agents which make treatment challenging. Improper antibiotic treatment of ventilated patients may lead to the emergence of multidrug resistant (MDR) pathogens.

Method

A prospective study was performed over a period of 20 months. Our study had two arms: the first, ‘Incidence and risk factors of VAP in a tertiary care hospital’ was the subject of an earlier publication; we therefore present the second investigative arm in this work. The aetiological agents of patients on mechanical ventilation (MV) were identified by standard bacteriological method. The susceptibility pattern was evaluated by Kirby-Bauer disc diffusion method. Extended spectrum beta lactamase (ESBL) testing was performed by combination disc method, and metallo-beta lactamase (MBL) testing was performed by EDTA disk synergy test (EDS).

Results

Late-onset VAP was associated with Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli, while early-onset VAP was commonly caused by members of Enterobacteriaceae, Candida albicans and Staphylococcus aureus. 72.2 per cent of VAP patients had monomicrobial and 27.8 per cent had polymicrobial infection. Out of the 24 isolates obtained from patients with VAP, seven (29.2 per cent) were MDR pathogens. ESBL and MBL production was detected in 40 per cent and 20 per cent of Klebsiella pneumoniae isolated in our study. Around 50 per cent of isolates associated with late-onset VAP were MDR, while 22.2 per cent isolates obtained from patients with earlyonset VAP were MDR.

Conclusion

VAP is a nosocomial pneumonia that is common among ventilated patients. The aetiological agents vary from common organisms to MDR pathogens that are difficult to treat. A proper knowledge of MDR pathogens and early isolation followed by prevention of prolonged antibiotic therapy can reduce the mortality of late onset VAP.