Culture and selection of viable blastocysts: a feasible proposition for human IVF?

In human in-vitro fertilization (IVF) embryos are routinelytransferred to the uterus on day 2 or day 3 of development.Resultant implantation and pregnancy rates are disappointinglylow, with only 10% of embryos transferred leading to a livebirth. The ability to culture embryos to the blastocyst stageshould help to resolve this problem by synchronizing the embryoswith the female reproductive tract, and by identifying thoseembryos with little developmental potential. Co-culture hasoffered a possible means of producing blastocysts capable ofhigh implantation rates. However, recent developments in thefield of embryo physiology and metabolism have led to the formulationof new sequential serum-free culture media capable of supportingthe development of viable blastocysts in several mammalian species,including the human. It is therefore proposed that blastocysttransfer should be considered for routine use in human IVF.The high viability of blastocysts cultured in the appropriatesequential media means that fewer embryos are required for transferto achieve a pregnancy, culminating in fewer multiple births.Furthermore, the development of suitable non-invasive testsof embryo viability should further increase the overall successof human IVF by the ability to select before transfer thoseblastocysts most able to establish a pregnancy.     

Exomphalos Defects : A Review of 15 Cases

Background

The outcome after repair of exomphalos defects has improved significantly with refinements in surgical techniques, multidisciplinary management and monitoring of intra-abdominal pressures.

Methods

A retrospective study of medical records of 15 cases with omphalocele was done.

Results

Antenatal diagnosis was available in six cases. There were eight females and seven males with a mean birth weight of 2.2 kg. Nine babies had associated anomalies. There were nine major (defect size> 5 cm) and six minor defects. Immediate closure in neonatal period was carried out in 12 cases. Urinary bladder pressure (UBP) was measured to assess intra-abdominal pressure in cases where primary closure was difficult. Primary closure was omitted in the event of intravesical pressures exceeding 20 mm Hg (~ 25 cms of water). Primary surgical closure was possible in five (56%) major cases. Two cases were subjected to silo repair followed by delayed primary closure whereas the other two required a Goretex mesh closure. Three minor defects could be repaired primarily whereas the remaining three were managed conservatively and closed at age of 9 to 12 months. There were no significant anaesthetic complications. Elective postoperative ventilation was required in one baby. There were three deaths at ages five, nine and ten months due to unrelated causes.

Conclusion

Satisfactory outcome is possible in cases with exomphalos defects with intra-operative intravesical pressure assessment forming a convenient method for excluding abdominal compartment syndrome.Key Words: Abdominal wall defects, Omphalocele, Intra-abdominal pressure, Intravesical pressure measurement     

Spectrum of Haemoglobinopathies in a Tertiary Care Hospital of Armed Forces

Background

Thalassaemia and other structural haemoglobinopathies are the major genetic disorders prevalent in certain parts of the world including India. This study presents the pattern of haemoglobinopathies amongst the referred patients of anaemia in a two-year period.

Methods

A total of 1032 patients were studied during a two-year period for anaemia investigation. Haematological indices, sickling test and haemoglobin electrophoresis with quantification of the bands was done in all cases.

Result

Out of 1032 cases, 774 (75%) were normal and 258 (25%) cases had abnormal haemoglobin pattern. Of the 258 abnormal cases, 136 (53%) were males and 122 (47%) were females. Of all cases of anaemia 370 (36%) were microcytic hypochromic, 237 (23%) macrocytic, 151 (15%) were dimorphic and the rest (26%) had normocytic normochromic picture. 82% of microcytic hypochromic anaemias had reduced serum iron and elevated total iron binding capacity (TIBC), whereas 85% had decreased serum ferritin levels. Spectrum of haemoglobinopathies prevalent were β-Thalassemia trait (17%), followed by sickle cell trait (2.3%). Other haemoglobinopathies in descending order of frequency were sickle cell disease (1.7%), Hb D trait (1%), Hb E trait (0.8%), sickle cell – β thalassemia, Hb E disease, E – β thalassemia (0.6% each) and thalassemia major (0.4%).

Conclusion

This study provides a comprehensive database on the spectrum of haemoglobinopathies in the Armed Forces. It is suggested that detection of HbA₂ should be carried out in all the high-risk groups with anaemia.Key Words: Haemoglobinopathies, Anaemia, Thalassemia     

Evaluation of diltiazem hydrochloride-loaded mucoadhesive microspheres prepared by emulsification-internal gelation technique

The problems of frequent administration and variable low bioavailability (40-60%) after oral administration of conventional dosage forms of diltiazem can be attenuated by designing it in the form of mucoadhesive microspheres which would prolong the residence time at the absorption site to facilitate intimate contact with the absorption surface and thereby improve and enhance the bioavailability. Diltiazem-loaded mucoadhesive microspheres were successfully prepared by emulsification-internal gelation technique with a maximum incorporation efficiency of 93.29 +/- 0.26%. The scanning electron microscopic study indicated that the microspheres were spherical in shape and the drug remained dispersed in the polymer matrix at amorphous state, which was further confirmed by x-ray diffraction analysis. The in vitro wash-off test indicated that the microspheres had good mucoadhesive properties. The wash-off was faster at simulated intestinal fluid (phosphate buffer, pH 7.4) than that at simulated gastric fluid (0.1 M HCl, pH 1.2). The in vitro drug release mechanism was non-fickian type controlled by swelling and relaxation of polymer. There was no significant change in drug content and cumulative drug release of drug-loaded microspheres stored at different storage condition after 8 weeks of study.     

The ancestral gene for transcribed, low-copy repeats in the Prader- Willi/Angelman region encodes a large protein implicated in protein trafficking, which is deficient in mice with neuromuscular and spermiogenic abnormalities

Transcribed, low-copy repeat elements are associated with the breakpoint regions of common deletions in Prader-Willi and Angelman syndromes. We report here the identification of the ancestral gene ( HERC2 ) and a family of duplicated, truncated copies that comprise these low-copy repeats. This gene encodes a highly conserved giant protein, HERC2, that is distantly related to p532 (HERC1), a guanine nucleotide exchange factor (GEF) implicated in vesicular trafficking. The mouse genome contains a single Herc2 locus, located in the jdf2 (juvenile development and fertility-2) interval of chromosome 7C. We have identified single nucleotide splice junction mutations in Herc2 in three independent N-ethyl-N-nitrosourea-induced jdf2 mutant alleles, each leading to exon skipping with premature termination of translation and/or deletion of conserved amino acids. Therefore, mutations in Herc2 lead to the neuromuscular secretory vesicle and sperm acrosome defects, other developmental abnormalities and juvenile lethality of jdf2 mice. Combined, these findings suggest that HERC2 is an important gene encoding a GEF involved in protein trafficking and degradation pathways in the cell.      

Osteomyelitis post acromioclavicular joint reconstruction

Acromioclavicular joint (ACJ) injury is a common shoulder injury. There are various techniques of ACJ reconstruction. Superficial infection after ACJ reconstruction is not an uncommon complication. However, osteomyelitis post ACJ reconstruction has never been highlighted as a possible complication. Our patient is a 31-year-old male who sustained a Rockwood 5 ACJ dislocation and had anatomical ACJ reconstruction with autogenous gracilis and semitendinosus graft. Our technique involved the anatomical reconstruction of the ACJ and the coracoclavicular ligament with the usage of two bioscrews and the temporary stabilisation of the ACJ with two k-wires. As in any orthopaedic surgery, infection is often disastrous especially when the surgery involves implants. It can be disastrous with high morbidity to the patient as well as a costly complication to treat. Therefore, we wish to highlight this case as despite its rarity, osteomyelitis can be devastating to the patient and should be prevented if possible.