Innate differences in the molecular signature of normal inferior & superior human parathyroid glands: potential implications for parathyroid adenoma.

Primary hyperparathyroidism is a common endocrine disorder. Interestingly, the majority (75%) of parathyroid tumors are localized to the inferior parathyroid glands. To date, the reason for this natural bias has not been investigated. We assessed the global gene expression profile of superior and inferior glands obtained from forensic autopsies. The genes with significant differential expression between superior and inferior parathyroids were further assessed by RT-PCR in 19 pairs. As an iterative approach, additional genes with an established role in parathyroid disorders, i.e., CASR, MAFB, PAX9, TBCE, TBX1, VDR, MEN1, CCND1, and CDC73 were also evaluated by RT-PCR in all 19 pairs of superior and inferior parathyroid glands. Seven homeobox genes, namely HOXA4, HOXA5, HOXBAS3, HOXB4, HOXB6, HOXB9, IRX1, and one encoding for ALDH1A2 showed a lower expression in the inferior parathyroid glands than in the superior. Conversely, SLC6A1 showed a higher expression in the inferior glands. Of the nine genes with significant differential mRNA expression among superior and inferior glands HOXB9, HOXB4 and IRX1 could be detected by western blotting/mass spectrometry. The study is the first to show the differential expression of nine genes HOXA4, HOXA5, HOXBAS3, HOXB4, HOXB6, HOXB9, IRX1, ALDH1A2, and SLC6A1 in inferior versus the superior parathyroid glands. This could have potential implications for the preferential localization of parathyroid tumors to the inferior parathyroid glands as observed in patients with primary hyperparathyroidism.

Osteogenic Mechanisms of Basal Ganglia Calcification and its ex vivo Model in the Hypoparathyroid Milieu.

CONTEXT: Basal-ganglia calcification (BGC) is common (70%) in patients with chronic hypoparathyroidism. Interestingly, cortical gray matter is spared from calcification. The mechanism of BGC, role of hyperphosphatemia, and modulation of osteogenic molecules by parathyroid hormone (PTH) in its pathogenesis is not clear. OBJECTIVE: We assessed the expression of a large repertoire of molecules with proosteogenic or antiosteogenic effects, including neuroprogenitor cells in caudate, dentate, and cortical gray matter from normal autopsy tissues. The effect of high phosphate and PTH was assessed in an ex vivo model of BGC using striatum tissue culture of the Sprague-Dawley rat. METHODS: The messenger RNA and protein expression of 39 molecules involved in multiple osteogenic pathways were assessed in 25 autopsy tissues using reverse-transcriptase polymerase chain reaction, Western blot, and immunofluorescence. The striatal culture was maintained in a hypoparathyroid milieu for 24 days with and without (a) high phosphate (10-mm ß-glycerophosphate) and (b) PTH(1-34) (50 ng/mL Dulbecco's modified Eagle's medium-F12 media) for their effect on striatal calcification and osteogenic molecules. RESULTS: Procalcification molecules (osteonectin, ß-catenin, klotho, FZD4, NT5E, LRP5, WNT3A, collagen-1α, and SOX2-positive neuroprogenitor stem cells) had significantly higher expression in the caudate than gray matter. Caudate nuclei also had higher expression of antiosteogenic molecules (osteopontin, carbonic anhydrase-II [CA-II], MGP, sclerostin, ISG15, ENPP1, and USP18). In an ex vivo model, striatum culture showed an increased propensity for calcified nodules with mineral deposition similar to that of bone tissue on Fourier-transformed infrared spectroscopy, alizarin, and von Kossa stain. Mineralization in striatal culture was enhanced by high phosphate and decreased by exogenous PTH through increased expression of CA-II. CONCLUSION: This study provides a conceptual advance on the molecular mechanisms of BGC and the possibility of PTH therapy to prevent this complication in a hypoparathyroid milieu.

Injuries and deaths caused by non-human primate attacks: Paediatric vulnerability.

A six-month-old girl was taken from her mother's lap by a macaque monkey (Macaca) that had entered the room through an open door. The monkey escaped with the infant but subsequently dropped her from a rooftop terrace. Death occurred soon after impact with the ground, and was found at autopsy to be due to blunt cranio-cerebral trauma with skull fractures and intracranial haemorrhage. Monkeys and apes may cause a variety of serious and potentially life-threatening injuries and may manifest quite unpredictable and aggressive behaviour around humans. The very young are particularly vulnerable to primate attacks, and 'baby theft' by apes and monkeys is an event that has now been reported, albeit rarely, in a number of different countries around the world. The fatally injured infants have died from bites, falls and drowning. Chimpanzees in the wild may also specifically hunt human infants for food.

Cadaveric Study of Topographic Anatomy of Temporal and Marginal Mandibular Branches of the Facial Nerve in Relation to Temporomandibular Joint Surgery.

PURPOSE: Detailed anatomy of the facial nerve, including the variations among different ethnic groups, is essential to prevent an iatrogenic injury. The purpose of the study was to document topographic anatomy of temporal and marginal mandibular (MM) branches of the facial nerve in relation to temporomandibular joint (TMJ) surgery. The specific aim was to demonstrate detailed course of temporal and MM nerves, their surgical implications, and to compare the results obtained with the previous studies. METHODS: The investigators implemented a prospective cadaveric study. A dissection was carried out on 52 facial halves. The facial nerve was dissected according to the instructions described in the Cunningham's dissection manual. Anatomic landmarks were selected as determined by Al-Kayat and Bramley, and results obtained were compared with previous published articles. RESULTS: The study sample was composed of 52 facial halves (males, n = 35; females, n = 17). The number of branches of temporal nerve varied in dissected facial halves from 3 (n = 37 [70%]), 2 (n = 14 [26%]), to 1 (n = 1 [2%]). The distance between the lowest concavity of the bony external auditory meatus to the point at which the facial nerve bifurcates (distance B) was considerably less in the study population (1.79 cm) when compared with the reported literature (2.3 cm). There was no significant influence of gender and cephalic index on distances measured. There was 1 branch in 15% of the dissected facial halves (1 in 52) and 2 branches in 85% (44 of 52). The MM nerve was seen coursing below the inferior border of the mandible, and in 44 (85%), the nerve was present above the inferior border of mandible all along the course. CONCLUSIONS: The topographic anatomy of the temporal and MM nerves is the same as reported in the literature. The only considerable difference was found in distance B; hence, surgical procedures involving the distance B require special consideration.

Fatal ovarian hyperstimulation syndrome in an anonymous egg donor.

Ovarian hyperstimulation syndrome is a rare, but potentially life-threatening iatrogenic disorder arising from ovulation induction or ovarian hyperstimulation for assisted reproduction techniques. We report a case of a 26-year-old multiparous woman, an anonymous egg donor, who died a few hours after undergoing a procedure to donate eggs at an in vitro fertilization clinic. Her husband alleged that medical negligence had led to her death. The autopsy confirmed death due to ovarian hyperstimulation syndrome. We know of no previous descriptions of fatal ovarian hyperstimulation syndrome in an anonymous egg donor in medico-legal literature.

Skin suicide note written in mehndi (henna).

Suicide messages on the skin are rare. Until now, in all reported cases, the writing tool used by the victims has been a pen. We report a suicide case by hanging in which the victim had written a note on her palm in mehndi, or henna, at a wedding ceremony three days before the fatal act. The note was discovered at autopsy.

Triple hanging in filicide-suicide: an unusual case report.

Filicide-suicide is a special category of homicide-suicide event where the victim(s) are children and the perpetrator is one of the parents or both. It is not extensively documented or adequately defined in literature. In developed countries, shooting is a common method of homicide and suicide. Uses of knives, blunt objects, strangulation, poisoning and drowning are other methods frequently employed by the perpetrator. Homicide by hanging in filicide-suicide is rarely reported in forensic literature. We present a rare case of filicide-suicide, where the mother killed both her children by hanging them one by one from a ceiling fan in the same room and later committed suicide by hanging in another room.

Quality and quantity of extracted deoxyribonucleic Acid (DNA) from preserved soft tissues of putrefied unidentifiable human corpse.

CONTEXT: The appropriate collection and preservation of soft tissues from putrefied unidentifiable human corpse for the purpose of identification using DNA profiling technique is critically important especially in developing countries like India having different levels of health-care set ups with largely varying facilities and varying climatic conditions. AIMS: The present study was carried out, mainly focusing on quality and quantity of extracted DNA from the soft tissues of putrefied unidentifiable human corpse stored upto 4 weeks at 4°C and at -80°C for DNA analysis. MATERIALS AND METHODS: The present study was conducted on 16 different putrefied unidentifiable human corpses after getting approval from institutional ethical committee. Around 2 g of four different tissues (brain, kidney, heart and muscle) were collected and preserved for one month followed by DNA extraction using the organic method, the quality and quantity of high molecular weight-DNA was estimated using the spectrophotometer and gel electrophoresis. Further, the amplification polymerase chain reaction (PCR) was also performed (AmpFLSTR(®) Indentifiler™ PCR Amplification kit for multiple loci, of Applied Biosystems, Lab India) and was checked using continuous PAGE. RESULTS: The yield of DNA was significantly higher at -80°C for all the four tissues collected and was best for brain followed by heart, kidney and worst for muscles in all cases. CONCLUSIONS: It is suggested that the brain tissue preserved at -80°C is the best among soft issues for DNA extraction. Refrigeration or deep freezing facility should be available at all the centers.

Expression of osteogenic molecules in the caudate nucleus and gray matter and their potential relevance for Basal Ganglia calcification in hypoparathyroidism.

BACKGROUND: Basal ganglia calcification (BGC) is an interesting example of ectopic calcification in patients with hypoparathyroidism. Its pathogenesis and reasons for predilection of calcification at basal ganglia are not clear. OBJECTIVE: To assess the expression of osteogenesis-related molecules in the caudate nucleus and surface gray matter (an area spared from calcification) and discuss potential relevance of the results in context of BGC in idiopathic hypoparathyroidism. METHODS: Caudate nucleus and gray matter were obtained from 14 autopsies performed in accidental deaths. The mRNA expression of bone transcription factors (RUNX2/osterix), bone morphogenetic proteins (BMPs) 2 and 4, osteonectin, osteopontin, osteocalcin, vitamin D receptor, calcium sensing-receptor, Na phosphate transporters (PiTs) 1 and 2, N-methyl-D-aspartate receptor 2B (NMDAR2B), carbonic anhydrase II (CA-II), PTH1 receptor (PTH1R), PTH2R, and PTHrP were assessed by RT-PCR. Western blot, spot densitometry, and immunohistochemistry were performed to assess protein expression of molecules showing differences in mRNA expression between caudate and gray tissues. RESULTS: The mean mRNA expression of PiT1 (11.0 ± 10.39 vs 32.9 ± 20.98, P = .003) and PTH2R (1.6 ± 1.47 vs 13.7 ± 6.11, P = .001) were significantly lower in the caudate nucleus than the gray matter. The expression of osteonectin, osteopontin, and CA-II were significantly higher in the caudate nucleus than the gray matter (P = .01, .001, and .04, respectively). The mRNA expression of other molecules was comparable in the 2 tissues. The protein expression of both CA-II and osteonectin was 24% higher and PiT1 17% lower in caudate than the gray matter. The differences in the PTH2R and osteopontin protein expression were not appreciable. CONCLUSIONS: The presence of several osteogenic molecules in caudate nucleus indicates that BGC would probably be the outcome of an active process. The differences in expression of these molecules in caudate over gray matter could favor BGC at this site in the unique biochemical milieu of hypoparathyroid state.