Experience on postmortem minimally invasive tissue sampling to ascertain the cause of death determination in South African children: A case for implementing as standard of care.

Determining the death burden for prioritising public health interventions necessitates detailed data on the causal pathways to death. Postmortem minimally invasive tissue sampling (MITS), incorporating histology, molecular and microbial culture diagnostics, enhances cause-of-death attribution, particularly for infectious deaths. MITS proves a valid alternative to full diagnostic autopsies, especially in low- and middle-income countries. In Soweto, South Africa (SA), the Child Health and Mortality Prevention Surveillance (CHAMPS) programme has delineated over 1 000 child and stillbirth deaths since 2017. This SA CHAMPS site supports advocating for the use of postmortem MITS as routine practice, for more granular insights into under-5 mortality causes. This knowledge is crucial for SA's pursuit of Sustainable Development Goal 3.2, targeting reduced neonatal and under-5 mortality rates. This commentary explores the public health advantages and ethicolegal considerations surrounding implementing MITS as standard of care for stillbirths, neonatal and paediatric deaths in SA. Furthermore, based on the data from CHAMPS, we present three pragmatic algorithmic approaches to the wide array of testing options for cost-effectiveness and scalability of postmortem MITS in South African state facilities.

Expanding the Klebsiella pneumoniae volatile metabolome using advanced analytical instrumentation for the detection of novel metabolites.

AIMS: The purpose of this study was to identify the volatile molecules produced by the pathogenic Gram-negative bacterium Klebsiella pneumoniae (ATCC 13883) during in vitro growth using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC×GC-TOFMS). METHODS AND RESULTS: Klebsiella pneumoniae ATCC 13883 was incubated in lysogeny broth to mid-exponential and stationary growth phases. Headspace volatile molecules from culture supernatants were concentrated using solid-phase microextraction (SPME) and analysed via GC×GC-TOFMS. Ninety-two K. pneumoniae-associated volatile molecules were detected, of which 78 (85%) were detected at both phases of growth and 14 (15%) were detected at either mid-exponential or stationary growth phases. CONCLUSIONS: This study has increased the total number of reported K. pneumoniae-associated volatile molecules from 77 to 150, demonstrating the sensitivity and resolution achieved by employing GC×GC-TOFMS for the analysis of bacterial headspace volatiles. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents an early-stage comprehensive volatile metabolomic analysis of an opportunistic bacterial pathogen. Characterizing the volatile molecules produced by K. pneumoniae during in vitro growth could provide us with a better understanding of this organisms' metabolism, an area that has not been extensively studied to date.

Understanding emotional abuse.

Emotional abuse lacks the public and political profile of physical and sexual abuse, despite being at their core and frequently their most damaging dimension. Difficulties in recognition, definition and legal proof put children at risk of remaining in damaging circumstances. Assessment of the emotional environment is necessary when interpreting possible physical or sexual abuse and balancing the risks and benefits of intervention. This article considers factors contributing to professional difficulty. It is suggested that understanding emotional abuse from the first principles of the causes and implications of the dysfunctional parent-child relationships it represents can help prevention, recognition and timely intervention. It may facilitate the professional communication needed to build up a picture of emotional abuse and of the emotional context of physical and sexual abuse. Doing so may contribute to the safety of child protection practice. The long-term cost of emotional abuse for individuals and society should be a powerful incentive for ensuring that development of services and clinical research are priorities, and that the false economy of short-term saving is avoided.

Non-infant adoption from care: lessons for safeguarding children.

AIM: To explore use of adoption in remedying abuse and neglect, to inform child protection practice and to identify professional responsibilities to adoptive families. METHOD: A cohort of 130 children was identified for whom adoption was recommended in 1991-1996 at a mean age of 5.7 years (range 3-11). All were in local authority care for child protection reasons. BACKGROUND: information was gathered from social work records. The children were traced between 6 and 11 years later and their adopters interviewed. The Strengths and Difficulties Questionnaire and the Parent-Child Communication Scale were completed. RESULTS: All but three birth parents had traumatic childhoods involving abuse, neglect and/or time in care. Eighty-six per cent had violent adult relationships. Mental illness, learning difficulties and substance abuse were prominent. Sixty-seven per cent of families were known to social services when children were born, but 98% experienced abuse or neglect. Most adopters found the first year challenging, though rewarding. Depression, anxiety and marital problems were common. Children's learning difficulties, conduct problems, emotional 'phoniness' and rejection affected closeness. At follow-up 28% described rewarding, happy placements, and 62% described continuing difficulties tempered by rewards. However, 10% reported no rewards. Hyperactivity and inattention frequently persisted despite stable adoption and were associated with conduct and attachment difficulties. Use of professional services was substantial. Thirty-eight per cent of children failed to achieve stable adoption. Later entry to care correlated with poorer outcome. CONCLUSIONS: This study highlights the importance in safeguarding children of considering the implications of parental childhood experiences, and indicates the risk of delay. The high prevalence of domestic violence in birth families indicates the need for better resources for managing emotional dysregulation. Adoption is a valuable therapeutic tool, but professional responsibilities in supporting it need to be acknowledged and adequately resourced. Dysfunctional hypothalamus-pituitary-adrenal axis programming may contribute to persisting difficulties. Supporting substitute care should be considered integral to safeguarding children.

Mind the gaps.

Future generations are likely to judge us as much by our management of what we do not know as of what we do. Trends towards increasing specialisation, guidelines, protocol and regulation, and devaluation of basic clinical skills of history taking and examination through over-reliance on clinical investigation, carry the risk that gaps in knowledge and services are overlooked and may be exacerbated. Unrecognised gaps are particularly hazardous. Professional practice and structures should give the best chance of eliminating blind spots, both to reduce scope for harm and to increase opportunity for progress, of which the uncharted territory should be a rich source. Safe practice should be as robust in addressing the implications of what we do not know as in applying knowledge prudently. The system itself needs to be evaluated as carefully as the practice it serves.

Skin surface lipids and skin and hair coat condition in dogs fed increased total fat diets containing polyunsaturated fatty acids.

It is generally believed that diets containing increased amounts of polyunsaturated fatty acids (PUFA) result in improved canine skin and hair coat (SHC). However, the extent to which dietary fat amount and type play a role remains to be systematically investigated. The objective of this study was to investigate the role of both increased dietary fat amount and type on SHC assessments of dogs. Improvements of SHC conditions were investigated after feeding three diets containing increased total dietary fat (i.e. 13% total fat) for 12 weeks in relation to a lower fat acclimation diet (i.e. 9% total fat). The higher fat diets varied in polyunsaturated and saturated fat types and amounts but total fat was kept constant. Skin and hair coat assessments were performed at selected intervals by a trained group of veterinarians and graduate students. In addition, hair lipids were fractionated by thin layer chromatography after extraction of plucked hair samples. Significant improvements were found in hair coat glossiness and softness in all dogs fed the higher fat diets in relation to the acclimation diet. Improvements as a result of fat type were also seen but only at 12 weeks. A parallel finding was a marked increase in hair cholesteryl ester content determined at the end of the study at which time SHC scores were significantly improved. Skin and hair coat condition improvements may thus be related to increased cholesteryl ester deposited on the hair shaft surface when high fat diets are fed. Whereas this finding is preliminary, hair lipid analysis may be a useful, non-invasive technique with which to help assess dietary effects on canine SHC.

Thinking about children's attachments.

Disordered parental attachment can commit children to lives characterised by relationship difficulties, behaviour problems, educational failure, and poor self-esteem. It is a major root of trans-generational neglect and abuse and frequently underlies mental health problems, drug and alcohol addiction, homelessness, and crime. Early childhood setting of hypothalamus-pituitary-adrenal axis function appears to contribute to these costly difficulties. More broadly, the concept of attachment can contribute to defining and managing the psychosocial dimension of routine paediatric care. The current under-representation of attachment in paediatric education, practice, and research needs to be rectified.

Effects of dietary flax seed and sunflower seed supplementation on normal canine serum polyunsaturated fatty acids and skin and hair coat condition scores.

This prospective study involved supplementing 18 normal dogs with flax seed (FLX) and sunflower seed (SUN) and evaluating their effects on skin and hair coat condition scores and serum polyunsaturated fatty acids (PUFA) concentrations. Skin and hair coat were evaluated in a double-blinded fashion using a numeric scoring system and serum PUFA concentrations were determined. Our hypothesis was that changes in serum PUFA concentrations are associated with improvements in skin and hair coat and that serum PUFA would provide an objective method for making dietary fatty acid supplement recommendations. Although a numerical improvement was found in hair coat quality in both groups, this improvement was not sustained beyond 28 days. The relative per cent of 18:3n-3 concentrations in serum phospholipids increased in the FLX treated dogs but these concentrations remained unchanged in the SUN treated dogs. Also, elevations in relative per cent of 18:2n-6 concentrations in serum phospholipids were seen in the FLX group. The ratio of serum polyunsaturated to saturated fatty acids also showed a transient increase. These increases preceded the peak skin condition score peak value by approximately 14 days. It was concluded that a 1-month supplementation with either flax seed or sunflower seed in dogs provides temporary improvement in skin and hair coat. These changes appeared to be associated with increased serum 18 carbon PUFA.

Response to immunotherapy in six related horses with urticaria secondary to atopy.

Urticaria secondary to atopy may be a familial problem in some horses. Immunotherapy using a vaccine containing antigens selected on the basis of history and results of intradermal testing can be an effective method of managing atopy in horses; a response to therapy may be seen within 2 months.

Canine and feline atopic dermatitis: a review of the diagnostic options.

Atopic dermatitis is an inherited pruritic skin disease in dogs and cats. This pruritic skin condition is due to the animal having an allergic reaction to environmental allergens. The environmental allergens that an individual dog or cat is allergic to are specific for that individual animal. Management options for affected dogs and cats include identification of the offending environmental allergens and subsequent avoidance of that allergen, or allergen-specific immunotherapy. Several diagnostic tests are available to veterinarians to try to identify these allergens. The pros and cons of each of these diagnostic tests will be addressed.

Small evolutionarily conserved RNA, resembling C/D box small nucleolar RNA, is transcribed from PWCR1, a novel imprinted gene in the Prader-Willi deletion region, which Is highly expressed in brain.

Prader-Willi syndrome is a complex neurodevelopmental disorder caused by the inactivation or deletion of imprinted, paternally expressed genes in chromosome band 15q11.2. We report the identification and characterization of PWCR1, a novel imprinted gene within that region, and its mouse orthologue, Pwcr1, which was mapped to the conserved syntenic region on mouse chromosome 7. Expressed only from the paternal allele, both genes require the imprinting-center regulatory element for expression and are transcribed from the same strand. They are intronless and do not appear to encode a protein product. High human/mouse sequence similarity (87% identity) is limited to a 99-bp region called "HMCR" (for "human-mouse conserved region"). The HMCR sequence has features of a C/D box small nucleolar RNA (snoRNA) and is represented in an abundant small transcript in both species. Located in nucleoli, snoRNAs serve as methylation guidance RNAs in the modification of ribosomal RNA and other small nuclear RNAs. In addition to the nonpolyadenylated small RNAs, larger polyadenylated PWCR1 transcripts are found in most human tissues, whereas expression of any Pwcr1 RNAs is limited to mouse brain. Genomic sequence analysis reveals the presence of multiple copies of PWCR1 and Pwcr1 that are organized within local tandem-repeat clusters. On a multispecies Southern blot, hybridization to an HMCR probe encoding the putative snoRNA is limited to mammals.

Molecular portraits of human breast tumours.

Human breast tumours are diverse in their natural history and in their responsiveness to treatments. Variation in transcriptional programs accounts for much of the biological diversity of human cells and tumours. In each cell, signal transduction and regulatory systems transduce information from the cell's identity to its environmental status, thereby controlling the level of expression of every gene in the genome. Here we have characterized variation in gene expression patterns in a set of 65 surgical specimens of human breast tumours from 42 different individuals, using complementary DNA microarrays representing 8,102 human genes. These patterns provided a distinctive molecular portrait of each tumour. Twenty of the tumours were sampled twice, before and after a 16-week course of doxorubicin chemotherapy, and two tumours were paired with a lymph node metastasis from the same patient. Gene expression patterns in two tumour samples from the same individual were almost always more similar to each other than either was to any other sample. Sets of co-expressed genes were identified for which variation in messenger RNA levels could be related to specific features of physiological variation. The tumours could be classified into subtypes distinguished by pervasive differences in their gene expression patterns.

New drugs in veterinary dermatology.

This article discusses several different new drugs currently being used in dermatology. Most of the drugs discussed showed some promise as being a useful therapy in veterinary medicine, but a few have questionable efficacy (nonsedating antihistamines). The majority of these drugs have not had any pharmacokinetic or clinical trials conducted on them in small animals. Hopefully, in the future, more studies are funded so that we can determine the clinical therapeutical efficacy and appropriate doses for these drugs.

Distinctive gene expression patterns in human mammary epithelial cells and breast cancers.

cDNA microarrays and a clustering algorithm were used to identify patterns of gene expression in human mammary epithelial cells growing in culture and in primary human breast tumors. Clusters of coexpressed genes identified through manipulations of mammary epithelial cells in vitro also showed consistent patterns of variation in expression among breast tumor samples. By using immunohistochemistry with antibodies against proteins encoded by a particular gene in a cluster, the identity of the cell type within the tumor specimen that contributed the observed gene expression pattern could be determined. Clusters of genes with coherent expression patterns in cultured cells and in the breast tumors samples could be related to specific features of biological variation among the samples. Two such clusters were found to have patterns that correlated with variation in cell proliferation rates and with activation of the IFN-regulated signal transduction pathway, respectively. Clusters of genes expressed by stromal cells and lymphocytes in the breast tumors also were identified in this analysis. These results support the feasibility and usefulness of this systematic approach to studying variation in gene expression patterns in human cancers as a means to dissect and classify solid tumors.

Cutaneous horn and squamous cell carcinoma in situ (Bowen's disease) in a cat.

Cutaneous horn and squamous cell carcinoma (SCC) in situ (i.e., Bowen's disease) were documented concurrently in a cat. The cat had multiple, crusted lesions and a cutaneous horn on the right dorsal lumbar area. All the crusted cutaneous lesions were diagnosed as SCC in situ. Other findings included the presence of a thymoma and hepatoma. This cat was tested, and results were negative for feline leukemia and feline immunodeficiency viruses. At necropsy (eight months after the initial diagnosis was made) the hepatoma had ruptured, resulting in exsanguination and death.

Heat Shock Protein Synthesis Induced by Methomyl in Maize (Zea mays L.) Seedlings.

Exposure of plumules of intact maize seedlings (Zea mays L.) to S-methyl-N-[(methylcarbamoyl)-oxy]thioacetimidate (methomyl) represses synthesis of several polypeptides normally made under control conditions and induces synthesis of polypeptides similar to maize heat shock polypeptides (HSPs). Three of the methomyl-induced polypeptides (18 kilodaltons) are recognized by antibodies raised against 18 kilodalton maize heat shock polypeptides.

Childhood lymphoblastic leukaemia: sex difference in 6-mercaptopurine utilization.

Twelve boys and 10 girls on similar long term remission maintenance treatment for lymphoblastic leukaemia had 79 random assays of their red cell 6 thioguanine nucleotide ( 6TGN ) concentrations performed as an index of cytotoxic activity generated by oral 6-mercaptopurine ( 6MP ). Correlation between the dose of 6MP and 6TGN was statistically significant in the girls (r = 0.58, P less than 0.001) but not in the boys (r = 0.15). Additionally, as a group the boys tolerated more 6MP (P less than 0.05), despite similar prescribing criteria, but this did not result in a higher mean 6TGN concentration or increased myelotoxicity. It appears that girls develop 6MP cytotoxicity at lower doses and more predictably than boys. If so, this may be relevant to the as yet unexplained but marked sex difference in prognosis apparent in some studies.

Disturbance of 6-mercaptopurine metabolism by cotrimoxazole in childhood lymphoblastic leukaemia.

The effect of cotrimoxazole on the utilization of 6-mercaptopurine (6MP) was studied in a group of children receiving remission maintenance treatment for lymphoblastic leukaemia (ALL). This was done by measuring the level of an active metabolite of 6MP, 6-thioguanine nucleotide (6TGN), and comparing it both with the drug dose and with subsequent neutropenia in the presence or absence of concurrent cotrimoxazole. In children who were not taken cotrimoxazole, the concentration of 6TGN showed a significant positive correlation with the dose and a significant negative correlation with the absolute neutrophil count 2 weeks later. In those who were taking the antibiotic both these relationships were lost. This suggests that cotrimoxazole can interfere with both the absorption and the cytotoxicity of 6MP and may, in turn, alter its antileukaemic effect.

Childhood leukaemia: a relationship between intracellular 6-mercaptopurine metabolites and neutropenia.

6-Mercaptopurine is extensively used in the treatment of childhood lymphoblastic leukaemia to prolong the duration of remission achieved with other drugs. The response to remission maintenance therapy varies widely. We investigated the relationship between red blood cell 6-thioguanine nucleotide, a metabolite of 6-mercaptopurine, and myelosuppression in 22 children with acute lymphoblastic leukaemia in remission. The peripheral neutrophil count was used as an index of myelosuppression. 6-Mercaptopurine dose was related to 6-thioguanine nucleotide concentration (r = 0.4; P less than 0.001; n = 90; y = 18.51 + 0.36 x). Large individual variations around the regression line are observed. Neither 6-mercaptopurine dose nor 6-thioguanine nucleotide concentration was related to the neutrophil count at the time of sampling (day 0) or 7 days later. Both 6-mercaptopurine dose and 6-thioguanine nucleotide concentration correlated with the neutrophil count at day 14 (r = -0.33; P less than 0.01; n = 90 and r = -0.3; P less than 0.01; n = 90 respectively). This delay is compatible with a cytotoxic action on bone marrow stem cells. Excluding children with other, uncontrolled, potentially myelosuppressive influences the correlation between 6-thioguanine nucleotide concentration and neutropenia improved (r = -0.6; P less than 0.001; n = 37). A significant degree of neutropenia was observed by day 14 if the 6-thioguanine nucleotide concentration (day 0) was greater than 210 pmol/8 X 10(8) RBCs. The assay of 6-thioguanine nucleotide may highlight those individuals with pharmacokinetic resistance. Two children on continuous high dose 6-mercaptopurine, had low red blood cell 6-thioguanine nucleotide concentrations and neutropenia was not observed.