Dealing with Death: Crisis and Resilience in Adolescent Boys

RESEARCH OBJECTIVE: The research investigates how young boys progress in life after the loss of their fathers. It explores the adjustments they make, the coping mechanisms they develop to navigate life, and the male roles they assume in the family. Additionally, it examines how family can aid and support their coping and adjustment process. THE RESEARCH PROBLEM AND METHODS: This study utilises a case study methodology focused on gaining an understanding of the lives of two adolescent boys who experienced the death of their fathers in the early formative years of their lives. THE PROCESS OF ARGUMENTATION: The paper focuses on five themes- family dynamics, life transitions with and without their fathers, post-loss reassignment of family roles, present circumstances, and future goals. RESEARCH RESULTS: A qualitative analysis of the data revealed that participants’ belief systems, family support, and striving toward future goals helped them to be resilient in this event. CONCLUSIONS RECOMMENDATIONS AND APPLICABLE VALUE OF RESEARCH: While most research typically emphasises the negative impacts of death on people, this study demonstrates how individuals’ can be resilient, dependent on both person-related factors and environmental factors. To help adolescents who are grieving become resilient, the paper offers intervention and coping strategies through counselling, the availability of support networks, and combined efforts by the state and educational institutions

Targeting the active site of the placental isozyme of alkaline phosphatase by phage-displayed scFv antibodies selected by a specific uncompetitive inhibitor

BACKGROUND: The isozymes of alkaline phosphatase, the tissue non-specific, intestinal and placental, have similar properties and a high degree of identity. The placental isozyme (PLAP) is an oncofetal antigen expressed in several malignancies including choriocarcinoma, seminoma and ovarian carcinoma. We had earlier attempted to isolate PLAP-specific scFv from a synthetic human immunoglobulin library but were unable to do so, presumably because of the similarity between the isozymes. In this work, we have employed a PLAP-specific uncompetitive inhibitor, L-Phe-Gly-Gly, to select isozyme specific scFvs. An uncompetitive inhibitor binds to the enzyme in the presence of substrate and stabilizes the enzyme-substrate complex. Several uncompetitive inhibitors have varying degrees of isozyme specificity for human alkaline phosphatase isozymes. A specific uncompetitive inhibitor would be able to unmask conformational differences between the otherwise very similar molecules. Also, such inhibitors would be directed to regions at/close to the active site of the enzyme. In this work, the library was first incubated with PLAP and the bound clones then eluted by incubation with L-Phe-Gly-Gly along with the substrate, para-nitro phenyl phosphate (pNPP). The scFvs were then studied with regard to the biochemical modulation of their binding, isozyme specificity and effect on enzyme activity. RESULTS: Of 13 clones studied initially, the binding of 9 was inhibited by L-Phe-Gly-Gly (with pNPP) and 2 clones were inhibited by pNPP alone. Two clones had absolute and 2 clones had partial specificity to PLAP. Two clones were cross-reactive with only one other isozyme. Three scFv clones, having an accessible His6-tag, were purified and studied for their modulation of enzyme activity. All the three scFvs inhibited PLAP activity with the kinetics of competitive inhibition. Cell ELISA could demonstrate binding of the specific scFvs to the cell surface expressed PLAP. CONCLUSION: The results demonstrate the biochemical modulation of scFv binding. Also, the scFvs bound to the active site and denied the access to the substrate. The selection strategy could generate specific anti-enzyme antibodies to PLAP that can potentially be used for targeting, for modulating enzyme activity in in vitro and in vivo and as probes for the active site. This strategy also has a general application in selecting antibodies from combinatorial libraries to closely related molecules and conformations

Single-tube Seminested PCR Assay for Detecting Human Papillomavirus in Clinical Samples

There is a growing appreciation of the potential value for routine screening for the presence of HPV not only for cervical specimens but also from oral cavity. The purpose of this study was to develop and clinically evaluate a single-tube seminested PCR assay for the detection of HPV. Several parameters such as PCR primers, primer annealing temperature, the number of PCR cycles and concentration of PCR components were optimized. The assay was evaluated using HPV inserts of type 6, 11, 16, 18, 31, 33, 38 and 51. Evaluation of seminested PCR assay was performed with cervical scrapings from 30 patients and buccal swabs from 30 head and neck cancer patients and results were compared with those of two-tube nested PCR. The results were found to be comparable with a total of 60% (36/60) of samples being positive for HPV using the single-tube assay, while 62% (37/60) positivity was found with two-tube PCR assay. We succeeded in developing a single-tube seminested PCR method for HPV DNA detection which is easier than the conventional nested PCR and can be further evaluated as a potential screening tool for detecting HPV in oral and cervical regions

Isolation of endophytic actinomycetes from Syzygium cumini and their antimicrobial activity against human pathogens

Isolation of endophytic actinomycetes is an important step to screen antimicrobial compounds to curb the threat of drug-resistant strains of human pathogens. Out of the 50 endophytic actinomycetes obtained from surface sterilized root, stem and leaf tissues of Syzygium cumini, 50 isolates (30%) exhibited antimicrobial activity. Antistaphylococcal activity was displayed by most of the isolates, with maximum percent inhibition by J-10 (Mean of Inhibition Factor=12.12 mm2). A total of 8 isolates (4 each) were able to hydrolyse protein (proteinase activity) and solubilize chitin (chitinase activity). Results of thin layer chromatography confirm the production of chloramphenicol family |antibiotic by the isolate J-5. This is the first report providing an insight into untapped endophytic actinomycete milieu of Syzygium cumini yet to be explored which might be a promising source for novel antimicrobial agents

Role of defensins in the pathogenesis of chronic lung allograft rejection

Chronic rejection predominantly manifested as bronchiolitis obliterans syndrome (BOS), still remains a major problem affecting long-term outcomes in human lung transplantation (LTx). Donor specific antibodies (DSA) and infiltration of neutrophils in the graft have been associated with the development of BOS. This study determines the role of defensins, produced by neutrophils, and its interaction with α-1-antitrypsin (AAT) towards induction of airway inflammation and fibrosis which are characteristic hallmarks of BOS. Bronchoalveolar lavage (BAL) and serum from LTx recipients, BOS+ (n=28), BOS− (n=26) and normal healthy controls (n=24) were analyzed. Our results show that BOS+ LTx recipients had higher α-defensins (HNP1–3) and β-defensin2 HBD2 concentration in BAL and serum compared to BOS-DSA-recipients and normal controls (p=0.03). BOS+ patients had significantly lower serum AAT along with higher circulating concentration of HNP–AAT complexes in BAL (p=0.05). Stimulation of primary small airway epithelial cells (SAECs) with HNPs induced expression of HBD2, adhesion molecules (ICAM and VCAM), cytokines (IL-6, IL-1β, IL-13, IL-8 and MCP-1) and growth-factor (VEGF and EGF). In contrast, anti-inflammatory cytokine, IL-10 expression decreased 2-fold (p=0.002). HNPs mediated SAEC activation was completely abrogated by AAT. In conclusion, our results demonstrates that neutrophil secretory product, α-defensins, stimulate β-defensin production by SAECs causing upregulation of pro-inflammatory and pro-fibrotic signaling molecules. Hence, chronic stimulation of airway epithelial cells by defensins can lead to inflammation and fibrosis the central events in the development of BOS following LTx

High transport spin polarization in the van der Waals ferromagnet Fe4_4GeTe2_2

The challenging task of scaling-down the size of the power saving electronic devices can be accomplished by exploiting the spin degree of freedom of the conduction electrons in van der Waals (vdW) spintronic architectures built with 2D materials. One of the key components of such a device is a near-room temperature 2D ferromagnet with good metallicity that can generate a highly spin-polarized electronic transport current. However, most of the known 2D ferromagnets have either a very low temperature ordering, poor conductivity, or low spin polarization. In this context, the Fe$_n$GeTe$_2$ (with $n\geq3$) family of ferromagnets stand out due to their near-room temperature ferromagnetism and good metallicity. We have performed spin-resolved Andreev reflection spectroscopy on Fe$_4$GeTe$_2$ ($T_{Curie} \sim$ 273 K) and demonstrated that the ferromagnet is capable of generating a very high transport spin polarization, exceeding 50$\%$. This makes Fe$_4$GeTe$_2$ a strong candidate for application in all-vdW power-saving spintronic devices.Comment: Accepted for publication in Physical Review

Biochemical Characterization of Chloromethane Emission from the Wood-Rotting Fungus Phellinus pomaceus

Many wood-rotting fungi, including Phellinus pomaceus, produce chloromethane (CH(3)Cl). P. pomaceus can be cultured in undisturbed glucose mycological peptone liquid medium to produce high amounts of CH(3)Cl. The biosynthesis of CH(3)Cl is catalyzed by a methyl chloride transferase (MCT), which appears to be membrane bound. The enzyme is labile upon removal from its natural location and upon storage at low temperature in its bound state. Various detergents failed to solubilize the enzyme in active form, and hence it was characterized by using a membrane fraction. The enzyme had a sharp pH optimum between 7 and 7.2. Its apparent K(m) for Cl(−) (ca. 300 mM) was much higher than that for I(−) (250 μM) or Br(−) (11 mM). A comparison of these K(m) values to the relative in vivo methylation rates for different halides suggests that the real K(m) for Cl(−) may be much lower, but the calculated value is high because the CH(3)Cl produced is used immediately in a coupled reaction. Among various methyl donors tested, S-adenosyl-l-methionine (SAM) was the only one that supported significant methylation by MCT. The reaction was inhibited by S-adenosyl-l-homocysteine, an inhibitor of SAM-dependent methylation, suggesting that SAM is the natural methyl donor. These findings advance our comprehension of a poorly understood metabolic sector at the origin of biogenic emissions of halomethanes, which play an important role in atmospheric chemistry