A Simple and User-Friendly Method for High-Quality Preparation of Pollen Grains for Scanning Electron Microscopy (SEM).

Pollen is becoming an increasingly important subject for molecular researchers in genetic engineering, plant breeding, and environmental monitoring. To broaden the scope of these studies, it is essential to develop accessible methods for scientists who are not specialized in palynology. The article presents a simplified technical procedure for preparing pollen grains for scanning electron microscopy (SEM). The protocol is convenient for any molecular laboratory due to its small set of reagents, ease of execution, low cost, does not require special equipment, and takes only one hour to complete. The high penetrating ability of formaldehyde and the final delicate dehydration using hexamethyldisilazane (HMDS) instead of critical point drying allow for sufficient preservation of the architecture of the aperture, which is considered a gateway for the passage of biomolecules. The method was successfully applied to pollen grains of representatives of dicotyledons (beetroot, petunia, radish, tomato and tobacco) and monocotyledons (lily, onion, corn, rye and wheat). Species studied included insect-pollinated (entomophilous) and wind-pollinated (anemophilous) species. A comparative analysis of the sizes of fresh living pollen grains under a light microscope and those prepared for SEM showed some shrinkage. Quantitative analysis of the degree of pollen grain shrinkage showed that this process depends on the initial shape of dry pollen grains, and the number and structure of apertures. The results support the theoretical model of the folding/unfolding pathways of pollen grains.

Advancing virus-induced gene silencing in sunflower: key factors of VIGS spreading and a novel simple protocol.

Virus-Induced Gene Silencing (VIGS) is a versatile tool in plant science, yet its application to non-model species like sunflower demands extensive optimization due to transformation challenges. In this study, we aimed to elucidate the factors that significantly affect the efficiency of Agrobacterium-VIGS in sunflowers. After testing a number of approaches, we concluded that the seed vacuum technique followed by 6 h of co-cultivation produced the most efficient VIGS results. Genotype-dependency analysis revealed varying infection percentages (62-91%) and silencing symptom spreading in different sunflower genotypes. Additionally, we explored the mobility of tobacco rattle virus (TRV) and phenotypic silencing manifestation (photo-bleaching) across different tissues and regions of VIGS-infected sunflower plants. We showed the presence of TRV is not necessarily limited to tissues with observable silencing events. Finally, time-lapse observation demonstrated a more active spreading of the photo-bleached spots in young tissues compared to mature ones. This study not only offers a robust VIGS protocol for sunflowers but also provides valuable insights into genotype-dependent responses and the dynamic nature of silencing events, shedding light on TRV mobility across different plant tissues.

Telemedicine-enabled biofeedback electropalatography rehabilitation (TEBER): A pilot study for patients treated with surgery for oral cavity carcinoma.

BACKGROUND: To address the rehabilitative barriers to frequency and precision of care, we conducted a pilot study of a biofeedback electropalatography (EPG) device paired with telemedicine for patients who underwent primary surgery +/- adjuvant radiation for oral cavity carcinoma. We hypothesized that lingual optimization followed by telemedicine-enabled biofeedback electropalatography rehabilitation (TEBER) would further improve speech and swallowing outcomes after "standard-of-care" SOC rehabilitation. METHOD: Pilot prospective 8-week (TEBER) program following 8 weeks of (SOC) rehabilitation. RESULTS: Twenty-seven patients were included and 11 completed the protocol. When examining the benefit of TEBER independent of standard of care, "range-of-liquids" improved by +0.36 [95% CI, 0.02-0.70, p = 0.05] and "range-of-solids" improved by +0.73 [95% CI, 0.12-1.34, p = 0.03]. There was a positive trend toward better oral cavity obliteration; residual volume decreased by -1.2 [95% CI, -2.45 to 0.053, p = 0.06], and "nutritional-mode" increased by +0.55 [95% CI, -0.15 to 1.24, p = 0.08]. CONCLUSION: This pilot suggests that TEBER bolsters oral rehabilitation after 8 weeks of SOC lingual range of motion.

Hidden Pitfalls of Using Onion Pollen in Molecular Research.

There is little information on the use of pollen in molecular research, despite the increased interest in genome editing by pollen-mediated transformation. This paper presents an essential toolbox of technical procedures and observations for molecular studies on onion (Allium cepa L.) pollen. PCR is a useful tool as an express method to evaluate editing results before pollination. A direct PCR protocol for pollen suspension has been adapted without needing DNA pre-extraction. We showed that the outer layer of lipids known as pollenkitt is a limiting factor for successful PCR on pollen. A simple pre-washing step of pollen suspension was able to eliminate the pollenkitt and enormously affect the PCR results. Additionally, our pollenkitt study helped us develop a simple and effective pollination method using wetted onion pollen grains. Classical manual pollination usually is conducted by intact pollen without wetting. Most existing methods of the editing system delivery into pollen are carried out in a wet medium with consequent drying before pollination, which adversely affects the viability of pollen. The optimal medium for wet pollination was 12% sucrose water solution. Our method of using wetted pollen grains for pollination might be very beneficial for pollen genetic manipulation.

Dataset for the content of bioactive components and phytonutrients of (Ocimum basilicum and Brassica rapa) microgreens.

The data provided in this article were obtained from fresh and dry samples of green Basilic (Ocimum Basillicum L.), red Basilic (Ocimum basilicum 'purpurascens'), green Mizuna (Brassica rapa var. niposinica), and red Mizuna (Brassica rapa var. pipposinica) microgreens grown in climatic chamber (phytotron) on jute mats for 15 days. Phytonutrients contents including chlorophylls, pheophytins, carotenoids pigments, total antioxidant capacity, total phenolic content, ascorbic acid, as well as organic acids contents varied between all cultivars. Spectrometry, electrophoresis, coulometric, and liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS) were the principal employed methods. Data of antioxidants and phytonutrients contents contribute to the understanding of the benefits of microgreens as a newly emerging product. Data of pigments content compares the difference of accumulation of chlorophylls, pheophytins, and carotenoids between red and green cultivars of the studied microgreens, and the variability of their concentrations along with the contents of organic acids provide insights to plants physiology during the differentiation phase.

The Power of Genomic in situ Hybridization (GISH) in Interspecific Breeding of Bulb Onion (Allium cepa L.) Resistant to Downy Mildew (Peronospora destructor [Berk.] Casp.).

We exploited the advantages of genomic in situ hybridization (GISH) to monitor the introgression process at the chromosome level using a simple and robust molecular marker in the interspecific breeding of bulb onion (Allium cepa L.) that is resistant to downy mildew. Downy mildew (Peronospora destructor [Berk.] Casp.) is the most destructive fungal disease for bulb onions. With the application of genomic in situ hybridization (GISH) and previously developed DMR1 marker, homozygous introgression lines that are resistant to downy mildew were successfully produced in a rather short breeding time. Considering that the bulb onion is a biennial plant, it took seven years from the F1 hybrid production to the creation of S2BC2 homozygous lines that are resistant to downy mildew. Using GISH, it was shown that three progeny plants of S2BC2 possessed an A. roylei homozygous fragment in the distal region of the long arm of chromosomes 3 in an A. cepa genetic background. Previously, it was hypothesized that a lethal gene(s) was linked to the downy mildew resistance gene. With the molecular cytogenetic approach, we physically mapped more precisely the lethal gene(s) using the homozygous introgression lines that differed in the size of the A. roylei fragments on chromosome 3.

Group 3 ITI Consensus Report: Patient-reported outcome measures associated with implant dentistry.

OBJECTIVES: The aim of Working Group 3 was to focus on three topics that were assessed using patient-reported outcome measures (PROMs). These topics included the following: (a) the aesthetics of tooth and implant-supported fixed dental prostheses focusing on partially edentulous patients, (b) a comparison of fixed and removable implant-retained prostheses for edentulous populations, and (c) immediate versus early/conventional loading of immediately placed implants in partially edentate patients. PROMs include ratings of satisfaction and oral health-related quality of life (QHRQoL), as well as other indicators, that is, pain, general health-related quality of life (e.g., SF-36). MATERIALS AND METHODS: The Consensus Conference Group 3 participants discussed the findings of the three systematic review manuscripts. Following comprehensive discussions, participants developed consensus statements and recommendations that were then discussed in larger plenary sessions. Following this, any necessary modifications were made and approved. RESULTS: Patients were very satisfied with the aesthetics of implant-supported fixed dental prostheses and the surrounding mucosa. Implant neck design, restorative material, or use of a provisional restoration did not influence patients' ratings. Edentulous patients highly rate both removable and fixed implant-supported prostheses. However, they rate their ability to maintain their oral hygiene significantly higher with the removable prosthesis. Both immediate provisionalization and conventional loading receive positive patient-reported outcomes. CONCLUSIONS: Patient-reported outcome measures should be gathered in every clinical study in which the outcomes of oral rehabilitation with dental implants are investigated. PROMs, such as patients' satisfaction and QHRQoL, should supplement other clinical parameters in our clinical definition of success.

Evaluation of a modular palatal lift prosthesis with a silicone velar lamina for hypernasal patients.

STATEMENT OF PROBLEM: Speech bulbs and palatal lift prostheses are used to improve oral-nasal balance in speakers with hypernasality resulting from velopharyngeal dysfunction. Fabricating such speech prostheses is often a protracted process, and the nasopharyngeal impression can be uncomfortable for the client. PURPOSE: The purpose of this study was to develop and test a modular palatal lift prosthesis with a silicone velar lamina that can be fabricated without a nasopharyngeal impression. MATERIAL AND METHODS: Six adult participants with different etiologies were treated with both a conventional palatal lift prosthesis and the new prosthesis. The outcome measures were nasalance scores, speech acceptability ratings, and participant responses on a questionnaire. Inferential statistical analyses were conducted with nonparametric Friedman tests and 2-tailed paired Wilcoxon signed ranks tests. The probability was set at P<.1. RESULTS: Among the 3 speaking conditions (no prosthesis, acrylic resin prosthesis, modular silicone palatal lift prosthesis), no differences were found in nasalance scores for the oral stimuli. For the nasal sentences, a numerically greater reduction was observed for the silicone than for the acrylic resin prosthesis. Speech acceptability was better with the modular silicone palatal lift prosthesis (z=2.032, P<.05) and the acrylic resin prosthesis (z=1.753, P<.1) than with no prosthesis. The questionnaire showed better subjective speech acceptability with the acrylic resin prosthesis (z=1.706, P<.05) and the modular silicone palatal lift prosthesis (z=1.706, P<.05) than with no prosthesis. Swallowing comfort was also numerically better for the acrylic resin prosthesis than for the modular silicone palatal lift prosthesis. CONCLUSIONS: This study demonstrates the feasibility of a new design for a flexible and modular palatal lift prosthesis. The functional outcomes were comparable to those of the traditional design. Although the overall results in this study favored the traditional prosthesis, the new design may be viable for patients who require alternative treatment solutions.

Prosthetic rehabilitation of the head and neck: the state of the art.

PURPOSE OF REVIEW: Cancer of the head and neck area is common worldwide. Rehabilitation of cancer patients can be done using a surgical and/or a prosthetic approach. This review presents the current trends in prosthetic rehabilitation of the head and neck area. RECENT FINDINGS: The literature is abundant with case reports describing prosthetic new techniques to rehabilitate patients with cancer of the head and neck area. However, limited evidence-based research exists on these techniques or on the science of materials used in prosthetic rehabilitation of patients with head and neck cancer. SUMMARY: The goals of the prosthetic rehabilitation of patients with head and neck cancer are management of speech and swallowing function, and the restoration of the original esthetics and functions of the bone and soft tissues. These goals remain a challenge. Dental implants are increasingly implemented to help in the prosthetic rehabilitation of patients with head and neck cancer.

Bisphosphonate associated osteonecrosis of the jaw.

In 2003, the first reports describing osteonecrosis of the jaw (ONJ) in patients receiving bisphosphonates (BP) were published. These cases occurred in patients with cancer receiving high-dose intravenous BP; however, 5% of the cases were in patients with osteoporosis receiving low-dose bisphosphonate therapy. We present the results of a systematic review of the incidence, risk factors, diagnosis, prevention, and treatment of BP associated ONJ. We conducted a comprehensive literature search for relevant studies on BP associated ONJ in oncology and osteoporosis patients published before February 2008.All selected relevant articles were sorted by area of focus. Data for each area were abstracted by 2 independent reviewers. The results showed that the diagnosis is made clinically. Prospective data evaluating the incidence and etiologic factors are very limited. In oncology patients receiving high-dose intravenous BP, ONJ appears to be dependent on the dose and duration of therapy, with an estimated incidence of 1%-12% at 36 months of exposure. In osteoporosis patients, it is rare, with an estimated incidence < 1 case per 100,000 person-years of exposure. The incidence of ONJ in the general population is not known. Currently, there is insufficient evidence to confirm a causal link between low-dose BP use in the osteoporosis patient population and ONJ. We concluded BP associated ONJ is associated with high-dose BP therapy primarily in the oncology patient population. Prevention and treatment strategies are currently based on expert opinion and focus on maintaining good oral hygiene and conservative surgical intervention.